@inbook {19138,
	title = {Mesenchymal Stem Cells},
	booktitle = {Principles of Regenerative Medicine},
	year = {In Press},
	publisher = {Elsevier},
	organization = {Elsevier},
	edition = {3},
	author = {Gazit, Zulma and Pelled, Gadi and Sheyn, Dmitriy and Cohn Yakubovich, Doron and Gazit, Dan}
}
@article {bez2019nonviral,
	title = {Nonviral ultrasound-mediated gene delivery in small and large animal models},
	journal = {Nature Protocols},
	year = {2019},
	pages = {1},
	publisher = {Nature Publishing Group},
	abstract = {Ultrasound-mediated gene delivery (sonoporation) is a minimally invasive, nonviral and clinically translatable method of gene therapy. This method offers a favorable safety profile over that of viral vectors and is less invasive as compared with other physical gene delivery approaches (e.g., electroporation). We have previously used sonoporation to overexpress transgenes in different skeletal tissues in order to induce tissue regeneration. Here, we provide a protocol that could easily be adapted to address various other targets of tissue regeneration or additional applications, such as cancer and neurodegenerative diseases. This protocol describes how to prepare, conduct and optimize ultrasound-mediated gene delivery in both a murine and a porcine animal model. The protocol includes the preparation of a microbubble-DNA mix and in vivo sonoporation under ultrasound imaging. Ultrasound-mediated gene delivery can be accomplished within 10 min. After DNA delivery, animals can be followed to monitor gene expression, protein secretion and other transgene-specific outcomes, including tissue regeneration. This procedure can be accomplished by a competent graduate student or technician with prior experience in ultrasound imaging or in performing in vivo procedures.},
	author = {Bez, Maxim and Foiret, Josquin and Shapiro, Galina and Pelled, Gadi and Ferrara, Katherine W. and Gazit, Dan}
}
@article {seguin2018latest,
	title = {Latest advances in intervertebral disc development and progenitor cells},
	journal = {JOR Spine},
	year = {2018},
	pages = {e1030},
	publisher = {Wiley Online Library},
	abstract = {This paper is a concise review aiming to assemble the most relevant topics presented by the authors at ORS-Philadelphia Spine Research Society\&nbsp;Fourth International Spine Research Symposium. It centers on the latest advances in disc development, its main structural entities, and the populating cells, with emphasis on the advances in pivotal molecular pathways responsible for forming the intervertebral discs (IVD). The objective of finding and emphasizing pathways and mechanisms that function to control tissue formation is to identify and to explore modifications occurring during normal aging, disease, and tissue repair. Thus, to comprehend that the cellular and molecular basis of tissue degeneration are crucial in the study of the dynamic interplay that includes cell-cell communication, gene regulation, and growth factors required to form a healthy and functional tissue during normal development.},
	url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/jsp2.1030},
	author = {S{\'e}guin, Cheryle A and Chan, Danny and Dahia, Chitra L and Gazit, Zulma}
}
@article {bez2018molecular,
	title = {Molecular pain markers correlate with pH-sensitive MRI signal in a pig model of disc degeneration},
	journal = {Scientific Reports},
	volume = {8},
	number = {1},
	year = {2018},
	pages = {17363},
	publisher = {Nature Publishing Group},
	abstract = {Intervertebral disc (IVD) degeneration is a leading cause of chronic low back pain that affects millions of people every year. Yet identification of the specific IVD causing this pain is based on qualitative visual interpretation rather than objective findings. One possible approach to diagnosing pain-associated IVD could be to identify acidic IVDs, as decreased pH within an IVD has been postulated to mediate discogenic pain. We hypothesized that quantitative chemical exchange saturation transfer (qCEST) MRI could detect pH changes in IVDs, and thence be used to diagnose pathologically painful IVDs objectively and noninvasively. To test this hypothesis, a surgical model of IVD degeneration in Yucatan minipigs was used. Direct measurement of pH inside the degenerated IVDs revealed a significant drop in pH after degeneration, which correlated with a significant increase in the qCEST signal. Gene analysis of harvested degenerated IVDs revealed significant upregulation of pain-, nerve- and inflammatory-related markers after IVD degeneration. A strong positive correlation was observed between the expression of pain markers and the increase in the qCEST signal. Collectively, these findings suggest that this approach might be used to identify which IVD is causing low back pain, thereby providing valuable guidance for pain and surgical management.},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/30478330},
	author = {Bez, Maxim and Zhou, Zhengwei and Sheyn, Dmitriy and Tawackoli, Wafa and Giaconi, Joseph C and Shapiro, Galina and David, Shiran Ben and Gazit, Zulma and Pelled, Gadi and Li, Debiao and others}
}
@article {21969,
	title = {Recent Advances and Future of Gene Therapy for Bone Regeneration},
	journal = {Current Osteoporosis Reports},
	year = {2018},
	abstract = {Purpose of Review The purpose of this review is to discuss the recent advances in gene therapy as a treatment for bone regeneration. While most fractures heal spontaneously, patients who present with fracture nonunion suffer from prolonged pain, disability, and often require additional operations to regain musculoskeletal function.Recent Findings In the last few years, BMP gene delivery by means of electroporation and sonoporation resulted in repair of nonunion bone defects in mice, rats, and minipigs. Ex vivo transfection of porcine mesenchymal stem cells (MSCs) resulted in bone regeneration following implantation in vertebral defects of minipigs. Sustained release of VEGF gene from a collagenhydroxyapatite scaffold to the mandible of a human patient was shown to be safe and osteoinductive.Summary In conclusion, gene therapy methods for bone regeneration are systematically becoming more efficient and show proof-of-concept in clinically relevant animal models. Yet, on the pathway to clinical use, more investigation is needed to determine the safety aspects of the various techniques in terms of biodistribution, toxicity, and tumorigenicity.},
	url = {https://doi.org/10.1007/s11914-018-0459-3},
	author = {Shapiro, Galina and Raphael Lieber and Gazit, Dan and Pelled, Gadi}
}
@article {21829,
	title = {Ultrasound-mediated Gene Delivery Enhances Tendon Allograft Integration in Mini-pig Ligament Reconstruction},
	journal = {Molecular Therapy},
	year = {2018},
	type = {Journal Article},
	abstract = {Ligament injuries occur frequently, substantially hindering routine daily activities and sports participation in patients. Surgical reconstruction using autogenous or allogeneic tissues is the gold standard treatment for ligament injuries. Although surgeons routinely perform ligament reconstructions, the integrity of these reconstructions largely depends on adequate biological healing of the interface between the\&nbsp;ligament graft and the bone. We hypothesized that localized ultrasound-mediated, microbubble-enhanced therapeutic gene delivery to endogenous stem cells would lead to significantly improved ligament graft integration. To test this hypothesis, an anterior cruciate ligament reconstruction procedure was performed in Yucatan mini-pigs. A collagen scaffold was implanted in the reconstruction sites to facilitate recruitment of endogenous mesenchymal stem cells. Ultrasound-mediated reporter gene delivery successfully transfected 40\% of cells recruited to the reconstruction sites. When BMP-6 encoding DNA was delivered, BMP-6 expression in the reconstruction sites was significantly enhanced. Micro-computed tomography and biomechanical analyses showed that ultrasound-mediated BMP-6 gene delivery led to significantly enhanced osteointegration in all animals 8\&nbsp;weeks after surgery. Collectively, these findings demonstrate that ultrasound-mediated gene delivery to endogenous mesenchymal progenitor cells can effectively improve ligament reconstruction in large animals, thereby addressing a major unmet orthopedic need and offering new possibilities for translation to the clinical setting.},
	issn = {1525-0016},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/29784586},
	author = {Bez, Maxim and Kremen, Thomas J and Tawackoli, Wafa and Avalos, Pablo and Sheyn, Dmitriy and Shapiro, Galina and Giaconi, Joseph C and David, Shiran Ben and Snedeker, Jess G and Gazit, Zulma and Ferrara, Katherine W. and Gazit, Dan and Pelled, Gadi}
}
@inbook {19137,
	title = {Consideration of Biological Sex in Translating Regenerative Stem Cell Therapies},
	booktitle = {Principles of Gender-Specific Medicine},
	volume = {1},
	year = {2017},
	pages = {443-458},
	publisher = {Elsevier},
	organization = {Elsevier},
	edition = {3},
	author = {Shapiro, Galina and Pelled, Gadi and Gazit, Dan}
}
@article {19118,
	title = {In situ bone tissue engineering via ultrasound-mediated gene delivery to endogenous progenitor cells in mini-pigs},
	journal = {Sci Transl Med},
	volume = {9},
	number = {390},
	year = {2017},
	note = {Bez, Maxim Sheyn, Dmitriy Tawackoli, Wafa Avalos, Pablo Shapiro, Galina Giaconi, Joseph C Da, Xiaoyu David, Shiran Ben Gavrity, Jayne Awad, Hani A Bae, Hyun W Ley, Eric J Kremen, Thomas J Gazit, Zulma Ferrara, Katherine W Pelled, Gadi Gazit, Dan eng P30 AR069655/AR/NIAMS NIH HHS/ P50 AR054041/AR/NIAMS NIH HHS/ R01 CA112356/CA/NCI NIH HHS/ Sci Transl Med. 2017 May 17;9(390). pii: eaal3128. doi: 10.1126/scitranslmed.aal3128.[["fid":"22192","view_mode":"default","type":"media","attributes":"height":"100","width":"100","class":"wysiwyg-placeholder media-element file-default"]]},
	type = {Journal Article},
	abstract = {More than 2 million bone-grafting procedures are performed each year using autografts or allografts. However, both options carry disadvantages, and there remains a clear medical need for the development of new therapies for massive bone loss and fracture nonunions. We hypothesized that localized ultrasound-mediated, microbubble-enhanced therapeutic gene delivery to endogenous stem cells would induce efficient bone regeneration and fracture repair. To test this hypothesis, we surgically created a critical-sized bone fracture in the tibiae of Yucatan mini-pigs, a clinically relevant large animal model. A collagen scaffold was implanted in the fracture to facilitate recruitment of endogenous mesenchymal stem/progenitor cells (MSCs) into the fracture site. Two weeks later, transcutaneous ultrasound-mediated reporter gene delivery successfully transfected 40\% of cells at the fracture site, and flow cytometry showed that 80\% of the transfected cells expressed MSC markers. Human bone morphogenetic protein-6 (BMP-6) plasmid DNA was delivered using ultrasound in the same animal model, leading to transient expression and secretion of BMP-6 localized to the fracture area. Micro-computed tomography and biomechanical analyses showed that ultrasound-mediated BMP-6 gene delivery led to complete radiographic and functional fracture healing in all animals 6 weeks after treatment, whereas nonunion was evident in control animals. Collectively, these findings demonstrate that ultrasound-mediated gene delivery to endogenous mesenchymal progenitor cells can effectively treat nonhealing bone fractures in large animals, thereby addressing a major orthopedic unmet need and offering new possibilities for clinical translation.},
	issn = {1946-6242 (Electronic) 1946-6234 (Linking)},
	doi = {10.1126/scitranslmed.aal3128},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/28515335},
	author = {Bez, M. and Sheyn, D. and Tawackoli, W. and Avalos, P. and Shapiro, G. and Giaconi, J. C. and Da, X. and David, S. B. and Gavrity, J. and Awad, H. A. and Bae, H. W. and Ley, E. J. and Kremen, T. J. and Gazit, Z. and Ferrara, K. W. and Pelled, G. and Gazit, D.}
}
@article {19121,
	title = {Isolation and characterization of mesenchymal stromal progenitors from the temporomandibular joint disc},
	journal = {J Tissue Eng Regen Med},
	volume = {11},
	number = {5},
	year = {2017},
	note = {Lavi, Amir Pelled, Gadi Tawackoli, Wafa Casap, Nardi Gazit, Dan Gazit, Zulma eng England J Tissue Eng Regen Med. 2017 May;11(5):1553-1561. doi: 10.1002/term.2055. Epub 2015 Jul 19.},
	pages = {1553-1561},
	type = {Journal Article},
	abstract = {Disorders of the temporomandibular joint (TMJ) complex affect 6-12\% of the population; the joint{\textquoteright}s disc is usually involved. Tissue engineering and regenerative medicine may constitute a promising therapeutic approach, with resident stromal progenitor cells a key factor in the process. We hypothesized that the TMJ disc (TMJD) contains multipotent stromal progenitors that may play an important role in regeneration of the disc. TMJD cells were cultured and evaluated for growth kinetics and colony-forming units (CFUs). Single cell-derived clones were isolated and induced to differentiate toward the osteogenic, adipogenic and chondrogenic lineages by culturing in various induction media. Flow cytometry was used to identify multipotent stromal cell surface markers in additional cell samples, and reverse transcription-polymerase chain reaction (RT-PCR) was used to determine gene expression patterns within isolated cells. High numbers of CFUs were observed, indicating cell self-renewal. Biochemical assays showed significantly higher alkaline phosphatase (ALP) activity, lipid droplet concentration and glycosaminoglycan levels in cells cultured in osteogenic, adipogenic and chondrogenic induction medium, respectively. Approximately 1\% of the total cell population demonstrated the capability to differentiate into all three mesenchymal lineages. Chondrogenic gene levels within TMJD-derived cells were significantly reduced in passaged culture. Our results support the hypothesis that multipotent stromal progenitor cells populate the TMJD and possess proliferation and differentiation capabilities. These cells may contribute to the regeneration potential of dysfunctional tissue and become the primary component in future attempts at tissue engineering or regeneration of this complex. Copyright (c) 2015 John Wiley \& Sons, Ltd.},
	keywords = {endogenous progenitors mesenchymal stem cells regenerative medicine stem cell differentiation temporomandibular joint disc tissue engineering},
	issn = {1932-7005 (Electronic) 1932-6254 (Linking)},
	doi = {10.1002/term.2055},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/26193866},
	author = {Lavi, A. and Pelled, G. and Tawackoli, W. and Casap, N. and Gazit, D. and Gazit, Z.}
}
@article {19122,
	title = {Semiautomated Longitudinal Microcomputed Tomography-based Quantitative Structural Analysis of a Nude Rat Osteoporosis-related Vertebral Fracture Model},
	journal = {J Vis Exp},
	number = {127},
	year = {2017},
	note = {Shapiro, Galina Bez, Maxim Tawackoli, Wafa Gazit, Zulma Gazit, Dan Pelled, Gadi eng J Vis Exp. 2017 Sep 28;(127). doi: 10.3791/55928.},
	type = {Journal Article},
	abstract = {Osteoporosis-related vertebral compression fractures (OVCFs) are a common and clinically unmet need with increasing prevalence as the world population ages. Animal OVCF models are essential to the preclinical development of translational tissue engineering strategies. While a number of models currently exist, this protocol describes an optimized method for inducing multiple highly reproducible vertebral defects in a single nude rat. A novel longitudinal semiautomated microcomputed tomography (microCT)-based quantitative structural analysis of the vertebral defects is also detailed. Briefly, rats were imaged at multiple time points post-op. The day 1 scan was reoriented to a standard position, and a standard volume of interest was defined. Subsequent microCT scans of each rat were automatically registered to the day 1 scan so the same volume of interest was then analyzed to assess for new bone formation. This versatile approach can be adapted to a variety of other models where longitudinal imaging-based analysis could benefit from precise 3D semiautomated alignment. Taken together, this protocol describes a readily quantifiable and easily reproducible system for osteoporosis and bone research. The suggested protocol takes 4 months to induce osteoporosis in nude ovariectomized rats and between 2.7 and 4 h to generate, image, and analyze two vertebral defects, depending on tissue size and equipment.},
	issn = {1940-087X (Electronic) 1940-087X (Linking)},
	doi = {10.3791/55928},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/28994771},
	author = {Shapiro, G. and Bez, M. and Tawackoli, W. and Gazit, Z. and Gazit, D. and Pelled, G.}
}
@article {19120,
	title = {Systemic administration of mesenchymal stem cells combined with parathyroid hormone therapy synergistically regenerates multiple rib fractures},
	journal = {Stem Cell Res Ther},
	volume = {8},
	number = {1},
	year = {2017},
	note = {Cohn Yakubovich, Doron Sheyn, Dmitriy Bez, Maxim Schary, Yeshai Yalon, Eran Sirhan, Afeef Amira, May Yaya, Alin De Mel, Sandra Da, Xiaoyu Ben-David, Shiran Tawackoli, Wafa Ley, Eric J Gazit, Dan Gazit, Zulma Pelled, Gadi eng R01 DE019902/DE/NIDCR NIH HHS/ England Stem Cell Res Ther. 2017 Mar 9;8(1):51. doi: 10.1186/s13287-017-0502-9.},
	pages = {51},
	type = {Journal Article},
	abstract = {BACKGROUND: A devastating condition that leads to trauma-related morbidity, multiple rib fractures, remain a serious unmet clinical need. Systemic administration of mesenchymal stem cells (MSCs) has been shown to regenerate various tissues. We hypothesized that parathyroid hormone (PTH) therapy would enhance MSC homing and differentiation, ultimately leading to bone formation that would bridge rib fractures. METHODS: The combination of human MSCs (hMSCs) and a clinically relevant PTH dose was studied using immunosuppressed rats. Segmental defects were created in animals{\textquoteright} fifth and sixth ribs. The rats were divided into four groups: a negative control group, in which animals received vehicle alone; the PTH-only group, in which animals received daily subcutaneous injections of 4 mug/kg teriparatide, a pharmaceutical derivative of PTH; the hMSC-only group, in which each animal received five injections of 2 x 106 hMSCs; and the hMSC + PTH group, in which animals received both treatments. Longitudinal in vivo monitoring of bone formation was performed biweekly using micro-computed tomography (muCT), followed by histological analysis. RESULTS: Fluorescently-dyed hMSCs were counted using confocal microscopy imaging of histological samples harvested 8 weeks after surgery. PTH significantly augmented the number of hMSCs that homed to the fracture site. Immunofluorescence of osteogenic markers, osteocalcin and bone sialoprotein, showed that PTH induced cell differentiation in both exogenously administered cells and resident cells. muCT scans revealed a significant increase in bone volume only in the hMSC + PTH group, beginning by the 4th week after surgery. Eight weeks after surgery, 35\% of ribs in the hMSC + PTH group had complete bone bridging, whereas there was complete bridging in only 6.25\% of ribs (one rib) in the PTH-only group and in none of the ribs in the other groups. Based on the muCT scans, biomechanical analysis using the micro-finite element method demonstrated that the healed ribs were stiffer than intact ribs in torsion, compression, and bending simulations, as expected when examining bone callus composed of woven bone. CONCLUSIONS: Administration of both hMSCs and PTH worked synergistically in rib fracture healing, suggesting this approach may pave the way to treat multiple rib fractures as well as additional fractures in various anatomical sites.},
	keywords = {Parathyroid hormone Rib fracture Systemic stem cell administration},
	issn = {1757-6512 (Electronic) 1757-6512 (Linking)},
	doi = {10.1186/s13287-017-0502-9},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/28279202},
	author = {Cohn Yakubovich, D. and Sheyn, D. and Bez, M. and Schary, Y. and Yalon, E. and Sirhan, A. and Amira, M. and Yaya, A. and de Mel, S. and Da, X. and Ben-David, S. and Tawackoli, W. and Ley, E. J. and Gazit, D. and Gazit, Z. and Pelled, G.}
}
@article {19119,
	title = {Teriparatide attenuates scarring around murine cranial bone allograft via modulation of angiogenesis},
	journal = {Bone},
	volume = {97},
	year = {2017},
	note = {Cohn Yakubovich, Doron Eliav, Uzi Yalon, Eran Schary, Yeshai Sheyn, Dmitriy Cook-Wiens, Galen Sun, Shuting McKenna, Charles E Lev, Shaya Binshtok, Alexander M Pelled, Gadi Navon, Gil Gazit, Dan Gazit, Zulma eng Bone. 2017 Apr;97:192-200. doi: 10.1016/j.bone.2017.01.020. Epub 2017 Jan 21.},
	pages = {192-200},
	type = {Journal Article},
	abstract = {Nearly all bone fractures in humans can deteriorate into a non-union fracture, often due to formation of fibrotic tissue. Cranial allogeneic bone grafts present a striking example: although seemingly attractive for craniofacial reconstructions, they often fail due to fibrosis at the host-graft junction, which physically prevents the desired bridging of bone between the host and graft and revitalization of the latter. In the present study we show that intermittent treatment with recombinant parathyroid hormone-analogue (teriparatide) modulates neovascularization feeding in the graft surroundings, consequently reducing fibrosis and scar tissue formation and facilitates osteogenesis. Longitudinal inspection of the vascular tree feeding the allograft has revealed that teriparatide induces formation of small-diameter vessels in the 1st week after surgery; by the 2nd week, abundant formation of small-diameter blood vessels was detected in untreated control animals, but far less in teriparatide-treated mice, although in total, more blood capillaries were detected in the animals that were given teriparatide. By that time point we observed expression of the profibrogenic mediator TGF-beta in untreated animals, but negligible expression in the teriparatide-treated mice. To evaluate the formation of scar tissue, we utilized a magnetization transfer contrast MRI protocol to differentiate osteoid tissue from scar tissue, based on the characterization of collagen fibers. Using this method we found that significantly more bone matrix was formed in animals given teriparatide than in control animals. Altogether, our findings show how teriparatide diminishes scarring, ultimately leading to superior bone graft integration.},
	keywords = {Bioengineering Bone anabolics Bone injury Fracture healing Magnetic resonance imaging Parathyroid hormone},
	issn = {1873-2763 (Electronic) 1873-2763 (Linking)},
	doi = {10.1016/j.bone.2017.01.020},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/28119180},
	author = {Cohn Yakubovich, D. and Eliav, U. and Yalon, E. and Schary, Y. and Sheyn, D. and Cook-Wiens, G. and Sun, S. and McKenna, C.E. and Lev, S. and Binshtok, A.M. and Pelled, G. and Navon, G. and Gazit, D. and Gazit, Z.}
}
@article {19125,
	title = {BMP6-Engineered MSCs Induce Vertebral Bone Repair in a Pig Model: A Pilot Study},
	journal = {Stem Cells Int},
	volume = {2016},
	year = {2016},
	note = {Pelled, Gadi Sheyn, Dmitriy Tawackoli, Wafa Jun, Deuk Soo Koh, Youngdo Su, Susan Cohn Yakubovich, Doron Kallai, Ilan Antebi, Ben Da, Xiaoyu Gazit, Zulma Bae, Hyun Gazit, Dan UL1 TR000124/TR/NCATS NIH HHS/United States Journal Article United States Stem Cells Int. 2016;2016:6530624. doi: 10.1155/2016/6530624. Epub 2015 Dec 7.},
	pages = {6530624},
	type = {Journal Article},
	abstract = {Osteoporotic patients, incapacitated due to vertebral compression fractures (VCF), suffer grave financial and clinical burden. Current clinical treatments focus on symptoms{\textquoteright} management but do not combat the issue at the source. In this pilot study, allogeneic, porcine mesenchymal stem cells, overexpressing the BMP6 gene (MSC-BMP6), were suspended in fibrin gel and implanted into a vertebral defect to investigate their effect on bone regeneration in a clinically relevant, large animal pig model. To check the effect of the BMP6-modified cells on bone regeneration, a fibrin gel only construct was used for comparison. Bone healing was evaluated in vivo at 6 and 12 weeks and ex vivo at 6 months. In vivo CT showed bone regeneration within 6 weeks of implantation in the MSC-BMP6 group while only minor bone formation was seen in the defect site of the control group. After 6 months, ex vivo analysis demonstrated enhanced bone regeneration in the BMP6-MSC group, as compared to control. This preclinical study presents an innovative, potentially minimally invasive, technique that can be used to induce bone regeneration using allogeneic gene modified MSCs and therefore revolutionize current treatment of challenging conditions, such as osteoporosis-related VCFs.},
	issn = {1687-966X (Print)},
	doi = {10.1155/2016/6530624},
	url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4685143/},
	author = {Pelled, G. and Sheyn, D. and Tawackoli, W. and Jun, D. S. and Koh, Y. and Su, S. and Cohn Yakubovich, D. and Kallai, I. and Antebi, B. and Da, X. and Gazit, Z. and Bae, H. and Gazit, D.}
}
@article {19123,
	title = {Controlling Arteriogenesis and Mast Cells Are Central to Bioengineering Solutions for Critical Bone Defect Repair Using Allografts},
	journal = {Bioengineering (Basel)},
	volume = {3},
	number = {1},
	year = {2016},
	note = {Antebi, Ben Zhang, Longze Sheyn, Dmitriy Pelled, Gadi Zhang, Xinping Gazit, Zulma Schwarz, Edward M Gazit, Dan P30 AR069655/AR/NIAMS NIH HHS/United States P50 AR054041/AR/NIAMS NIH HHS/United States R01 DE019902/DE/NIDCR NIH HHS/United States P30 AR061307/AR/NIAMS NIH HHS/United States R01 AR067859/AR/NIAMS NIH HHS/United States R01 AR066517/AR/NIAMS NIH HHS/United States S10 RR027340/RR/NCRR NIH HHS/United States Journal Article Switzerland Bioengineering (Basel). 2016 Mar;3(1). pii: 6.},
	type = {Journal Article},
	abstract = {Although most fractures heal, critical defects in bone fail due to aberrant differentiation of mesenchymal stem cells towards fibrosis rather than osteogenesis. While conventional bioengineering solutions to this problem have focused on enhancing angiogenesis, which is required for bone formation, recent studies have shown that fibrotic non-unions are associated with arteriogenesis in the center of the defect and accumulation of mast cells around large blood vessels. Recently, recombinant parathyroid hormone (rPTH; teriparatide; Forteo) therapy have shown to have anti-fibrotic effects on non-unions and critical bone defects due to inhibition of arteriogenesis and mast cell numbers within the healing bone. As this new direction holds great promise towards a solution for significant clinical hurdles in craniofacial reconstruction and limb salvage procedures, this work reviews the current state of the field, and provides insights as to how teriparatide therapy could be used as an adjuvant for healing critical defects in bone. Finally, as teriparatide therapy is contraindicated in the setting of cancer, which constitutes a large subset of these patients, we describe early findings of adjuvant therapies that may present future promise by directly inhibiting arteriogenesis and mast cell accumulation at the defect site.},
	keywords = {arteriogenesis critical bone defect fibrosis mast cells osteogenesis recombinant parathyroid hormone (rPTH teriparatide Forteo)},
	issn = {2306-5354 (Print) 2306-5354},
	doi = {10.3390/bioengineering3010006},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/27141513},
	author = {Antebi, B. and Zhang, L. and Sheyn, D. and Pelled, G. and Zhang, X. and Gazit, Z. and Schwarz, E. M. and Gazit, D.}
}
@article {19127,
	title = {Human Induced Pluripotent Stem Cells Differentiate Into Functional Mesenchymal Stem Cells and Repair Bone Defects},
	journal = {Stem Cells Transl Med},
	volume = {5},
	number = {11},
	year = {2016},
	note = {Sheyn, Dmitriy Ben-David, Shiran Shapiro, Galina De Mel, Sandra Bez, Maxim Ornelas, Loren Sahabian, Anais Sareen, Dhruv Da, Xiaoyu Pelled, Gadi Tawackoli, Wafa Liu, Zhenqiu Gazit, Dan Gazit, Zulma P30 DK063491/DK/NIDDK NIH HHS/United States Journal Article United States Stem Cells Transl Med. 2016 Nov;5(11):1447-1460. doi: 10.5966/sctm.2015-0311. Epub 2016 Jul 11.},
	pages = {1447-1460},
	type = {Journal Article},
	abstract = {: Mesenchymal stem cells (MSCs) are currently the most established cells for skeletal tissue engineering and regeneration; however, their availability and capability of self-renewal are limited. Recent discoveries of somatic cell reprogramming may be used to overcome these challenges. We hypothesized that induced pluripotent stem cells (iPSCs) that were differentiated into MSCs could be used for bone regeneration. Short-term exposure of embryoid bodies to transforming growth factor-beta was used to direct iPSCs toward MSC differentiation. During this process, two types of iPSC-derived MSCs (iMSCs) were identified: early (aiMSCs) and late (tiMSCs) outgrowing cells. The transition of iPSCs toward MSCs was documented using MSC marker flow cytometry. Both types of iMSCs differentiated in vitro in response to osteogenic or adipogenic supplements. The results of quantitative assays showed that both cell types retained their multidifferentiation potential, although aiMSCs demonstrated higher osteogenic potential than tiMSCs and bone marrow-derived MSCs (BM-MSCs). Ectopic injections of BMP6-overexpressing tiMSCs produced no or limited bone formation, whereas similar injections of BMP6-overexpressing aiMSCs resulted in substantial bone formation. Upon orthotopic injection into radial defects, all three cell types regenerated bone and contributed to defect repair. In conclusion, MSCs can be derived from iPSCs and exhibit self-renewal without tumorigenic ability. Compared with BM-MSCs, aiMSCs acquire more of a stem cell phenotype, whereas tiMSCs acquire more of a differentiated osteoblast phenotype, which aids bone regeneration but does not allow the cells to induce ectopic bone formation (even when triggered by bone morphogenetic proteins), unless in an orthotopic site of bone fracture. SIGNIFICANCE: Mesenchymal stem cells (MSCs) are currently the most established cells for skeletal tissue engineering and regeneration of various skeletal conditions; however, availability of autologous MSCs is very limited. This study demonstrates a new method to differentiate human fibroblast-derived induced pluripotent stem cells (iPSCs) to cells with MSC properties, which we comprehensively characterized including differentiation potential and transcriptomic analysis. We showed that these iPS-derived MSCs are able to regenerate nonunion bone defects in mice more efficiently than bone marrow-derived human MSCs when overexpressing BMP6 using a nonviral transfection method.},
	keywords = {Bone regeneration Human induced pluripotent stem cells Mesenchymal stem cells Transforming growth factor-beta},
	issn = {2157-6564 (Print) 2157-6564},
	doi = {10.5966/sctm.2015-0311},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/27400789},
	author = {Sheyn, D. and Ben-David, S. and Shapiro, G. and de Mel, S. and Bez, M. and Ornelas, L. and Sahabian, A. and Sareen, D. and Da, X. and Pelled, G. and Tawackoli, W. and Liu, Z. and Gazit, D. and Gazit, Z.}
}
@article {19131,
	title = {Detection of low back pain using pH level-dependent imaging of the intervertebral disc using the ratio of R1rho dispersion and -OH chemical exchange saturation transfer (RROC)},
	journal = {Magn Reson Med},
	volume = {73},
	number = {3},
	year = {2015},
	note = {1522-2594 Liu, Qi Tawackoli, Wafa Pelled, Gadi Fan, Zhaoyang Jin, Ning Natsuaki, Yutaka Bi, Xiaoming Gart, Avrom Bae, Hyun Gazit, Dan Li, Debiao R01 AR066517/AR/NIAMS NIH HHS/United States Journal Article United States Magn Reson Med. 2015 Mar;73(3):1196-205. doi: 10.1002/mrm.25186. Epub 2014 Apr 2.},
	pages = {1196-205},
	type = {Journal Article},
	abstract = {PURPOSE: Low pH is associated with intervertebral disc (IVD)-generated low back pain (LBP). The purpose of this work was to develop an in vivo pH level-dependent magnetic resonance imaging (MRI) method for detecting discogenic LBP, without using exogenous contrast agents. METHODS: The ratio of R1rho dispersion and chemical exchange saturation transfer (CEST) (RROC) was used for pH-level dependent imaging of the IVD while eliminating the effect of labile proton concentration. The technique was validated by numerical simulations and studies on phantoms and ex vivo porcine spines. Four male (ages 42.8 +/- 18.3) and two female patients (ages 55.5 +/- 2.1) with LBP and scheduled for discography were examined with the method on a 3.0 Tesla MR scanner. RROC measurements were compared with discography outcomes using paired t-test. RESULTS: Simulation and phantom results indicated RROC is a concentration independent and pH level-dependent technique. Porcine spine study results found higher RROC value was related to lower pH level. Painful discs based on discography had significant higher RROC values than those with negative diagnosis (P \&lt; 0.05). CONCLUSION: RROC imaging is a promising pH level dependent MRI technique that has the potential to be a noninvasive imaging tool to detect painful IVDs in vivo.},
	keywords = {*Algorithms Animals Biomarkers/chemistry Hydrogen-Ion Concentration Hydroxides Image Interpretation, Computer-Assisted/*methods Intervertebral Disc/*chemistry Intervertebral Disc Degeneration/complications/*diagnosis Low Back Pain/*diagnosis/etiology Magnetic Resonance Imaging/*methods Phantoms, Imaging Reproducibility of Results Sensitivity and Specificity Swine Cest Intervertebral disc R1rho dispersion Rroc pH imaging},
	issn = {0740-3194},
	doi = {10.1002/mrm.25186},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/24700573},
	author = {Liu, Q. and Tawackoli, W. and Pelled, G. and Fan, Z. and Jin, N. and Natsuaki, Y. and Bi, X. and Gart, A. and Bae, H. and Gazit, D. and Li, D.}
}
@article {19132,
	title = {Matrix stiffness determines the fate of nucleus pulposus-derived stem cells},
	journal = {Biomaterials},
	volume = {49},
	year = {2015},
	note = {1878-5905 Navaro, Yosi Bleich-Kimelman, Nadav Hazanov, Lena Mironi-Harpaz, Iris Shachaf, Yonatan Garty, Shai Smith, Yoav Pelled, Gadi Gazit, Dan Seliktar, Dror Gazit, Zulma R03 AR057143/AR/NIAMS NIH HHS/United States Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t Netherlands Biomaterials. 2015 May;49:68-76. doi: 10.1016/j.biomaterials.2015.01.021. Epub 2015 Feb 14.},
	pages = {68-76},
	type = {Journal Article},
	abstract = {Intervertebral disc (IVD) degeneration and consequent low-back pain present a major medical challenge. Nucleus pulposus-derived stem cells (NP-SCs) may lead to a novel therapy for this severe disease. It was recently shown that survival and function of mature NP cells are regulated in part by tissue stiffness. We hypothesized that modification of matrix stiffness will influence the ability of cultured NP-SCs to proliferate, survive, and differentiate into mature NP cells. NP-SCs were subcultured in three-dimensional matrices of varying degrees of stiffness as measured by the material{\textquoteright}s shear storage modulus. Cell survival, activity, and rate of differentiation toward the chondrogenic or osteogenic lineage were analyzed. NP-SCs were found to proliferate and differentiate in all matrices, irrespective of matrix stiffness. However, matrices with a low shear storage modulus (G{\textquoteright} = 1 kPa) promoted significantly more proliferation and chondrogenic differentiation, whereas matrices with a high modulus (G{\textquoteright} = 2 kPa) promoted osteogenic differentiation. Imaging performed via confocal and scanning electron microscopes validated cell survival and highlighted stiffness-dependent cell-matrix interactions. These results underscore the effect of the matrix modulus on the fate of NP-SCs. This research may facilitate elucidation of the complex cross-talk between NP-SCs and their surrounding matrix in healthy as well as pathological conditions.},
	keywords = {Animals Biomechanical Phenomena/drug effects Cell Differentiation/drug effects *Cell Lineage/drug effects Cell Shape/drug effects Cell Survival/drug effects Cell-Matrix Junctions/drug effects/metabolism Cells, Cultured Chondrogenesis/drug effects Elastic Modulus/drug effects Ethylenediamines/pharmacology Extracellular Matrix/drug effects/*physiology Hydrogels/pharmacology Intervertebral Disc/*cytology Rheology/drug effects Stem Cells/*cytology Sus scrofa Elast},
	issn = {0142-9612},
	doi = {10.1016/j.biomaterials.2015.01.021},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/25725556},
	author = {Navaro, Y. and Bleich-Kimelman, N. and Hazanov, L. and Mironi-Harpaz, I. and Shachaf, Y. and Garty, S. and Smith, Y. and Pelled, G. and Gazit, D. and Seliktar, D. and Gazit, Z.}
}
@article {19134,
	title = {Stem cell therapy for osteoporosis},
	journal = {Curr Osteoporos Rep},
	volume = {12},
	number = {1},
	year = {2014},
	note = {1544-2241 Antebi, Ben Pelled, Gadi Gazit, Dan Journal Article Research Support, Non-U.S. Gov{\textquoteright}t Review United States Curr Osteoporos Rep. 2014 Mar;12(1):41-7. doi: 10.1007/s11914-013-0184-x.},
	pages = {41-7},
	type = {Journal Article},
	abstract = {Osteoporosis is a debilitating disease that affects millions of people worldwide. Current osteoporosis treatments are predominantly bone-resorbing drugs that are associated with several side effects. The use of stem cells for tissue regeneration has raised great hope in various fields of medicine, including musculoskeletal disorders. Stem cell therapy for osteoporosis could potentially reduce the susceptibility of fractures and augment lost mineral density by either increasing the numbers or restoring the function of resident stem cells that can proliferate and differentiate into bone-forming cells. Such osteoporosis therapies can be carried out by exogenous introduction of mesenchymal stem cells (MSCs), typically procured from bone marrow, adipose, and umbilical cord blood tissues or through treatments with drugs or small molecules that recruit endogenous stem cells to osteoporotic sites. The main hurdle with cell-based osteoporosis therapy is the uncertainty of stem cell fate and biodistribution following cell transplantation. Therefore, future advancements will focus on long-term engraftment and differentiation of stem cells at desired bone sites for tangible clinical outcome.},
	keywords = {Humans Mesenchymal Stem Cell Transplantation/*methods Osteoporosis/*therapy},
	issn = {1544-1873},
	doi = {10.1007/s11914-013-0184-x},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/24407712},
	author = {Antebi, B. and Pelled, G. and Gazit, D.}
}
@article {shapiro2014ultrasound,
	title = {Ultrasound-mediated transgene expression in endogenous stem cells recruited to bone injury sites},
	journal = {Polymers for Advanced Technologies},
	volume = {25},
	number = {5},
	year = {2014},
	pages = {525{\textendash}531},
	publisher = {Wiley Online Library},
	url = {http://onlinelibrary.wiley.com/doi/10.1002/pat.3297/abstract},
	author = {Shapiro, Galina and Kallai, Ilan and Sheyn, Dmitriy and Tawackoli, Wafa and Koh, Young Do and Bae, Hyun and Trietel, Tamar and Goldbart, Riki and Kost, Joseph and Gazit, Zulma and others}
}
@article {17128,
	title = {BMP-6 is more efficient in bone formation than BMP-2 when overexpressed in mesenchymal stem cells},
	journal = {Gene Ther},
	volume = {20},
	number = {4},
	year = {2013},
	note = {Mizrahi, O Sheyn, D Tawackoli, W Kallai, I Oh, A Su, S Da, X Zarrini, P Cook-Wiens, G Gazit, D Gazit, Z eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. England 2012/06/22 06:00 Gene Ther. 2013 Apr;20(4):370-7. doi: 10.1038/gt.2012.45. Epub 2012 Jun 21.},
	pages = {370-7},
	type = {Journal Article},
	abstract = {Bone regeneration achieved using mesenchymal stem cells (MSCs) and nonviral gene therapy holds great promise for patients with fractures seemingly unable to heal. Previously, MSCs overexpressing bone morphogenetic proteins (BMPs) were shown to differentiate into the osteogenic lineage and induce bone formation. In the present study, we evaluated the potential of osteogenic differentiation in porcine adipose tissue- and bone marrow-derived MSCs (ASCs and BMSCs, respectively) in vitro and in vivo when induced by nucleofection with rhBMP-2 or rhBMP-6. Our assessment of the in vivo efficiency of this procedure was made using quantitative micro-computed tomography (micro-CT). Nucleofection efficiency and cell viability were similar in both cell types; however, the micro-CT analyses demonstrated that in both ASCs and BMSCs, nucleofection with rhBMP-6 generated bone tissue faster and of higher volumes than nucleofection with rhBMP-2. RhBMP-6 induced more efficient osteogenic differentiation in vitro in BMSCs, and in fact, greater osteogenic potential was identified in BMSCs both in vitro and in vivo than in ASCs. On the basis of our findings, we conclude that BMSCs nucleofected with rhBMP-6 are superior at inducing bone formation in vivo than all other groups studied.},
	keywords = {Adipose Tissue/cytology Animals Bone Morphogenetic Protein 2/genetics/*metabolism Bone Morphogenetic Protein 6/genetics/*metabolism Cell Differentiation Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells/cytology/*metabolism *Osteogenesis Swi, genetic transfection},
	issn = {1476-5462 (Electronic) 0969-7128 (Linking)},
	doi = {10.1038/gt.2012.45},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/22717741},
	author = {Mizrahi, O. and Sheyn, D. and Tawackoli, W. and Kallai, I. and Oh, A. and Su, S. and Da, X. and Zarrini, P. and Cook-Wiens, G. and Gazit, D. and Gazit, Z.}
}
@article {17127,
	title = {Nucleus pulposus degeneration alters properties of resident progenitor cells},
	journal = {Spine J},
	volume = {13},
	number = {7},
	year = {2013},
	note = {Mizrahi, Olga Sheyn, Dmitriy Tawackoli, Wafa Ben-David, Shiran Su, Susan Li, Ning Oh, Anthony Bae, Hyun Gazit, Dan Gazit, Zulma eng R03 AR057143/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2013/04/13 06:00 Spine J. 2013 Jul;13(7):803-14. doi: 10.1016/j.spinee.2013.02.065. Epub 2013 Apr 9.},
	pages = {803-14},
	type = {Journal Article},
	abstract = {BACKGROUND CONTEXT: The intervertebral disc (IVD) possesses a minimal capability for self-repair and regeneration. Changes in the differentiation of resident progenitor cells can represent diminished tissue regeneration and a loss of homeostasis. We previously showed that progenitor cells reside in the nucleus pulposus (NP). The effect of the degenerative process on these cells remains unclear. PURPOSE: We sought to explore the effect of IVD degeneration on the abundance of resident progenitor cells in the NP, their differentiation potential, and their ability to give rise to NP-like cells. We hypothesize that disc degeneration affects those properties. STUDY DESIGN: Nucleus pulposus cells derived from healthy and degenerated discs were methodically compared for proliferation, differentiation potential, and ability to generate NP-like cells. METHODS: Intervertebral disc degeneration was induced in 10 skeletally, mature mini pigs using annular injury approach. Degeneration was induced in three target discs, whereas intact adjacent discs served as controls. The disc degeneration was monitored using magnetic resonance imaging for 6 to 8 weeks. After there was a clear evidence of degeneration, we isolated and compared cells from degenerated discs (D-NP cells [NP-derived cells from porcine degenerated discs]) with cells isolated from healthy discs (H-NP cells) obtained from the same animal. RESULTS: The comparison showed that D-NP cells had a significantly higher colony-forming unit rate and a higher proliferation rate in vitro. Our data also indicate that although both cell types are able to differentiate into mesenchymal lineages, H-NP cells exhibit significantly greater differentiation toward the chondrogenic lineage and NP-like cells than D-NP cells, displaying greater production of glycosaminoglycans and higher gene expression of aggrecan and collagen IIa. CONCLUSIONS: Based on these findings, we conclude that IVD degeneration has a meaningful effect on the cells in the NP. D-NP cells clearly go through the regenerative process; however, this process is not powerful enough to facilitate full regeneration of the disc and reverse the degenerative course. These findings facilitate deeper understanding of the IVD degeneration process and trigger further studies that will contribute to development of novel therapies for IVD degeneration.},
	keywords = {Adipogenesis/*physiology Adult Stem Cells/*pathology Animals Disease Models, Animal Intervertebral Disc/*pathology Intervertebral Disc Degeneration/*pathology Osteogenesis/*physiology Swine Differentiation Intervertebral disc degeneration Nucleus pulposus Progenitor cells Proliferation},
	issn = {1878-1632 (Electronic) 1529-9430 (Linking)},
	doi = {10.1016/j.spinee.2013.02.065},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/23578990},
	author = {Mizrahi, O. and Sheyn, D. and Tawackoli, W. and Ben-David, S. and Su, S. and Li, N. and Oh, A. and Bae, H. and Gazit, D. and Gazit, Z.}
}
@article {17124,
	title = {Oxygenated environment enhances both stem cell survival and osteogenic differentiation},
	journal = {Tissue Eng Part A},
	volume = {19},
	number = {5-6},
	year = {2013},
	note = {Benjamin, Shimon Sheyn, Dmitriy Ben-David, Shiran Oh, Anthony Kallai, Ilan Li, Ning Gazit, Dan Gazit, Zulma eng 2012/12/12 06:00 Tissue Eng Part A. 2013 Mar;19(5-6):748-58. doi: 10.1089/ten.TEA.2012.0298. Epub 2013 Jan 4.},
	pages = {748-58},
	type = {Journal Article},
	abstract = {Osteogenesis of mesenchymal stem cells (MSCs) is highly dependent on oxygen supply. We have shown that perfluorotributylamine (PFTBA), a synthetic oxygen carrier, enhances MSC-based bone formation in vivo. Exploring this phenomenon{\textquoteright}s mechanism, we hypothesize that a transient increase in oxygen levels using PFTBA will affect MSC survival, proliferation, and differentiation, thus increasing bone formation. To test this hypothesis, MSCs overexpressing bone morphogenetic protein 2 were encapsulated in alginate beads that had been supplemented with an emulsion of PFTBA or phosphate-buffered saline. Oxygen measurements showed that supplementation of PFTBA significantly increased the available oxygen level during a 96-h period. PFTBA-containing beads displayed an elevation in cell viability, which was preserved throughout 2 weeks, and a significantly lower ratio of dead cells throughout the experiment. Furthermore, the cells from the control group expressed significantly more hypoxia-related genes such as VEGF, DDIT3, and PKG1. Additionally, PFTBA supplementation led to an increase in the osteogenic differentiation and to a decrease in chondrogenic differentiation of MSCs. In conclusion, PFTBA increases the oxygen availability in the vicinity of the MSCs, which suffer oxygen exhaustion shortly after encapsulation in alginate beads. Consequently, cell survival is increased, and hypoxia-related genes are downregulated. In addition, PFTBA promotes osteogenic differentiation over chondrogeneic differentiation, and thereby can accelerate MSC-based bone regeneration.},
	keywords = {Animals Bone Morphogenetic Protein 2/pharmacology *Cell Differentiation/drug effects Cell Line Cell Proliferation/drug effects Cell Survival/drug effects Chondrogenesis/drug effects/genetics Down-Regulation/drug effects/genetics Female Fluorocarbons/pharm},
	issn = {1937-335X (Electronic) 1937-3341 (Linking)},
	doi = {10.1089/ten.TEA.2012.0298},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/23215901},
	author = {Benjamin, S. and Sheyn, D. and Ben-David, S. and Oh, A. and Kallai, I. and Li, N. and Gazit, D. and Gazit, Z.}
}
@article {17129,
	title = {PTH promotes allograft integration in a calvarial bone defect},
	journal = {Mol Pharm},
	volume = {10},
	number = {12},
	year = {2013},
	note = {Sheyn, Dmitriy Cohn Yakubovich, Doron Kallai, Ilan Su, Susan Da, Xiaoyu Pelled, Gadi Tawackoli, Wafa Cook-Weins, Galen Schwarz, Edward M Gazit, Dan Gazit, Zulma eng P50 AR054041/AR/NIAMS NIH HHS/ R01 DE019902/DE/NIDCR NIH HHS/ DE019902/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2013/10/18 06:00 Mol Pharm. 2013 Dec 2;10(12):4462-71. doi: 10.1021/mp400292p. Epub 2013 Nov 8.[["fid":"22196","view_mode":"default","type":"media","attributes":"height":"100","width":"100","class":"wysiwyg-placeholder media-element file-default"]]},
	pages = {4462-71},
	type = {Journal Article},
	abstract = {Allografts may be useful in craniofacial bone repair, although they often fail to integrate with the host bone. We hypothesized that intermittent administration of parathyroid hormone (PTH) would enhance mesenchymal stem cell recruitment and differentiation, resulting in allograft osseointegration in cranial membranous bones. Calvarial bone defects were created in transgenic mice, in which luciferase is expressed under the control of the osteocalcin promoter. The mice were given implants of allografts with or without daily PTH treatment. Bioluminescence imaging (BLI) was performed to monitor host osteprogenitor differentiation at the implantation site. Bone formation was evaluated with the aid of fluorescence imaging (FLI) and microcomputed tomography (muCT) as well as histological analyses. Reverse transcription polymerase chain reaction (RT-PCR) was performed to evaluate the expression of key osteogenic and angiogenic genes. Osteoprogenitor differentiation, as detected by BLI, in mice treated with an allograft implant and PTH was over 2-fold higher than those in mice treated with an allograft implant without PTH. FLI also demonstrated that the bone mineralization process in PTH-treated allografts was significantly higher than that in untreated allografts. The muCT scans revealed a significant increase in bone formation in allograft + PTH treated mice comparing to allograft + PBS treated mice. The osteogenic genes osteocalcin (Oc/Bglap) and integrin binding sialoprotein (Ibsp) were upregulated in the allograft + PTH treated animals. In summary, PTH treatment enhances osteoprogenitor differentiation and augments bone formation around structural allografts. The precise mechanism is not clear, but we show that infiltration pattern of mast cells, associated with the formation of fibrotic tissue, in the defect site is significantly affected by the PTH treatment.},
	keywords = {Allografts/drug effects/physiology Animals Bone Transplantation/methods Bone and Bones/*drug effects/*physiology Cell Differentiation/drug effects/genetics/physiology Gene Expression/drug effects/genetics/physiology Mast Cells/drug effects/physiology Mese, Genetic/genetics Sialoglycoproteins/genetics Transplantation, Homologous/methods, Physiologic/drug effects/genetics/physiology Osteocalcin/genetics Osteogenesis/*drug effects/genetics/*physiology Parathyroid Hormone/*pharmacology Promoter Regions, Transgenic Neovascularization},
	issn = {1543-8392 (Electronic) 1543-8384 (Linking)},
	doi = {10.1021/mp400292p},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/24131143},
	author = {Sheyn, D. and Cohn Yakubovich, D. and Kallai, I. and Su, S. and Da, X. and Pelled, G. and Tawackoli, W. and Cook-Weins, G. and Schwarz, E. M. and Gazit, D. and Gazit, Z.}
}
@article {17125,
	title = {Real-time bioluminescence functional imaging for monitoring tissue formation and regeneration},
	journal = {Methods Mol Biol},
	volume = {1048},
	year = {2013},
	note = {Kimelman, Nadav Bleich Kallai, Ilan Sheyn, Dmitriy Tawackoli, Wafa Gazit, Zulma Pelled, Gadi Gazit, Dan eng R01AR056694/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 2013/08/10 06:00 Methods Mol Biol. 2013;1048:181-93. doi: 10.1007/978-1-62703-556-9_14.},
	pages = {181-93},
	type = {Journal Article},
	abstract = {Real-time bioluminescence functional imaging holds great promise for regenerative medicine because it improves the researcher{\textquoteright}s ability to analyze and understand the healing process. Using transgenic mice coupled with gene-modified cells, one can employ this method to monitor host and graft activity in various models of tissue regeneration. We implemented real-time bioluminescence functional imaging to analyze bone formation by following a unique protocol in which the luciferase reporter gene, driven by an osteocalcin promoter, is used to visualize host and graft activity during bone formation. Real-time bioluminescence functional imaging can be used to assess the "host reaction" in transgenic mice models; it can also be used to assess "graft activity" in other animals in which genetically labeled stem cells have been implanted or direct gene delivery has been applied. The suggested imaging protocol requires 25 min per sample. However, special attention must be given to the layout of the experimental design, which determines the specific activity that will be analyzed.},
	keywords = {Animals *Bone Regeneration Gene Transfer Techniques Guided Tissue Regeneration/*methods Luciferases/genetics *Luminescent Measurements Mice Mice, Genetic/genetics Tissue Engineering/*methods, Transgenic Osteocalcin/genetics Osteogenesis/physiology Promoter Regions},
	issn = {1940-6029 (Electronic) 1064-3745 (Linking)},
	doi = {10.1007/978-1-62703-556-9_14},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/23929106},
	author = {Kimelman, N. B. and Kallai, I. and Sheyn, D. and Tawackoli, W. and Gazit, Z. and Pelled, G. and Gazit, D.}
}
@article {17126,
	title = {Reliable chemical exchange saturation transfer imaging of human lumbar intervertebral discs using reduced-field-of-view turbo spin echo at 3.0 T},
	journal = {NMR Biomed},
	volume = {26},
	number = {12},
	year = {2013},
	note = {Liu, Qi Jin, Ning Fan, Zhaoyang Natsuaki, Yutaka Tawackoli, Wafa Pelled, Gadi Bae, Hyun Gazit, Dan Li, Debiao eng England 2013/07/31 06:00 NMR Biomed. 2013 Dec;26(12):1672-9. doi: 10.1002/nbm.3001. Epub 2013 Jul 25.},
	pages = {1672-9},
	type = {Journal Article},
	abstract = {The reduced field-of-view (rFOV) turbo-spin-echo (TSE) technique, which effectively suppresses bowel movement artifacts, is developed for the purpose of chemical exchange saturation transfer (CEST) imaging of the intervertebral disc (IVD) in vivo. Attempts to quantify IVD CEST signals in a clinical setting require high reliability and accuracy, which is often compromised in the conventionally used technique. The proposed rFOV TSE CEST method demonstrated significantly superior reproducibility when compared with the conventional technique on healthy volunteers, implying it is a more reliable measurement. Phantom study revealed a linear relation between CEST signal and glycosaminoglycan (GAG) concentration. The feasibility of detecting IVD degeneration was demonstrated on a healthy volunteer, indicating that the proposed method is a promising tool to quantify disc degeneration.},
	keywords = {Adult Female Glycosaminoglycans/metabolism Healthy Volunteers Humans Intervertebral Disc/*pathology Intervertebral Disc Degeneration/pathology Lumbar Vertebrae/*pathology *Magnetic Resonance Imaging Male Phantoms, Imaging *Spin Labels Cest bowel movement intervertebral disc reduced FOV},
	issn = {1099-1492 (Electronic) 0952-3480 (Linking)},
	doi = {10.1002/nbm.3001},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/23893565},
	author = {Liu, Q. and Jin, N. and Fan, Z. and Natsuaki, Y. and Tawackoli, W. and Pelled, G. and Bae, H. and Gazit, D. and Li, D.}
}
@article {17130,
	title = {Transient overexpression of Ppargamma2 and C/ebpalpha in mesenchymal stem cells induces brown adipose tissue formation},
	journal = {Regen Med},
	volume = {8},
	number = {3},
	year = {2013},
	note = {Sheyn, Dmitriy Pelled, Gadi Tawackoli, Wafa Su, Susan Ben-David, Shiran Gazit, Dan Gazit, Zulma eng Research Support, Non-U.S. Gov{\textquoteright}t England 2013/05/01 06:00 Regen Med. 2013 May;8(3):295-308. doi: 10.2217/rme.13.25.},
	pages = {295-308},
	type = {Journal Article},
	abstract = {BACKGROUND: Brown adipose tissue plays a pivotal role in mammal metabolism and thermogenesis. It has a great therapeutic potential in several metabolic disorders such as obesity and diabetes. Mesenchymal stem cells (MSCs) are suitable candidates for brown adipose tissue formation de novo. Ppargamma2 and C/ebpalpha are nucleic receptors known to mediate adipogenic differentiation. We hypothesized that overexpression of the Ppargamma2 and C/ebpalpha genes in MSCs would lead to the formation of adipose tissue. MATERIALS \& METHODS: MSCs bearing the Luc reporter gene were transfected to overexpress Ppargamma2 and C/ebpalpha. Differentiation of nucleofected cells was evaluated in vitro and in vivo following ectopic implantation of the cells in C3H/HeN mice. RESULTS: After implantation, the engineered cells survived for 5 weeks and brown adipose-like tissue was observed in histological samples. Immunostaining and bioluminescent imaging showed new adipocytes expressing Luc and the brown adipose tissue marker, UCP1, in vitro and in vivo. CONCLUSION: We show that gene delivery of transcription factors into MSCs generates brown adipose tissue in vitro and in vivo.},
	keywords = {Adipogenesis/genetics Adipose Tissue, Brown/*metabolism Animals Biomarkers/metabolism CCAAT-Enhancer-Binding Protein-alpha/*metabolism Cell Differentiation/genetics Cell Line Cell Survival/genetics Gene Expression Profiling Genetic Engineering HEK293 Cells Humans Ion Channels/metabolism Lumi},
	issn = {1746-076X (Electronic) 1746-0751 (Linking)},
	doi = {10.2217/rme.13.25},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/23627824},
	author = {Sheyn, D. and Pelled, G. and Tawackoli, W. and Su, S. and Ben-David, S. and Gazit, D. and Gazit, Z.}
}
@article {17123,
	title = {Smad8/BMP2-engineered mesenchymal stem cells induce accelerated recovery of the biomechanical properties of the Achilles tendon},
	journal = {J Orthop Res},
	volume = {30},
	number = {12},
	year = {2012},
	note = {Pelled, Gadi Snedeker, Jess G Ben-Arav, Ayelet Rigozzi, Samuela Zilberman, Yoram Kimelman-Bleich, Nadav Gazit, Zulma Muller, Ralph Gazit, Dan eng R03 TW006923/TW/FIC NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2012/06/15 06:00 J Orthop Res. 2012 Dec;30(12):1932-9. doi: 10.1002/jor.22167. Epub 2012 Jun 13.},
	pages = {1932-9},
	type = {Journal Article},
	abstract = {Tendon tissue regeneration is an important goal for orthopedic medicine. We hypothesized that implantation of Smad8/BMP2-engineered MSCs in a full-thickness defect of the Achilles tendon (AT) would induce regeneration of tissue with improved biomechanical properties. A 2 mm defect was created in the distal region of murine ATs. The injured tendons were then sutured together or given implants of genetically engineered MSCs (GE group), non-engineered MSCs (CH3 group), or fibrin gel containing no cells (FG group). Three weeks later the mice were killed, and their healing tendons were excised and processed for histological or biomechanical analysis. A biomechanical analysis showed that tendons that received implants of genetically engineered MSCs had the highest effective stiffness (>70\% greater than natural healing, p < 0.001) and elastic modulus. There were no significant differences in either ultimate load or maximum stress among the treatment groups. Histological analysis revealed a tendon-like structure with elongated cells mainly in the GE group. ATs that had been implanted with Smad8/BMP2-engineered stem cells displayed a better material distribution and functional recovery than control groups. While additional study is required to determine long-term effects of GE MSCs on tendon healing, we conclude that genetically engineered MSCs may be a promising therapeutic tool for accelerating short-term functional recovery in the treatment of tendon injuries.},
	keywords = {Achilles Tendon/pathology Animals Biomechanical Phenomena Bone Morphogenetic Protein 2/*metabolism Elastic Modulus Female Fibrin/metabolism *Gene Expression Regulation Genetic Engineering/methods Mesenchymal Stromal Cells/*cytology Mice Mice, Inbred C3H Smad8 Protein/*metabolism Tendon Injuries/pathology Tendons/pathology Tissue Engineering/*methods Wound Healing},
	issn = {1554-527X (Electronic) 0736-0266 (Linking)},
	doi = {10.1002/jor.22167},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/22696396},
	author = {Pelled, G. and Snedeker, J. G. and Ben-Arav, A. and Rigozzi, S. and Zilberman, Y. and Kimelman-Bleich, N. and Gazit, Z. and Muller, R. and Gazit, D.}
}
@article {17112,
	title = {Microcomputed tomography-based structural analysis of various bone tissue regeneration models},
	journal = {Nat Protoc},
	volume = {6},
	number = {1},
	year = {2011},
	note = {Kallai, Ilan Mizrahi, Olga Tawackoli, Wafa Gazit, Zulma Pelled, Gadi Gazit, Dan eng R01AR056694/AR/NIAMS NIH HHS/ R01DE019902/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. England 2011/01/08 06:00 Nat Protoc. 2011 Jan;6(1):105-10. doi: 10.1038/nprot.2010.180. Epub 2011 Jan 6.},
	pages = {105-10},
	type = {Journal Article},
	abstract = {Microcomputed tomography (microCT) analysis is a powerful tool for the evaluation of bone tissue because it provides access to the 3D microarchitecture of the bone. It is invaluable for regenerative medicine as it provides the researcher with the opportunity to explore the skeletal system both in vivo and ex vivo. The quantitative assessment of macrostructural characteristics and microstructural features may improve our ability to estimate the quality of newly formed bone. We have developed a unique procedure for analyzing data from microCT scans to evaluate bone structure and repair. This protocol describes the procedures for microCT analysis of three main types of mouse bone regeneration models (ectopic administration of bone-forming mesenchymal stem cells, and administration of cells after both long bone defects and cranial segmental bone defects) that can be easily adapted for a variety of other models. Precise protocols are crucial because the system is extremely user sensitive and results can be easily biased if standardized methods are not applied. The suggested protocol takes 1.5-3.5 h per sample, depending on bone tissue sample size, the type of equipment used, variables of the scanning protocol and the operator{\textquoteright}s experience.},
	keywords = {Animals *Bone Regeneration Bone and Bones/pathology/*physiology/ultrastructure Mice Models, Biological X-Ray Microtomography/*methods},
	issn = {1750-2799 (Electronic) 1750-2799 (Linking)},
	doi = {10.1038/nprot.2010.180},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/21212786},
	author = {Kallai, I. and Mizrahi, O. and Tawackoli, W. and Gazit, Z. and Pelled, G. and Gazit, D.}
}
@article {17117,
	title = {Ovariectomy and genes encoding core circadian regulatory proteins in murine bone},
	journal = {Osteoporos Int},
	volume = {22},
	number = {5},
	year = {2011},
	note = {Smith, B J Sutton, G M Wu, X Yu, G Goh, B C Hebert, T Pelled, G Gazit, Z Gazit, D Butler, A A Gimble, J M eng MYNCBI/CB/NCI NIH HHS/ P30 DK072476/DK/NIDDK NIH HHS/ 1P30 DK072476/DK/NIDDK NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t England 2010/07/02 06:00 Osteoporos Int. 2011 May;22(5):1633-9. doi: 10.1007/s00198-010-1325-z. Epub 2010 Jul 1.},
	pages = {1633-9},
	type = {Journal Article},
	abstract = {SUMMARY: This study investigated the influence of ovarian hormone deficiency on core circadian regulatory protein (CCRP) in the context of bone loss. Our data suggest that ovarian hormone deficiency disrupts diurnal rhythmicity and CCRP expression in bone. Further studies should determine if chronobiology provides a novel therapeutic target for osteoporosis intervention. INTRODUCTION: CCRP synchronize metabolic activities and display an oscillatory expression profile in murine bone. In vitro studies using bone marrow mesenchymal stromal/stem cells have demonstrated that the CCRP is present and can be regulated within osteoblast progenitors. In vivo studies have shown that the CCRP regulates bone mass via leptin/neuroendocrine pathways. The current study used an ovariectomized murine model to test the hypothesis that ovarian hormone deficiency is associated with either an attenuation and/or temporal phase shift of the CCRP oscillatory expression in bone and that these changes are correlated with the onset of osteoporosis. METHODS: Sham-operated controls and ovariectomized female C57BL/6 mice were euthanized at 4-h intervals 2 weeks post-operatively. RESULTS: Ovariectomy attenuated the oscillatory expression of CCRP mRNAs in the femur and vertebra relative to the controls and reduced the wheel-running activity profile. CONCLUSION: Ovarian hormone deficiency modulates the expression profile of the CCRP with potential impact on bone marrow mesenchymal stem cell lineage commitment.},
	keywords = {Animal Estrogens/deficiency/*physiology Female Femur/metabolism Gene Expression Profiling/methods Gene Expression Regulation/physiology Lumbar Vertebrae/metabolism Mice Mice, Animals Circadian Rhythm/genetics/*physiology Circadian Rhythm Signaling Peptides and Proteins/*biosynthesis/genetics Disease Models, Inbred C57BL Motor Activity/physiology Osteoporosis/genetics/metabolism/*physiopathology Ovariectomy RNA, Mechanical X-Ray Microtomography/methods, Messenger/genetics Stress},
	issn = {1433-2965 (Electronic) 0937-941X (Linking)},
	doi = {10.1007/s00198-010-1325-z},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20593165},
	author = {Smith, B. J. and Sutton, G. M. and Wu, X. and Yu, G. and Goh, B. C. and Hebert, T. and Pelled, G. and Gazit, Z. and Gazit, D. and Butler, A. A. and Gimble, J. M.}
}
@article {17119,
	title = {Patterning osteogenesis by inducible gene expression in microfluidic culture systems},
	journal = {Integr Biol (Camb)},
	volume = {3},
	number = {1},
	year = {2011},
	note = {Zhang, Yue Gazit, Zulma Pelled, Gadi Gazit, Dan Vunjak-Novakovic, Gordana eng P41 EB002520/EB/NIBIB NIH HHS/ R01 DE016525/DE/NIDCR NIH HHS/ DE161525/DE/NIDCR NIH HHS/ EB02520/EB/NIBIB NIH HHS/ Research Support, N.I.H., Extramural England 2010/10/07 06:00 Integr Biol (Camb). 2011 Jan;3(1):39-47. doi: 10.1039/c0ib00053a. Epub 2010 Oct 5.},
	pages = {39-47},
	type = {Journal Article},
	abstract = {The development of transitional interfacial zones between adjacent tissues remains a significant challenge for developing tissue engineering and regenerative medicine strategies. Using osteogenic differentiation as a model, we describe a novel approach to spatially regulate expression and secretion of the bone morphogenetic protein (BMP-2) in a two-dimensional field of cultured cells, by flow patterning the modulators of inducible BMP-2 gene expression. We first demonstrate control of gene expression, and of osteogenic differentiation of the cell line with inducible expression of BMP-2. Then we design laminar flow systems, with patterned delivery of Doxycycline (Dox), the expression modulator of BMP-2. The patterned concentration profiles were verified by computational simulation and dye separation experiments. Patterned differentiation experiments conducted in the flow systems for a period of three weeks showed the Dox concentration dependent osteogenic differentiation, as evidenced by mineral deposition. In summary, by combining inducible gene expression with laminar flow technologies, this study provided an innovative way to engineer tissue interfaces.},
	keywords = {Animals Bone Morphogenetic Protein 2/biosynthesis/genetics Cell Differentiation Cell Line Doxycycline/pharmacology Gene Expression/drug effects Humans Mesenchymal Stromal Cells/cytology/drug effects/metabolism Mice *Microfluidic Analytical Techniques Mode, Biological Osteogenesis/drug effects/*genetics/physiology Recombinant Proteins/biosynthesis/genetics Systems Biology Tissue Engineering Transfection},
	issn = {1757-9708 (Electronic) 1757-9694 (Linking)},
	doi = {10.1039/c0ib00053a},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20924519},
	author = {Zhang, Y. and Gazit, Z. and Pelled, G. and Gazit, D. and Vunjak-Novakovic, G.}
}
@article {17113,
	title = {Targeted gene-and-host progenitor cell therapy for nonunion bone fracture repair},
	journal = {Mol Ther},
	volume = {19},
	number = {1},
	year = {2011},
	note = {Kimelman-Bleich, Nadav Pelled, Gadi Zilberman, Yoram Kallai, Ilan Mizrahi, Olga Tawackoli, Wafa Gazit, Zulma Gazit, Dan eng R01 AR056694/AR/NIAMS NIH HHS/ R01 DE019902/DE/NIDCR NIH HHS/ R01AR056694-01A1/AR/NIAMS NIH HHS/ R01DE019902-01/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2010/09/23 06:00 Mol Ther. 2011 Jan;19(1):53-9. doi: 10.1038/mt.2010.190. Epub 2010 Sep 21.},
	pages = {53-9},
	type = {Journal Article},
	abstract = {Nonunion fractures present a challenge to orthopedics with no optimal solution. In-vivo DNA electroporation is a gene-delivery technique that can potentially accelerate regenerative processes. We hypothesized that in vivo electroporation of an osteogenic gene in a nonunion radius bone defect site would induce fracture repair. Nonunion fracture was created in the radii of C3H/HeN mice, into which a collagen sponge was placed. To allow for recruitment of host progenitor cells (HPCs) into the implanted sponge, the mice were housed for 10 days before electroporation. Mice were electroporated with either bone morphogenetic protein 9 (BMP-9) plasmid, Luciferase plasmid or injected with BMP-9 plasmid but not electroporated. In vivo bioluminescent imaging indicated that gene expression was localized to the defect site. Microcomputed tomography (microCT) and histological analysis of murine radii electroporated with BMP-9 demonstrated bone formation bridging the bone gap, whereas in the control groups the defect remained unbridged. Population of the implanted collagen sponge by HPCs transfected with the injected plasmid following electroporation was noted. Our data indicate that regeneration of nonunion bone defect can be attained by performing in vivo electroporation with an osteogenic gene combined with recruitment of HPCs. This gene therapy approach may pave the way for regeneration of other skeletal tissues.},
	keywords = {Animals Bone Regeneration/*genetics Collagen/administration \& dosage Electroporation/*methods Female Fractures, Inbred C3H Osteogenesis/*genetics Plasmids/genetics Stem Cells/*physiology Wound Healing/genetics, Ununited/pathology/*therapy Gene Transfer Techniques Genetic Therapy/*methods Genetic Vectors/administration \& dosage/genetics Growth Differentiation Factor 2/biosynthesis/*genetics Luciferases/biosynthesis/genetics Mice Mice},
	issn = {1525-0024 (Electronic) 1525-0016 (Linking)},
	doi = {10.1038/mt.2010.190},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20859259},
	author = {Kimelman-Bleich, N. and Pelled, G. and Zilberman, Y. and Kallai, I. and Mizrahi, O. and Tawackoli, W. and Gazit, Z. and Gazit, D.}
}
@article {17106,
	title = {Direct gene therapy for bone regeneration: gene delivery, animal models, and outcome measures},
	journal = {Tissue Eng Part B Rev},
	volume = {16},
	number = {1},
	year = {2010},
	note = {Pelled, Gadi Ben-Arav, Ayelet Hock, Colleen Reynolds, David G Yazici, Cemal Zilberman, Yoram Gazit, Zulma Awad, Hani Gazit, Dan Schwarz, Edward M eng R01 DE019902-01/DE/NIDCR NIH HHS/ P50 AR054041/AR/NIAMS NIH HHS/ R01 DE019902/DE/NIDCR NIH HHS/ R21 DE017096/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Review 2010/02/11 06:00 Tissue Eng Part B Rev. 2010 Feb;16(1):13-20. doi: 10.1089/ten.teb.2009.0156.},
	pages = {13-20},
	type = {Journal Article},
	abstract = {While various problems with bone healing remain, the greatest clinical change is the absence of an effective approach to manage large segmental defects in limbs and craniofacial bones caused by trauma or cancer. Thus, nontraditional forms of medicine, such as gene therapy, have been investigated as a potential solution. The use of osteogenic genes has shown great potential in bone regeneration and fracture healing. Several methods for gene delivery to the fracture site have been described. The majority of them include a cellular component as the carrying vector, an approach known as cell-mediated gene therapy. Yet, the complexity involved with cell isolation and culture emphasizes the advantages of direct gene delivery as an alternative strategy. Here we review the various approaches of direct gene delivery for bone repair, the choice of animal models, and the various outcome measures required to evaluate the efficiency and safety of each technique. Special emphasis is given to noninvasive, quantitative, in vivo monitoring of gene expression and biodistribution in live animals. Research efforts should aim at inducing a transient, localized osteogenic gene expression within a fracture site to generate an effective therapeutic approach that would eventually lead to clinical use.},
	keywords = {Animal Treatment Outcome, Animals Bone Regeneration/*physiology Fracture Healing/physiology *Gene Transfer Techniques Genetic Therapy/*methods *Models},
	issn = {1937-3376 (Electronic) 1937-3368 (Linking)},
	doi = {10.1089/ten.teb.2009.0156},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20143927},
	author = {Pelled, G. and Ben-Arav, A. and Hock, C. and Reynolds, D. G. and Yazici, C. and Zilberman, Y. and Gazit, Z. and Awad, H. and Gazit, D. and Schwarz, E. M.}
}
@article {17109,
	title = {The effect of simulated microgravity on human mesenchymal stem cells cultured in an osteogenic differentiation system: a bioinformatics study},
	journal = {Tissue Eng Part A},
	volume = {16},
	number = {11},
	year = {2010},
	note = {Sheyn, Dima Pelled, Gadi Netanely, Dvir Domany, Eytan Gazit, Dan eng R01AR056694-01A1/AR/NIAMS NIH HHS/ R01DE019902-01/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2010/09/03 06:00 Tissue Eng Part A. 2010 Nov;16(11):3403-12. doi: 10.1089/ten.tea.2009.0834. Epub 2010 Aug 31.},
	pages = {3403-12},
	type = {Journal Article},
	abstract = {One proposed strategy for bone regeneration involves ex vivo tissue engineering, accomplished using bone-forming cells, biodegradable scaffolds, and dynamic culture systems, with the goal of three-dimensional tissue formation. Rotating wall vessel bioreactors generate simulated microgravity conditions ex vivo, which lead to cell aggregation. Human mesenchymal stem cells (hMSCs) have been extensively investigated and shown to possess the potential to differentiate into several cell lineages. The goal of the present study was to evaluate the effect of simulated microgravity on all genes expressed in hMSCs, with the underlying hypothesis that many important pathways are affected during culture within a rotating wall vessel system. Gene expression was analyzed using a whole genome microarray and clustering with the aid of the National Institutes of Health{\textquoteright}s Database for Annotation, Visualization and Integrated Discovery database and gene ontology analysis. Our analysis showed 882 genes that were downregulated and 505 genes that were upregulated after exposure to simulated microgravity. Gene ontology clustering revealed a wide variety of affected genes with respect to cell compartment, biological process, and signaling pathway clusters. The data sets showed significant decreases in osteogenic and chondrogenic gene expression and an increase in adipogenic gene expression, indicating that ex vivo adipose tissue engineering may benefit from simulated microgravity. This finding was supported by an adipogenic differentiation assay. These data are essential for further understanding of ex vivo tissue engineering using hMSCs.},
	keywords = {Adipogenesis/genetics Aged Bioreactors *Cell Differentiation/genetics Cell Survival Cells, Cultured Cluster Analysis *Computational Biology Down-Regulation/genetics Female Humans Male Mesenchymal Stromal Cells/*cytology/metabolism Mesoderm/metabolism Middle Aged Organ Specificity/genetics *Osteogenesis/genetics Up-Regulation/genetics *Weightle},
	issn = {1937-335X (Electronic) 1937-3341 (Linking)},
	doi = {10.1089/ten.tea.2009.0834},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20807102},
	author = {Sheyn, D. and Pelled, G. and Netanely, D. and Domany, E. and Gazit, D.}
}
@article {17108,
	title = {Genetically modified cells in regenerative medicine and tissue engineering},
	journal = {Adv Drug Deliv Rev},
	volume = {62},
	number = {7-8},
	year = {2010},
	note = {Sheyn, Dima Mizrahi, Olga Benjamin, Shimon Gazit, Zulma Pelled, Gadi Gazit, Dan eng R01 AR056694/AR/NIAMS NIH HHS/ R03 AR057143/AR/NIAMS NIH HHS/ R43 AR057587/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. Review Netherlands 2010/02/02 06:00 Adv Drug Deliv Rev. 2010 Jun 15;62(7-8):683-98. doi: 10.1016/j.addr.2010.01.002. Epub 2010 Jan 28.},
	pages = {683-98},
	type = {Journal Article},
	abstract = {Regenerative medicine appears to take as its patron, the Titan Prometheus, whose liver was able to regenerate daily, as the field attempts to restore lost, damaged, or aging cells and tissues. The tremendous technological progress achieved during the last decade in gene transfer methods and imaging techniques, as well as recent increases in our knowledge of cell biology, have opened new horizons in the field of regenerative medicine. Genetically engineered cells are a tool for tissue engineering and regenerative medicine, albeit a tool whose development is fraught with difficulties. Gene-and-cell therapy offers solutions to severe problems faced by modern medicine, but several impediments obstruct the path of such treatments as they move from the laboratory toward the clinical setting. In this review we provide an overview of recent advances in the gene-and-cell therapy approach and discuss the main hurdles and bottlenecks of this approach on its path to clinical trials and prospective clinical practice.},
	keywords = {Animals Cell Transplantation/methods Clinical Trials as Topic Genetic Engineering/*methods Genetic Therapy/methods Humans Regenerative Medicine/*methods Stem Cell Transplantation/methods Tissue Engineering/*methods},
	issn = {1872-8294 (Electronic) 0169-409X (Linking)},
	doi = {10.1016/j.addr.2010.01.002},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20114067},
	author = {Sheyn, D. and Mizrahi, O. and Benjamin, S. and Gazit, Z. and Pelled, G. and Gazit, D.}
}
@article {17110,
	title = {Genetically modified mesenchymal stem cells induce mechanically stable posterior spine fusion},
	journal = {Tissue Eng Part A},
	volume = {16},
	number = {12},
	year = {2010},
	note = {Sheyn, Dima Ruthemann, Martin Mizrahi, Olga Kallai, Ilan Zilberman, Yoram Tawackoli, Wafa Kanim, Linda E A Zhao, Li Bae, Hyun Pelled, Gadi Snedeker, Jess G Gazit, Dan eng R01AR056694-01A1/AR/NIAMS NIH HHS/ R01DE019902-01/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural 2010/07/14 06:00 Tissue Eng Part A. 2010 Dec;16(12):3679-86. doi: 10.1089/ten.TEA.2009.0786. Epub 2010 Sep 28.},
	pages = {3679-86},
	type = {Journal Article},
	abstract = {Most spine fusion procedures involve the use of prosthetic fixation devices combined with autologous bone grafts rather than biological treatment. We had shown that spine fusion could be achieved by injection of bone morphogenetic protein-2 (BMP-2)-expressing mesenchymal stem cells (MSCs) into the paraspinal muscle. In this study, we hypothesized that posterior spinal fusion achieved using genetically modified MSCs would be mechanically comparable to that realized using a mechanical fixation. BMP-2-expressing MSCs were injected bilaterally into paravertebral muscles of the mouse lumbar spine. In one control group BMP-2 expression was inhibited. Microcomputed tomography and histological analyses were used to evaluate bone formation. For comparison, a group of mouse spines were bilaterally fused with stainless steel pins. The harvested spines were later tested using a custom four-point bending apparatus and structural bending stiffness was estimated. To assess the degree to which MSC vertebral fusion was targeted and to quantify the effects of fusion on adjacent spinal segments, images of the loaded spine curvature were analyzed to extract rigidity of the individual spinal segments. Bone bridging of the targeted vertebrae was observed in the BMP-2-expressing MSC group, whereas no bone formation was noted in any control group. The biomechanical tests showed that MSC-mediated spinal fusion was as effective as stainless steel pin-based fusion and significantly more rigid than the control groups. Local analysis showed that the distribution of stiffness in the MSC-based fusion group was similar to that in the steel pin fusion group, with the majority of spinal stiffness contributed by the targeted fusion at L3-L5. Our findings demonstrate that MSC-induced spinal fusion can convey biomechanical rigidity to a targeted segment that is comparable to that achieved using an instrumental fixation.},
	keywords = {Animals Biomechanical Phenomena Cell Line Female Immunohistochemistry Mesenchymal Stromal Cells/*cytology/*metabolism/physiology Mice Osteogenesis/genetics/physiology *Spinal Fusion Spine/cytology/surgery X-Ray Microtomography},
	issn = {1937-335X (Electronic) 1937-3341 (Linking)},
	doi = {10.1089/ten.TEA.2009.0786},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20618082},
	author = {Sheyn, D. and Ruthemann, M. and Mizrahi, O. and Kallai, I. and Zilberman, Y. and Tawackoli, W. and Kanim, L. E. and Zhao, L. and Bae, H. and Pelled, G. and Snedeker, J. G. and Gazit, D.}
}
@article {17107,
	title = {Monitoring of the effect of intervertebral disc nucleus pulposus ablation by MRI},
	journal = {NMR Biomed},
	volume = {23},
	number = {6},
	year = {2010},
	note = {Saar, Galit Zilberman, Yoram Shinar, Hadassah Keinan-Adamsky, Keren Pelled, Gadi Gazit, Dan Navon, Gil eng Research Support, Non-U.S. Gov{\textquoteright}t England 2010/02/23 06:00 NMR Biomed. 2010 Jul;23(6):554-62. doi: 10.1002/nbm.1493.},
	pages = {554-62},
	type = {Journal Article},
	abstract = {In order to investigate intervertebral disc (IVD) degeneration and repair, a quantitative non-invasive tool is needed. Various MRI methods including qCPMG, which yields dipolar echo relaxation time (T(DE)), magnetization transfer contrast (MTC), and (1)H and (2)H double quantum filtered (DQF) MRI were used in the present work to monitor changes in rat IVD after ablation of the nucleus pulposus (NP), serving as a model of severe IVD degeneration. In the intact IVD, a clear distinction between the annulus fibrosus (AF) and the NP is obtained on T(2) and T(DE) weighted images as well as on MTC maps, reflecting the high concentration of ordered collagen fibers in the AF. After ablation of the NP, the distinction between the compartments is lost. T(2) and T(DE) relaxation times are short throughout the disc and MTC is high. (1)H and (2)H DQF signal, which in intact discs is obtained only for the AF, is now observable throughout the tissue. These results indicate that after ablation, there is an ingression of collagen fibers from the AF into the area that was previously occupied by the NP, as was confirmed by histology.},
	keywords = {Animals Collagen/metabolism Female Humans Intervertebral Disc/metabolism/*pathology Intervertebral Disc Degeneration/*pathology Magnetic Resonance Imaging/*methods Rats Rats, Wistar},
	issn = {1099-1492 (Electronic) 0952-3480 (Linking)},
	doi = {10.1002/nbm.1493},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20175140},
	author = {Saar, G. and Zilberman, Y. and Shinar, H. and Keinan-Adamsky, K. and Pelled, G. and Gazit, D. and Navon, G.}
}
@article {17104,
	title = {Peptide-modified "smart" hydrogels and genetically engineered stem cells for skeletal tissue engineering},
	journal = {Biomacromolecules},
	volume = {11},
	number = {6},
	year = {2010},
	note = {Garty, Shai Kimelman-Bleich, Nadav Hayouka, Zvi Cohn, Daniel Friedler, Assaf Pelled, Gadi Gazit, Dan eng R01AR056694/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 2010/05/14 06:00 Biomacromolecules. 2010 Jun 14;11(6):1516-26. doi: 10.1021/bm100157s.},
	pages = {1516-26},
	type = {Journal Article},
	abstract = {Stimuli responsive or "smart" hydrogels are of interest for tissue-engineering applications, featuring the advantages of minimally invasive application. Currently, these materials have yet to be used as a biological replacement in restoring the function of damaged tissues or organs. The aim of this study was to demonstrate the advantages of thermoresponsive, peptide-containing hydrogels as a supportive matrix for genetically engineered stem cells. We used injectable hydrogels, enabling cell delivery to the desired site and providing adequate scaffolding postimplantation. Thermoresponsive hydrogels were developed based on amphiphilic block copolymers of polyethylene-oxide and polypropylene-oxide end-capped with methacrylate or maleimide entities and further reacted with RGD-containing peptides. Cell metabolic activity and survival within those hydrogels was studied, illustrating that the stable peptide-polymer conjugate is required for prolonged cell support. The unique polymer characteristics, combined with its enhanced cell interactions, suggest the potential use of these biomaterials in various tissue engineering applications.},
	keywords = {Animals Bone Morphogenetic Protein 2/*genetics Bone and Bones/*cytology/metabolism Cell Culture Techniques Cell Line Cell Proliferation Cell Survival DNA/metabolism Humans Hydrogels/*chemistry Mesenchymal Stromal Cells/*cytology/metabolism Microscopy, Electron, Scanning Oligopeptides/*chemistry Rheology Solubility Spectrometry, X-Ray Emission Surface Properties Temperature Tissue Engineering/*methods Tissue Scaffolds/*chemistry Viscosity},
	issn = {1526-4602 (Electronic) 1525-7797 (Linking)},
	doi = {10.1021/bm100157s},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20462241},
	author = {Garty, S. and Kimelman-Bleich, N. and Hayouka, Z. and Cohn, D. and Friedler, A. and Pelled, G. and Gazit, D.}
}
@article {17111,
	title = {Quantitative microstructural studies of the armor of the marine threespine stickleback (Gasterosteus aculeatus)},
	journal = {J Struct Biol},
	volume = {171},
	number = {3},
	year = {2010},
	note = {Song, Juha Reichert, Steffen Kallai, Ilan Gazit, Dan Wund, Matthew Boyce, Mary C Ortiz, Christine eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 2010/05/04 06:00 J Struct Biol. 2010 Sep;171(3):318-31. doi: 10.1016/j.jsb.2010.04.009. Epub 2010 Apr 28.},
	pages = {318-31},
	type = {Journal Article},
	abstract = {In this study, a quantitative investigation of the microstructure and composition of field-caught marine Gasterosteus aculeatus (threespine stickleback) armor is presented, which provides useful phylogenetic information and insights into biomechanical function. Micro-computed tomography (microCT) was employed to create full three-dimensional images of the dorsal spines and basal plate, lateral plates, pelvic girdle and spines and to assess structural and compositional properties such as the spatial distribution of thickness (approximately 100-300 microm), the heterogeneous cross-sectional geometry (centrally thickened), plate-to-plate juncture and overlap (approximately 50\% of the plate width), and bone mineral density (634-748 HA/cm(3)). The convolution of plate geometry in conjunction with plate-to-plate overlap allows a relatively constant armor thickness to be maintained throughout the assembly, promoting spatially homogeneous protection and thereby avoiding weakness at the armor unit interconnections. Plate-to-plate junctures act to register and join the plates while permitting compliance in sliding and rotation in selected directions. Mercury porosimetry was used to determine the pore size distribution and volume percent porosity of the lateral plates (20-35 vol.\%) and spines (10-15 vol.\%). SEM and microCT revealed a porous, sandwich-like cross-section beneficial for bending stiffness and strength at minimum weight. Back-scattered electron microscopy and energy dispersive X-ray analysis were utilized to quantify the weight percent mineral content (58-68\%). Scanning electron microscopy and surface profilometry were used to characterize the interior and exterior surface topography (tubercles) of the lateral plates. The results obtained in this study are discussed in the context of mechanical function, performance, fitness, and survivability.},
	keywords = {Animal Structures/*anatomy \& histology/chemistry/ultrastructure Animals Microscopy, Electron, Fourier Transform Infrared X-Ray Diffraction X-Ray Microtomography, Scanning Smegmamorpha/*anatomy \& histology Spectrometry, X-Ray Emission Spectroscopy},
	issn = {1095-8657 (Electronic) 1047-8477 (Linking)},
	doi = {10.1016/j.jsb.2010.04.009},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20433929},
	author = {Song, J. and Reichert, S. and Kallai, I. and Gazit, D. and Wund, M. and Boyce, M. C. and Ortiz, C.}
}
@article {17105,
	title = {Quantitative, structural, and image-based mechanical analysis of nonunion fracture repaired by genetically engineered mesenchymal stem cells},
	journal = {J Biomech},
	volume = {43},
	number = {12},
	year = {2010},
	note = {Kallai, Ilan van Lenthe, G Harry Ruffoni, Davide Zilberman, Yoram Muller, Ralph Pelled, Gadi Gazit, Dan eng R01DE019902-01/DE/NIDCR NIH HHS/ R01 DE019902-01/DE/NIDCR NIH HHS/ R01 AR056694-01A1/AR/NIAMS NIH HHS/ R01AR056694-01A1/AR/NIAMS NIH HHS/ R01 DE019902/DE/NIDCR NIH HHS/ R01 AR056694/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2010/05/18 06:00 J Biomech. 2010 Aug 26;43(12):2315-20. doi: 10.1016/j.jbiomech.2010.04.031. Epub 2010 May 14.},
	pages = {2315-20},
	type = {Journal Article},
	abstract = {Stem cell-mediated gene therapy for fracture repair, utilizes genetically engineered mesenchymal stem cells (MSCs) for the induction of bone growth and is considered a promising approach in skeletal tissue regeneration. Previous studies have shown that murine nonunion fractures can be repaired by implanting MSCs over-expressing recombinant human bone morphogenetic protein-2 (rhBMP-2). Nanoindentation studies of bone tissue induced by MSCs in a radius fracture site indicated similar elastic modulus compared to intact murine bone, eight weeks post-treatment. In the present study we sought to investigate temporal changes in microarchitecture and biomechanical properties of repaired murine radius bones, following the implantation of MSCs. High-resolution micro-computed tomography (micro-CT) was performed 10 and 35 weeks post MSC implantation, followed by micro-finite element (micro-FE) analysis. The results have shown that the regenerated bone tissue remodels over time, as indicated by a significant decrease in bone volume, total volume, and connectivity density combined with an increase in mineral density. In addition, the axial stiffness of limbs repaired with MSCs was 2-1.5 times higher compared to the contralateral intact limbs, at 10 and 35 weeks post-treatment. These results could be attributed to the fusion that occurred in between the ulna and radius bones. In conclusion, although MSCs induce bone formation, which exceeds the fracture site, significant remodeling of the repair callus occurs over time. In addition, limbs treated with an MSC graft demonstrated superior biomechanical properties, which could indicate the clinical benefit of future MSC application in nonunion fracture repair.},
	keywords = {Animal Elastic Modulus Female Finite Element Analysis Fractures, Animals Biomechanical Phenomena Bone Density Bone Morphogenetic Protein 2/genetics Bone Regeneration Bone Remodeling Disease Models, Inbred C3H Radius Fractures/diagnostic imaging/physiopathology/therapy Recombinant Proteins/genetics X-Ray Microtomography, Ununited/diagnostic imaging/physiopathology/*therapy Genetic Engineering Genetic Therapy/*methods Humans *Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells/metabolism Mice Mice},
	issn = {1873-2380 (Electronic) 0021-9290 (Linking)},
	doi = {10.1016/j.jbiomech.2010.04.031},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/20471652},
	author = {Kallai, I. and van Lenthe, G. H. and Ruffoni, D. and Zilberman, Y. and Muller, R. and Pelled, G. and Gazit, D.}
}
@article {17101,
	title = {An analytical model for elucidating tendon tissue structure and biomechanical function from in vivo cellular confocal microscopy images},
	journal = {Cells Tissues Organs},
	volume = {190},
	number = {2},
	year = {2009},
	note = {Snedeker, J G Pelled, G Zilberman, Y Ben Arav, A Huber, E Muller, R Gazit, D eng Research Support, Non-U.S. Gov{\textquoteright}t Switzerland 2009/01/06 09:00 Cells Tissues Organs. 2009;190(2):111-9. doi: 10.1159/000189211. Epub 2008 Dec 22.},
	pages = {111-9},
	type = {Journal Article},
	abstract = {Fibered confocal laser scanning microscopes have given us the ability to image fluorescently labeled biological structures in vivo and at exceptionally high spatial resolutions. By coupling this powerful imaging modality with classic optical elastography methods, we have developed novel techniques that allow us to assess functional mechanical integrity of soft biological tissues by measuring the movements of cells in response to externally applied mechanical loads. Using these methods we can identify minute structural defects, monitor the progression of certain skeletal tissue disease states, and track subsequent healing following therapeutic intervention in the living animal. Development of these methods using a murine Achilles tendon model has revealed that the hierarchical and composite anatomical structure of the tendon presents various technical challenges that can confound a mechanical analysis of local material properties. Specifically, interfascicle gliding can yield complex cellular motions that must be interpreted within the context of an appropriate anatomical model. In this study, we explore the various classes of cellular images that may result from fibered confocal microscopy of the murine Achilles tendon, and introduce a simple two-fascicle model to interpret the images in terms of mechanical strains within the fascicles, as well as the relative gliding between fascicles.},
	keywords = {*Achilles Tendon/cytology/physiology Animals Biomechanical Phenomena Female Imaging, Confocal/methods *Models, Structural, Three-Dimensional/methods Mice Microscopy},
	issn = {1422-6421 (Electronic) 1422-6405 (Linking)},
	doi = {10.1159/000189211},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19122452},
	author = {Snedeker, J. G. and Pelled, G. and Zilberman, Y. and Ben Arav, A. and Huber, E. and Muller, R. and Gazit, D.}
}
@article {17098,
	title = {Circadian rhythm of osteocalcin in the maxillomandibular complex},
	journal = {J Dent Res},
	volume = {88},
	number = {1},
	year = {2009},
	note = {Gafni, Y Ptitsyn, A A Zilberman, Y Pelled, G Gimble, J M Gazit, D eng R21 DE016371/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2009/01/10 09:00 J Dent Res. 2009 Jan;88(1):45-50. doi: 10.1177/0022034508328012.},
	pages = {45-50},
	type = {Journal Article},
	abstract = {The human body displays central circadian rhythms of activity. Recent findings suggest that peripheral tissues, such as bone, possess their own circadian clocks. Studies have shown that osteocalcin protein levels oscillate over a 24-hour period, yet the specific skeletal sites involved and its transcriptional profile remain unknown. The current study aimed to test the hypothesis that peripheral circadian mechanisms regulate transcription driven by the osteocalcin promoter. Transgenic mice harboring the human osteocalcin promoter linked to a luciferase reporter gene were used. Mice of both genders and various ages were analyzed non-invasively at sequential times throughout 24-hour periods. Statistical analyses of luminescent signal intensity of osteogenic activity from multiple skeletal sites indicated a periodicity of   24 hrs. The maxillomandibular complex displayed the most robust oscillatory pattern. These findings have implications for dental treatments in orthodontics and maxillofacial surgery, as well as for the mechanisms underlying bone remodeling in the maxillomandibular complex.},
	keywords = {Animal Osteocalcin/*genetics Osteogenesis/genetics Promoter Regions, Animals Carpal Bones/anatomy \& histology/metabolism Circadian Rhythm/*genetics Female Gene Expression Regulation/genetics Half-Life Humans Image Processing, Computer-Assisted/methods Luciferases Luminescence Male Mandible/anatomy \& histology/*metabolism Maxilla/anatomy \& histology/*metabolism Mice Mice, Genetic/genetics, Genetic/genetics Sex Factors Skull/anatomy \& histology/metabolism Tail/anatomy \& histology/metabolism Tarsal Bones/anatomy \& histology/metabolism Transcription, Transgenic Models},
	issn = {1544-0591 (Electronic) 0022-0345 (Linking)},
	doi = {10.1177/0022034508328012},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19131316},
	author = {Gafni, Y. and Ptitsyn, A. A. and Zilberman, Y. and Pelled, G. and Gimble, J. M. and Gazit, D.}
}
@article {17097,
	title = {Expression of Brachyury in mesenchymal progenitor cells leads to cartilage-like tissue that is resistant to the destructive effect of rheumatoid arthritis synovial fibroblasts},
	journal = {J Tissue Eng Regen Med},
	volume = {3},
	number = {2},
	year = {2009},
	note = {Dinser, Robert Pelled, Gadi Muller-Ladner, Ulf Gazit, Dan Neumann, Elena eng Research Support, Non-U.S. Gov{\textquoteright}t England 2009/01/22 09:00 J Tissue Eng Regen Med. 2009 Feb;3(2):124-8. doi: 10.1002/term.148.},
	pages = {124-8},
	type = {Journal Article},
	abstract = {Our objectives were to determine the chondrogenic potential of a murine Brachyury-transformed mesenchymal progenitor cell line in the presence of rheumatoid arthritis-activated synovial fibroblasts (RASFs). Brachyury-transformed mesenchymal progenitor cells were implanted alone or combined with RASFs isolated from diseased human joints in each of six immunodeficient SCID mice. De novo tissue formation was analysed by histology and immunohistochemistry after 60 days. Spheroid nodules resembling cartilage morphologically and by the expression of proteoglycans and collagen II developed in four of six implants in the absence and in five of six implants in the presence of RASFs. No evidence for hypertrophic differentiation could be observed. Mesenchymal progenitor cells transformed with Brachyury are able to produce a cartilage like tissue in vivo over an extended period of time that is resistant to the destructive effect of RASF. This observation may provide opportunities for a cell-based reconstructive treatment in joint disease.},
	keywords = {Animals Arthritis, Rheumatoid/*pathology Cartilage/*cytology Female Fetal Proteins/*metabolism Fibroblasts/pathology Immunohistochemistry Mesenchymal Stromal Cells/*metabolism Mice Mice, SCID Synovial Membrane/*pathology T-Box Domain Proteins/*metabolism},
	issn = {1932-7005 (Electronic) 1932-6254 (Linking)},
	doi = {10.1002/term.148},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19156687},
	author = {Dinser, R. and Pelled, G. and Muller-Ladner, U. and Gazit, D. and Neumann, E.}
}
@article {17100,
	title = {Functional fibered confocal microscopy: a promising tool for assessing tendon regeneration},
	journal = {Tissue Eng Part C Methods},
	volume = {15},
	number = {3},
	year = {2009},
	note = {Snedeker, Jess G Ben Arav, Ayelet Zilberman, Yoram Pelled, Gadi Gazit, Dan eng Evaluation Studies 2009/04/16 09:00 Tissue Eng Part C Methods. 2009 Sep;15(3):485-91. doi: 10.1089/ten.tec.2008.0612.},
	pages = {485-91},
	type = {Journal Article},
	abstract = {This work advances fibered confocal microscopy (FCM) as a functional imaging platform for in vivo assessment of tissue mechanics. Building on our earlier studies demonstrating proof of principle and introducing an analytical framework for FCM image processing, here we present data that improve and validate several critical aspects of FCM. Specifically, we have considerably reduced the invasiveness of the imaging procedure, and verified that endoscopic imaging through a transcutaneous access point does not induce functional changes in passive ankle joint biomechanics. We have also verified that periodic (weekly) measurements on uninjured tendons are reproducible. Importantly, we have further proven that the method can sensitively detect and quantify compromised tendon mechanics in injured tendons. These incremental but essential developments further push FCM measurement of tissue mechanics from a novel concept to a usable tool that fills an important niche by functionally imaging living tissue at the highest available spatial resolution of any currently available in vivo imaging method. It is expected that functional FCM imaging will eventually enable accelerated screening of preclinical therapies, and allow researchers to quantifiably relate implanted cell behavior with resulting changes in tissue structure and function.},
	keywords = {Animals Elastic Modulus Equipment Design Equipment Failure Analysis Female Fiber Optic Technology/*instrumentation Image Enhancement/*instrumentation Mice Mice, Confocal/*instrumentation Regeneration/*physiology Reproducibility of Results Sensitivity and Specificity Stress, Inbred C3H Microscopy, Mechanical Tendons/*cytology/*physiology},
	issn = {1937-3392 (Electronic) 1937-3384 (Linking)},
	doi = {10.1089/ten.tec.2008.0612},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19366317},
	author = {Snedeker, J. G. and Ben Arav, A. and Zilberman, Y. and Pelled, G. and Gazit, D.}
}
@article {17095,
	title = {Genetically engineered mesenchymal stem cells: applications in spine therapy},
	journal = {Regen Med},
	volume = {4},
	number = {1},
	year = {2009},
	note = {Aslan, Hadi Sheyn, Dima Gazit, Dan eng Review England 2008/12/25 09:00 Regen Med. 2009 Jan;4(1):99-108. doi: 10.2217/17460751.4.1.99.},
	pages = {99-108},
	type = {Journal Article},
	abstract = {Spine disorders and intervertebral disc degeneration are considered the main causes for the clinical condition commonly known as back pain. Spinal fusion by implanting autologous bone to produce bony bridging between the two vertebrae flanking the degenerated-intervertebral disc is currently the most efficient treatment for relieving the symptoms of back pain. However, donor-site morbidity, complications and the long healing time limit the success of this approach. Novel developments undertaken by regenerative medicine might bring more efficient and available treatments. Here we discuss the pros and cons of utilizing genetically engineered mesenchymal stem cells for inducing spinal fusion. The combination of the stem cells, gene and carrier are crucial elements for achieving optimal spinal fusion in both small and large animal models, which hopefully will lead to the development of clinical applications.},
	keywords = {Animals Genetic Engineering Humans Mesenchymal Stem Cell Transplantation/*methods Mesenchymal Stromal Cells/*cytology/metabolism Models, Biological Osteogenesis/genetics Regenerative Medicine Spinal Diseases/*therapy Spinal Fusion/methods Spine/cytology/metabolism},
	issn = {1746-076X (Electronic) 1746-0751 (Linking)},
	doi = {10.2217/17460751.4.1.99},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19105619},
	author = {Aslan, H. and Sheyn, D. and Gazit, D.}
}
@article {17103,
	title = {Micro-electroporation of mesenchymal stem cells with alternating electrical current pulses},
	journal = {Biomed Microdevices},
	volume = {11},
	number = {1},
	year = {2009},
	note = {Ziv, Roee Steinhardt, Yair Pelled, Gadi Gazit, Dan Rubinsky, Boris eng Research Support, Non-U.S. Gov{\textquoteright}t 2008/09/26 09:00 Biomed Microdevices. 2009 Feb;11(1):95-101. doi: 10.1007/s10544-008-9213-4.},
	pages = {95-101},
	type = {Journal Article},
	abstract = {Micro-electroporation is an electroporation technology in which the electrical field that induces cell membrane poration is focused onto a single cell contained in a micro-electromechanical structure. Micro-electroporation has many unique attributes including that it facilitates real time control over the process of electroporation at the single cell level. Flow-through micro-electroporation expands on this principle and was developed to facilitate electroporation of a large numbers of cells with control over the electroporation of every single cell. However, our studies show that when electroporation employs conventional direct current (DC) electrical pulses the micro-electroporation system fails, because of electrolysis induced gas bubble formation. We report in this study that when certain alternating currents (AC) electrical pulses are used for micro-electroporation it becomes possible to avoid electrolytic gas bubble formation in a micro-electroporation flow-through system. The effect of AC micro-electroporation on electrolysis was found to depend on the AC frequency used. This concept was tested with mesenchymal stem cells and preliminary results show successful electroporation using this system.},
	keywords = {Animals Cell Line Electroporation/*instrumentation/*methods Mesenchymal Stromal Cells/*cytology Mice Microfluidic Analytical Techniques/*instrumentation/*methods},
	issn = {1572-8781 (Electronic) 1387-2176 (Linking)},
	doi = {10.1007/s10544-008-9213-4},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18815886},
	author = {Ziv, R. and Steinhardt, Y. and Pelled, G. and Gazit, D. and Rubinsky, B.}
}
@article {17096,
	title = {Promoter hypermethylation of mismatch repair genes, hMLH1 and hMSH2 in oral squamous cell carcinoma},
	journal = {Oral Dis},
	volume = {15},
	number = {3},
	year = {2009},
	note = {Czerninski, R Krichevsky, S Ashhab, Y Gazit, D Patel, V Ben-Yehuda, D eng Research Support, Non-U.S. Gov{\textquoteright}t Denmark 2009/02/12 09:00 Oral Dis. 2009 Apr;15(3):206-13. doi: 10.1111/j.1601-0825.2008.01510.x. Epub 2009 Jan 16.},
	pages = {206-13},
	type = {Journal Article},
	abstract = {OBJECTIVES: Major risk factors of oral squamous cell carcinoma (OSCC) are environmental and can lead to DNA mutagenesis. Mismatch repair (MMR) system functions to repair small DNA lesions, which can be targeted for promoter hypermethylation. We therefore wanted to test whether hypermethylation of MMR genes (hMLH1, hMSH2) could contribute to oral carcinogenesis by correlating the information to patient clinical data. METHODS: Genomic DNA was extracted from 28 OSCC and six normal oral epithelium samples. The methylation status of the two MMR genes was assessed using Methylation Specific PCR after DNA modification with sodium bisulfite. Serial sections of the same tissues were immunostained with antibodies against hMLH1 and hMSH2 protein. RESULTS: Promoter hypermethylation was observed in 14/28 OSCC cases. Remarkably, 100\% of patients with multiple oral malignancies showed hypermethylation in hMLH1 or hMSH2 compared with 31.5\% of single tumor patients. In 10 cancer cases, expression of the hMLH1 and hMSH2 genes by immunostaining showed reduced or absence of expression of one of the genes, although some did not reflect the methylation status. CONCLUSIONS: Hypermethylation of hMLH1 and hMSH2 might play a role in oral carcinogenesis and may be correlated with a tendency to develop multiple oral malignancies.},
	keywords = {80 and over Carcinoma, Adaptor Proteins, Genetic/physiology, Multiple Primary/genetics/metabolism Nuclear Proteins/*genetics/metabolism Promoter Regions, Signal Transducing/*genetics/metabolism Adult Aged Aged, Squamous Cell/*genetics/metabolism DNA Mismatch Repair/*genetics DNA Repair Enzymes/genetics/metabolism Female Humans Male Methylation Middle Aged Mouth Neoplasms/*genetics/metabolism MutL Protein Homolog 1 MutS Homolog 2 Protein/*genetics/metabolism Neo},
	issn = {1601-0825 (Electronic) 1354-523X (Linking)},
	doi = {10.1111/j.1601-0825.2008.01510.x},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19207881},
	author = {Czerninski, R. and Krichevsky, S. and Ashhab, Y. and Gazit, D. and Patel, V. and Ben-Yehuda, D.}
}
@article {17102,
	title = {Revealing the interplay of bone and cartilage in osteoarthritis through multimodal imaging of murine joints},
	journal = {Bone},
	volume = {45},
	number = {3},
	year = {2009},
	note = {Stok, Kathryn S Pelled, Gadi Zilberman, Yoram Kallai, Ilan Goldhahn, Jorg Gazit, Dan Muller, Ralph eng Research Support, Non-U.S. Gov{\textquoteright}t 2009/06/02 09:00 Bone. 2009 Sep;45(3):414-22. doi: 10.1016/j.bone.2009.05.017. Epub 2009 May 28.},
	pages = {414-22},
	type = {Journal Article},
	abstract = {Osteoarthritis (OA) affects both cartilage and bone tissues, and the subsequent breakdown of the two tissues appears to be interrelated. The interest in the role of subchondral bone changes with OA is growing, and one suggestion is that a simple inverse correlation exists between the cartilage loss and increased bone mineral density. In this work the STR/ort mouse is used as a model for human OA, in order to investigate disease progression. The aim of the work is to elucidate the tempero-spatial relationships between bone and cartilage architecture and determine whether a simple inverse correlation is satisfactory. We employ 3D whole joint quantitative imaging techniques for assessment of subchondral bone and articular cartilage. The knee joints of mice aged 3, 4, 7 and 10 months are scanned with muCT and then the tibial plateaus are scanned with CLSM. The results show that depending on site (medial and lateral), compartment (epiphyseal, metaphyseal, cortical), and age (3, 4, 7, 10 months), the subchondral bone undergoes changes that lead to an altered architecture. This is primarily seen as densification of the cortex and epiphysis in the STR/ort mice, with a significant change occurring between 7 and 10 months, while the medial cartilage thickness is significantly reduced after 7 months. Using a novel multimodal imaging approach, morphometric changes in the murine osteoarthritic knee joint are elucidated. It is seen that a complex interplay of events - both spatially and temporally - is involved in OA onset and progression. The initial measured differences between the two strains suggest a possible morphological phenotype involved in OA resistance/vulnerability. Temporally the changes have a strong strain:age dependence, although no separate timeline of events between the two tissues could be discerned. Spatially, the changes to medial and lateral morphometry across the cartilage and bone, indicate a relationship to altered joint mechanics.},
	keywords = {Aging/pathology Animals Bone and Bones/diagnostic imaging/*pathology Cartilage, Articular/diagnostic imaging/*pathology Imaging, Confocal Osteoarthritis/diagnostic imaging/*pathology X-Ray Microtomography, Three-Dimensional Knee Joint/pathology Male Mice Microscopy},
	issn = {1873-2763 (Electronic) 1873-2763 (Linking)},
	doi = {10.1016/j.bone.2009.05.017},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19481620},
	author = {Stok, K. S. and Pelled, G. and Zilberman, Y. and Kallai, I. and Goldhahn, J. and Gazit, D. and Muller, R.}
}
@article {17099,
	title = {The use of a synthetic oxygen carrier-enriched hydrogel to enhance mesenchymal stem cell-based bone formation in vivo},
	journal = {Biomaterials},
	volume = {30},
	number = {27},
	year = {2009},
	note = {Kimelman-Bleich, Nadav Pelled, Gadi Sheyn, Dima Kallai, Ilan Zilberman, Yoram Mizrahi, Olga Tal, Yamit Tawackoli, Wafa Gazit, Zulma Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t Netherlands 2009/06/23 09:00 Biomaterials. 2009 Sep;30(27):4639-48. doi: 10.1016/j.biomaterials.2009.05.027. Epub 2009 Jun 21.},
	pages = {4639-48},
	type = {Journal Article},
	abstract = {A major hurdle to surmount in bone-tissue engineering is ensuring a sufficient oxygen supply to newly forming tissue to avoid cell death or delayed development of osteogenic features. We hypothesized that an oxygen-enriched hydrogel scaffold would enhance tissue-engineered bone formation in vivo. To test this, we used a well-characterized mesenchymal stem cell (MSC) line, Tet-off BMP2 MSC, whose cells were engineered to express recombinant human bone morphogenetic protein-2. Cells were suspended in hydrogel supplemented with perfluorotributylamine (PFTBA) and implanted subcutaneously in an ectopic site, a radial bone defect, or a lumbar paravertebral muscle (mouse model of spinal fusion) in C3H/HeN mice. For controls, we used cells suspended in the same gel without PFTBA. In the ectopic site, there were significant increases in bone formation (2.5-fold increase), cell survival, and osteocalcin activity in the PFTBA-supplemented groups. PFTBA supplementation significantly increased structural parameters of bone in radial bone defects and triggered a significant 1.4-fold increase in bone volume in the spinal fusion model. We conclude that synthetic oxygen carrier supplementation of tissue-engineered implants enhances ectopic bone formation and yields better bone quality and volume in bone-repair and spinal fusion models, probably due to increased cell survival.},
	keywords = {Animals Bone Morphogenetic Protein 2/metabolism Bone Regeneration/drug effects Cell Survival/drug effects Fluorocarbons/*pharmacology Gene Expression Regulation/drug effects Humans Hydrogels/*chemistry Implants, Experimental Mesenchymal Stromal Cells/*cytology/drug effects/metabolism Mice Neovascularization, Physiologic/drug effects Osteocalcin/genetics/metabolism Osteogenesis/*drug effects Oxygen/*metabolism Paracrine Communication/drug effects Radius/drug effects/pathology Spinal Fusion Tetracycline/pharmacology Wound Healing/drug effects},
	issn = {1878-5905 (Electronic) 0142-9612 (Linking)},
	doi = {10.1016/j.biomaterials.2009.05.027},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/19540585},
	author = {Kimelman-Bleich, N. and Pelled, G. and Sheyn, D. and Kallai, I. and Zilberman, Y. and Mizrahi, O. and Tal, Y. and Tawackoli, W. and Gazit, Z. and Gazit, D.}
}
@article {17093,
	title = {Bioluminescent imaging in bone},
	journal = {Methods Mol Biol},
	volume = {455},
	year = {2008},
	note = {Zilberman, Yoram Gafni, Yossi Pelled, Gadi Gazit, Zulma Gazit, Dan eng 2008/05/09 09:00 Methods Mol Biol. 2008;455:261-72. doi: 10.1007/978-1-59745-104-8_18.},
	pages = {261-72},
	type = {Journal Article},
	abstract = {Monitoring gene expression in vitro and in vivo, is crucial when analyzing osteogenesis and developing effective bone gene therapy protocols. Until recently, molecular analytical tools were only able to detect protein expression either in vitro or in vivo. These systems include histology and immunohistochemistry, fluorescent imaging, PET (micro-PET), CT (micro-CT), and bioluminescent imaging. The last is the only system to date that can enable efficient quantitative monitoring of gene expression both in vitro and in vivo. Effective bioluminescent imaging in bone can be achieved by using transgenic mice harboring the luciferase reporter gene, downstream of an osteogenesis specific promoter. The aim of this chapter is to comprehensively describe the various protocols needed for the detection of bioluminescence in bone development and repair.},
	keywords = {Animals Bone and Bones/*cytology/metabolism Humans Immunohistochemistry/methods Luminescent Proteins/genetics/*metabolism Mice Mice, Fluorescence/instrumentation/methods Osteocalcin/genetics Recombinant Fusion Proteins/genetics/metabolism, Transgenic *Microscopy},
	issn = {1064-3745 (Print) 1064-3745 (Linking)},
	doi = {10.1007/978-1-59745-104-8_18},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18463824},
	author = {Zilberman, Y. and Gafni, Y. and Pelled, G. and Gazit, Z. and Gazit, D.}
}
@article {17092,
	title = {Circadian mechanisms in murine and human bone marrow mesenchymal stem cells following dexamethasone exposure},
	journal = {Bone},
	volume = {42},
	number = {5},
	year = {2008},
	note = {Wu, Xiying Yu, Gang Parks, Helen Hebert, Teddi Goh, Brian C Dietrich, Marilyn A Pelled, Gadi Izadpanah, Reza Gazit, Dan Bunnell, Bruce A Gimble, Jeffrey M eng P30 DK072476-03/DK/NIDDK NIH HHS/ P30 DK072476/DK/NIDDK NIH HHS/ R21 DE016371/DE/NIDCR NIH HHS/ DE016371/DE/NIDCR NIH HHS/ DK072476/DK/NIDDK NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2008/02/28 09:00 Bone. 2008 May;42(5):861-70. doi: 10.1016/j.bone.2007.12.226. Epub 2008 Jan 26.},
	pages = {861-70},
	type = {Journal Article},
	abstract = {A core group of regulatory factors control circadian rhythms in mammalian cells. While the suprachiasmatic nucleus in the brain serves as the central core circadian oscillator, circadian clocks also exist within peripheral tissues and cells. A growing body of evidence has demonstrated that >20\% of expressed mRNAs in bone and adipose tissues oscillate in a circadian manner. The current manuscript reports evidence of the core circadian transcriptional apparatus within primary cultures of murine and human bone marrow-derived mesenchymal stem cells (BMSCs). Exposure of confluent, quiescent BMSCs to dexamethasone synchronized the oscillating expression of the mRNAs encoding the albumin D binding protein (dbp), brain-muscle arnt-like 1 (bmal1), period 3 (per3), rev-erb alpha (Rev A), and rev-erb beta (Rev B). The genes displayed a mean oscillatory period of 22.2 to 24.3 h. The acrophase or peak expression of mRNAs encoding "positive" (bmal1) and "negative" (per3) components of the circadian regulatory apparatus were out of phase with each other by approximately 8-12 h, consistent with in vivo observations. In vivo, phosphyrylation by glycogen synthase kinase 3beta (GSK3beta) is known to regulate the turnover of per3 and components of the core circadian regulatory apparatus. In vitro addition of lithium chloride, a GSK3beta inhibitor, significantly shifted the acrophase of all genes by 4.2-4.7 h oscillation in BMSCs; however, only the male murine BMSCs displayed a significant increase in the length of the period of oscillation. We conclude that human and murine BMSCs represent a valid in vitro model for the analysis of circadian mechanisms in bone metabolism and stem cell biology.},
	keywords = {ARNTL Transcription Factors Adult Animals Antigens, CD/analysis Basic Helix-Loop-Helix Transcription Factors/genetics Bone Marrow Cells/cytology/drug effects/metabolism Cell Cycle Proteins/genetics Circadian Rhythm/*physiology DNA-Binding Proteins/genetics Dexamethasone/*pharmacology Female Gene Expressio, Cytoplasmic and Nuclear/genetics Repressor Proteins/genetics Reverse Transcriptase Polymerase Chain Reaction Transcription Factors/genetics, Group D, Inbred C57BL Nuclear Proteins/genetics Nuclear Receptor Subfamily 1, Member 1 Period Circadian Proteins Receptors},
	issn = {8756-3282 (Print) 1873-2763 (Linking)},
	doi = {10.1016/j.bone.2007.12.226},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18302991},
	author = {Wu, X. and Yu, G. and Parks, H. and Hebert, T. and Goh, B. C. and Dietrich, M. A. and Pelled, G. and Izadpanah, R. and Gazit, D. and Bunnell, B. A. and Gimble, J. M.}
}
@article {17094,
	title = {Fluorescence molecular tomography enables in vivo visualization and quantification of nonunion fracture repair induced by genetically engineered mesenchymal stem cells},
	journal = {J Orthop Res},
	volume = {26},
	number = {4},
	year = {2008},
	note = {Zilberman, Yoram Kallai, Ilan Gafni, Yossi Pelled, Gadi Kossodo, Sylvie Yared, Wael Gazit, Dan eng 2007/11/07 09:00 J Orthop Res. 2008 Apr;26(4):522-30.},
	pages = {522-30},
	type = {Journal Article},
	abstract = {Fluorescence molecular tomography (FMT) is a novel tomographic near-infrared (NIR) imaging modality that enables 3D quantitative determination of fluorochrome distribution in tissues of live small animals at any depth. This study demonstrates a noninvasive, quantitative method of monitoring engineered bone remodeling via FMT. Murine mesenchymal stem cells overexpressing the osteogenic gene BMP2 (mMSCs-BMP2) were implanted into the thigh muscle and into a radial nonunion bone defect model in C3H/HeN mice. Real-time imaging of bone formation was performed following systemic administration of the fluorescent bisphosphonate imaging agent OsteoSense, an hydroxyapatite-directed bone-imaging probe. The mice underwent imaging on days 7, 14, and 21 postimplantation. New bone formation at the implantation sites was quantified using micro-computed tomography (micro-CT) imaging. A higher fluorescent signal occurred at the site of the mMSC-BMP2 implants than that found in controls. Micro-CT imaging revealed a mass of mature bone formed in the implantation sites on day 21, a finding also confirmed by histology. These findings highlight the effectiveness of FMT as a functional platform for molecular imaging in the field of bone regeneration and tissue engineering.},
	keywords = {Animals Female Fluorescent Dyes Fracture Healing/*physiology Fractures, Inbred C3H Osteogenesis/*physiology Radius Fractures/diagnostic imaging/pathology/*therapy Tomography, Ununited/diagnostic imaging/pathology/*therapy Genetic Engineering *Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells/*physiology Mice Mice, X-Ray Computed},
	issn = {1554-527X (Electronic) 0736-0266 (Linking)},
	doi = {10.1002/jor.20518},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17985393},
	author = {Zilberman, Y. and Kallai, I. and Gafni, Y. and Pelled, G. and Kossodo, S. and Yared, W. and Gazit, D.}
}
@article {17090,
	title = {Maxillofacial-derived stem cells regenerate critical mandibular bone defect},
	journal = {Tissue Eng Part A},
	volume = {14},
	number = {11},
	year = {2008},
	note = {Steinhardt, Yair Aslan, Hadi Regev, Eran Zilberman, Yoram Kallai, Ilan Gazit, Dan Gazit, Zulma eng Research Support, Non-U.S. Gov{\textquoteright}t 2008/07/19 09:00 Tissue Eng Part A. 2008 Nov;14(11):1763-73. doi: 10.1089/ten.tea.2008.0007.},
	pages = {1763-73},
	type = {Journal Article},
	abstract = {Stem cell-based bone tissue regeneration in the maxillofacial complex is a clinical necessity. Genetic engineering of mesenchymal stem cells (MSCs) to follow specific differentiation pathways may enhance the ability of these cells to regenerate and increase their clinical relevance. MSCs isolated from maxillofacial bone marrow (BM) are good candidates for tissue regeneration at sites of damage to the maxillofacial complex. In this study, we hypothesized that MSCs isolated from the maxillofacial complex can be engineered to overexpress the bone morphogenetic protein-2 gene and induce bone tissue regeneration in vivo. To demonstrate that the cells isolated from the maxillofacial complex were indeed MSCs, we performed a flow cytometry analysis, which revealed a high expression of mesenchyme-related markers and an absence of non-mesenchyme-related markers. In vitro, the MSCs were able to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Gene delivery of the osteogenic gene BMP2 via an adenoviral vector revealed high expression levels of BMP2 protein that induced osteogenic differentiation of these cells in vitro and induced bone formation in an ectopic site in vivo. In addition, implantation of genetically engineered maxillofacial BM-derived MSCs into a mandibular defect led to regeneration of tissue at the site of the defect; this was confirmed by performing micro-computed tomography analysis. Histological analysis of the mandibles revealed osteogenic differentiation of implanted cells as well as bone tissue regeneration. We conclude that maxillofacial BM-derived MSCs can be genetically engineered to induce bone tissue regeneration in the maxillofacial complex and that this finding may be clinically relevant.},
	keywords = {80 and over Animals Bone Marrow Cells/cytology Bone Regeneration/*physiology Cell Differentiation Cells, Adolescent Adult Aged Aged, Cultured Child Child, Inbred NOD Mice, Preschool Facial Bones/cytology Female Humans Infant Male Mandibular Diseases/physiopathology/*surgery Maxilla/cytology Mesenchymal Stem Cell Transplantation/*methods Mesenchymal Stromal Cells/*cytology Mice Mice, SCID Middle Aged Young Adult},
	issn = {1937-3341 (Print) 1937-3341 (Linking)},
	doi = {10.1089/ten.tea.2008.0007},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18636943},
	author = {Steinhardt, Y. and Aslan, H. and Regev, E. and Zilberman, Y. and Kallai, I. and Gazit, D. and Gazit, Z.}
}
@article {17087,
	title = {Molecular targets for tendon neoformation},
	journal = {J Clin Invest},
	volume = {118},
	number = {2},
	year = {2008},
	note = {Aslan, Hadi Kimelman-Bleich, Nadav Pelled, Gadi Gazit, Dan eng Review 2008/02/05 09:00 J Clin Invest. 2008 Feb;118(2):439-44. doi: 10.1172/JCI33944.},
	pages = {439-44},
	type = {Journal Article},
	abstract = {Tendons and ligaments are unique forms of connective tissue that are considered an integral part of the musculoskeletal system. The ultimate function of tendon is to connect muscles to bones and to conduct the forces generated by muscle contraction into movements of the joints, whereas ligaments connect bone to bone and provide joint stabilization. Unfortunately, the almost acellular and collagen I-rich structure of tendons and ligaments makes them very poorly regenerating tissues. Injured tendons and ligaments are considered a major clinical challenge in orthopedic and sports medicine. This Review discusses the several factors that might serve as molecular targets that upon activation can enhance or lead to tendon neoformation.},
	keywords = {Animals Biomarkers/metabolism Cell Line Humans Mice Morphogenesis *Regeneration Tendon Injuries/*metabolism Tendons/embryology/*physiology},
	issn = {0021-9738 (Print) 0021-9738 (Linking)},
	doi = {10.1172/JCI33944},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18246194},
	author = {Aslan, H. and Kimelman-Bleich, N. and Pelled, G. and Gazit, D.}
}
@article {17091,
	title = {Nanobiomechanics of repair bone regenerated by genetically modified mesenchymal stem cells},
	journal = {Tissue Eng Part A},
	volume = {14},
	number = {10},
	year = {2008},
	note = {Tai, Kuangshin Pelled, Gadi Sheyn, Dima Bershteyn, Anna Han, Lin Kallai, Ilan Zilberman, Yoram Ortiz, Christine Gazit, Dan eng Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 2008/07/16 09:00 Tissue Eng Part A. 2008 Oct;14(10):1709-20. doi: 10.1089/ten.tea.2007.0241.},
	pages = {1709-20},
	type = {Journal Article},
	abstract = {Genetically modified mesenchymal stem cells (MSCs), overexpressing a BMP gene, have been previously shown to be potent inducers of bone regeneration. However, little was known of the chemical and intrinsic nanomechanical properties of this engineered bone. A previous study utilizing microcomputed tomography, back-scattered electron microscopy, energy-dispersive X-ray, nanoindentation, and atomic force microscopy showed that engineered ectopic bone, although similar in chemical composition and topography, demonstrated an elastic modulus range (14.6-22.1 GPa) that was less than that of the native bone (16.6-38.5 GPa). We hypothesized that these results were obtained due to the specific conditions that exist in an intramuscular ectopic implantation site. Here, we implanted MSCs overexpressing BMP-2 gene in an orthotopic site, a nonunion radial bone defect, in mice. The regenerated bone tissue was analyzed using the same methods previously utilized. The samples revealed high similarity between the engineered and native radii in chemical structure and elemental composition. In contrast to the previous study, nanoindentation data showed that, in general, the native bone exhibited a statistically similar elastic modulus values compared to that of the engineered bone, while the hardness was found to be marginally statistically different at 1000 muN and statistically similar at 7000 muN. We hypothesize that external loading, osteogenic cytokines and osteoprogenitors that exist in a fracture site could enhance the maturation of engineered bone derived from BMP-modified MSCs. Further studies should determine whether longer duration periods postimplantation would lead to increased bone adaptation.},
	keywords = {Animals Biomechanical Phenomena Bone Morphogenetic Protein 2/genetics/metabolism Bone Regeneration/*physiology Bone and Bones/*physiopathology/surgery Cell Line Female Mesenchymal Stem Cell Transplantation/methods Mesenchymal Stromal Cells/*cytology/metab, Atomic force microscopy, Electron, Raman Tissue Engineering/methods Tomography Scanners, Scanning Spectrum Analysis, X-Ray Computed},
	issn = {1937-3341 (Print) 1937-3341 (Linking)},
	doi = {10.1089/ten.tea.2007.0241},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18620480},
	author = {Tai, K. and Pelled, G. and Sheyn, D. and Bershteyn, A. and Han, L. and Kallai, I. and Zilberman, Y. and Ortiz, C. and Gazit, D.}
}
@article {17089,
	title = {Nonvirally engineered porcine adipose tissue-derived stem cells: use in posterior spinal fusion},
	journal = {Stem Cells},
	volume = {26},
	number = {4},
	year = {2008},
	note = {Sheyn, Dima Pelled, Gadi Zilberman, Yoram Talasazan, Farahnaz Frank, Jonathan M Gazit, Dan Gazit, Zulma eng Comparative Study 2008/01/26 09:00 Stem Cells. 2008 Apr;26(4):1056-64. doi: 10.1634/stemcells.2007-0858. Epub 2008 Jan 24.},
	pages = {1056-64},
	type = {Journal Article},
	abstract = {Multiple factors alter intervertebral disc volume, structure, shape, composition, and biomechanical properties, often leading to low back pain. Spinal fusion is frequently performed to treat this problem. We recently published results of our investigation of a novel system of in vivo bone formation, in which we used nonvirally nucleofected human mesenchymal stem cells that overexpress a bone morphogenetic protein gene. We hypothesized that primary porcine adipose tissue-derived stem cells (ASCs) nucleofected with plasmid containing recombinant human bone morphogenetic protein-6 (rhBMP-6) could induce bone formation and achieve spinal fusion in vivo. Primary ASCs were isolated from freshly harvested porcine adipose tissue. Overexpression of rhBMP-6 was achieved ex vivo by using a nucleofection technique. Transfection efficiency was monitored by assessing a parallel transfection involving an enhanced green fluorescent protein reporter gene and flow cytometry analysis. rhBMP-6 protein secreted by the cells was measured by performing an enzyme-linked immunosorbent assay. Genetically engineered cells were injected into the lumbar paravertebral muscle in immunodeficient mice. In vivo bone formation was monitored by a quantitative microcomputed tomography (muCT). The animals were euthanized 5 weeks postinjection, and spinal fusion was evaluated using in vitro muCT and histological analysis. We found formation of a large bone mass adjacent to the lumbar area, which produced posterior spinal fusion of two to four vertebrae. Our data demonstrate that efficient bone formation and spinal fusion can be achieved using ex vivo, nonvirally transfected primary ASCs. These results could pave the way to a novel biological solution for spine treatment.},
	keywords = {Adipose Tissue/cytology/physiology/*transplantation Animals Cells, Cultured Female Mice Mice, Inbred NOD Mice, SCID Spinal Fusion/*methods Stem Cell Transplantation/*methods Stem Cells/cytology/physiology Swine Tissue Engineering/*methods},
	issn = {1549-4918 (Electronic) 1066-5099 (Linking)},
	doi = {10.1634/stemcells.2007-0858},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18218819},
	author = {Sheyn, D. and Pelled, G. and Zilberman, Y. and Talasazan, F. and Frank, J. M. and Gazit, D. and Gazit, Z.}
}
@article {17088,
	title = {Ultrasound-based nonviral gene delivery induces bone formation in vivo},
	journal = {Gene Ther},
	volume = {15},
	number = {4},
	year = {2008},
	note = {Sheyn, D Kimelman-Bleich, N Pelled, G Zilberman, Y Gazit, D Gazit, Z eng England 2007/11/23 09:00 Gene Ther. 2008 Feb;15(4):257-66. Epub 2007 Nov 22.},
	pages = {257-66},
	type = {Journal Article},
	abstract = {Nonviral gene delivery is a promising, safe, therapeutic tool in regenerative medicine. This study is the first to achieve nonviral, ultrasound-based, osteogenic gene delivery that leads to bone tissue formation, in vivo. We hypothesized that direct in vivo sonoporation of naked DNA encoding for the osteogenic gene, recombinant human bone morphogenetic protein-9 (rhBMP-9) would induce bone formation. A luciferase plasmid (Luc), encoding rhBMP-9 or empty pcDNA3 vector mixed with microbubbles, was injected into the thigh muscles of mice. After injection, noninvasive sonoporation was applied. Luc activity was monitored noninvasively, and quantitatively using bioluminescence imaging in vivo, and found for 14 days with a peak expression on day 7. To examine osteogenesis in vivo, rhBMP-9 plasmid was sonoporated into the thigh muscles of transgenic mice that express the Luc gene under the control of a human osteocalcin promoter. Following rhBMP-9 sonoporation, osteocalcin-dependent Luc expression lasted for 24 days and peaked on day 10. Bone tissue was formed in the site of rhBMP-9 delivery, as was shown by micro-computerized tomography and histology. The sonoporation method was also compared with previously developed electrotransfer-based gene delivery and was found significantly inferior in its efficiency of gene delivery. We conclude that ultrasound-mediated osteogenic gene delivery could serve as a therapeutic solution in conditions requiring bone tissue regeneration after further development that will increase the transfection efficiency.},
	keywords = {Animals Bone Development/*genetics Bone Morphogenetic Proteins/administration \& dosage/genetics Electroporation Gene Expression Profiling *Gene Transfer Techniques Genes, Inbred C3H Recombinant Proteins/administration \& dosage/genetics *Ultrasonics, Reporter Growth Differentiation Factor 2 Growth Differentiation Factors Immunohistochemistry Luciferases/genetics Mice Mice},
	issn = {1476-5462 (Electronic) 0969-7128 (Linking)},
	doi = {10.1038/sj.gt.3303070},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/18033309},
	author = {Sheyn, D. and Kimelman-Bleich, N. and Pelled, G. and Zilberman, Y. and Gazit, D. and Gazit, Z.}
}
@article {17086,
	title = {Circadian oscillation of gene expression in murine calvarial bone},
	journal = {J Bone Miner Res},
	volume = {22},
	number = {3},
	year = {2007},
	note = {Zvonic, Sanjin Ptitsyn, Andrey A Kilroy, Gail Wu, Xiying Conrad, Steven A Scott, L Keith Guilak, Farshid Pelled, Gadi Gazit, Dan Gimble, Jeffrey M eng DK072476/DK/NIDDK NIH HHS/ R21 DE016371/DE/NIDCR NIH HHS/ Comparative Study Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2006/12/06 09:00 J Bone Miner Res. 2007 Mar;22(3):357-65.},
	pages = {357-65},
	type = {Journal Article},
	abstract = {UNLABELLED: The genes encoding the core circadian transcription factors display an oscillating expression profile in murine calvarial bone. More than 26\% of the calvarial bone transcriptome exhibits a circadian rhythm, comparable with that observed in brown and white adipose tissues and liver. Thus, circadian mechanisms may directly modulate oxidative phosphorylation and multiple metabolic pathways in bone homeostasis. INTRODUCTION: Although circadian rhythms have been associated historically with central regulatory mechanisms, there is emerging evidence that the circadian transcriptional apparatus exists in peripheral tissues. The aim of this study was to determine the presence and extent of circadian oscillation in the transcriptome of murine calvarial bone. MATERIALS AND METHODS: Cohorts of 8-week-old male AKR/J mice were maintained in a controlled 12-h light:12-h dark cycle on an ad libitum diet for 2 weeks. Groups of three mice were killed every 4 h over a 48-h period. The level of gene expression at successive times-points was determined by quantitative RT-PCR and Affymetrix microarray. Data were analyzed using multiple statistical time series algorithms, including Cosinor, Fisher g-test, and the permutation time test. RESULTS: Both the positive (Bmal1, Npas2) and negative (Cry1, Cry2, Per1, Per2, Per3) elements of the circadian transcriptional apparatus and their immediate downstream targets and mediators (Dbp, Rev-erbalpha, Rev-erbbeta) exhibited oscillatory expression profiles. Consistent with findings in other tissues, the positive and negative elements were in antiphase relative to each other. More than 26\% of the genes present on the microarray displayed an oscillatory profile in calvarial bone, comparable with the levels observed in brown and white adipose tissues and liver; however, only a subset of 174 oscillating genes were shared among all four tissues. CONCLUSIONS: Our findings show that the components of the circadian transcriptional apparatus are represented in calvarial bone and display coordinated oscillatory behavior. However, these are not the only genes to display an oscillatory expression profile, which is seen in multiple pathways involving oxidative phosphorylation and lipid, protein, and carbohydrate metabolism.},
	keywords = {Adipose Tissue, Brown/metabolism Adipose Tissue, White/metabolism Animals Circadian Rhythm/*physiology Gene Expression Profiling Gene Expression Regulation/*physiology Liver/metabolism Male Mice Oligonucleotide Array Sequence Analysis Organ Specificity/physiology Skull/*metabolism Transcription Factors},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1359/jbmr.061114},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17144790},
	author = {Zvonic, S. and Ptitsyn, A. A. and Kilroy, G. and Wu, X. and Conrad, S. A. and Scott, L. K. and Guilak, F. and Pelled, G. and Gazit, D. and Gimble, J. M.}
}
@article {17084,
	title = {Evidence for skeletal progenitor cells in the degenerate human intervertebral disc},
	journal = {Spine (Phila Pa 1976)},
	volume = {32},
	number = {23},
	year = {2007},
	note = {Risbud, Makarand V Guttapalli, Asha Tsai, Tsung-Ting Lee, Joon Y Danielson, Keith G Vaccaro, Alexander R Albert, Todd J Gazit, Zulma Gazit, Dan Shapiro, Irving M eng AR050087/AR/NIAMS NIH HHS/ Research Support, N.I.H., Extramural 2007/11/06 09:00 Spine (Phila Pa 1976). 2007 Nov 1;32(23):2537-44.},
	pages = {2537-44},
	type = {Journal Article},
	abstract = {STUDY DESIGN: To identify and characterize endogenous progenitor cell population from intervertebral disc. OBJECTIVE: To determine if progenitor cells exist in degenerate human discs. SUMMARY OF BACKGROUND DATA: Back pain, a significant source of morbidity in our society, is directly linked to the pathology of the intervertebral disc. Because disc disease is accompanied by a loss of cellularity, there is considerable interest in regeneration of cells of both the anulus fibrosus (AF) and nucleus pulposus (NP). METHODS: To determine if skeletal progenitor cells are present in the disc, samples were obtained from the degenerate AF and NP of 5 patients (Thompson grade 2 and 3, mean age 34 +/- 7.6 years) undergoing anterior cervical discectomy and fusion procedures as well as adult rat lumbar spine. RESULTS: Cells isolated from degenerate human tissues expressed CD105, CD166, CD63, CD49a, CD90, CD73, p75 low affinity nerve growth factor receptor, and CD133/1, proteins that are characteristic of marrow mesenchymal stem cells. In osteogenic media, there was an induction of alkaline phosphatase activity and expression of alkaline phosphatase, osteocalcin, and Runx-2 mRNA. When maintained in adipogenic media, a small percentage of cells displayed evidence of adipogenic differentiation: accumulation of cytosolic lipid droplets and increased expression of peroxisome proliferator-activated receptor-gamma2 and lipoprotein lipase mRNA. AF- and NP-derived cells also evidenced chondrogenic differentiation. CD133 (+) cells in the AF were able to commit to either the chondrogenic or adipogenic lineages. The results of the human disc studies were confirmed using cell derived from the NP and AF tissue of the mature rat disc. CONCLUSION: The analytical data indicated that the pathologically degenerate human disc contained populations of skeletal progenitor cells. These findings suggest that these endogenous progenitors may be used to orchestrate the repair of the intervertebral disc.},
	keywords = {Adipocytes/cytology/metabolism Adult Alkaline Phosphatase/biosynthesis/genetics Animals Antigens, CD/biosynthesis/genetics Bone and Bones/cytology/metabolism Cell Differentiation/drug effects Cell Separation/methods Cells, Cultured/cytology/drug effects Cervical Vertebrae/*pathology Chondrocytes/cytology/metabolism Core Binding Factor Alpha 1 Subunit/biosynthesis/genetics Humans Intervertebral Disc/*pathology Lipids/analysis Lipoprotein Lipase/biosynthesis/genetics Lumbar, Nerve Growth Factor/biosynthesis/genetics Spinal Diseases/*pathology, Wistar Receptors},
	issn = {1528-1159 (Electronic) 0362-2436 (Linking)},
	doi = {10.1097/BRS.0b013e318158dea6},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17978651},
	author = {Risbud, M. V. and Guttapalli, A. and Tsai, T. T. and Lee, J. Y. and Danielson, K. G. and Vaccaro, A. R. and Albert, T. J. and Gazit, Z. and Gazit, D. and Shapiro, I. M.}
}
@article {17082,
	title = {Review: gene- and stem cell-based therapeutics for bone regeneration and repair},
	journal = {Tissue Eng},
	volume = {13},
	number = {6},
	year = {2007},
	note = {Kimelman, Nadav Pelled, Gadi Helm, Gregory A Huard, J Schwarz, Edward M Gazit, Dan eng Review 2007/05/23 09:00 Tissue Eng. 2007 Jun;13(6):1135-50.},
	pages = {1135-50},
	type = {Journal Article},
	abstract = {Many clinical conditions require regeneration or implantation of bone. This is one focus shared by neurosurgery and orthopedics. Current therapeutic options (bone grafting and protein-based therapy) do not provide satisfying solutions to the problem of massive bone defects. In the past few years, gene- and stem cell-based therapy has been extensively studied to achieve a viable alternative to current solutions offered by modern medicine for bone-loss repair. The use of adult stem cells for bone regeneration has gained much focus. This unique population of multipotential cells has been isolated from various sources, including bone marrow, adipose, and muscle tissues. Genetic engineering of adult stem cells with potent osteogenic genes has led to fracture repair and rapid bone formation in vivo. It is hypothesized that these genetically modified cells exert both an autocrine and a paracrine effects on host stem cells, leading to an enhanced osteogenic effect. The use of direct gene delivery has also shown much promise for in vivo bone repair. Several viral and nonviral methods have been used to achieve substantial bone tissue formation in various sites in animal models. To advance these platforms to the clinical setting, it will be mandatory to overcome specific hurdles, such as control over transgene expression, viral vector toxicity, and prolonged culture periods of therapeutic stem cells. This review covers a prospect of cell and gene therapy for bone repair as well as some very recent advancements in stem cell isolation, genetic engineering, and exogenous control of transgene expression.},
	keywords = {Animals Bone Diseases/*genetics/*therapy Bone Regeneration/*genetics Genetic Therapy/*trends Humans Osteogenesis/*genetics Stem Cell Transplantation/*trends Tissue Engineering/*trends},
	issn = {1076-3279 (Print) 1076-3279 (Linking)},
	doi = {10.1089/ten.2007.0096},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17516852},
	author = {Kimelman, N. and Pelled, G. and Helm, G. A. and Huard, J. and Schwarz, E. M. and Gazit, D.}
}
@article {17083,
	title = {Structural and nanoindentation studies of stem cell-based tissue-engineered bone},
	journal = {J Biomech},
	volume = {40},
	number = {2},
	year = {2007},
	note = {Pelled, Gadi Tai, Kuangshin Sheyn, Dima Zilberman, Yoram Kumbar, Sangamesh Nair, Lakshmi S Laurencin, Cato T Gazit, Dan Ortiz, Christine eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 2006/03/10 09:00 J Biomech. 2007;40(2):399-411. Epub 2006 Mar 9.},
	pages = {399-411},
	type = {Journal Article},
	abstract = {Stem cell-based gene therapy and tissue engineering have been shown to be an efficient method for the regeneration of critical-sized bone defects. Despite being an area of active research over the last decade, no knowledge of the intrinsic ultrastructural and nanomechanical properties of such bone tissue exists. In this study, we report the nanomechanical properties of engineered bone tissue derived from genetically modified mesenchymal stem cells (MSCs) overexpressing the rhBMP2 gene, grown in vivo in the thigh muscle of immunocompetent mice for 4 weeks, compared to femoral bone adjacent to the transplantation site. The two types of bone had similar mineral contents (61 and 65 wt\% for engineered and femoral bone, respectively), overall microstructures showing lacunae and canaliculi (both measured by back-scattered electron microscopy), chemical compositions (measured by energy dispersive X-ray analysis), and nanoscale topographical morphologies (measured by tapping-mode atomic force microscopy imaging or TMAFM). Nanoindentation experiments revealed that the small length scale mechanical properties were statistically different with the femoral bone (indented parallel to the bone long axis) being stiffer and harder (apparent elastic modulus, E approximately 27.3+/-10.5 GPa and hardness, H approximately 1.0+/-0.7G Pa) than the genetically engineered bone (E approximately 19.8+/-5.6 GPa, H approximately 0.9+/-0.4G Pa). TMAFM imaging showed clear residual indents characteristic of viscoelastic plastic deformation for both types of bone. However, fine differences in the residual indent area (smaller for the engineered bone), pile up (smaller for the engineered bone), and fracture mechanisms (microcracks for the engineered bone) were observed with the genetically engineered bone behaving more brittle than the femoral control.},
	keywords = {Atomic Force *Nanotechnology *Stem Cells *Tissue Engineering, Biomechanical Phenomena *Bone Substitutes/chemistry Bone and Bones/pathology/*physiology Microscopy},
	issn = {0021-9290 (Print) 0021-9290 (Linking)},
	doi = {10.1016/j.jbiomech.2005.12.012},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/16524583},
	author = {Pelled, G. and Tai, K. and Sheyn, D. and Zilberman, Y. and Kumbar, S. and Nair, L. S. and Laurencin, C. T. and Gazit, D. and Ortiz, C.}
}
@article {17085,
	title = {Structural bone allograft combined with genetically engineered mesenchymal stem cells as a novel platform for bone tissue engineering},
	journal = {Tissue Eng},
	volume = {13},
	number = {3},
	year = {2007},
	note = {Xie, Chao Reynolds, David Awad, Hani Rubery, Paul T Pelled, Gadi Gazit, Dan Guldberg, Robert E Schwarz, Edward M O{\textquoteright}Keefe, Regis J Zhang, Xinping eng R01 AR046545/AR/NIAMS NIH HHS/ R01 AR051469/AR/NIAMS NIH HHS/ AR051469/AR/NIAMS NIH HHS/ R01 AR051469-03/AR/NIAMS NIH HHS/ R21 DE017096/DE/NIDCR NIH HHS/ P50 AR054041/AR/NIAMS NIH HHS/ AR46545/AR/NIAMS NIH HHS/ AR054041/AR/NIAMS NIH HHS/ DE017096/DE/NIDCR NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov{\textquoteright}t 2007/05/24 09:00 Tissue Eng. 2007 Mar;13(3):435-45.},
	pages = {435-45},
	type = {Journal Article},
	abstract = {The presence of live periosteal progenitor cells on the surface of bone autografts confers better healing than devitalized allograft. We have previously demonstrated in a murine 4 mm segmental femoral bone-grafting model that live periosteum produces robust endochondral and intramembraneous bone formation that is essential for effective healing and neovascularization of structural bone grafts. To the end of engineering a live pseudo-periosteum that could induce a similar response onto devitalized bone allograft, we seeded a mesenchymal stem cell line stably transfected with human bone morphogenic protein-2/beta-galactosidase (C9) onto devitalized bone allografts or onto a membranous small intestinal submucosa scaffold that was wrapped around the allograft. Histology showed that C9-coated allografts displayed early cartilaginous tissue formation at day 7. By 6 and 9 weeks, a new cortical shell was found bridging the segmental defect that united the host bones. Biomechanical testing showed that C9-coated allografts displayed torsional strength and stiffness equivalent to intact femurs at 6 weeks and superior to live isografts at 9 weeks. Volumetric and histomorphometric micro-computed tomography analyses demonstrated a 2-fold increase in new bone formation around C9-coated allografts, which resulted in a substantial increase in polar moment of inertia (pMOI) due to the formation of new cortical shell around the allografts. Positive correlations between biomechanics and new bone volume and pMOI were found, suggesting that the biomechanical function of the grafted femur relates to both morphological parameters. C9-coated allograft also exhibited slower resorption of the graft cortex at 9 weeks than live isograft. Both new bone formation and the persistent allograft likely contributed to the improved biomechanics of C9-coated allograft. Taken together, we propose a novel strategy to combine structural bone allograft with genetically engineered mesenchymal stem cells as a novel platform for bone tissue engineering.},
	keywords = {Animals *Bone Transplantation/methods/pathology *Bone and Bones Cattle Cell Line *Genetic Engineering *Mesenchymal Stem Cell Transplantation/methods *Mesenchymal Stromal Cells/cytology Mice Mice, Homologous, Inbred C3H Mice, Inbred C57BL *Tissue Engineering Transplantation},
	issn = {1076-3279 (Print) 1076-3279 (Linking)},
	doi = {10.1089/ten.2006.0182},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17518596},
	author = {Xie, C. and Reynolds, D. and Awad, H. and Rubery, P. T. and Pelled, G. and Gazit, D. and Guldberg, R. E. and Schwarz, E. M. and O{\textquoteright}Keefe, R. J. and Zhang, X.}
}
@article {17075,
	title = {Advanced molecular profiling in vivo detects novel function of dickkopf-3 in the regulation of bone formation},
	journal = {J Bone Miner Res},
	volume = {21},
	number = {12},
	year = {2006},
	note = {Aslan, Hadi Ravid-Amir, Osnat Clancy, Brian M Rezvankhah, Saeid Pittman, Debra Pelled, Gadi Turgeman, Gadi Zilberman, Yoram Gazit, Zulma Hoffmann, Andrea Gross, Gerhard Domany, Eytan Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t 2006/09/28 09:00 J Bone Miner Res. 2006 Dec;21(12):1935-45.},
	pages = {1935-45},
	type = {Journal Article},
	abstract = {UNLABELLED: A bioinformatics-based analysis of endochondral bone formation model detected several genes upregulated in this process. Among these genes the dickkopf homolog 3 (Dkk3) was upregulated and further studies showed that its expression affects in vitro and in vivo osteogenesis. This study indicates a possible role of Dkk3 in regulating bone formation. INTRODUCTION: Endochondral bone formation is a complex biological process involving numerous chondrogenic, osteogenic, and angiogenic proteins, only some of which have been well studied. Additional key genes may have important roles as well. We hypothesized that to identify key genes and signaling pathways crucial for bone formation, a comprehensive gene discovery strategy should be applied to an established in vivo model of osteogenesis. MATERIALS AND METHODS: We used in vivo implanted C3H10T1/2 cells that had been genetically engineered to express human bone morphogenetic protein-2 (BMP2) in a tetracycline-regulated system that controls osteogenic differentiation. Oligonucleotide microarray data from the implants (n = 4 repeats) was analyzed using coupled two-way clustering (CTWC) and statistical methods. For studying the effects of dickkopf homolog 3 (Dkk3) in chondrogenesis and osteogenesis, C3H10T1/2 mesenchymal progenitors were used. RESULTS: The CTWC revealed temporal expression of Dkk3 with other chondrogenesis-, osteogenesis-, and Wnt-related genes. Quantitative RT-PCR confirmed the expression of Dkk3 in the implants. C3H10T1/2 cells that expressed Dkk3 in the presence of BMP2 displayed lower levels of alkaline phosphatase and collagen I mRNA expression than control C3H10T1/2 cells that did not express Dkk3. Interestingly, the levels of collagen II mRNA expression, Alcian blue staining, and glucose aminoglycans (GAGs) production were not influenced by Dkk3 expression. In vivo microCT and bioluminescence imaging revealed that co-expression of Dkk3 and BMP2 by implanted C3H10T1/2 cells induced the formation of significantly lower quantities of bone than cells expressing only BMP2. CONCLUSIONS: A bioinformatics analysis enabled the identification of Dkk3 as a pivotal gene with a novel function in endochondral bone formation. Our results showed that Dkk3 might have inhibitory effects on osteogenesis, but no effect on chondrogenesis, indicating that Dkk3 plays a regulatory role in endochondral bone formation. Further mechanistic studies are required to reveal the mechanism of action of Dkk3 in endochondral bone formation.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/*biosynthesis/genetics Cell Line Chondrogenesis/physiology Computational Biology/methods Gene Expression Profiling/methods Humans Intercellular Signaling Peptides and Proteins/*biosynthesis/},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1359/jbmr.060819},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17002559},
	author = {Aslan, H. and Ravid-Amir, O. and Clancy, B. M. and Rezvankhah, S. and Pittman, D. and Pelled, G. and Turgeman, G. and Zilberman, Y. and Gazit, Z. and Hoffmann, A. and Gross, G. and Domany, E. and Gazit, D.}
}
@article {17080,
	title = {Applications of gene therapy and adult stem cells in bone bioengineering},
	journal = {Regen Med},
	volume = {1},
	number = {4},
	year = {2006},
	note = {Kimelman, N Pelled, G Gazit, Zul Gazit, D eng Review England 2007/05/01 09:00 Regen Med. 2006 Jul;1(4):549-61.},
	pages = {549-61},
	type = {Journal Article},
	abstract = {Bone tissue engineering is an emerging field, that could become a main therapeutic strategy in orthopedics in coming years. While bone has regenerative abilities that enable the self repair and regeneration of fractures, there are extreme situations in which the extent of bone loss is too large for complete regeneration to occur. In order to achieve bone regeneration, osteogenic genes (mainly from the bone morphogenetic protein family) can be delivered either directly into the target tissue, or by using adult stem cells, which are later implanted into the target site. Engineered adult stem cells combined with biodegradable polymeric scaffolds can be implanted into target sites, with or without ex vivo culture period. Several important factors influence the success of bone engineering approaches including: choice of cell and scaffold, the vector used in order to deliver the osteogenic gene, and the osteogenic gene itself. Cutting-edge imaging technologies, bioinformatics-based analysis of gene expression and exogenous regulation of transgene expression are among the tools that are being used to optimize and control bone formation in vivo. In this review we have attempted to provide an overview of the main factors that should be considered when utilizing adult stem cells and gene therapy strategies to regenerate bone defects or to promote new bone formation in vivo.},
	keywords = {Adult Animals Bone Morphogenetic Proteins/genetics Bone and Bones/*physiology *Genetic Therapy Humans *Regeneration Stem Cells/*cytology *Tissue Engineering},
	issn = {1746-076X (Electronic) 1746-0751 (Linking)},
	doi = {10.2217/17460751.1.4.549},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17465849},
	author = {Kimelman, N. and Pelled, G. and Gazit, Z. and Gazit, D.}
}
@article {17078,
	title = {Design of a filamentous polymeric scaffold for in vivo guided angiogenesis},
	journal = {Tissue Eng},
	volume = {12},
	number = {11},
	year = {2006},
	note = {Gafni, Yossi Zilberman, Yoram Ophir, Zohar Abramovitch, Rinat Jaffe, Michael Gazit, Zulma Domb, Avraham Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t 2007/05/24 09:00 Tissue Eng. 2006 Nov;12(11):3021-34.},
	pages = {3021-34},
	type = {Journal Article},
	abstract = {Angiogenesis is mandatory for reperfusion of viable tissues, and lack of vascularization may cause ischemia. The increasing disparity between the demand and availability of adequate substitutes for small-diameter human blood vessels has prompted an intensive search for artificial materials or biological allograft tissues, both of which usually fail in the long term. The objective of this study was to pioneer a novel model for in vivo guided angiogenesis based on a specific design process of a filamentous polymeric scaffold with endothelial cells in a 3-dimensional culture system. To our knowledge, this is the first report of an in vivo guided angiogenesis approach based on a 2-step model, composed of endothelial cells and a filamentous polymeric scaffold framework. Endothelial cells that had been cultured on a specifically designed filamentous polymeric scaffold within a regulated dynamic tissue culture system were shown in vivo to induce guided angiogenesis. Cells seeded on a biodegradable polymeric scaffold were implanted into mice. On day 28 after implantation, analysis revealed a guided angiogenic process along the path of the implanted polymeric scaffold as well as initial evidence for early maturation of engineered vessels, allowing red blood cells to flow through the forming lumina of new vessels as the polymer degraded. The authors conclude that in vivo guided angiogenesis can be achieved by combining endothelial cells with biodegradable filamentous polymeric scaffolds and that this model can lay the cornerstone for vascular engineering and future development of clinically available protocols aimed to treat life-threatening cardiovascular conditions.},
	keywords = {Actins/metabolism Animals Antibodies, Biological Molecular Weight Muscle, Cultured Endothelial Cells/chemistry/*cytology/transplantation Genes, Differential Scanning Caproates/chemistry Cells, Monoclonal/metabolism Bioreactors Brain Neoplasms/pathology Calorimetry, Nude Models, Physiologic Organ Culture Techniques Polyethylene Glycols/chemistry Polymers/*chemistry Substrate Specificity Time Factors Tin Tissue Engineering/*methods, Reporter Green Fluorescent Proteins/metabolism Immunohistochemistry Luciferases/metabolism Luminescent Measurements Magnetic Resonance Imaging Materials Testing Mice Mice, Smooth/metabolism *Neovascularization},
	issn = {1076-3279 (Print) 1076-3279 (Linking)},
	doi = {10.1089/ten.2006.12.3021},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17518618},
	author = {Gafni, Y. and Zilberman, Y. and Ophir, Z. and Abramovitch, R. and Jaffe, M. and Gazit, Z. and Domb, A. and Gazit, D.}
}
@article {17081,
	title = {Endoscopic cellular microscopy for in vivo biomechanical assessment of tendon function},
	journal = {J Biomed Opt},
	volume = {11},
	number = {6},
	year = {2006},
	note = {Snedeker, Jess G Pelled, Gadi Zilberman, Yoram Gerhard, Friederike Muller, Ralph Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t 2007/01/11 09:00 J Biomed Opt. 2006 Nov-Dec;11(6):064010.},
	pages = {064010},
	type = {Journal Article},
	abstract = {This study explores a novel method to quantify in vivo soft tissue biomechanics from endoscopic confocal fluorescence microscope images of externally loaded biological tissues. A custom algorithm based on normalized cross-correlation is used to track fluorescently labeled cells within soft tissue structures as they deform. Cellular displacements are subsequently reduced to tissue strains by deriving the spatial gradient of the spline smoothed cellular displacement field. The relative performance of the tracking method is verified using a synthetic dataset with known underlying deformation. In biological application of the method, tissue strains are measured in the Achilles tendon of an anesthetized mouse. Over repeated trials, structural strain in the tendon (i.e., the relative change in distance between cells located at view field extremes) is 20.3+/-3.1\%, thus establishing the reproducibility of the loading protocol. Analysis of local tendon tissue strains reveal primary engineering strains in the tissue to range from 5 to 55\%, signifying a highly inhomogeneous strain state, with complex relative motions of neighboring tendon substructures. In summary, the current work establishes a baseline for a promising experimental method, and demonstrates its technical feasibility.},
	keywords = {Animals Biomechanical Phenomena/methods Cell Movement/physiology Cell Size Elasticity Endoscopy/*methods Female Image Interpretation, Computer-Assisted/*methods Mice Mice, Confocal/*methods Microscopy, Fluorescence/*methods Microscopy, Inbred C3H Micromanipulation/*methods Microscopy, Mechanical Tendons/*cytology/*physiology, Video/methods Stress},
	issn = {1083-3668 (Print) 1083-3668 (Linking)},
	doi = {10.1117/1.2393153},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/17212533},
	author = {Snedeker, J. G. and Pelled, G. and Zilberman, Y. and Gerhard, F. and Muller, R. and Gazit, D.}
}
@article {17079,
	title = {Neotendon formation induced by manipulation of the Smad8 signalling pathway in mesenchymal stem cells},
	journal = {J Clin Invest},
	volume = {116},
	number = {4},
	year = {2006},
	note = {Hoffmann, Andrea Pelled, Gadi Turgeman, Gadi Eberle, Peter Zilberman, Yoram Shinar, Hadassah Keinan-Adamsky, Keren Winkel, Andreas Shahab, Sandra Navon, Gil Gross, Gerhard Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t 2006/04/06 09:00 J Clin Invest. 2006 Apr;116(4):940-52.},
	pages = {940-52},
	type = {Journal Article},
	abstract = {Tissue regeneration requires the recruitment of adult stem cells and their differentiation into mature committed cells. In this study we describe what we believe to be a novel approach for tendon regeneration based on a specific signalling molecule, Smad8, which mediates the differentiation of mesenchymal stem cells (MSCs) into tendon-like cells. A biologically active Smad8 variant was transfected into an MSC line that coexpressed the osteogenic gene bone morphogenetic protein 2 (BMP2). The engineered cells demonstrated the morphological characteristics and gene expression profile of tendon cells both in vitro and in vivo. In addition, following implantation in an Achilles tendon partial defect, the engineered cells were capable of inducing tendon regeneration demonstrated by double quantum filtered MRI. The results indicate what we believe to be a novel mechanism in which Smad8 inhibits the osteogenic pathway in MSCs known to be induced by BMP2 while promoting tendon differentiation. These findings may have considerable importance for the therapeutic replacement of tendons or ligaments and for engineering other tissues in which BMP plays a pivotal developmental role.},
	keywords = {Achilles Tendon/*metabolism Animals Biomarkers Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics/*metabolism/pharmacology Cell Differentiation Cells, Cultured Female Histocytochemistry Mesenchymal Stromal Cells/cytology/*metabolism Mice Mice, Inbred C3H Microscopy, Nude Recombinant Proteins/genetics/metabolism *Signal Transduction Smad8 Protein/genetics/*metabolism Stem Cells Tissue Engineering Transcriptional Activation Transfection Transforming Growth Factor beta/genetics/*metabolism/pharmacology, Phase-Contrast Protein Structure, Tertiary Rats Rats},
	issn = {0021-9738 (Print) 0021-9738 (Linking)},
	doi = {10.1172/JCI22689},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/16585960},
	author = {Hoffmann, A. and Pelled, G. and Turgeman, G. and Eberle, P. and Zilberman, Y. and Shinar, H. and Keinan-Adamsky, K. and Winkel, A. and Shahab, S. and Navon, G. and Gross, G. and Gazit, D.}
}
@article {17076,
	title = {Nucleofection-based ex vivo nonviral gene delivery to human stem cells as a platform for tissue regeneration},
	journal = {Tissue Eng},
	volume = {12},
	number = {4},
	year = {2006},
	note = {Aslan, Hadi Zilberman, Yoram Arbeli, Vered Sheyn, Dima Matan, Yoav Liebergall, Meir Li, Jin Zhong Helm, Gregory A Gazit, Dan Gazit, Zulma eng Research Support, Non-U.S. Gov{\textquoteright}t 2006/05/06 09:00 Tissue Eng. 2006 Apr;12(4):877-89.},
	pages = {877-89},
	type = {Journal Article},
	abstract = {There are several gene therapy approaches to tissue regeneration. Although usually efficient, virusbased approaches may elicit an immune response against the viral proteins. An alternative approach, nonviral transfer, is safer, and can be controlled and reproduced. We hypothesized that in vivo bone formation could be achieved using human mesenchymal stem cells (hMSCs) nonvirally transfected with the human bone morphogenetic protein-2 (hBMP-2) or -9 (hBMP-9) gene. Human MSCs were transfected using nucleofection, a unique electropermeabilization-based technique. Postnucleofection, cell viability was 53.6 +/- 2.5\% and gene delivery efficiency was 51\% to 88\% (mean 68.2 +/- 4.1\%), as demonstrated by flow cytometry in enhanced green fluorescent protein (EGFP)-nucleofected hMSCs. Transgene expression lasted longer than 14 days and was very low 21 days postnucleofection. Both hBMP-2- and hBMP-9-nucleofected hMSCs in culture demonstrated a significant increase in calcium deposition compared with EGFP-nucleofected hMSCs. Human BMP-2- and hBMP-9-nucleofected hMSCs transplanted in ectopic sites in NOD/SCID mice induced bone formation 4 weeks postinjection. We conclude that in vivo bone formation can be achieved by using nonvirally nucleofected hMSCs. This could lead to a breakthrough in the field of regenerative medicine, in which safer, nonviral therapeutic strategies present a very attractive alternative.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/*genetics/metabolism Bone Regeneration/genetics/*physiology Calcium/analysis Cell Survival Cells, Cultured Gene Transfer Techniques *Genetic Therapy Green Fluorescent Proteins/metabolism Growth Differentiation Factor 2 Growth Differentiation Factors Humans Mesenchymal Stem Cell Transplantation/methods Mesenchymal Stromal Cells/cytology/*metabolism Mi, Heterologous, Inbred NOD Mice, SCID Recombinant Proteins/secretion Time Factors Tissue Engineering/*methods Transforming Growth Factor beta/genetics/metabolism Transgenes Transplantation},
	issn = {1076-3279 (Print) 1076-3279 (Linking)},
	doi = {10.1089/ten.2006.12.877},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/16674300},
	author = {Aslan, H. and Zilberman, Y. and Arbeli, V. and Sheyn, D. and Matan, Y. and Liebergall, M. and Li, J. Z. and Helm, G. A. and Gazit, D. and Gazit, Z.}
}
@article {17077,
	title = {Osteogenic differentiation of noncultured immunoisolated bone marrow-derived CD105+ cells},
	journal = {Stem Cells},
	volume = {24},
	number = {7},
	year = {2006},
	note = {Aslan, Hadi Zilberman, Yoram Kandel, Leonid Liebergall, Meir Oskouian, Rod J Gazit, Dan Gazit, Zulma eng Research Support, Non-U.S. Gov{\textquoteright}t 2006/04/08 09:00 Stem Cells. 2006 Jul;24(7):1728-37. Epub 2006 Apr 6.},
	pages = {1728-37},
	type = {Journal Article},
	abstract = {The culture expansion of human mesenchymal stem cells (hMSCs) may alter their characteristics and is a costly and time-consuming stage. This study demonstrates for the first time that immunoisolated noncultured CD105-positive (CD105(+)) hMSCs are multipotent in vitro and exhibit the capacity to form bone in vivo. hMSCs are recognized as promising tools for bone regeneration. However, the culture stage is a limiting step in the clinical setting. To establish a simple, efficient, and fast method for applying these cells for bone formation, a distinct population of CD105(+) hMSCs was isolated from bone marrow (BM) by using positive selection based on the expression of CD105 (endoglin). The immunoisolated CD105(+) cell fraction represented 2.3\% +/- 0.45\% of the mononuclear cells (MNCs). Flow cytometry analysis of freshly immunoisolated CD105(+) cells revealed a purity of 79.7\% +/- 3.2\%. In vitro, the CD105(+) cell fraction displayed significantly more colony-forming units-fibroblasts (CFU-Fs; 6.3 +/- 1.4) than unseparated MNCs (1.1 +/- 0.3; p < .05). Culture-expanded CD105(+) cells expressed CD105, CD44, CD29, CD90, and CD106 but not CD14, CD34, CD45, or CD31 surface antigens, and these cells were able to differentiate into osteogenic, chondrogenic, and adipogenic lineages. In addition, freshly immunoisolated CD105(+) cells responded in vivo to recombinant bone morphogenetic protein-2 by differentiating into chondrocytes and osteoblasts. Genetic engineering of freshly immunoisolated CD105(+) cells was accomplished using either adenoviral or lentiviral vectors. Based on these findings, it is proposed that noncultured BM-derived CD105(+) hMSCs are osteogenic cells that can be genetically engineered to induce tissue generation in vivo.},
	keywords = {80 and over Antigens, Adolescent Adult Aged Aged, CD/isolation \& purification/*metabolism Bone Marrow Cells/*metabolism Cartilage/cytology *Cell Differentiation Endoglin Female Flow Cytometry Genetic Engineering Humans Immunoassay Male Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells/*met, Cell Surface/isolation \& purification/*metabolism},
	issn = {1066-5099 (Print) 1066-5099 (Linking)},
	doi = {10.1634/stemcells.2005-0546},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/16601078},
	author = {Aslan, H. and Zilberman, Y. and Kandel, L. and Liebergall, M. and Oskouian, R. J. and Gazit, D. and Gazit, Z.}
}
@article {17074,
	title = {Murine spinal fusion induced by engineered mesenchymal stem cells that conditionally express bone morphogenetic protein-2},
	journal = {J Neurosurg Spine},
	volume = {3},
	number = {1},
	year = {2005},
	note = {Hasharoni, Amir Zilberman, Yoram Turgeman, Gadi Helm, Gregory A Liebergall, Meir Gazit, Dan eng 2005/08/27 09:00 J Neurosurg Spine. 2005 Jul;3(1):47-52.},
	pages = {47-52},
	type = {Journal Article},
	abstract = {OBJECT: The authors hypothesized that spinal fusion can be achieved and monitored by using cell-mediated gene therapy. Mesenchymal stem cells (MSCs) genetically engineered to express recombinant human bone morphogenetic protein-2 (rhBMP-2) conditionally, were implanted into the paraspinal muscles of mice to establish spinal fusion. The goal was to demonstrate an MSC-based gene therapy platform in which controlled gene expression is used to obtain spinal fusion in a murine model. METHODS: Mesenchymal stem cells expressing the rhBMP-2 gene were injected into the paravertebral muscle in mice. Bone formation in the paraspinal region was longitudinally followed by performing micro-computerized tomography scanning, histological studies, and an analysis of osteocalcin expression to demonstrate the presence of engrafted engineered MSCs. The minimal period of rhBMP-2 expression by the engineered MSCs required to induce fusion was determined. The results of this study demonstrate that genetically engineered MSCs induce bone formation in areas adjacent to and touching the posterior elements of the spine. This newly formed bone fuses the spine, as demonstrated by radiological and histological studies. The authors demonstrate that injected cells induce active osteogenesis at the site of implantation for up to 4 weeks postinjection. They found that a 7-day induction of rhBMP-2 expression in genetically engineered MSCs was sufficient to form new bone tissue, although the quantity of this bone increased as longer expression periods were implemented. CONCLUSIONS: After their injection genetically engineered MSCs can efficiently form new bone in the paraspinal muscle of the mouse to obtain spinal fusion. The extent and quantity of this newly formed bone can be monitored by controlling the duration of rhBMP-2 gene expression.},
	keywords = {Animal Osteogenesis/*physiology Spinal Fusion/*methods Time Factors Transforming Growth Factor beta/*physiology, Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/*physiology Feasibility Studies Female *Genetic Engineering *Mesenchymal Stem Cell Transplantation Mesenchymal Stromal Cells/*physiology Mice Mice, Inbred C3H Models},
	issn = {1547-5654 (Print) 1547-5646 (Linking)},
	doi = {10.3171/spi.2005.3.1.0047},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/16122022},
	author = {Hasharoni, A. and Zilberman, Y. and Turgeman, G. and Helm, G. A. and Liebergall, M. and Gazit, D.}
}
@article {17073,
	title = {Novel dextran-spermine conjugates as transfecting agents: comparing water-soluble and micellar polymers},
	journal = {Gene Ther},
	volume = {12},
	number = {6},
	year = {2005},
	note = {Eliyahu, H Makovitzki, A Azzam, T Zlotkin, A Joseph, A Gazit, D Barenholz, Y Domb, A J eng Comparative Study Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. England 2004/11/27 09:00 Gene Ther. 2005 Mar;12(6):494-503.},
	pages = {494-503},
	type = {Journal Article},
	abstract = {Recently, a novel cationic polymer, dextran-spermine (D-SPM) was developed for gene delivery. An efficient transfection was obtained using this polycation for a variety of genes and cell lines in serum-free or serum-poor medium. However, transfection using the water-soluble D-SPM-based polyplexes decreased with increasing serum concentration in cell culture in a concentration-dependent manner, reaching 95\% inhibition at 50\% serum in the cell growth medium. In order to overcome this obstacle, oleyl derivatives of D-SPM (which form micelles in aqueous phase) were synthesized at 1, 10, and 20 mol\% of oleyl moiety to polymer epsilon-NH2 to form N-oleyl-D-SPM (ODS). Polyplexes based on ODS transfected well in medium containing 50\% serum. Comparison with polyplexes based on well-established polymers (branched and linear polyethyleneimine) and with DOTAP/Cholesterol lipoplexes showed that regarding beta-galactosidase transgene expression level and cytotoxicity in tissue culture, the D-SPM and ODS compare well with the above polyplexes and lipoplexes. Intracellular trafficking using FITC-labeled ODS and Rhodamine-labeled pGeneGrip plasmid cloned with hBMP2 monitored by confocal microscopy revealed that during the transfection process the fluorescent-labeled polymer concentrates in the Golgi apparatus and around the nucleus, while the cell cytoplasm was free of fluorescent particles, suggesting that the polyplexes move in the cell toward the nucleus by vesicular transport through the cytoplasm and not by a random diffusion. We found that the plasmids penetrate the cell nucleus without the polymer. Preliminary results in zebra fish and mice demonstrate the potential of ODS to serve as an efficient nonviral vector for in vivo transfection.},
	keywords = {Animals Cations Cell Culture Techniques Cell Line Cell Nucleus/metabolism Culture Media Culture Media, Confocal NIH 3T3 Cells Plasmids/*administration \& dosage Polyethyleneimine Polymers Quaternary Ammonium Compounds Spermine Transfection/*methods Transgenes Zebrafish, Inbred C3H Micelles Microscopy, Monounsaturated Female Flow Cytometry Gene Expression Genetic Therapy/*methods Golgi Apparatus/metabolism Humans Injections Luciferases/genetics Mice Mice, Serum-Free Cytoplasm/metabolism Dextrans Fatty Acids},
	issn = {0969-7128 (Print) 0969-7128 (Linking)},
	doi = {10.1038/sj.gt.3302395},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/15565162},
	author = {Eliyahu, H. and Makovitzki, A. and Azzam, T. and Zlotkin, A. and Joseph, A. and Gazit, D. and Barenholz, Y. and Domb, A. J.}
}
@article {17070,
	title = {Distinct roles of BMP receptors Type IA and IB in osteo-/chondrogenic differentiation in mesenchymal progenitors (C3H10T1/2)},
	journal = {Biofactors},
	volume = {20},
	number = {2},
	year = {2004},
	note = {Kaps, Christian Hoffmann, Andrea Zilberman, Yoram Pelled, Gadi Haupl, Thomas Sittinger, Michael Burmester, Gerd Gazit, Dan Gross, Gerhard eng Research Support, Non-U.S. Gov{\textquoteright}t Netherlands 2004/08/24 05:00 Biofactors. 2004;20(2):71-84.},
	pages = {71-84},
	type = {Journal Article},
	abstract = {The functional roles of BMP type IA and IB receptors mediating differentiation into the osteogenic and chondrogenic lineage were investigated in the mesenchymal progenitor line C3H10T1/2 in vitro. The capacity of type IA and IB BMP receptors was assessed by the forced expression of the wild-type (wtBMPR-IA or IB) and of the kinase-deficient, dominant-negative form (dnBMPR-IA or -IB) in parental C3H10T1/2 progenitors as well as in C3H10T1/2 progenitors which recombinantly express BMP2 (C3H10T1/2-BMP2) or GDF5 (C3H10T1/2-GDF5). Consistent with the higher endogenous expression rate of BMPR-IA in comparison with BMPR-IB, BMPR-IA plays the dominant role in BMP2-mediated osteo-/chondrogenic development. BMPR-IB moderately influences osteogenic and hardly chondrogenic development. BMPR-IB seems to be unable to efficiently activate downstream signaling pathways upon forced expression. However, a mutation conferring constitutive activity to the BMPR-IB receptor indicates that this receptor possesses the capacity to activate downstream signaling cascades. These results suggest that in mesenchymal progenitors C3H10T1/2 BMPR-IA is responsible for the initiation of the osteogenic as well as chondrogenic development and that BMPR-IA and -IB receptor pathways are well separated in this mesenchymal progenitor line and may not substitute each other. In addition this indicates that type IB and IA BMP receptors may transmit different signals during the specification and differentiation of mesenchymal lineages.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Protein Receptors, Growth Factor/genetics/*physiology Recombinant Fusion Proteins Recombinant Proteins Reverse Transcriptase Polymerase Chain Reaction Transfection Transforming Growth Factor beta/genetics/physiology, Inbred C3H Osteoblasts/*cytology Osteogenesis Protein-Serine-Threonine Kinases/genetics/*physiology RNA, Messenger/analysis Receptors, Type I Bone Morphogenetic Proteins/genetics/physiology Cell Differentiation/*physiology Cell Line Chondrocytes/*cytology Gene Expression Green Fluorescent Proteins/genetics Humans Mesenchymal Stromal Cells/*cytology Mice Mice},
	issn = {0951-6433 (Print) 0951-6433 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/15322331},
	author = {Kaps, C. and Hoffmann, A. and Zilberman, Y. and Pelled, G. and Haupl, T. and Sittinger, M. and Burmester, G. and Gazit, D. and Gross, G.}
}
@article {17067,
	title = {Gene therapy platform for bone regeneration using an exogenously regulated, AAV-2-based gene expression system},
	journal = {Mol Ther},
	volume = {9},
	number = {4},
	year = {2004},
	note = {Gafni, Yossi Pelled, Gadi Zilberman, Yoram Turgeman, Gadi Apparailly, Florence Yotvat, Hagit Galun, Eithan Gazit, Zulma Jorgensen, Christian Gazit, Dan eng 2004/04/20 05:00 Mol Ther. 2004 Apr;9(4):587-95.},
	pages = {587-95},
	type = {Journal Article},
	abstract = {Viral delivery of the therapeutic gene bone morphogenetic protein-2 (BMP-2) is a promising approach for bone regeneration. The human parvovirus adeno-associated virus (AAV) type 2 is considered one of the most encouraging viral vector systems because of its high transduction rates and biosafety ratings. Bone morphogenetic protein-2 is a highly potent osteoinductive protein, which induces bone formation in vivo and osteogenic differentiation in vitro. The exogenous regulation of BMP-2 expression in bone-regenerating sites is required to control BMP-2 protein secretion, thus promoting safe and controlled bone formation and regeneration. We have therefore constructed a dual-construct vector for the recombinant AAV (rAAV)-based recombinant human BMP-2 (rhBMP-2) gene delivery system, which is regulated by the tetracycline-sensitive promoter (TetON). Each vector was encapsidated separately, yielding two recombinant viruses. We evaluated the efficiency of rAAV-hBMP-2 to induce bone formation in ectopic and orthotopic sites. Doxycycline (Dox), an analogue of tetracycline, was orally administered to mice via their drinking water to induce rhBMP-2 expression. Bone formation was measured using quantitative imaging-microcomputerized tomography and cooled charge-coupled device imaging-to detect osteogenic activity at the cellular level, detecting osteocalcin expression. The rAAV-hBMP-2-treated mice that were given Dox demonstrated bone formation in both in vivo models compared to none in mice prevented from receiving Dox. Thus, the Tet-regulated rAAV-hBMP-2 vector is an effective means of induction and regulation of bone regeneration and repair.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/metabolism *Bone Regeneration Cell Line Dependovirus/*genetics Doxycycline/metabolism/pharmacology *Gene Expression Regulation Genetic Therapy/*methods Genetic Vectors Humans Mice Mice, Genetic Recombinant Proteins Stem Cells/cytology Tetracycline/pharmacology Tomography, Inbred C3H Mice, Transgenic Muscles/metabolism Osteocalcin/metabolism Promoter Regions, X-Ray Computed Transforming Growth Factor beta/metabolism},
	issn = {1525-0016 (Print) 1525-0016 (Linking)},
	doi = {10.1016/j.ymthe.2003.12.009},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/15093189},
	author = {Gafni, Y. and Pelled, G. and Zilberman, Y. and Turgeman, G. and Apparailly, F. and Yotvat, H. and Galun, E. and Gazit, Z. and Jorgensen, C. and Gazit, D.}
}
@article {17072,
	title = {Imaging using osteocalcin-luciferase},
	journal = {J Musculoskelet Neuronal Interact},
	volume = {4},
	number = {4},
	year = {2004},
	note = {Pelled, G Gazit, D eng Review Greece 2005/03/11 09:00 J Musculoskelet Neuronal Interact. 2004 Dec;4(4):362-3.},
	pages = {362-3},
	type = {Journal Article},
	keywords = {Animal Osteocalcin/*genetics Osteogenesis/*genetics Promoter Regions, Animals Biomarkers/analysis/metabolism Bone and Bones/*metabolism Gene Expression Regulation, Computer-Assisted/instrumentation/methods Luciferases/biosynthesis/genetics Luminescent Measurements Mice Mice, Developmental/*physiology Humans Image Processing, Genetic/genetics, Transgenic/*genetics/growth \& development/metabolism Models},
	issn = {1108-7161 (Print) 1108-7161 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/15758260},
	author = {Pelled, G. and Gazit, D.}
}
@article {17069,
	title = {Overexpression of CXCR4 on human CD34+ progenitors increases their proliferation, migration, and NOD/SCID repopulation},
	journal = {Blood},
	volume = {103},
	number = {8},
	year = {2004},
	note = {Kahn, Joy Byk, Tamara Jansson-Sjostrand, Lottie Petit, Isabelle Shivtiel, Shoham Nagler, Arnon Hardan, Izhar Deutsch, Varda Gazit, Zulma Gazit, Dan Karlsson, Stefan Lapidot, Tsvee eng Research Support, Non-U.S. Gov{\textquoteright}t 2004/04/09 05:00 Blood. 2004 Apr 15;103(8):2942-9. Epub 2003 Dec 24.},
	pages = {2942-9},
	type = {Journal Article},
	abstract = {A major limitation to clinical stem cell-mediated gene therapy protocols is the low levels of engraftment by transduced progenitors. We report that CXCR4 overexpression on human CD34+ progenitors using a lentiviral gene transfer technique helped navigate these cells to the murine bone marrow and spleen in response to stromal-derived factor 1 (SDF-1) signaling. Cells overexpressing CXCR4 exhibited significant increases in SDF-1-mediated chemotaxis and actin polymerization compared with control cells. A major advantage of CXCR4 overexpression was demonstrated by the ability of transduced CD34+ cells to respond to lower, physiologic levels of SDF-1 when compared to control cells, leading to improved SDF-1-induced migration and proliferation/survival, and finally resulting in significantly higher levels of in vivo repopulation of nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice including primitive CD34+/CD38(-/low) cells. Importantly, no cellular transformation was observed following transduction with the CXCR4 vector. Unexpectedly, we documented lack of receptor internalization in response to high levels of SDF-1, which can also contribute to increased migration and proliferation by the transduced CD34+ cells. Our results suggest CXCR4 overexpression for improved definitive human stem cell motility, retention, and multilineage repopulation, which could be beneficial for in vivo navigation and expansion of hematopoietic progenitors.},
	keywords = {Animals Antigens, CD34/metabolism Cell Differentiation Cell Division Cell Movement Chemokine CXCL12 Chemokines, CXC/pharmacology Gene Expression Gene Transfer Techniques Genetic Vectors Graft Survival Hematopoietic Stem Cells/*cytology/*immunology Humans Lentivirus/genetics Mice Mice, CXCR4/*genetics Stem Cell Transplantation Transplantation, Heterologous, Inbred NOD Mice, SCID Receptors},
	issn = {0006-4971 (Print) 0006-4971 (Linking)},
	doi = {10.1182/blood-2003-07-2607},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/15070669},
	author = {Kahn, J. and Byk, T. and Jansson-Sjostrand, L. and Petit, I. and Shivtiel, S. and Nagler, A. and Hardan, I. and Deutsch, V. and Gazit, Z. and Gazit, D. and Karlsson, S. and Lapidot, T.}
}
@article {17071,
	title = {Short-term BMP-2 expression is sufficient for in vivo osteochondral differentiation of mesenchymal stem cells},
	journal = {Stem Cells},
	volume = {22},
	number = {1},
	year = {2004},
	note = {Noel, Daniele Gazit, Dan Bouquet, Celine Apparailly, Florence Bony, Claire Plence, Pascale Millet, Virginie Turgeman, Gadi Perricaudet, Michel Sany, Jacques Jorgensen, Christian eng Research Support, Non-U.S. Gov{\textquoteright}t 2003/12/23 05:00 Stem Cells. 2004;22(1):74-85.},
	pages = {74-85},
	type = {Journal Article},
	abstract = {Currently available murine models to evaluate mesenchymal stem cell (MSC) differentiation are based on cell injection at ectopic sites such as muscle or skin. Due to the importance of environmental factors on the differentiation capacities of stem cells in vivo, we investigated whether the peculiar synovial/cartilaginous environment may influence the lineage specificity of bone morphogenetic protein (BMP)-2-engineered MSCs. To this aim, we used the C3H10T1/2-derived C9 MSCs that express BMP-2 under control of the doxycycline (Dox)-repressible promoter, Tet-Off, and showed in vitro, using the micropellet culture system that C9 MSCs kept their potential to differentiate toward chondrocytes. Implantation of C9 cells, either into the tibialis anterior muscles or into the joints of CB17-severe combined immunodeficient bg mice led to the formation of cartilage and bone filled with bone marrow as soon as day 10. However, no differentiation was observed after injection of naive MSCs or C9 cells that were repressed to secrete BMP-2 by Dox addition. The BMP-2-induced differentiation of adult MSCs is thus independent of soluble factors present in the local environment of the synovial/cartilaginous tissues. Importantly, we demonstrated that a short-term expression of the BMP-2 growth factor is necessary and sufficient to irreversibly induce bone formation, suggesting that a stable genetic modification of MSCs is not required for stem cell-based bone/cartilage engineering.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/*biosynthesis/genetics Bone and Bones/cytology/embryology/metabolism Cartilage/*cytology/embryology/metabolism Cell Communication/physiology Cell Differentiation/genetics/*physiology Cell Li, Developmental/genetics Growth Substances/metabolism/secretion Joints/cytology/growth \& development/surgery Mesenchymal Stromal Cells/*cytology/*metabolism Mice Mice, Genetic/drug effects/genetics Stem Cell Transplantation *Transforming Growth Factor beta, Inbred C3H Muscle, Skeletal/cytology/growth \& development/surgery NIH 3T3 Cells Osteogenesis/genetics/*physiology Promoter Regions},
	issn = {1066-5099 (Print) 1066-5099 (Linking)},
	doi = {10.1634/stemcells.22-1-74},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/14688393},
	author = {Noel, D. and Gazit, D. and Bouquet, C. and Apparailly, F. and Bony, C. and Plence, P. and Millet, V. and Turgeman, G. and Perricaudet, M. and Sany, J. and Jorgensen, C.}
}
@article {17068,
	title = {Stem cells as vehicles for orthopedic gene therapy},
	journal = {Gene Ther},
	volume = {11},
	number = {4},
	year = {2004},
	note = {Gafni, Y Turgeman, G Liebergal, M Pelled, G Gazit, Z Gazit, D eng Review England 2004/01/16 05:00 Gene Ther. 2004 Feb;11(4):417-26.},
	pages = {417-26},
	type = {Journal Article},
	abstract = {Adult stem cells reside in adult tissues and serve as the source for their specialized cells. In response to specific factors and signals, adult stem cells can differentiate and give rise to functional tissue specialized cells. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into various mesenchymal lineages such as muscle, bone, cartilage, fat, tendon and ligaments. Adult MSCs can be relatively easily isolated from different tissues such as bone marrow, fat and muscle. Adult MSCs are also easy to manipulate and expand in vitro. It is these properties of adult MSCs that have made them the focus of cell-mediated gene therapy for skeletal tissue regeneration. Adult MSCs engineered to express various factors not only deliver them in vivo, but also respond to these factors and differentiate into skeletal specialized cells. This allows them to actively participate in the tissue regeneration process. In this review, we examine the recent achievements and developments in stem-cell-based gene therapy approaches and their applications to bone, cartilage, tendon and ligament tissues that are the current focus of orthopedic medicine.},
	keywords = {Adult Bone Regeneration Genetic Therapy/*methods Genetic Vectors Humans Musculoskeletal Diseases/*therapy Stem Cell Transplantation/*methods Tissue Engineering/methods},
	issn = {0969-7128 (Print) 0969-7128 (Linking)},
	doi = {10.1038/sj.gt.3302197},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/14724684},
	author = {Gafni, Y. and Turgeman, G. and Liebergal, M. and Pelled, G. and Gazit, Z. and Gazit, D.}
}
@article {17066,
	title = {Estrogens activate bone morphogenetic protein-2 gene transcription in mouse mesenchymal stem cells},
	journal = {Mol Endocrinol},
	volume = {17},
	number = {1},
	year = {2003},
	note = {Zhou, Shuanhu Turgeman, Gadi Harris, Stephen E Leitman, Dale C Komm, Barry S Bodine, Peter V N Gazit, Dan eng Research Support, Non-U.S. Gov{\textquoteright}t 2003/01/04 04:00 Mol Endocrinol. 2003 Jan;17(1):56-66.},
	pages = {56-66},
	type = {Journal Article},
	abstract = {Estrogens exert their physiological effects on target tissues by interacting with the estrogen receptors, ERalpha and ERbeta. Estrogen replacement is one the most common and effective strategies used to prevent osteoporosis in postmenopausal women. Whereas it was thought that estrogens work exclusively by inhibiting bone resorption, our previous results show that 17beta-estradiol (E2) increases mouse bone morphogenetic protein (BMP)-2 mRNA, suggesting that estrogens may also enhance bone formation. In this study, we used quantitative real-time RT-PCR analysis to demonstrate that estrogens increase BMP-2 mRNA in mouse mesenchymal stem cells. The selective ER modulators, tamoxifen, raloxifene, and ICI-182,780 (ICI), failed to enhance BMP-2 mRNA, whereas ICI inhibited E2 stimulation of expression. To investigate if estrogens increase BMP-2 expression by transcriptional mechanisms and if the response is mediated by ERalpha and/or ERbeta, we studied the effects of estrogens on BMP-2 promoter activity in transient transfected C3H10T1/2 cells. E2 produced a dose-dependent induction of the mouse -2712 BMP-2 promoter activity in cells cotransfected with ERalpha and ERbeta. At a dose of 10 nM E2, ERalpha induced mouse BMP-2 promoter activity 9-fold, whereas a 3-fold increase was observed in cells cotransfected with ERbeta. Tamoxifen and raloxifene were weak activators of the mouse BMP-2 promoter via ERalpha, but not via ERbeta. ICI blocked the activation of BMP-2 promoter activity by E2 acting via both ERalpha and ERbeta, indicating that mouse BMP-2 promoter activation is ER dependent. In contrast to E2 and selective ER modulators, the phytoestrogen, genistein was more effective at activating the mouse BMP-2 promoter with ERbeta, compared with ERalpha. Using a deletion series of the BMP-2 promoter, we determined that AP-1 or Sp1 sites are not required for E2 activation. A mutation in a sequence at -415 to -402 (5{\textquoteright}-GGGCCActcTGACCC-3{\textquoteright}) that resembles the classical estrogen-responsive element abolished the activation of the BMP-2 promoter in response to E2. Our studies demonstrate that E2 activation of mouse BMP-2 gene transcription requires ERalpha or ERbeta acting via a variant estrogen-responsive element binding site in the promoter, with ERalpha being the more efficacious regulator. Estrogenic compounds may enhance bone formation by increasing the transcription of the BMP-2 gene.},
	keywords = {Animals Bone Marrow Cells/metabolism Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics/*metabolism Cells, Cultured Dose-Response Relationship, Drug Estradiol/genetics/*physiology Female Gene Expression Regulation Genistein/pharmacology Mesoderm/cytology/physiology Mice RNA, Estrogen/classification/genetics/metabolism Response Elements/physiology Reverse Transcriptase Polymerase Chain Reaction Selective Estrogen Receptor Modulators/pharmacology Stem Cells/*metabolism Transcription, Genetic Transcriptional Activation *Transforming Growth Factor beta, Messenger/metabolism Receptors},
	issn = {0888-8809 (Print) 0888-8809 (Linking)},
	doi = {10.1210/me.2002-0210},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12511606},
	author = {Zhou, S. and Turgeman, G. and Harris, S. E. and Leitman, D. C. and Komm, B. S. and Bodine, P. V. and Gazit, D.}
}
@article {17065,
	title = {Molecular imaging of the skeleton: quantitative real-time bioluminescence monitoring gene expression in bone repair and development},
	journal = {J Bone Miner Res},
	volume = {18},
	number = {3},
	year = {2003},
	note = {Iris, Bar Zilberman, Yoram Zeira, Eveline Galun, Eithan Honigman, Alik Turgeman, Gadi Clemens, Thomas Gazit, Zulma Gazit, Dan eng 2003/03/07 04:00 J Bone Miner Res. 2003 Mar;18(3):570-8.},
	pages = {570-8},
	type = {Journal Article},
	abstract = {Monitoring gene expression in vivo, noninvasively, is a critical issue in effective gene therapy systems. To date, there are no adequate molecular imaging techniques, which quantitatively monitor gene expression in vivo in skeletal development and repair. The aim of this study was to monitor gene expression in skeletal development and repair, using a real-time molecular imaging system, which quantitatively and noninvasively detects bioluminescence in vivo. Our experimental model consisted of transgenic mice harboring the luciferase marker gene under the regulation of the human osteocalcin (hOC) promoter. A new light detection cooled charge coupled device (CCCD) camera was applied to monitor luciferase expression. In vitro, mesenchymal stem cells (MSCs) isolated from bone marrow of transgenic mice exhibited hOC promoter regulation, detected by luciferase expression that correlated with their osteogenic differentiation. During development from 1 week to 1.5 years, transgenic mice exhibited transgene expression in a wide spectrum of skeletal organs, including calvaria, vertebra, tail, and limbs, reaching a peak at 1 week in most of the skeletal organs. In two skeletal repair models, bone fracture and marrow ablation, the noninvasive CCCD system revealed a peak of luciferase expression at 6 days postsurgery. All quantitative, noninvasive, real-time CCCD measurements correlated with a luciferase biochemical assay and luciferase immunohistochemistry, which demonstrated luciferase expression in hypertrophic chondrocytes and trabecular osteoblasts. Our studies show for the first time (1) the CCCD detection system is a reliable quantitative gene detection tool for the skeleton in vivo, (2) expression of luciferase regulated by the hOC promoter is significantly decreased with age in most skeletal sites, and (3) the dynamics of hOC regulation during mice skeletal development and repair in real time, quantitatively and noninvasively.},
	keywords = {Animals Base Sequence *Bone Development *Bone Remodeling DNA Primers *Gene Expression Humans Luciferases Luminescent Measurements Mice Mice, Transgenic Osteocalcin/genetics},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1359/jbmr.2003.18.3.570},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12619943},
	author = {Iris, B. and Zilberman, Y. and Zeira, E. and Galun, E. and Honigman, A. and Turgeman, G. and Clemens, T. and Gazit, Z. and Gazit, D.}
}
@article {17063,
	title = {Cell-mediated gene therapy for bone formation and regeneration},
	journal = {Curr Opin Mol Ther},
	volume = {4},
	number = {4},
	year = {2002},
	note = {Turgeman, Gadi Aslan, Hadi Gazit, Zulma Gazit, Dan eng Review England 2002/09/12 10:00 Curr Opin Mol Ther. 2002 Aug;4(4):390-4.},
	pages = {390-4},
	type = {Journal Article},
	abstract = {Cell-mediated gene therapy is one of the new modalities branching out from the wide-ranging field of gene transfer and therapy. When applied to bone formation and regeneration, it has particular advantages depending on the type of cell used as a platform for gene delivery. When utilizing adult mesenchymal stem cells or osteoprogenitor cells for the expression of bone-promoting osteogenic factors, the cells not only express the factors promoting bone growth, but can respond, differentiate and participate in the bone formation process. The ability of engineered cells to respond to the transgene, as well as to other local signals in vivo, confers on them special properties that enable the formation and regeneration of large-scale bone tissue. This approach is a paradigm for the development of gene therapy strategies for other skeletal tissues. Here, we review the most recent studies related to cell-mediated gene therapy for bone formation and regeneration.},
	keywords = {Animals Bone Development/*genetics Bone Regeneration/*genetics *Genetic Therapy Genetic Vectors Humans Mesoderm/physiology Mice Osteogenesis Imperfecta/genetics/therapy Osteopetrosis/genetics/therapy Osteoporosis/genetics/therapy Promoter Regions, Genetic Stem Cells/physiology},
	issn = {1464-8431 (Print) 1464-8431 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12222877},
	author = {Turgeman, G. and Aslan, H. and Gazit, Z. and Gazit, D.}
}
@article {17062,
	title = {Mesenchymal stem cells for bone gene therapy and tissue engineering},
	journal = {Curr Pharm Des},
	volume = {8},
	number = {21},
	year = {2002},
	note = {Pelled, G G, Turgeman Aslan, H Gazit, Z Gazit, D eng Review Netherlands 2002/08/13 10:00 Curr Pharm Des. 2002;8(21):1917-28.},
	pages = {1917-28},
	type = {Journal Article},
	abstract = {Mesenchymal Stem Cells (MSCs) are adult stem cells that constitute a variety of adult tissues. MSCs maintain self-renewal ability with the ability to give rise to different mesenchymal cells, and are therefore responsible in part, for the regenerative capacity of mesenchymal tissues. MSCs throughout a variety of species were found to be able to differentiate to several mesenchymal tissues including: bone, cartilage, stroma, adipose, connective tissue, muscle and tendon. MSCs are relatively easily isolated from the bone marrow and expanded in vitro. It was found that MSCs play an important role in bone physiology and hematopoiesis, and in part participate in the pathophysiology related to bone diseases, mainly osteoporosis. MSCs were widely used in experimental studies in vivo, and were shown to form mesenchymal tissues. These discovered features have made MSCs good candidates for the development of various therapeutic modalities aimed to regenerate mesenchymal tissues, mainly bone. The more important approaches currently utilizing MSCs are gene therapy and tissue engineering. Both exploit the current knowledge in molecular biology and biomaterial science in order to direct MSCs to differentiate in vivo to desired lineages and tissues. Better understanding of the molecular mechanism directing the differentiation of MSCs, will eventually allow us to properly manipulate MSCs both in vivo and ex vivo to allow the regeneration of complex tissues and organs.},
	keywords = {Animal Osteogenesis Imperfecta/pathology/therapy Osteoporosis/pathology/therapy Rats *Stem Cell Transplantation Stem Cells/*cytology Tissue Engineering/*methods, Animals Bone Marrow Cells/*cytology Bone Neoplasms/pathology/therapy Genetic Therapy/*methods Humans Mesoderm/*cytology Models},
	issn = {1381-6128 (Print) 1381-6128 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12171527},
	author = {Pelled, G. and G, T. and Aslan, H. and Gazit, Z. and Gazit, D.}
}
@article {17060,
	title = {Synthesis and biodegradation of arabinogalactan sponges prepared by reductive amination},
	journal = {Biomaterials},
	volume = {23},
	number = {23},
	year = {2002},
	note = {Ehrenfreund-Kleinman, T Gazit, Z Gazit, D Azzam, T Golenser, J Domb, A J eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. Netherlands 2002/09/27 06:00 Biomaterials. 2002 Dec;23(23):4621-31.},
	pages = {4621-31},
	type = {Journal Article},
	abstract = {The synthesis of polysaccharide-based sponges for the use in tissue engineering was systematically investigated. A comparison study of the branched polysaccharide arabinogalactan (AG) and the linear polysaccharide dextran in the formation of sponges by the reaction with diamines or polyamines was conducted. Three AG-based sponges were synthesized from the crosslinking reaction with different amine molecules. The sponges obtained were highly porous, rapidly swelled in water, and were stable in vitro for at least 11 weeks in aqueous media at 37 degrees C. AG-chitosan sponges were chosen as most suitable to serve as scaffolds for cell growth in tissue engineering. The biocompatibility in vivo of these sponges was evaluated by histological staining and non-invasive MRI technique after implantation in BALB/c mice. The sponge evoked an inflammatory response with vascularization of the implant. The inflammatory reaction decreased with time, indicating a healing process.},
	keywords = {Animals *Biocompatible Materials Carbohydrate Sequence Chitin/*analogs \& derivatives/chemistry/pharmacology Chitosan Female Galactans/*chemistry/*pharmacology Glutamic Acid/chemistry Hydrogen-Ion Concentration Lysine/chemistry Mice Mice, Bacterial/*chemistry Temperature Time Factors Tissue Engineering, Electron, Inbred BALB C Microscopy, Scanning Molecular Sequence Data Molecular Weight Polysaccharides/chemistry Polysaccharides},
	issn = {0142-9612 (Print) 0142-9612 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12322984},
	author = {Ehrenfreund-Kleinman, T. and Gazit, Z. and Gazit, D. and Azzam, T. and Golenser, J. and Domb, A. J.}
}
@article {17064,
	title = {Systemically administered rhBMP-2 promotes MSC activity and reverses bone and cartilage loss in osteopenic mice},
	journal = {J Cell Biochem},
	volume = {86},
	number = {3},
	year = {2002},
	note = {Turgeman, Gadi Zilberman, Yoram Zhou, Shuanhu Kelly, Pam Moutsatsos, Ioannis K Kharode, Yogendra P Borella, Luis E Bex, Frederick J Komm, Barry S Bodine, Peter V N Gazit, Dan eng 2002/09/05 10:00 J Cell Biochem. 2002;86(3):461-74.},
	pages = {461-74},
	type = {Journal Article},
	abstract = {Osteoporosis is a disease manifested in drastic bone loss resulting in osteopenia and high risk for fractures. This disease is generally divided into two subtypes. The first, post-menopausal (type I) osteoporosis, is primarily related to estrogen deficiency. The second, senile (type II) osteoporosis, is mostly related to aging. Decreased bone formation, as well as increased bone resorption and turnover, are thought to play roles in the pathophysiology of both types of osteoporosis. In this study, we demonstrate in murine models for both type I (estrogen deficiency) and type II (senile) osteopenia/osteoporosis that reduced bone formation is related to a decrease in adult mesenchymal stem cell (AMSC) number, osteogenic activity, and proliferation. Decreased proliferation is coupled with increased apoptosis in AMSC cultures obtained from osteopenic mice. Recombinant human bone morphogenetic protein (rhBMP-2) is a highly osteoinductive protein, promoting osteogenic differentiation of AMSCs. Systemic intra-peritoneal (i.p.) injections of rhBMP-2 into osteopenic mice were able to reverse this phenotype in the bones of these animals. Moreover, this change in bone mass was coupled to an increase in AMSCs numbers, osteogenic activity, and proliferation as well as a decrease in apoptosis. Bone formation activity was increased as well. However, the magnitude of this response to rhBMP-2 varied among different stains of mice. In old osteopenic BALB/c male mice (type II osteoporosis model), rhBMP-2 systemic treatment also restored both articular and epiphyseal cartilage width to the levels seen in young mice. In summary, our study shows that AMSCs are a good target for systemically active anabolic compounds like rhBMP-2.},
	keywords = {Aging/physiology Alkaline Phosphatase/metabolism Animals Apoptosis/drug effects Bone Diseases, Cultured Dose-Response Relationship, Drug Enzyme Activation Female Humans Male Mesoderm/*cytology Mice Mice, Inbred BALB C Mice, Inbred ICR Osteogenesis/*drug effects Osteoporosis/drug therapy/pathology Ovariectomy Recombinant Proteins/administration \& dosage/pharmacology/therapeutic use Stem Cells/*drug effects *Transforming Growth Factor beta, Metabolic/*drug therapy/pathology Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/administration \& dosage/*pharmacology/therapeutic use Bone and Bones/*drug effects/pathology Cartilage/*drug effects/growth \& development/pathology Cells},
	issn = {0730-2312 (Print) 0730-2312 (Linking)},
	doi = {10.1002/jcb.10231},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/12210753},
	author = {Turgeman, G. and Zilberman, Y. and Zhou, S. and Kelly, P. and Moutsatsos, I. K. and Kharode, Y. P. and Borella, L. E. and Bex, F. J. and Komm, B. S. and Bodine, P. V. and Gazit, D.}
}
@article {17061,
	title = {The T-box transcription factor Brachyury mediates cartilage development in mesenchymal stem cell line C3H10T1/2},
	journal = {J Cell Sci},
	volume = {115},
	number = {Pt 4},
	year = {2002},
	note = {Hoffmann, Andrea Czichos, Stefan Kaps, Christian Bachner, Dietmar Mayer, Hubert Kurkalli, Basan Gowda Zilberman, Yoram Turgeman, Gadi Pelled, Gadi Gross, Gerhard Gazit, Dan eng England 2002/02/28 10:00 J Cell Sci. 2002 Feb 15;115(Pt 4):769-81.},
	pages = {769-81},
	type = {Journal Article},
	abstract = {The BMP2-dependent onset of osteo/chondrogenic differentiation in the acknowledged pluripotent murine mesenchymal stem cell line (C3H10T1/2) is accompanied by the immediate upregulation of Fibroblast Growth Factor Receptor 3 (FGFR3) and a delayed response by FGFR2. Forced expression of FGFR3 in C3H10T1/2 is sufficient for chondrogenic differentiation, indicating an important role for FGF-signaling during the manifestation of the chondrogenic lineage in this cell line. Screening for transcription factors exhibiting a chondrogenic capacity in C3H10T1/2 identified that the T-box containing transcription factor Brachyury is upregulated by FGFR3-mediated signaling. Forced expression of Brachyury in C3H10T1/2 was sufficient for differentiation into the chondrogenic lineage in vitro and in vivo after transplantation into muscle. A dominant-negative variant of Brachyury, consisting of its DNA-binding domain (T-box), interferes with BMP2-mediated cartilage formation. These studies indicate that BMP-initiated FGF-signaling induces a novel type of transcription factor for the onset of chondrogenesis in a mesenchymal stem cell line. A potential role for this T-box factor in skeletogenesis is further delineated from its expression profile in various skeletal elements such as intervertebral disks and the limb bud at late stages (18.5 d.p.c.) of murine embryonic development.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics/metabolism Cartilage/*embryology Cell Differentiation Cell Line Chondrocytes/cytology/*physiology/transplantation *Chondrogenesis Female *Fetal Proteins Fibroblast Growth Factor 3 F, Fibroblast Growth Factor/metabolism Recombinant Proteins/metabolism Signal Transduction Stem Cells/physiology T-Box Domain Proteins/genetics/*metabolism Transcription Factors/genetics/*metabolism *Transforming Growth Factor beta Up-Regulation, Nude Proto-Oncogene Proteins/metabolism Receptors},
	issn = {0021-9533 (Print) 0021-9533 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11865033},
	author = {Hoffmann, A. and Czichos, S. and Kaps, C. and Bachner, D. and Mayer, H. and Kurkalli, B. G. and Zilberman, Y. and Turgeman, G. and Pelled, G. and Gross, G. and Gazit, D.}
}
@article {17058,
	title = {Engineered human mesenchymal stem cells: a novel platform for skeletal cell mediated gene therapy},
	journal = {J Gene Med},
	volume = {3},
	number = {3},
	year = {2001},
	note = {Turgeman, G Pittman, D D Muller, R Kurkalli, B G Zhou, S Pelled, G Peyser, A Zilberman, Y Moutsatsos, I K Gazit, D eng Research Support, Non-U.S. Gov{\textquoteright}t England 2001/07/05 10:00 J Gene Med. 2001 May-Jun;3(3):240-51.},
	pages = {240-51},
	type = {Journal Article},
	abstract = {BACKGROUND: Human mesenchymal stem cells (hMSCs) are pluripotent cells that can differentiate to various mesenchymal cell types. Recently, a method to isolate hMSCs from bone marrow and expand them in culture was described. Here we report on the use of hMSCs as a platform for gene therapy aimed at bone lesions. METHODS: Bone marrow derived hMSCs were expanded in culture and infected with recombinant adenoviral vector encoding the osteogenic factor, human BMP-2. The osteogenic potential of genetically engineered hMSCs was assessed in vitro and in vivo. RESULTS: Genetically engineered hMSCs displayed enhanced proliferation and osteogenic differentiation in culture. In vivo, transplanted genetically engineered hMSCs were able to engraft and form bone and cartilage in ectopic sites, and regenerate bone defects (non-union fractures) in mice radius bone. Importantly, the same results were obtained with hMSCs isolated from a patient suffering from osteoporosis. CONCLUSIONS: hMSCs represent a novel platform for skeletal gene therapy and the present results suggest that they can be genetically engineered to express desired therapeutic proteins inducing specific differentiation pathways. Moreover, hMSCs obtained from osteoporotic patients can restore their osteogenic activity following human BMP-2 gene transduction, an important finding in the future planning of gene therapy treatment for osteoporosis.},
	keywords = {Adenoviridae/*genetics Animals Bone Development/genetics Bone Diseases/genetics/*therapy Bone Marrow Cells/*cytology Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/*genetics Bone Regeneration/genetics Cells, Cultured Genetic Therapy/*methods Genetic Vectors Humans Mesoderm/*cytology Mice Recombinant Proteins *Stem Cells Transduction, Genetic *Transforming Growth Factor beta},
	issn = {1099-498X (Print) 1099-498X (Linking)},
	doi = {10.1002/1521-2254(200105/06)3:3<240::AID-JGM181>3.0.CO;2-A},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11437329},
	author = {Turgeman, G. and Pittman, D. D. and Muller, R. and Kurkalli, B. G. and Zhou, S. and Pelled, G. and Peyser, A. and Zilberman, Y. and Moutsatsos, I. K. and Gazit, D.}
}
@article {17059,
	title = {Estrogen modulates estrogen receptor alpha and beta expression, osteogenic activity, and apoptosis in mesenchymal stem cells (MSCs) of osteoporotic mice},
	journal = {J Cell Biochem Suppl},
	volume = {Suppl 36},
	year = {2001},
	note = {Zhou, S Zilberman, Y Wassermann, K Bain, S D Sadovsky, Y Gazit, D eng Research Support, Non-U.S. Gov{\textquoteright}t 2001/07/17 10:00 J Cell Biochem Suppl. 2001;Suppl 36:144-55.},
	pages = {144-55},
	type = {Journal Article},
	abstract = {In the mouse, ovariectomy (OVX) leads to significant reductions in cancellous bone volume while estrogen (17beta-estradiol, E2) replacement not only prevents bone loss but can increase bone formation. As the E2-dependent increase in bone formation would require the proliferation and differentiation of osteoblast precursors, we hypothesized that E2 regulates mesenchymal stem cells (MSCs) activity in mouse bone marrow. We therefore investigated proliferation, differentiation, apoptosis, and estrogen receptor (ER) alpha and beta expression of primary culture MSCs isolated from OVX and sham-operated mice. MSCs, treated in vitro with 10(-7) M E2, displayed a significant increase in ERalpha mRNA and protein expression as well as alkaline phosphatase (ALP) activity and proliferation rate. In contrast, E2 treatment resulted in a decrease in ERbeta mRNA and protein expression as well as apoptosis in both OVX and sham mice. E2 up-regulated the mRNA expression of osteogenic genes for ALP, collagen I, TGF-beta1, BMP-2, and cbfa1 in MSCs. In a comparison of the relative mRNA expression and protein levels for two ER isoforms, ERalpha was the predominant form expressed in MSCs obtained from both OVX and sham-operated mice. Cumulatively, these results indicate that estrogen in vitro directly augments the proliferation and differentiation, ERalpha expression, osteogenic gene expression and, inhibits apoptosis and ERbeta expression in MSCs obtained from OVX and sham-operated mice. Co-expression of ERalpha, but not ERbeta, and osteogenic differentiation markers might indicate that ERalpha function as an activator and ERbeta function as a repressor in the osteogenic differentiation in MSCs. These results suggest that mouse MSCs are anabolic targets of estrogen action, via ERalpha activation. J. Cell. Biochem. Suppl. 36: 144-155, 2001.},
	keywords = {Alkaline Phosphatase/genetics/metabolism Animals *Apoptosis Bone Marrow Cells/metabolism/pathology Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics/metabolism Cell Differentiation Cell Division Cells, Cultured Collagen/genetics/metabolism *Collagen Type I Core Binding Factor Alpha 1 Subunit Estradiol/*pharmacology Estrogen Receptor alpha Estrogen Receptor beta Female Immunohistochemistry Mesoderm/pathology Mice *Neoplasm Proteins Osteogenesis/*drug ef, Estrogen/genetics/*metabolism Reverse Transcriptase Polymerase Chain Reaction Stem Cells/metabolism/*pathology Transcription Factors/genetics/metabolism Transforming Growth Factor beta/genetics/metabolism Transforming Growth Factor beta1, Messenger/metabolism Receptors},
	issn = {0733-1959 (Print) 0733-1959 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11455579},
	author = {Zhou, S. and Zilberman, Y. and Wassermann, K. and Bain, S. D. and Sadovsky, Y. and Gazit, D.}
}
@article {17057,
	title = {Exogenously regulated stem cell-mediated gene therapy for bone regeneration},
	journal = {Mol Ther},
	volume = {3},
	number = {4},
	year = {2001},
	note = {Moutsatsos, I K Turgeman, G Zhou, S Kurkalli, B G Pelled, G Tzur, L Kelley, P Stumm, N Mi, S Muller, R Zilberman, Y Gazit, D eng Research Support, Non-U.S. Gov{\textquoteright}t 2001/04/26 10:00 Mol Ther. 2001 Apr;3(4):449-61.},
	pages = {449-61},
	type = {Journal Article},
	abstract = {Regulated expression of transgene production and function is of great importance for gene therapy. Such regulation can potentially be used to monitor and control complex biological processes. We report here a regulated stem cell-based system for controlling bone regeneration, utilizing genetically engineered mesenchymal stem cells (MSCs) harboring a tetracycline-regulated expression vector encoding the osteogenic growth factor human BMP-2. We show that doxycycline (a tetracycline analogue) is able to control hBMP-2 expression and thus control MSC osteogenic differentiation both in vitro and in vivo. Following in vivo transplantation of genetically engineered MSCs, doxycycline administration controlled both bone formation and bone regeneration. Moreover, our findings showed increased angiogenesis accompanied by bone formation whenever genetically engineered MSCs were induced to express hBMP-2 in vivo. Thus, our results demonstrate that regulated gene expression in mesenchymal stem cells can be used as a means to control bone healing.},
	keywords = {Animals Anti-Bacterial Agents/pharmacology Bone Development/genetics Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics Bone Regeneration/*genetics Cell Differentiation Cell Line Chick Embryo Doxycycline/pharmacology Female Fracture Healing, Fluorescence Neovascularization, Inbred BALB C Mice, Inbred C3H Microscopy, Physiologic Recombinant Proteins/genetics Reverse Transcriptase Polymerase Chain Reaction Stem Cells/*metabolism *Transforming Growth Factor beta Transgenes},
	issn = {1525-0016 (Print) 1525-0016 (Linking)},
	doi = {10.1006/mthe.2001.0291},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11319905},
	author = {Moutsatsos, I. K. and Turgeman, G. and Zhou, S. and Kurkalli, B. G. and Pelled, G. and Tzur, L. and Kelley, P. and Stumm, N. and Mi, S. and Muller, R. and Zilberman, Y. and Gazit, D.}
}
@article {17055,
	title = {Human parathyroid hormone 1-34 reverses bone loss in ovariectomized mice},
	journal = {J Bone Miner Res},
	volume = {16},
	number = {9},
	year = {2001},
	note = {Alexander, J M Bab, I Fish, S Muller, R Uchiyama, T Gronowicz, G Nahounou, M Zhao, Q White, D W Chorev, M Gazit, D Rosenblatt, M eng R01 DK47940/DK/NIDDK NIH HHS/ Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, P.H.S. 2001/09/08 10:00 J Bone Miner Res. 2001 Sep;16(9):1665-73.},
	pages = {1665-73},
	type = {Journal Article},
	abstract = {The experimental work characterizing the anabolic effect of parathyroid hormone (PTH) in bone has been performed in nonmurine ovariectomized (OVX) animals, mainly rats. A major drawback of these animal models is their inaccessibility to genetic manipulations such as gene knockout and overexpression. Therefore, this study on PTH anabolic activity was carried out in OVX mice that can be manipulated genetically in future studies. Adult Swiss-Webster mice were OVX, and after the fifth postoperative week were treated intermittently with human PTH(1-34) [hPTH(1-34)] or vehicle for 4 weeks. Femoral bones were evaluated by microcomputed tomography (microCT) followed by histomorphometry. A tight correlation was observed between trabecular density (BV/TV) determinations made by both methods. The BV/TV showed >60\% loss in the distal metaphysis in 5-week and 9-week post-OVX, non-PTH-treated animals. PTH induced a approximately 35\% recovery of this loss and a approximately 40\% reversal of the associated decreases in trabecular number (Tb.N) and connectivity. PTH also caused a shift from single to double calcein-labeled trabecular surfaces, a significant enhancement in the mineralizing perimeter and a respective 2- and 3-fold stimulation of the mineral appositional rate (MAR) and bone formation rate (BFR). Diaphyseal endosteal cortical MAR and thickness also were increased with a high correlation between these parameters. These data show that OVX osteoporotic mice respond to PTH by increased osteoblast activity and the consequent restoration of trabecular network. The Swiss-Webster mouse model will be useful in future studies investigating molecular mechanisms involved in the pathogenesis and treatment of osteoporosis, including the mechanisms of action of known and future bone antiresorptive and anabolic agents.},
	keywords = {Animal Female Femur/*drug effects/pathology/physiopathology Humans Mice Osteoporosis, Animals Disease Models, Postmenopausal/drug therapy/*pathology/physiopathology Ovariectomy Teriparatide/administration \& dosage/*pharmacology/therapeutic use},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1359/jbmr.2001.16.9.1665},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11547836},
	author = {Alexander, J. M. and Bab, I. and Fish, S. and Muller, R. and Uchiyama, T. and Gronowicz, G. and Nahounou, M. and Zhao, Q. and White, D. W. and Chorev, M. and Gazit, D. and Rosenblatt, M.}
}
@article {17056,
	title = {Imaging transgene expression in live animals},
	journal = {Mol Ther},
	volume = {4},
	number = {3},
	year = {2001},
	note = {Honigman, A Zeira, E Ohana, P Abramovitz, R Tavor, E Bar, I Zilberman, Y Rabinovsky, R Gazit, D Joseph, A Panet, A Shai, E Palmon, A Laster, M Galun, E eng Research Support, Non-U.S. Gov{\textquoteright}t 2001/09/08 10:00 Mol Ther. 2001 Sep;4(3):239-49.},
	pages = {239-49},
	type = {Journal Article},
	abstract = {Monitoring the expression of therapeutic genes in targeted tissues in disease models is important to assessing the effectiveness of systems of gene therapy delivery. We applied a new light-detection cooled charged-coupled device (CCCD) camera for continuous in vivo assessment of commonly used gene therapy delivery systems (such as ex vivo manipulated cells, viral vectors, and naked DNA), without the need to kill animals. We examined a variety of criteria related to real-time monitoring of luciferase (luc) gene expression in tissues including bone, muscle, salivary glands, dermis, liver, peritoneum, testis, teeth, prostate, and bladder in living mice and rats. These criteria included determination of the efficiency of infection/transfection of various viral and nonviral delivery systems, promoter specificity, and visualization of luciferase activity, and of the ability of luciferin to reach various organs. The exposure time for detection of luc activity by the CCCD camera is relatively short (approximately 2 minutes) compared with the intensified CCD camera photon-counting method (approximately 15 minutes). Here we transduce a variety of vectors (such as viruses, transfected cells, and naked DNA) by various delivery methods, including electroporation, systemic injection of viruses, and tail-vein, high-velocity-high-volume administration of DNA plasmids. The location, intensity, and duration of luc expression in different organs were determined. The distribution of luciferin is most probably not a barrier for the detection of in vivo luciferase activity. We showed that the CCCD photon detection system is a simple, reproducible, and applicable method that enables the continuous monitoring of a gene delivery system in living animals.},
	keywords = {Adenoviridae/metabolism Animals Diagnostic Imaging/*instrumentation/*methods Firefly Luciferin/metabolism *Gene Expression Genes, Reporter/genetics Genetic Therapy/*methods Liver/metabolism Luciferases/genetics/metabolism Male Mice Muscles/metabolism Organ Specificity Photography/methods Prostate/metabolism Rats Spleen/metabolism Time Factors Tooth/metabolism Transgenes/*genetics},
	issn = {1525-0016 (Print) 1525-0016 (Linking)},
	doi = {10.1006/mthe.2001.0437},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/11545615},
	author = {Honigman, A. and Zeira, E. and Ohana, P. and Abramovitz, R. and Tavor, E. and Bar, I. and Zilberman, Y. and Rabinovsky, R. and Gazit, D. and Joseph, A. and Panet, A. and Shai, E. and Palmon, A. and Laster, M. and Galun, E.}
}
@article {17053,
	title = {Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy},
	journal = {J Gene Med},
	volume = {1},
	number = {2},
	year = {1999},
	note = {Gazit, D Turgeman, G Kelley, P Wang, E Jalenak, M Zilberman, Y Moutsatsos, I eng England 2000/03/30 09:00 J Gene Med. 1999 Mar-Apr;1(2):121-33.},
	pages = {121-33},
	type = {Journal Article},
	abstract = {BACKGROUND: Among the approximately 6.5 million fractures suffered in the United States every year, about 15\% are difficult to heal. As yet, for most of these difficult cases there is no effective therapy. We have developed a mouse radial segmental defect as a model experimental system for testing the capacity of Genetically Engineered Pluripotent Mesenchymal Cells (GEPMC, C3H10T1/2 clone expressing rhBMP-2), for gene delivery, engraftment, and induction of bone growth in regenerating bone. METHODS: Transfected GEPMC expressing rhBMP-2 were further infected with a vector carrying the lacZ gene, that encodes for beta-galactosidase (beta-gal). In vitro levels of rhBMP-2 expression and function were confirmed by immunohistochemistry, and bioassay. Differentiation was assayed using alkaline phosphatase staining. GEPMC were transplanted in vivo into a radial segmental defect. The main control groups included lacZ clones of WT-C3H10T1/2-LacZ, and CHO-rhBMP-2 cells. New bone formation was measured quantitatively via fluorescent labeling, X-ray analysis and histomorphometry. Engrafted mesenchymal cells were localized in vivo by beta-gal expression, and double immunofluorescence. RESULTS: In vitro, GEPMC expressed rhBMP-2, beta-gal and spontaneously differentiated into osteogenic cells expressing alkaline phosphatase. Detection of transplanted cells revealed engrafted cells that had differentiated into osteoblasts and co-expressed beta-gal and rhBMP-2. Analysis of new bone formation revealed that at four to eight week post-transplantation, GEPMS significantly enhanced segmental defect repair. CONCLUSIONS: Our study shows that cell-mediated gene transfer can be utilized for growth factor delivery to signaling receptors of transplanted cells (autocrine effect) and host mesenchymal cells (paracrine effect) suggesting the ability of GEPMC to engraft, differentiate, and stimulate bone growth. We suggest that our approach should lead to the designing of mesenchymal stem cell based gene therapy strategies for bone lesions as well as other tissues.},
	keywords = {Animals Bone Morphogenetic Protein 2 Bone Morphogenetic Proteins/genetics Bone and Bones/cytology/*physiology CHO Cells Cell Differentiation Cell Line Cell Transplantation Cricetinae Female Fracture Healing/genetics/physiology Genetic Engineering *Genetic, Biological Radius Fractures/pathology/physiopathology/therapy Regeneration/*genetics/*physiology Stem Cells/cytology/metabolism Transfection *Transforming Growth Factor beta, Inbred C3H Models},
	issn = {1099-498X (Print) 1099-498X (Linking)},
	doi = {10.1002/(SICI)1521-2254(199903/04)1:2<121::AID-JGM26>3.0.CO;2-J},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/10738576},
	author = {Gazit, D. and Turgeman, G. and Kelley, P. and Wang, E. and Jalenak, M. and Zilberman, Y. and Moutsatsos, I.}
}
@article {17054,
	title = {Recombinant TGF-beta1 stimulates bone marrow osteoprogenitor cell activity and bone matrix synthesis in osteopenic, old male mice},
	journal = {J Cell Biochem},
	volume = {73},
	number = {3},
	year = {1999},
	note = {Gazit, D Zilberman, Y Turgeman, G Zhou, S Kahn, A eng AR37716/AR/NIAMS NIH HHS/ AR41328/AR/NIAMS NIH HHS/ Comparative Study Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, P.H.S. 1999/05/13 02:05 J Cell Biochem. 1999 Jun 1;73(3):379-89.},
	pages = {379-89},
	type = {Journal Article},
	abstract = {We have previously hypothesized that the osteopenic changes seen in the skeletons of old male BALB/c mice are due to reductions in the availability and/or synthesis of bone TGF-beta which results in fewer, less osteogenic marrow osteoprogenitor cells (CFU-f; OPCs) and lower levels of bone formation. Among other things, this hypothesis would predict that introducing exogenous TGF-beta into old mice (growth factor replacement) should stimulate marrow CFU-f and increase bone formation. In the present study, we have tested this prediction and, indirectly the hypothesis, by injecting human recombinant TGF-beta1, i.p., into both young adult (4 month) and old mice (24 month). The effects of the growth factor on the skeleton were then assessed by measurements of trabecular bone volume, bone formation, fracture healing, and the number, proliferative, apoptotic, and alkaline phosphatase activity of marrow CFU-f/OPCs. Our data show that the introduction of 0.5 or 5.0 ug/day of TGF-beta1 into old mice for 20 days 1) increases trabecular bone volume, bone formation and the mineral apposition rate, 2) augments fracture healing, 3) increases the number and size of CFU-f colonies, and 4) increases proliferation and diminishes apoptosis of CFU-f in primary bone marrow cultures. Importantly, these stimulatory effects of injected growth factor are apparently age-specific, i.e., they are either not seen in young animals or, if seen, are found at much lower levels. While these observations do not exclude other possible mechanisms for the osteopenia of old mice, they provide further support for the hypothesis that, with age, diminished TGF-beta synthesis or availability results in a reduction in the marrow osteoprogenitor pool and bone formation. The findings also demonstrate that the latter changes can be reversed, at least transiently, by introducing exogenous TGF-beta1.},
	keywords = {Age Factors Alkaline Phosphatase/metabolism Animals Bone Diseases, Cultured Femur/anatomy \& histology Fracture Healing Male Mice Mice, Inbred BALB C Recombinant Proteins Ribs/anatomy \& histology Stem Cells/*metabolism Tibia/anatomy \& histology Transforming Growth Factor beta/*physiology, Metabolic/*metabolism Bone Marrow Cells/*metabolism Bone Matrix/*metabolism Cell Division Cells},
	issn = {0730-2312 (Print) 0730-2312 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/10321837},
	author = {Gazit, D. and Zilberman, Y. and Turgeman, G. and Zhou, S. and Kahn, A.}
}
@article {17052,
	title = {Bone loss (osteopenia) in old male mice results from diminished activity and availability of TGF-beta},
	journal = {J Cell Biochem},
	volume = {70},
	number = {4},
	year = {1998},
	note = {Gazit, D Zilberman, Y Ebner, R Kahn, A eng AR37716/AR/NIAMS NIH HHS/ AR41328/AR/NIAMS NIH HHS/ Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, P.H.S. 1998/08/26 02:22 J Cell Biochem. 1998 Sep 15;70(4):478-88.},
	pages = {478-88},
	type = {Journal Article},
	abstract = {One of the universal characteristics of the long bones and spines of middle-age and older mammals is a loss in bone mass (osteopenia). In humans, if this bone loss is severe enough, it results in osteoporosis, a skeletal disorder characterized by a markedly increased incidence of fractures with sequelae that may include pain, loss of mobility, and in the event of hip fracture, even death within a relatively few months of injury. An important contributing factor to the development of osteoporosis appears to be a diminution in the number and activity of osteoblasts responsible for synthesizing new bone matrix. The findings in the present and other similar studies suggest that this reduction in osteoblast number and activity is due to an age-related diminution in the size and osteogenic potential of the bone marrow osteoblast progenitor cell (OPC or CFU-f) compartment. We previously postulated that these regressive changes in the OPC/CFU-f compartment occurred in old animals because of a reduction in the amount and/or activity of TGF-beta1, an autocrine growth factor important in the promotion of OPC/CFU-f proliferation and differentiation. In support of this hypothesis, we now report that (1) the osteogenic capacity of the bone marrow of 24-month-old BALB/c mice, as assessed in vivo, is markedly reduced relative to that of 3-4-month-old animals, (2) that the matrix of the long bones of old mice contains significantly less TGF-beta than that of young mice, (3) that OPC{\textquoteright}s/CFU-f{\textquoteright}s isolated from old mice produce less TGF-beta in vitro than those recovered from young mice, and (4) that OPC{\textquoteright}s/CFU-f{\textquoteright}s from old mice express significantly more TGF-beta receptor (Types I, II, and III) than those of young animals and that such cells are more responsive in vitro to exogenous recombinant TGF-beta1. We also find that colony number and proliferative activity of OPC{\textquoteright}s/CFU-f{\textquoteright}s of young mice and old mice, respectively, are significantly reduced when incubated in the presence of neutralizing TGF-beta1 antibody. Collectively, these data are consistent with the hypothesis that in old male mice the reduction in the synthesis and, perhaps, availability from the bone matrix of TGF-beta1 contributes to a diminution in the size and development potential of the bone marrow osteoprogenitor pool.},
	keywords = {Aging/*metabolism Animals Bone Development/drug effects Bone Diseases, Inbred BALB C Recombinant Proteins/pharmacology Transforming Growth Factor beta/*metabolism/pharmacology, Metabolic/*metabolism Bone Marrow Cells/cytology/metabolism Male Mice Mice},
	issn = {0730-2312 (Print) 0730-2312 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/9712146},
	author = {Gazit, D. and Zilberman, Y. and Ebner, R. and Kahn, A.}
}
@article {17051,
	title = {Early root resorption of maxillary primary first molars in a child with severe congenital heart disease},
	journal = {J Clin Pediatr Dent},
	volume = {21},
	number = {2},
	year = {1997},
	note = {Peretz, B Gleicher, H Gazit, D Eidelman, E eng Case Reports 1997/01/01 00:00 J Clin Pediatr Dent. 1997 Winter;21(2):163-6.},
	pages = {163-6},
	type = {Journal Article},
	keywords = {Bottle Feeding/adverse effects Child, Congenital/surgery Heart Septal Defects, Deciduous/*pathology Truncus Arteriosus, Intratracheal/adverse effects Molar Root Resorption/*etiology/therapy Tooth, Persistent/surgery, Preschool *Dental Care for Chronically Ill Dental Caries/complications/etiology Female *Heart Defects, Ventricular/surgery Humans Intubation},
	issn = {1053-4628 (Print) 1053-4628 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/9484110},
	author = {Peretz, B. and Gleicher, H. and Gazit, D. and Eidelman, E.}
}
@article {17050,
	title = {Internal coronal resorption of a permanent molar: a conservative approach for treatment},
	journal = {J Clin Pediatr Dent},
	volume = {21},
	number = {4},
	year = {1997},
	note = {Eidelman, E Rotstein, I Gazit, D eng Case Reports 1997/07/01 00:00 J Clin Pediatr Dent. 1997 Summer;21(4):287-90.},
	pages = {287-90},
	type = {Journal Article},
	abstract = {A case of internal coronal resorption in a maxillary first permanent molar of a young adult is described. Conservative treatment was done, which included endodontic therapy followed by coronal restoration with bonded composites. The importance of early diagnosis and treatment of the resorptive defect is stressed.},
	keywords = {Adolescent Composite Resins Dentin-Bonding Agents Female Humans Maxilla Molar/*pathology *Root Canal Therapy Tooth Crown/*pathology Tooth Movement Techniques/adverse effects Tooth Resorption/diagnosis/etiology/*therapy},
	issn = {1053-4628 (Print) 1053-4628 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/9484113},
	author = {Eidelman, E. and Rotstein, I. and Gazit, D.}
}
@article {17039,
	title = {Assessment of combined 24,25(OH)2D3 and 1 alpha (OH)D3 therapy for bone disease in dialysis patients},
	journal = {Bone},
	volume = {13},
	number = {5},
	year = {1992},
	note = {Popovtzer, M M Levi, J Bar-Khayim, Y Shasha, S M Boner, G Bernheim, J Chaimovitz, C Rubinger, D Gafter, U Gazit, D eng Comparative Study 1992/01/01 Bone. 1992;13(5):369-77.},
	pages = {369-77},
	type = {Journal Article},
	abstract = {An increasing body of experimental data suggests a role for 24,25(OH)2D3 in bone metabolism. The present study was carried out to assess a possible therapeutic role of this vitamin D metabolite in renal osteodystrophy. Twenty-two chronic dialysis patients, most of whom were previously maintained on 1 alpha (OH)D3 therapy, received additional treatment with 10 micrograms/day 24,25(OH)2D3 and were compared to 19 patients receiving 1 alpha (OH)D3 alone. Analysis of transiliac bone biopsies obtained at study entry and following 10-16 months of treatment revealed that the combined therapy produced a decrease in bone turnover. Specifically, the addition of 24,25(OH)2D3 inhibited an increase in trabecular bone volume (BV/TV) and suppressed osteoclastic parameters. Thus BV/TV increased from 26.2 +/- 8.6 to 32.1 +/- 7.5\% (p < 0.01) in the 1 alpha (OH)D3 group, but it remained unchanged in the combined therapy group. In contrast, the eroded surface (ES/BS), the osteoclast surface (Oc.S/BS), and the osteoclast numbers were significantly suppressed in patients receiving both 24,25(OH)2D3 and 1 alpha (OH)D3, as compared with those receiving 1 alpha (OH)D3 alone (p < 0.01, p < 0.01, and p < 0.001, respectively). These improvements were independent of changes in 1 alpha (OH)D3 dosage. The extent of bone aluminium deposits was unrelated to the administration of 24,25(OH)2D3 or to its effect. 24,25(OH)2D3 therapy was not associated with any adverse effects.},
	keywords = {24, 25-Dihydroxyvitamin D 3/blood/*therapeutic use Adult Aged Alkaline Phosphatase/blood *Bone Density Calcium/blood Chronic Kidney Disease-Mineral and Bone Disorder/blood/*drug therapy/etiology Creatinine/blood Drug Therapy, Combination Female Humans Hydroxycholecalciferols/blood/*therapeutic use Male Middle Aged Phosphates/blood Renal Dialysis/*adverse effects},
	issn = {8756-3282 (Print) 1873-2763 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1419378},
	author = {Popovtzer, M. M. and Levi, J. and Bar-Khayim, Y. and Shasha, S. M. and Boner, G. and Bernheim, J. and Chaimovitz, C. and Rubinger, D. and Gafter, U. and Gazit, D. and et al.}
}
@article {17038,
	title = {Cancrum oris-like lesions},
	journal = {Br J Oral Maxillofac Surg},
	volume = {30},
	number = {3},
	year = {1992},
	note = {Nahlieli, O Neder, A Ulmansky, M Gazit, D eng Comment Letter Scotland 1992/06/01 Br J Oral Maxillofac Surg. 1992 Jun;30(3):202.},
	pages = {202},
	type = {Journal Article},
	keywords = {Cutaneous/pathology, Humans Mouth Diseases/*pathology Noma/*pathology Purpura, Leukocytoclastic, Schoenlein-Henoch/*pathology Vasculitis},
	issn = {0266-4356 (Print) 0266-4356 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1622972},
	author = {Nahlieli, O. and Neder, A. and Ulmansky, M. and Gazit, D.}
}
@article {17037,
	title = {Expression of helix-loop-helix regulatory genes during differentiation of mouse osteoblastic cells},
	journal = {J Bone Miner Res},
	volume = {7},
	number = {10},
	year = {1992},
	note = {Murray, S S Glackin, C A Winters, K A Gazit, D Kahn, A J Murray, E J eng 5T32GM07616/GM/NIGMS NIH HHS/ AR 38168/AR/NIAMS NIH HHS/ Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. Research Support, U.S. Gov{\textquoteright}t, P.H.S. 1992/10/01 J Bone Miner Res. 1992 Oct;7(10):1131-8.},
	pages = {1131-8},
	type = {Journal Article},
	abstract = {Although much is known about the hormonal regulation of osteoblastic cell differentiation, much less is known about the nuclear regulatory molecules that affect this process. We analyzed the expression of several regulatory molecules of the helix-loop-helix (H-L-H) group in primary mouse calvarial cells and in MC3T3-E1 mouse osteoblastic cells in situations representing different degrees of cellular differentiation. H-L-H class regulators are known to participate directly in directing cell fate and differentiation decisions in other mesodermal lineages. Two of the molecules that we studied, Id and E12, have well-established roles in this process. The other, mTwi, the murine homolog of the Drosophila twist gene, is a newly cloned mammalian H-L-H gene. Levels of E12 RNA remained unchanged during differentiation. On the other hand, in both primary osteoblastic cells and MC3T3-E1 cells, the abundance of Id and mTwi declined with cell maturation; mTwi less dramatically than Id. That Id expression is causally related to differentiation is suggested by the finding that MC3T3-E1 cells transfected with an Id-expression plasmid fail to undergo differentiation. We conclude that helix-loop-helix regulatory genes are expressed in mouse osteoblastic cells, where they are likely to participate in differentiation. The E12 gene product is likely to function as a positive modulating factor. In contrast, Id inhibits differentiation, probably by sequestering other H-L-H gene regulators, including E12, in inactive complexes. The precise role of mTwi is more speculative at this time, but the observed pattern of expression is consistent with a role in early and midmesodermal specification that is terminated as cells differentiate.},
	keywords = {3T3 Cells Alkaline Phosphatase/analysis Animals Autoradiography Cell Differentiation/genetics Cells, Cultured DNA-Binding Proteins/*genetics *Gene Expression *Genes, Regulator Inhibitor of Differentiation Protein 1 Mice Nucleic Acid Hybridization Osteoblasts/cytology/*metabolism Plasmids RNA/analysis/genetics *Repressor Proteins TCF Transcription Factors Transcription Factor 7-Like 1 Protein *Transcription Factors Tr},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1002/jbmr.5650071004},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1280901},
	author = {Murray, S. S. and Glackin, C. A. and Winters, K. A. and Gazit, D. and Kahn, A. J. and Murray, E. J.}
}
@article {17035,
	title = {Histone H4-related osteogenic growth peptide (OGP): a novel circulating stimulator of osteoblastic activity},
	journal = {EMBO J},
	volume = {11},
	number = {5},
	year = {1992},
	note = {Bab, I Gazit, D Chorev, M Muhlrad, A Shteyer, A Greenberg, Z Namdar, M Kahn, A eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. England 1992/05/01 EMBO J. 1992 May;11(5):1867-73.},
	pages = {1867-73},
	type = {Journal Article},
	abstract = {It has been established that regenerating marrow induces an osteogenic response in distant skeletal sites and that this activity is mediated by factors released into the circulation by the healing tissue. In the present study we have characterized one of these factors, a 14 amino acid peptide named osteogenic growth peptide (OGP). Synthetic OGP, identical in structure to the native molecule, stimulates the proliferation and alkaline phosphatase activity of osteoblastic cells in vitro and increases bone mass in rats when injected in vivo. Immunoreactive OGP in high abundance is present physiologically in the serum, mainly in the form of an OGP-OGP binding protein complex. A marked increase in serum bound and unbound OGP accompanies the osteogenic phase of post-ablation marrow regeneration and associated systemic osteogenic response. Authentic OGP is identical to the C-terminus of histone H4 and shares a five residue motif with a T-cell receptor beta-chain V-region and the Bacillus subtilis outB locus. Since these latter proteins have not been implicated previously in the control of cell proliferation or differentiation, OGP may belong to a novel, heretofore unrecognized family of regulatory peptides. Perhaps more importantly, OGP appears to represent a new class of molecules involved in the systemic control of osteoblast proliferation and differentiation.},
	keywords = {Amino Acid Sequence Animals Autoradiography Bone Marrow/growth \& development Bone Marrow Cells Cells, Cultured Chromatography, Liquid Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Growth Substances/genetics/isolation \& purification/*metabolism Histones/metabolism *Intercellular Signaling Peptides and Proteins Molecular Sequence Data Osteoblasts/*metabolism Peptides/genetics/isol},
	issn = {0261-4189 (Print) 0261-4189 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1582415},
	author = {Bab, I. and Gazit, D. and Chorev, M. and Muhlrad, A. and Shteyer, A. and Greenberg, Z. and Namdar, M. and Kahn, A.}
}
@article {17036,
	title = {Unique congenital features in tuberous sclerosis},
	journal = {Clin Pediatr (Phila)},
	volume = {31},
	number = {6},
	year = {1992},
	note = {Gross-Tsur, V Klar, A Gazit, D Shalev, R S eng Case Reports 1992/06/01 Clin Pediatr (Phila). 1992 Jun;31(6):364-5.},
	pages = {364-5},
	type = {Journal Article},
	keywords = {Cheek Child, Preschool Female Fibrous Dysplasia of Bone/*congenital/diagnostic imaging/pathology Humans Maxillary Diseases/*congenital/diagnostic imaging Pigmentation Disorders/*congenital Tomography, X-Ray Computed Tuberous Sclerosis/*congenital/diagnostic imaging},
	issn = {0009-9228 (Print) 0009-9228 (Linking)},
	doi = {10.1177/000992289203100610},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1628471},
	author = {Gross-Tsur, V. and Klar, A. and Gazit, D. and Shalev, R. S.}
}
@article {17032,
	title = {Leukocytoclastic vasculitis (anaphylactoid purpura): a unique occurrence in the oral cavity},
	journal = {J Oral Pathol Med},
	volume = {20},
	number = {10},
	year = {1991},
	note = {Gazit, D Nahlieli, O Neder, A Berstein, I Ulmansky, M eng Case Reports Denmark 1991/11/01 J Oral Pathol Med. 1991 Nov;20(10):509-11.},
	pages = {509-11},
	type = {Journal Article},
	abstract = {Leukocytoclastic vasculitis, immune complex disorder (type III), is a skin disease with both an acute form characterized by bullae, vesicles and ulcerations, and a chronic form characterized by petechiae, macules and ulcerations. The disease presents certain systemic features including diffuse or focal glomerulonephritis and renal failure. The histopathologic characteristics of leukocytoclastic vasculitis in the skin appear primarily in small blood vessels and consist of an infiltration of inflammatory cells, leukoclasis, swelling of endothelial cells, occlusion of blood vessels, accumulation of fibrin and fibrinoid degeneration, as well as the presence of immune complexes in and around blood vessel walls. Although leukocytoclastic vasculitis is described as several diseases which can spread systemically, including the gastrointestinal tract and the kidneys, the manifestations of the disease in the oral cavity have not yet been reported. The present paper reports unique oral lesions in a 38-yr-old woman, diagnosed as leukocytoclastic vasculitis, without any accompanying skin or systemic lesions.},
	keywords = {Adult Female *Gingival Diseases/pathology Humans Ulcer/pathology *Vasculitis, Cutaneous/pathology, Leukocytoclastic},
	issn = {0904-2512 (Print) 0904-2512 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1753354},
	author = {Gazit, D. and Nahlieli, O. and Neder, A. and Berstein, I. and Ulmansky, M.}
}
@article {17033,
	title = {Osteomalacia in hereditary hypophosphatemic rickets with hypercalciuria: a correlative clinical-histomorphometric study},
	journal = {J Clin Endocrinol Metab},
	volume = {72},
	number = {1},
	year = {1991},
	note = {Gazit, D Tieder, M Liberman, U A Passi-Even, L Bab, I A eng 1991/01/01 J Clin Endocrinol Metab. 1991 Jan;72(1):229-35.},
	pages = {229-35},
	type = {Journal Article},
	abstract = {We characterized the bone disease of transilial biopsy specimens from children with hereditary hypophosphatemic rickets with hypercalciuria (HHRH) and genetically related asymptomatic hypercalciuric subjects. All HHRH patients showed irregular mineralization fronts, markedly elevated osteoid surface and seam width, increased number of osteoid lamellae, and prolonged mineralization lag time. These findings are consistent with a mineralization defect and indicate unambiguously that the bone disease in HHRH is osteomalacia. The only abnormality seen in the asymptomatic hypercalciuric subjects was slightly extended osteoid surface. Parametric and nonparametric statistical analyses performed on a pooled sample of HHRH patients and asymptomatic hypercalciuric subjects revealed a very high inverse correlation and a tight linear relationship between serum phosphorus and osteoid parameters. Serum 1,25-dihydroxyvitamin D, which is low in other forms of hereditary hypophosphatemia and osteomalacia, is elevated in HHRH and correlated positively with osteoid parameters and the mineralization lag time. Serum alkaline phosphatase showed similar relationships. These results as well as the clinical, biochemical, and radiological remission of bone disease consequent to phosphate therapy strongly suggest that in HHRH 1) hypophosphatemia alone is sufficient to cause osteomalacia; and 2) the elevation of 1,25-dihydroxyvitamin D reflects the degree of the primary renal phosphate leak, but is not involved in the pathogenesis of the bone disease.},
	keywords = {Absorption Adolescent Alkaline Phosphatase/blood Bone and Bones/*pathology Calcification, Physiologic Calcitriol/blood Calcium/blood/*urine Child Child, Preschool Female Glomerular Filtration Rate Humans Kidney Tubules/metabolism Male Osteoblasts/pathology Osteoclasts/pathology Osteomalacia/complications/*pathology/physiopathology Phosphates/*blood Phosphorus/blood/metabolism Rickets/complications/*genet},
	issn = {0021-972X (Print) 0021-972X (Linking)},
	doi = {10.1210/jcem-72-1-229},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/1986023},
	author = {Gazit, D. and Tieder, M. and Liberman, U. A. and Passi-Even, L. and Bab, I. A.}
}
@article {17034,
	title = {Successful high-dose calcium treatment of aluminum-induced metabolic bone disease in long-term home parenteral nutrition},
	journal = {JPEN J Parenter Enteral Nutr},
	volume = {15},
	number = {2},
	year = {1991},
	note = {Lidor, C Schwartz, I Freund, U Gazit, D eng Case Reports 1991/03/01 JPEN J Parenter Enteral Nutr. 1991 Mar-Apr;15(2):202-6.},
	pages = {202-6},
	type = {Journal Article},
	abstract = {A patient who developed severe metabolic bone disease is presented. He had received long-term home parenteral nutrition (HPN) following extensive small bowel resection after mesenteric vein thrombosis. Bone disease caused by aluminum intoxication had components of osteomalacia and low-turnover osteoporosis. Aluminum was detected at the surface of mineralized bone and was elevated in the serum, resulting in a positive deferoxamine infusion test. One year of treatment with high doses of calcium (up to 24 mEq per day) significantly diminished the patient{\textquoteright}s bone pain, increased the serum levels of calcium, abolished aluminum deposits in the mineralized trabecula, improved bone formation, and increased trabecular bone volume as assessed by repeated histomorphometric analysis.},
	keywords = {Aluminum/*adverse effects/blood/metabolism Bone Development Bone Diseases, Metabolic/*chemically induced/drug therapy/physiopathology Calcification, Physiologic Calcium/administration \& dosage/blood/*therapeutic use Deferoxamine Humans Male Middle Aged},
	issn = {0148-6071 (Print) 0148-6071 (Linking)},
	doi = {10.1177/0148607191015002202},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2051559},
	author = {Lidor, C. and Schwartz, I. and Freund, U. and Gazit, D.}
}
@article {17031,
	title = {Acromegaly and proximal femoral fracture in an aged person: bone histomorphometry, vitamin D metabolites and parathormone},
	journal = {Isr J Med Sci},
	volume = {26},
	number = {5},
	year = {1990},
	note = {Lowenthal, M N Galinsky, D Fried, V Castel, H Shany, S Gazit, D eng Case Reports Israel 1990/05/01 Isr J Med Sci. 1990 May;26(5):280-3.},
	pages = {280-3},
	type = {Journal Article},
	abstract = {Osteoporosis and fractures are rare in acromegaly. An 84-year-old acromegalic woman sustained a fractured neck of femur in a fall. Histomorphometric analysis of an iliac crest biopsy showed marked osteoporosis and augmented resorption parameters. Cortical plates were very thin and bone volume was 8.5\%; 12.5\% is the reference value for women at this age. The total resorption surfaces were 23.7\% compared with the reference value of 8\%. We conjecture that postmenopausal and involutional osteoporosis were far advanced before the development of acromegaly, explaining the coexistence of the two conditions. Parathormone (65 pmol/l) and 24,25-dihydroxyvitamin D (1.34 ng/ml) levels were within normal limits, but those of 25-hydroxyvitamin D (5.6 ng/ml) and 1,25-dihydroxyvitamin D (15 pg/ml) were markedly decreased.},
	keywords = {80 and over Biopsy Bone Resorption/pathology Calcitriol/blood Female Femoral Neck Fractures/*complications Humans Hydroxycholecalciferols/blood Ilium/pathology Osteoporosis, Acromegaly/*complications Aged Aged, Postmenopausal/blood/complications/pathology Parathyroid Hormone/*blood Vitamin D/*blood},
	issn = {0021-2180 (Print) 0021-2180 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2380027},
	author = {Lowenthal, M. N. and Galinsky, D. and Fried, V. and Castel, H. and Shany, S. and Gazit, D.}
}
@article {17028,
	title = {Osteochondral differentiation and the emergence of stage-specific osteogenic cell-surface molecules by bone marrow cells in diffusion chambers},
	journal = {Bone Miner},
	volume = {11},
	number = {2},
	year = {1990},
	note = {Bruder, S P Gazit, D Passi-Even, L Bab, I Caplan, A I eng GMO-7250-13/GM/NIGMS NIH HHS/ Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, P.H.S. Ireland 1990/11/01 Bone Miner. 1990 Nov;11(2):141-51.},
	pages = {141-51},
	type = {Journal Article},
	abstract = {The osteochondral potential and emergence of osteogenic cell-surface molecules by avian marrow cells was evaluated in in vivo diffusion chamber cultures. The chambers were inoculated with unselected marrow cells from young chick tibiae and implanted intraperitoneally into athymic mice. At the light microscopic level, morphologic evidence of de novo bone and cartilage formation, including specific immunostaining by antibody probes, was observed in 14 out of 16 chambers incubated for 20 days or longer. In order to monitor the osteogenic differentiation of the marrow-derived cells, indirect immunofluorescence was performed with monoclonal antibodies against stage-specific cell surface antigens on cells of the embryonic osteogenic lineage. The binding of these and other specific monoclonal antibodies in the developing tissue indicates that the cell surface and extracellular matrix molecules expressed by descendants of marrow-derived mesenchymal progenitor cells are indistinguishable from their in vivo counterparts found in embryonic skeletal structures. Furthermore, the experiments reported here describe the first molecular identification of osteogenic cells by probes which are selective for stage-specific surface antigens on cells of the osteogenic lineage. Importantly, bone formation by these marrow-derived cells appears to occur through a lineage progression which is similar to that observed for embryonic tibial osteoblasts. In summary, these data support the use of diffusion chambers inoculated with avian marrow to study aspects of osteogenic and chondrogenic differentiation.},
	keywords = {Animals Bone Development/physiology *Bone Marrow Cells Cartilage/cytology/*growth \& development Cell Membrane/physiology Chickens Diffusion Chambers, Culture Hematopoietic Stem Cells/cytology Osteogenesis/*physiology},
	issn = {0169-6009 (Print) 0169-6009 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2268743},
	author = {Bruder, S. P. and Gazit, D. and Passi-Even, L. and Bab, I. and Caplan, A. I.}
}
@article {17030,
	title = {Osteoporosis as the sole presentation of bone marrow mastocytosis},
	journal = {J Bone Miner Res},
	volume = {5},
	number = {8},
	year = {1990},
	note = {Lidor, C Frisch, B Gazit, D Gepstein, R Hallel, T Mekori, Y A eng 1990/08/01 J Bone Miner Res. 1990 Aug;5(8):871-6.},
	pages = {871-6},
	type = {Journal Article},
	abstract = {Three (young) adults with severe generalized osteopenia and vertebral compression fractures were studied. Extensive clinical and laboratory investigations were not contributory. Undecalcified bone biopsies demonstrated multiple mast cell granulomas in the marrow in two patients and numerous mast cells diffusely distributed throughout the bone marrow in the third patient. Mast cells may serve as a pathogenic agent in osteoporosis. Therefore, we conclude that isolated skeletal mastocytosis without clinical evidence of mast cell mediator release should be sought in the evaluation of a patient with unexplained severe bone loss.},
	keywords = {Adult Biopsy Bone Density/physiology Bone Marrow Diseases/*complications/diagnosis/pathology Bone and Bones/pathology Female Humans Male Mastocytosis/*complications/diagnosis/pathology Middle Aged Osteoporosis/diagnostic imaging/*etiology/pathology Radiog},
	issn = {0884-0431 (Print) 0884-0431 (Linking)},
	doi = {10.1002/jbmr.5650050811},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2239371},
	author = {Lidor, C. and Frisch, B. and Gazit, D. and Gepstein, R. and Hallel, T. and Mekori, Y. A.}
}
@article {17029,
	title = {Regenerating marrow induces systemic increase in osteo- and chondrogenesis},
	journal = {Endocrinology},
	volume = {126},
	number = {5},
	year = {1990},
	note = {Gazit, D Karmish, M Holzman, L Bab, I eng 1990/05/01 Endocrinology. 1990 May;126(5):2607-13.},
	pages = {2607-13},
	type = {Journal Article},
	abstract = {Marrow ablation in long bones induces an increase in osteogenesis in distant skeletal sites. To test the role of marrow regeneration in this phenomenon, rat mandibular condyles were evaluated histomorphometrically during postablation healing of tibial marrow and after inhibition of healing. Ten days after removal of tibial marrow all bone formation parameters in the condylar subchondral bone were markedly elevated, indicating an enhanced osteoblastic activity. The thickness of the cartilaginous zone of calcification was also augmented. These changes were absent when postablation healing was inhibited in the tibia and after massive liver injury. Extensive periosteal injury induced only a slight increase in osteoblast activity. Except for a fall on day 7, the [methyl-3H]thymidine labeling index in the condylar cartilage and oral mucosa remained at control levels 3-18 days after ablation. These findings imply that stimulation of cell proliferation has only a secondary role in the skeletal response to marrow ablation. It is concluded that the systemic increase in osteogenesis occurs preferentially during marrow regeneration and is not a nonspecific skeletal reaction to tissue injury. Apparently, the systemic osteogenic response is mediated by circulating factors produced by the healing marrow; conceptually it is related to other instances where local repair in extraskeletal sites is accompanied by generalized alterations in respective tissues.},
	keywords = {Animals Bone Marrow/*physiology Bone Marrow Cells Cartilage/cytology/*physiology Cell Count Cell Division Epithelial Cells Fibroblasts/cytology Male Mandibular Condyle Osteoblasts/cytology/physiology *Osteogenesis Rats Regeneration/*physiology Tibia},
	issn = {0013-7227 (Print) 0013-7227 (Linking)},
	doi = {10.1210/endo-126-5-2607},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2328701},
	author = {Gazit, D. and Karmish, M. and Holzman, L. and Bab, I.}
}
@article {17025,
	title = {Effect of verapamil on bone resorption and formation in uremic rats},
	journal = {Miner Electrolyte Metab},
	volume = {15},
	number = {5},
	year = {1989},
	note = {Chagnac, A Gazit, D Zahavi, I Sela, J Levi, J eng Switzerland 1989/01/01 Miner Electrolyte Metab. 1989;15(5):291-4.},
	pages = {291-4},
	type = {Journal Article},
	abstract = {It has been shown in vitro that verapamil inhibits parathyroid hormone-induced bone resorption. To determine the effect of verapamil administration on bone histology in rats with chronic renal failure, male Wistar rats were divided into three groups: subtotally nephrectomized (SNX), SNX treated with verapamil, 8 mg/kg/day p.o. (SNX-V) and control (C). Thirteen weeks later, the mandibular condyle bone was studied by histomorphometry. When compared to C rats, SNX rats had active bone disease, with increased resorption parameters (resorption surface, active resorption surface, osteoclast number) and accelerated mineral appositional rate. These parameters improved with verapamil administration. When compared to C rats, SNX-V rats had decreased osteoid seam width and mineral appositional rate. Serum parathyroid hormone was similarly elevated in both uremic groups. These data suggest that verapamil administration improves active bone disease in rats with chronic renal failure and decreases bone formation.},
	keywords = {Animals *Bone Resorption Bone and Bones/drug effects Calcium/blood Creatinine/metabolism Hyperparathyroidism/*complications Kidney Failure, Chronic/*physiopathology Osteogenesis/*drug effects Parathyroid Hormone/blood Phosphorus/blood Rats Rats, Inbred Strains Uremia/*complications Verapamil/*pharmacology},
	issn = {0378-0392 (Print) 0378-0392 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2811787},
	author = {Chagnac, A. and Gazit, D. and Zahavi, I. and Sela, J. and Levi, J.}
}
@article {17026,
	title = {Further characterization of osteogenic-cell growth promoting activity derived from healing bone marrow},
	journal = {Connect Tissue Res},
	volume = {23},
	number = {2-3},
	year = {1989},
	note = {Gazit, D Shteyer, A Bab, I eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. England 1989/01/01 Connect Tissue Res. 1989;23(2-3):153-61.},
	pages = {153-61},
	type = {Journal Article},
	abstract = {During its osteogenic phase, post-ablation regenerating bone marrow produces bone promoting activity to osteogenic cells. In the experiments reported, activity derived from (rat) healing bone marrow conditioned medium (HBMCM) after boiling was analyzed using chromatography on heparin-Sepharose. The activity in HBMCM was shown to be divided among at least six independent activities that stimulated DNA synthesis rates is osteogenic rat osteosarcoma (ROS) cells. Three activities resolved when heparin-Sepharose was washed isocratically with phosphate buffered saline. Two of these were resistant to reduction and acidification and their effect was considerably more potent in osteogenic than non-osteogenic ROS cells. Three additional activity peaks recovered when the heparin-Sepharose column was pumped with an NaCl gradient. Two of them eluted at 0.3 and 0.65 M NaCl, affected osteogenic and non-osteogenic ROS cells to a similar extent and may be attributed to platelet-derived growth factor. A third peak, resolved at 1.2 M NaCl, implies the residual activity of acidic fibroblast growth factor that persisted after boiling of the conditioned medium. It is concluded that the activity profile of HBMCM reflects the in vivo situation where the osteogenic phase of marrow regeneration is probably regulated by multiple growth factor species.},
	keywords = {Affinity Culture Media/pharmacology Electrophoresis, Alkaline Phosphatase/metabolism Animals Bone Marrow/*analysis/physiology Bone Marrow Cells Bone and Bones/cytology/drug effects/metabolism Cell Division/drug effects Cells, Cultured Chromatography, Cultured/drug effects/metabolism/pathology Wound Healing/*physiology, Polyacrylamide Gel Growth Substances/analysis/*pharmacology/physiology Osteogenesis/drug effects/*physiology Osteosarcoma/metabolism/pathology Rats Tumor Cells},
	issn = {0300-8207 (Print) 0300-8207 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2630168},
	author = {Gazit, D. and Shteyer, A. and Bab, I.}
}
@article {17027,
	title = {Oral clinicopathologic manifestations in maple syrup urine disease},
	journal = {Oral Surg Oral Med Oral Pathol},
	volume = {67},
	number = {6},
	year = {1989},
	note = {Gazit, D Zeltser, R Galili, D Kaufman, E eng Case Reports 1989/06/01 Oral Surg Oral Med Oral Pathol. 1989 Jun;67(6):694-7.},
	pages = {694-7},
	type = {Journal Article},
	abstract = {Maple syrup urine disease is a rare inborn error of metabolism that manifests mainly by neurologic deterioration, coma, and death in early childhood. Oral manifestations described in this article were mostly gingival enlargement and rampant caries, while histopathologic findings revealed a picture compatible osteomyelitis.},
	keywords = {Adolescent Gingival Hyperplasia/pathology Humans Jaw Abnormalities/pathology Male *Maple Syrup Urine Disease/pathology *Mouth Abnormalities/pathology *Tooth Abnormalities/pathology},
	issn = {0030-4220 (Print) 0030-4220 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2740092},
	author = {Gazit, D. and Zeltser, R. and Galili, D. and Kaufman, E.}
}
@article {17022,
	title = {99mTc-MDP uptake and histological changes during rat bone marrow regeneration},
	journal = {Int J Rad Appl Instrum B},
	volume = {15},
	number = {4},
	year = {1988},
	note = {Chisin, R Gazit, D Ulmansky, M Laron, A Atlan, H Sela, J eng England 1988/01/01 Int J Rad Appl Instrum B. 1988;15(4):469-76.},
	pages = {469-76},
	type = {Journal Article},
	abstract = {An established experimental model of tibial bone regeneration in rats was used in order to try to provide further information on the binding site of 99mTc-MDP, which is still not clearly defined. Four groups of rats on which surgical tibial bone marrow evacuation was performed and two control groups (nonoperated animals and sham-operated animals) underwent bone scan during the different stages of marrow regeneration; they were killed immediately after, and histological examination carried out. The correlation between the scintigraphic and the histological findings suggests that 99mTc-MDP binds primarily to calcification sites in young bone trabecules.},
	keywords = {Animals Bone Marrow/*physiology Bone and Bones/*diagnostic imaging/metabolism Male Radionuclide Imaging Rats *Regeneration Technetium Tc 99m Medronate/metabolism/*pharmacokinetics},
	issn = {0883-2897 (Print) 0883-2897 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3255743},
	author = {Chisin, R. and Gazit, D. and Ulmansky, M. and Laron, A. and Atlan, H. and Sela, J.}
}
@article {17024,
	title = {Changes in teeth and gingiva of dogs following laser surgery: a block surface light microscope study},
	journal = {Lasers Surg Med},
	volume = {8},
	number = {4},
	year = {1988},
	note = {Goultschin, J Gazit, D Bichacho, N Bab, I eng 1988/01/01 Lasers Surg Med. 1988;8(4):402-8.},
	pages = {402-8},
	type = {Journal Article},
	abstract = {The effect of laser surgery on tissues of the periodontal apparatus was studied histologically in dogs using block surface light microscopy, a novel microscopical method. With this approach, changes in the hard and soft tissue components were concomitantly demonstrated; the method enabled preservation of the in situ relationship between these components. Following laser surgery, healing in the gingiva was delayed as suggested by the presence of epithelial ulcerations and dense inflammatory infiltrate. In the enamel and cementum the application of laser resulted in crater-like defects that could be avoided only partially by insertion of a tinfoil shield into the gingival sulcus. In the vicinity of the cementoenamel junction these defects were filled with epithelium or periodontal ligament fibers; the close proximity of the hard and soft tissues at the defect sites suggested occurrence of new attachment. Enamel defects located coronal to the gingiva contained bacterial plaque. These histologic results do not demonstrate any substantial advantage of laser over conventional knife gingivectomy. Such advantage may be accomplished with the design of a special intraoral handpiece and further experiments.},
	keywords = {Animals Dogs Gingiva/*injuries Gingivectomy/*methods Laser Therapy/*adverse effects *Tooth Injuries Wound Healing},
	issn = {0196-8092 (Print) 0196-8092 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2902501},
	author = {Goultschin, J. and Gazit, D. and Bichacho, N. and Bab, I.}
}
@article {17021,
	title = {Gingivitis in the human deciduous dentition. A correlative clinical and block surface light microscopic (BSLM) study},
	journal = {J Clin Periodontol},
	volume = {15},
	number = {9},
	year = {1988},
	note = {Bimstein, E Soskolne, W A Lustmann, J Gazit, D Bab, I eng 1988/10/01 J Clin Periodontol. 1988 Oct;15(9):575-80.},
	pages = {575-80},
	type = {Journal Article},
	abstract = {This study examined the relationship between clinical and histomorphometric parameters in the human deciduous dentition. Clinical parameters including plaque index, gingival swelling, gingival color, tooth mobility and degree of root resorption were determined prior to the extraction of teeth. The teeth were extracted with their surrounding gingiva in order to preserve the in situ relationship between the hard and soft tissues. Histomorphometric analysis was carried out on 55 sites, using block surface light microscopy (BSLM). Apical migration of the junctional epithelium was found at 53\% (29) of the sites. The gingival sulcus was shallow (0.3 +/- 0.19 mm) and coronal to the cemento-enamel junction at 84\% (46) of the sites. Junctional epithelium with retepegs was present at 89\% (49) of the sites, whilst an inflammatory cell infiltrate (ICI) was present at all sites examined. The ICI was located opposite to the junctional epithelium and cementum at 80\% (44) of the sites. The extent of ICI correlated positively with the patients{\textquoteright} age and was significantly increased when clinical evidence of gingival swelling or redness was present.},
	keywords = {Adolescent Cell Movement Child Connective Tissue/pathology Dental Cementum/pathology Dental Enamel/pathology Epithelial Attachment/pathology Female Gingiva/pathology Gingivitis/*pathology Humans Male Microscopy/methods Tooth Extraction/methods *Tooth, Deciduous},
	issn = {0303-6979 (Print) 0303-6979 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3198785},
	author = {Bimstein, E. and Soskolne, W. A. and Lustmann, J. and Gazit, D. and Bab, I.}
}
@article {17020,
	title = {Osteogenesis in in vivo diffusion chamber cultures of human marrow cells},
	journal = {Bone Miner},
	volume = {4},
	number = {4},
	year = {1988},
	note = {Bab, I Passi-Even, L Gazit, D Sekeles, E Ashton, B A Peylan-Ramu, N Ziv, I Ulmansky, M eng Research Support, Non-U.S. Gov{\textquoteright}t Ireland 1988/09/01 Bone Miner. 1988 Sep;4(4):373-86.},
	pages = {373-86},
	type = {Journal Article},
	abstract = {The osteogenic diffusion chamber culture of rodent marrow cells is a well established system. In the present study, marrow cells from children and adult human donors were incubated in diffusion chambers implanted intraperitoneally in athymic mice. After 4 or 8 weeks, the chamber content was examined by light and electron microscopy. Child-cell cultures showed osteogenic tissue consisting of a mineralizing fibrous component and cartilage. Ultrastructurally, the fibrous tissue was similar to osteoid and exhibited osteoblast-like cells and mineralizing nodules. Mineral aggregates were also found in the cartilage. These features in child-cell chambers were similar to those found in control chambers of rabbit marrow cells. Adult-cell chambers showed only unmineralized fibrous tissue. These results render previous findings in animal-cell diffusion chamber systems relevant to the understanding of bone formation in man. It is suggested that the difference between child- and adult-cell chambers reflects an age-related decline in the number of marrow osteoprogenitor cells or their potential to undergo terminal osteogenic differentiation.},
	keywords = {Adult Animals *Bone Marrow Cells Calcification, Culture Humans Mice Mice, Cultured Child *Diffusion Chambers, Electron Minerals/metabolism *Osteogenesis Peritoneal Cavity Rabbits, Inbred BALB C Mice, Nude Microscopy, Physiologic Cartilage/cytology/physiology Cells},
	issn = {0169-6009 (Print) 0169-6009 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3191291},
	author = {Bab, I. and Passi-Even, L. and Gazit, D. and Sekeles, E. and Ashton, B. A. and Peylan-Ramu, N. and Ziv, I. and Ulmansky, M.}
}
@article {17019,
	title = {Regenerating bone marrow produces a potent growth-promoting activity to osteogenic cells},
	journal = {Endocrinology},
	volume = {123},
	number = {1},
	year = {1988},
	note = {Bab, I Gazit, D Muhlrad, A Shteyer, A eng Research Support, Non-U.S. Gov{\textquoteright}t Research Support, U.S. Gov{\textquoteright}t, Non-P.H.S. 1988/07/01 Endocrinology. 1988 Jul;123(1):345-52.},
	pages = {345-52},
	type = {Journal Article},
	abstract = {It is well documented that injury to bone marrow is followed by an osteogenic phase that precedes the complete tissue regeneration. We have recently shown that postablation healing of bone marrow in rat tibiae is associated with a systemic increase in osteogenesis. It was hypothesized that a growth factor(s) with an effect on osteogenic cells is produced in the healing limb, is transferred to the blood circulation, and enhances osteogenesis systemically. To test growth factor production, healing bone marrow-conditioned medium was prepared with tissue separated from rat tibias during the osteogenic phase and assayed for enhancement of mitogenic activity in culture of osteogenic rat osteosarcoma cells (ROS 17/2). Partial purification of healing bone marrow-conditioned medium-derived growth factor(s) consisted of gel filtration on Sephadex G-25, boiling, chromatography on heparin-Sepharose, and gel filtration on Sephadex G-75. Mitogenic activity eluted in the void volume of the Sephadex G-25 column (mol wt greater than 5,000). Potent activity resolved from heparin-Sepharose with PBS, and on filtration by Sephadex G-75 this activity recovered in 3 peaks with mol wt estimates of 35,000, 19,000, and less than 10,000. The partially purified factor also showed considerable stimulatory effect on DNA synthesis in osteoblastic fetal rat calvarial cells and on in vitro elongation of fetal long bone; it had only a small effect on nonosteoblastic ROS and fetal rat calvarial cells. These data indicate that healing bone marrow produces growth factor activity with a preferential effect on osteogenic cells. It is suggested that local growth factors have a role as mediators in the sequence of events whereby bone marrow expresses its osteogenic potential. During postablation healing of bone marrow these factors may also function as systemic promoters to osteogenic cells.},
	keywords = {Animals *Bone Development Bone Marrow/*physiology Cell Line DNA Replication Growth Substances/isolation \& purification/*physiology Male Molecular Weight Organ Culture Techniques Osteoblasts/cytology Osteosarcoma Rats Rats, Inbred Strains *Regeneration Tibia},
	issn = {0013-7227 (Print) 0013-7227 (Linking)},
	doi = {10.1210/endo-123-1-345},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3164264},
	author = {Bab, I. and Gazit, D. and Muhlrad, A. and Shteyer, A.}
}
@article {17023,
	title = {Tooth-germ sequestration as a sequela of chronic periapical inflammation of the primary predecessor},
	journal = {ASDC J Dent Child},
	volume = {55},
	number = {6},
	year = {1988},
	note = {Goodman-Topper, E D Gazit, D Eidelman, E eng Case Reports 1988/11/01 ASDC J Dent Child. 1988 Nov-Dec;55(6):455-8.},
	pages = {455-8},
	type = {Journal Article},
	abstract = {In this case the permanent successor was so radiographically indistinct due to the inflammatory process that this three-year-old Arab boy might have been classified as having congenital absence of the mandibular left first premolar, if the mass had not been sent for histological section. The clinical implications are identical.},
	keywords = {Bicuspid/*pathology Child, Deciduous/*pathology, Preschool Chronic Disease Humans Male Periapical Periodontitis/complications/*pathology Tooth Germ/*pathology Tooth},
	issn = {1945-1954 (Print) 1945-1954 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3198836},
	author = {Goodman-Topper, E. D. and Gazit, D. and Eidelman, E.}
}
@article {17016,
	title = {Aluminum-related osteomalacia: clinical and histological improvement following treatment with desferrioxamine (DFO)},
	journal = {Isr J Med Sci},
	volume = {23},
	number = {12},
	year = {1987},
	note = {Rapoport, J Chaimovitz, C Abulfil, A Mostovlavsky, M Gazit, D Bab, I eng Case Reports Israel 1987/12/01 Isr J Med Sci. 1987 Dec;23(12):1242-6.},
	pages = {1242-6},
	type = {Journal Article},
	abstract = {Aluminum-related osteomalacia has been a topic of major interest for some time. Patients on dialysis who are affected exhibit reduced parathyroid function and osteomalacia associated with high levels of aluminum in the extracellular fluid and tissues. In bone, aluminum is found in the interface between mineralized and nonmineralized osteoid. In the present study we attempted treatment of the aluminum-related bone disease with DFO, a potent chelating agent. A thyroparathyroidectomized patient on hemodialysis with proven osteomalacia and aluminum deposits in bone received DFO 3 times/week for 1 year. Consequently, there was a marked improvement in his skeletal status, both clinically and histologically. This was associated with aluminum deposits within the mineralized component of bone as well as elevated parathyroid function measured in blood and bone. These findings suggest that calcification may occur despite the presence of aluminum in the mineralizing front. The main beneficial effect of DFO could be relief of the inhibitory action of aluminum on parathyroid cells and osteoblasts.},
	keywords = {Adult Aluminum/*adverse effects Biopsy Bone and Bones/pathology Deferoxamine/*therapeutic use Humans Male Osteomalacia/drug therapy/*etiology/pathology Renal Dialysis/adverse effects},
	issn = {0021-2180 (Print) 0021-2180 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3440748},
	author = {Rapoport, J. and Chaimovitz, C. and Abulfil, A. and Mostovlavsky, M. and Gazit, D. and Bab, I.}
}
@article {17015,
	title = {Effect of occlusal (mechanical) stimulus on bone remodelling in rat mandibular condyle},
	journal = {J Oral Pathol},
	volume = {16},
	number = {8},
	year = {1987},
	note = {Gazit, D Ehrlich, J Kohen, Y Bab, I eng Denmark 1987/09/01 J Oral Pathol. 1987 Sep;16(8):395-8.},
	pages = {395-8},
	type = {Journal Article},
	abstract = {Mechanical load influences the remodelling of skeletal tissues. In the mandibular condyle, occlusal alterations and the consequent mechanical stimulus induce changes in chondrocytes and cartilage mineralization. In the present study we quantified in the mandibular condyle the effect of occlusal interference on remodelling of the subchondral bone. Computerized histomorphometry after 5-21-day exposure to the influence of a unilateral occlusal splint revealed an increased rate of trabecular remodelling, consisting of enhancement in osteoblast and osteoclast numbers and activities. The bone formation parameters reached their high values on Days 5 or 9 and remained stable thereafter. Bone resorption showed a gradual increase throughout the experimental period. These results further characterize the temporomandibular joint reaction to occlusal alterations. It is suggested that the present increase in bone turnover together with the known enhancement in chondrogenesis are part of a process of functional adaptation in response to mechanical stimulus.},
	keywords = {Animals Bone Resorption/pathology/physiopathology Cartilage/pathology/*physiopathology Dental Occlusion, Inbred Strains Stress, Mechanical Time Factors, Traumatic/pathology/*physiopathology Male Mandibular Condyle/pathology/*physiopathology Osteogenesis Rats Rats},
	issn = {0300-9777 (Print) 0300-9777 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3121824},
	author = {Gazit, D. and Ehrlich, J. and Kohen, Y. and Bab, I.}
}
@article {17017,
	title = {Hormone-responsive cells derived from human dental papilla: characterization in vitro and in vivo in diffusion chambers},
	journal = {In Vitro Cell Dev Biol},
	volume = {23},
	number = {1},
	year = {1987},
	note = {Shteyer, A Gazit, D Binderman, I Bab, I A eng Comparative Study 1987/01/01 In Vitro Cell Dev Biol. 1987 Jan;23(1):15-20.},
	pages = {15-20},
	type = {Journal Article},
	abstract = {Cells of the dental papilla are capable of odontoblastic, fibroblastic, and endothelial differentiation and formation of dentin and the dental pulp. In the present study dental papilla cells, obtained from human tooth buds (HDP cells), were cultured in vitro through 3 to 7 passages. After exposure to prostaglandin E2 there was a marked decrease in intracellular cyclic AMP (cAMP) levels as compared to hormone-free controls. Parathyroid hormone and calcitonin had stimulatory effects with 1 and 2 log increases in cAMP, respectively. The HDP cells showed moderate activity of alkaline phosphatase, 1 log higher than that of hamster kidney fibroblasts (BHK 13) and 1 log lower than that of osteoblastic osteosarcoma cells (ROS 17/2). When cultured for 4 or 8 wk in diffusion chambers (DC) implanted in athymic mice, many of the HDP cells underwent odontoblastic morphodifferentiation with very long, single processes extending into the matrix. This matrix contained banded and unbanded collagen fibers. Neither light nor electron microscopy of the DC content revealed mineral deposits. These results suggest that HDP cells have an intrinsic potential for partial odontoblastic differentiation; inductive signals like those originating from odontogenic epithelium are probably essential for the completion of hard tissue formation.},
	keywords = {Alkaline Phosphatase/analysis Animals Calcitonin/*pharmacology Cattle Cell Differentiation/drug effects Child Cyclic AMP/analysis Dental Papilla/*cytology/drug effects/transplantation Dinoprostone Extracellular Matrix/metabolism Humans Male Mice Mice, Chemical Tooth Germ/*cytology, Nude Odontoblasts/*cytology/metabolism Parathyroid Hormone/*pharmacology Prostaglandins E/chemical synthesis/*pharmacology Stimulation},
	issn = {0883-8364 (Print) 0883-8364 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3027024},
	author = {Shteyer, A. and Gazit, D. and Binderman, I. and Bab, I. A.}
}
@article {17018,
	title = {Long-term intracaval calcium infusion therapy in end-organ resistance to 1,25-dihydroxyvitamin D},
	journal = {Am J Med},
	volume = {83},
	number = {5},
	year = {1987},
	note = {Weisman, Y Bab, I Gazit, D Spirer, Z Jaffe, M Hochberg, Z eng Case Reports 1987/11/01 Am J Med. 1987 Nov;83(5):984-90.},
	pages = {984-90},
	type = {Journal Article},
	abstract = {Two boys aged six and four with the syndrome of hereditary resistance to 1,25-dihydroxyvitamin D3 with rickets alopecia and growth retardation are presented. After unsuccessful therapeutic trials with pharmacologic doses of vitamin D or its active metabolites, the patients were treated by long-term intracaval infusions of calcium through an implantable catheter. A total of 0.5 to 0.9 g of elemental calcium was infused daily for 18 months and the serum calcium concentration was maintained at 9 to 10 mg/dl. Bone pain subsided within one week of treatment. Serum phosphorus, immunoreactive parathyroid hormone, and 1,25-dihydroxyvitamin D concentrations and alkaline phosphatase activity were normalized within four to nine months. Radiographs of the knees and hands revealed progressive healing of rickets with complete resolution after one year of treatment. The patients gained 12 cm and 8 cm per year in height as compared with 3 cm and 2 cm, respectively, in the previous year. A transilial bone biopsy obtained from one patient prior to treatment revealed severe osteomalacia associated with osteitis fibrosa. A follow-up biopsy examined after 12 months of therapy showed almost complete healing of osteomalacia and normal mineralization. These observations indicate the following: (1) Long-term intracaval calcium infusions are an effective mode of therapy for these patients, and (2) When adequate serum calcium and phosphorus concentrations are maintained, healing of rickets and normal growth rate could be achieved even in the absence of a normal 1,25-dihydroxyvitamin D3 receptor-effector system.},
	keywords = {Biopsy Bone and Bones/pathology *Calcitriol Calcium/*administration \& dosage/therapeutic use Child Child, Familial/*drug therapy/pathology Infusions, Intravenous Male Time Factors, Preschool Growth Disorders/*drug therapy/etiology Humans Hypophosphatemia},
	issn = {0002-9343 (Print) 0002-9343 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/2823606},
	author = {Weisman, Y. and Bab, I. and Gazit, D. and Spirer, Z. and Jaffe, M. and Hochberg, Z.}
}
@article {17013,
	title = {Capsule around silicone implants in diabetic rats: histological and biochemical study},
	journal = {Ann Plast Surg},
	volume = {17},
	number = {4},
	year = {1986},
	note = {Peled, I J Wexler, M R Shoshan, S Moshe, S B Gazit, D Bab, I Ticher, S eng Comparative Study 1986/10/01 Ann Plast Surg. 1986 Oct;17(4):288-91.},
	pages = {288-91},
	type = {Journal Article},
	abstract = {Polydimethylsiloxane (silicone) implants were subcutaneously placed in the back of diabetic and normal rats. After three months the rats were killed and the fibrous capsule around the implants was histologically and biochemically examined. A significant quantitative difference (p less than 0.001) was found in the thickness of the capsules, which were two to three times thicker in the diabetic animals. The biochemistry showed an increase of neutral salt-soluble collagen in the diabetic group; electrophoresis revealed only type I collagen in the diabetic and type I and III in the normal rats. From this experimental trial it seems that diabetes mellitus is another factor in formation of a thick capsule around silicone implants.},
	keywords = {Animal Glucose Tolerance Test Prostheses and Implants/*adverse effects Prosthesis Failure Rats Silicone Elastomers/*adverse effects Skin/drug effects/metabolism/pathology, Animals Blood Glucose/analysis *Breast Diabetes Mellitus, Experimental/metabolism/*pathology Dimethylpolysiloxanes/adverse effects Disease Models},
	issn = {0148-7043 (Print) 0148-7043 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3273108},
	author = {Peled, I. J. and Wexler, M. R. and Shoshan, S. and Moshe, S. B. and Gazit, D. and Bab, I. and Ticher, S.}
}
@article {17010,
	title = {Chondromyxoid fibroma of the jaws: a clinicopathological study},
	journal = {J Oral Pathol},
	volume = {15},
	number = {6},
	year = {1986},
	note = {Lustmann, J Gazit, D Ulmansky, M Lewin-Epstein, J eng Case Reports Denmark 1986/07/01 J Oral Pathol. 1986 Jul;15(6):343-6.},
	pages = {343-6},
	type = {Journal Article},
	abstract = {Chondromyxoid fibroma is a benign skeletal tumor which rarely affects the jaws. Only 10 cases have been found in the literature, all of them located in the mandible. In the present articles, 2 additional cases are described, one of them being the first reported case located in the maxilla. Up-to-date clinical and pathological data of 2 reported cases and a review of the literature are presented.},
	keywords = {Adult Chondroma/*pathology Female Humans Male Mandibular Neoplasms/*pathology Palatal Neoplasms/*pathology},
	issn = {0300-9777 (Print) 0300-9777 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3093652},
	author = {Lustmann, J. and Gazit, D. and Ulmansky, M. and Lewin-Epstein, J.}
}
@article {17011,
	title = {A correlative assessment of photon interaction and histomorphometric measurements of bone density},
	journal = {Arch Orthop Trauma Surg},
	volume = {105},
	number = {4},
	year = {1986},
	note = {Margulies, J Y Leichter, I Robin, G C Gazit, D Bab, I eng Comparative Study Germany 1986/01/01 Arch Orthop Trauma Surg. 1986;105(4):239-42.},
	pages = {239-42},
	type = {Journal Article},
	abstract = {Thirty-four femoral necks from human cadavers were measured by techniques assessing bone density and bone mineral density, and by the Singh index. These methods are based on photon interaction with biological components and can be applied noninvasively for clinical evaluation of changes in skeletal status. Trabecular bone volume, mineralized bone volume, and relative osteoid volume were evaluated histomorphometrically using undecalcified histologic sections obtained from the same samples. The trabecular and mineralized bone volumes showed significant correlations with the bone density and mineral density. These results enhance the validity of recently developed photon-interaction techniques for evaluating bone properties.},
	keywords = {Adult Aged Bone and Bones/analysis/*anatomy \& histology/*diagnostic imaging Female Humans Male Middle Aged Minerals/*analysis Radionuclide Imaging Spectrum Analysis},
	issn = {0344-8444 (Print) 0344-8444 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3753180},
	author = {Margulies, J. Y. and Leichter, I. and Robin, G. C. and Gazit, D. and Bab, I.}
}
@article {17014,
	title = {Formation of calcifying matrix by osteosarcoma cells in diffusion chambers in vivo},
	journal = {Calcif Tissue Int},
	volume = {39},
	number = {1},
	year = {1986},
	note = {Shteyer, A Gazit, D Passi-Even, L Bab, I Majeska, R Gronowicz, G Lurie, A Rodan, G eng 1986/07/01 Calcif Tissue Int. 1986 Jul;39(1):49-54.},
	pages = {49-54},
	type = {Journal Article},
	abstract = {The potential of clonal rat osteosarcoma (ROS) cell lines to form mineralized matrices was assessed in diffusion chambers in vivo. Diffusion chambers were inoculated with osteoblastic (ROS 17/2 and its subclone ROS 17/2.8) and non-osteoblastic (ROS 24/1) clonal lines and implanted either intraperitoneally into athymic mice or subcutaneously into syngeneic ACI rats. Control chamber cultures of rabbit marrow or spleen cells were also incubated in athymic mice. Light and electron microscopy of chambers with ROS 17/2 and ROS 17/2.8 cells revealed production of mineralized matrices typical of osteosarcoma and characterized by abundance of collagen fibrils and associated mineralizing nodules. ROS 24/1 cells produced similar collagenous matrices, but these were devoid of mineral. The present experiments, carried out independently in two different laboratories, demonstrate the potential of ROS cells to produce a mineralized matrix. This corroborates previous studies on other osteoblastic features of these cell lines.},
	keywords = {Animals *Calcinosis Clone Cells Culture Techniques/methods Diffusion Mice Microscopy, Electron Osteosarcoma/*pathology Rats},
	issn = {0171-967X (Print) 0171-967X (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3089564},
	author = {Shteyer, A. and Gazit, D. and Passi-Even, L. and Bab, I. and Majeska, R. and Gronowicz, G. and Lurie, A. and Rodan, G.}
}
@article {17009,
	title = {Kinetics and differentiation of marrow stromal cells in diffusion chambers in vivo},
	journal = {J Cell Sci},
	volume = {84},
	year = {1986},
	note = {Bab, I Ashton, B A Gazit, D Marx, G Williamson, M C Owen, M E eng Research Support, Non-U.S. Gov{\textquoteright}t England 1986/08/01 J Cell Sci. 1986 Aug;84:139-51.},
	pages = {139-51},
	type = {Journal Article},
	abstract = {Rabbit marrow cells inoculated into diffusion chambers (10(7) cells/chamber) were implanted intraperitoneally into athymic mouse hosts and cultured in vivo for 20 days. A connective tissue consisting of bone, cartilage and fibrous tissues is formed by the stromal fibroblastic cells of marrow within the chambers. Cell kinetics and tissue differentiation have been studied using histomorphometric and biochemical analyses. Haemopoietic cell numbers decrease to less than 0.05\% of the initial inoculum during the 20-day period. At 3 days an average of 15 stromal fibroblastic cells only are identifiable within the chambers. After 3 days there is a high rate of stromal cell proliferation with a doubling time of 14.5 h during the period from 3 to 8 days and an increase in the total stromal cell population by more than six orders of magnitude from 3 to 20 days. Thirteen to fourteen population doublings occur before expression of the first observable differentiation parameter, alkaline phosphatase activity. The data demonstrate that the mixture of connective tissues formed within the chamber is generated by a small number of cells with high capacity for proliferation and differentiation. This is consistent with the current hypothesis that stromal stem cells are present in bone marrow.},
	keywords = {Alkaline Phosphatase/metabolism Animals Bone Marrow/*physiology Calcium/metabolism Cell Differentiation Cell Line Kinetics Male Micropore Filters Rabbits Stem Cells/cytology/*physiology},
	issn = {0021-9533 (Print) 0021-9533 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3805151},
	author = {Bab, I. and Ashton, B. A. and Gazit, D. and Marx, G. and Williamson, M. C. and Owen, M. E.}
}
@article {17012,
	title = {MRI and CT scan compared with microscopic histopathology in osteogenic sarcoma of the proximal tibia},
	journal = {Orthop Rev},
	volume = {15},
	number = {3},
	year = {1986},
	note = {Milgrom, C Sigal, R Robin, G C Gazit, D Fields, S Benmair, J Caine, Y Atlan, H eng Case Reports Comparative Study 1986/03/01 Orthop Rev. 1986 Mar;15(3):165-9.},
	pages = {165-9},
	type = {Journal Article},
	abstract = {The authors report the case of a 15-year-old female who presented with a history of vague but constant pain about the medial aspect of her right knee. X-ray established the presence of an expanding lesion in the medial tibial plateau. Computerized axial tomography (CT) and magnetic resonance imaging (MRI) were used in the evaluation of the lesion. The authors compare the preoperative CT and MRI findings with the microscopic histopathology of the amputation specimen and note that the CT scan underestimated the extent of the microscopic tumor boundaries, whereas MRI showed altered activity beyond these boundaries.},
	keywords = {Adolescent Bone Neoplasms/diagnosis/*pathology Female Humans *Magnetic Resonance Imaging Osteosarcoma/diagnosis/*pathology Tibia/diagnostic imaging/*pathology *Tomography, X-Ray Computed},
	issn = {0094-6591 (Print) 0094-6591 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3483188},
	author = {Milgrom, C. and Sigal, R. and Robin, G. C. and Gazit, D. and Fields, S. and Benmair, J. and Caine, Y. and Atlan, H.}
}
@article {17007,
	title = {Calcification of non-collagenous matrix in human gingiva: a light and electron microscopic study},
	journal = {J Oral Pathol},
	volume = {14},
	number = {7},
	year = {1985},
	note = {Bab, I Lustmann, J Azaz, B Gazit, D Garfunkel, A eng Denmark 1985/08/01 J Oral Pathol. 1985 Aug;14(7):573-80.},
	pages = {573-80},
	type = {Journal Article},
	abstract = {Tissue samples obtained from human gingiva with soft tissue calcification were processed for light and transmission electron microscopy. The stroma in these specimens revealed numerous foci of calcification in a matrix that consisted of closely packed branching microfibrils, 12 nm in diameter and a maximum of 2.5 micron in length. Calcospherites, 0.25-1.11 micron in diameter, were present within the matrix. They were constructed of needle-like units shown by high resolution electron microscopy and energy dispersive X-ray microanalysis to be apatite. Larger calcified masses were composed of calcospherites which were fused together. Matrix vesicles or other forms of membraneous material could not be found. The relationship between the mineral and non-collagenous microfibrils may suggest a role for the latter in the onset of calcification in this ectopic site.},
	keywords = {Amelogenesis Imperfecta/pathology Calcinosis/metabolism/*pathology Electron Probe Microanalysis Gingiva/analysis/*ultrastructure Gingival Diseases/metabolism/*pathology Humans Microscopy, Electron},
	issn = {0300-9777 (Print) 0300-9777 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3928852},
	author = {Bab, I. and Lustmann, J. and Azaz, B. and Gazit, D. and Garfunkel, A.}
}
@article {17008,
	title = {Childhood odontogenic myxoma: report of two cases},
	journal = {Pediatr Dent},
	volume = {7},
	number = {2},
	year = {1985},
	note = {Nitzan, D W Gazit, D Azaz, B eng Case Reports 1985/06/01 Pediatr Dent. 1985 Jun;7(2):140-4.},
	pages = {140-4},
	type = {Journal Article},
	keywords = {Child Female Humans Mandibular Neoplasms/*diagnosis Odontogenic Tumors/*diagnosis},
	issn = {0164-1263 (Print) 0164-1263 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3859846},
	author = {Nitzan, D. W. and Gazit, D. and Azaz, B.}
}
@article {17006,
	title = {Removal of tibial marrow induces increased formation of bone and cartilage in rat mandibular condyle},
	journal = {Calcif Tissue Int},
	volume = {37},
	number = {5},
	year = {1985},
	note = {Bab, I Gazit, D Massarawa, A Sela, J eng Research Support, Non-U.S. Gov{\textquoteright}t 1985/09/01 Calcif Tissue Int. 1985 Sep;37(5):551-5.},
	pages = {551-5},
	type = {Journal Article},
	abstract = {The present paper reports alterations in osteogenesis recorded in mandibular condyles 3-18 days after the removal of marrow tissue from tibial bones. Computerized histomorphometric evaluation of undecalcified condyles revealed a considerable increase in the thickness of condylar cartilage, in particular, the zone of provisional calcification. Measurements of the subcartilaginous trabecular bone suggested an increase in the number of osteoblasts and in their activity. The systemic enhancement of osteogenesis may be initiated by circulating factors released at the affected limb during regeneration of the marrow.},
	keywords = {Animals *Bone Development Bone Marrow/*physiopathology Bone Resorption Cartilage/*growth \& development Male Mandibular Condyle/*growth \& development Minerals/metabolism Organ Size Rats Rats, Inbred Strains Tibia Time Factors},
	issn = {0171-967X (Print) 0171-967X (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/3933795},
	author = {Bab, I. and Gazit, D. and Massarawa, A. and Sela, J.}
}
@article {17005,
	title = {Multifocal cystic hygroma: report of case},
	journal = {ASDC J Dent Child},
	volume = {51},
	number = {6},
	year = {1984},
	note = {Kischinovsky, D Gazit, D Azaz, B eng Case Reports 1984/11/01 ASDC J Dent Child. 1984 Nov-Dec;51(6):444-7.},
	pages = {444-7},
	type = {Journal Article},
	keywords = {Child Child, Multiple Primary/*pathology Salivary Gland Neoplasms/*pathology Submandibular Gland Neoplasms/*pathology Tongue Neoplasms/*pathology, Preschool Humans Lymphangioma/*pathology Male Mouth Floor/pathology Mouth Neoplasms/*pathology Neoplasms},
	issn = {1945-1954 (Print) 1945-1954 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/6594361},
	author = {Kischinovsky, D. and Gazit, D. and Azaz, B.}
}
@article {17004,
	title = {A study of a sample of oral cancer in Israel},
	journal = {Oral Surg Oral Med Oral Pathol},
	volume = {57},
	number = {1},
	year = {1984},
	note = {Gazit, D Ulmansky, M Fischman, S Bab, I Sela, J eng 1984/01/01 Oral Surg Oral Med Oral Pathol. 1984 Jan;57(1):118-21.},
	pages = {118-21},
	type = {Journal Article},
	abstract = {A review of data on oral cancer in Israel revealed a lower incidence and a later onset age than in other countries. These data are reviewed and consideration is given to the high prevalence of smoking and the low incidence of alcoholism in Israel.},
	keywords = {Adolescent Adult Age Factors Aged Child Female Humans Israel Male Middle Aged Mouth Neoplasms/*epidemiology/pathology Smoking},
	issn = {0030-4220 (Print) 0030-4220 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/6582424},
	author = {Gazit, D. and Ulmansky, M. and Fischman, S. and Bab, I. and Sela, J.}
}
@article {17003,
	title = {Correlative clinico-pathological evaluation of oral premalignancy},
	journal = {J Oral Pathol},
	volume = {11},
	number = {4},
	year = {1982},
	note = {Fischman, S L Ulmansky, M Sela, J Bab, I Gazit, D eng Research Support, Non-U.S. Gov{\textquoteright}t Denmark 1982/08/01 J Oral Pathol. 1982 Aug;11(4):283-9.},
	pages = {283-9},
	type = {Journal Article},
	abstract = {In 1972, the World Health Organization{\textquoteright}s "Meeting of Investigators on the Histological Definitions in Precancerous Lesions" defined a precancerous lesion as a "morphologically altered tissue in which cancer is more likely to occur than in its apparently normal counter part" (Pindborg 1980). There are two generally accepted precancerous lesions in the oral cavity, leukoplakia and erythroplakia (Pindborg 1980). Leukoplakia is currently defined as "a white patch or plaque that cannot be characterized clinically or pathologically as any other disease" (WHO 1978). This definition has no histological connotation and is used in a strictly clinical sense (Pindborg 1980, Banoczy 1977). Erythroplakia is defined as a "bright red velvety plaque which cannot be characterized clinically or pathologically as being due to any other condition" (Pindborg 1980).},
	keywords = {Diagnosis, Differential Erythroplasia/pathology Humans Leukoplakia, Oral/pathology Mouth Neoplasms/*pathology Precancerous Conditions/*pathology World Health Organization},
	issn = {0300-9777 (Print) 0300-9777 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/6809916},
	author = {Fischman, S. L. and Ulmansky, M. and Sela, J. and Bab, I. and Gazit, D.}
}
