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Neural crest-related stem cells of oral origins in vitro and used in osteoporotic sheep model for being investigated due to therapeutic effects in alveolar bone regeneration

[Experimental medicine]
Wolf-Dieter Grimm; Tilman Fritsch; Bernd Giesenhagen; Sergey Sirak; Alexandr Sletov; Magomet Aybazov; S. Tebyakina; Evgeny Shchetinin; Alexander Vukovic; Ivan Koshel;

Neural crest-related stem cells (NCSC) have great promise in the field of regenerative medicine due to their differentiation potential into several lineages. Besides the periodontium, and root surfaces of extracted teeth, NCSC can be obtained from the palate, which represents a large stem cell reservoir in the mucosa. Osteoporotic Sheep provide an appropriate large animal model for preclinical studies. In this initial study, we focused on approving the Osteoporotic Sheep Model, the isolation and characterization of NCSC from osteoporotic sheep palate as an alternative to other stem cell sources and on appropriate X-ray and histological methods for evaluation of the alveolar bone regeneration process. Primary ovine neural crest-related stem cells were obtained from the palate for comparison with human derived NCSC. The cultured ovine NCSC, which were sorted by different methods of characterization were examined for morphology, proliferation and bone-regenerative activity in our osteoporotic sheep model. Density expression of alveolar bone was measured by using CT images. Osteogenic potentials of ovine NCSCs were evaluated by different histological measurements on non-demineralized and demineralized alveolar bone tissues. The in vitro cell assays demonstrated the osteoinductive potential of ovine NCSC as typical sphere-forming pure stem cell culture and its proliferation, and differentiation as well as initial bone formation in a large-animal model. The different histological measurements on non-demineralized and demineralized alveolar bone tissues used in our proof of principle study have showed a high potential for evaluating the regenerative bone processes taking place in our Osteoporotic Sheep Model. Based on these findings, ovine NCSC might serve as an appropriate alternative cell source in pre-clinical research using a large animal model for evaluating the regenerative bone processes taking place in the Osteoporotic Sheep Model developed by our interdisciplinary and international research group.

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Keywords: NCSC, osteoporotic sheep model, CT, 3D-reconstruction, density measurements, histological analysis, stem cell


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Stavropol State Medical Academy
Pyatigorsk State Research Institute of Balneotherapeutics
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