310 Mira Street, Stavropol, Russia, 355017
+7 8652 352524; +7 8652 353229.
+7 8652 352524.
The journal is included into The list of leading scientific periodicals.
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.
1. Arnold W. H., Becher S., Dannan A. Morphological characterization of periodontium-derived human stem cells. Ann Anat. 2010;192(4):215-219.
2. Chen G., Sun Q., Xie L. Comparison of the Odontogenic Differentiation Potential of Dental Follicle, Dental Papilla, and Cranial Neural Crest Cells. J Endod. 2015;41:1091-1099.
3. Grimm W.-D., Giesenhagen B., Hakki S. S. Translational Research and Therapeutic Applications of Neural Crest-Derived Stem Cells in Regenerative Periodontology. Curr Oral Health Rep. 2015.
4. Grimm W.-D., Dannan A., Becher S. et al. The ability of human periodontium-derived stem cells to regenerate periodontal tissues: a preliminary in vivo investigation. Int J Periodontics Restorative Dent. 2011;31(6):94-101.
5. Grimm W.-D., Plöger M., Schau I. Complex, threedimensional reconstruction of critical size defects following delayed implant placement using stem cellcontaining subepithelial connective tissue graft and allogenic human bone blocks for horizontal alveolar bone augmentation: A case report as proof of clinical study principles. Medical News of North Caucasus. 2014;9(2):131-133.
6. Grimm W.-D., Dannan A., Giesenhagen B. Translational Research: Palatal-derived Ecto-mesenchymal Stem Cells from Human Palate: A New Hope for Alveolar Bone and Cranio-Facial Bone Reconstruction. Int J Stem Cells. 2014;7(1):23-29.
7. Grimm W.-D., Plöger M., Schau I. Prefabricated 3D allogenic bone block in conjunction with stem cellcontaining subepithelial connective tissue graft for horizontal alveolar bone augmentation: A case report as proof of clinical study principles. Medical News of North Caucasus. 2014;9(2):175-178.
8. Handa K., Saito M., Yamauchi M. Cementum matrix formation in vivo by cultured dental follicle cells. Bone. 2002;31:606-611. 9. Keeve P. L., Dittmar T., Gassmann G., Grimm W.-D. Characterization and analysis of migration patterns of dentospheres derived from periodontal tissue and the palate. J Periodontal Res. 2013;48(3):276-285.
10. Kharazi A., Levy M. L., Visperas M. C. Chicken embryonic brain: an in vivo model for verifying neural stem cell potency. Laboratory investigation. 2013.
11. Kharzinova V. R., Sermyagin A. A., Gladyr E. A. A Study of Applicability of SNP Chips Developed for Bovine and Ovine Species to Whole-Genome Analysis of Reindeer Rangifer tarandus. J Hered. 2015;106(6):758-761.
12. Király M., Porcsalmy B., Pataki A. Simultaneous PKC and cAMP activation induces differentiation of human dental pulp stem cells into functionally active neurons. Neurochem Int. 2009;55(5):323-332.
13. Miura M., Gronthos S., Zhao M. SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA, 2003;100:5807-5812.
14. Miletich I., Sharpe P. T. Neural crest contribution to mammalian tooth formation. Birth Defects Res C Embryo Today, 2004;72:200-212.
15. Mitsiadis T. A., Cheraud Y., Sharpe P., Fontaine-Perus J. Development of teeth in chick embryos after mouse neural crest transplantations. Proc Natl Acad Sci USA, 2003;100:6541-6545.
16. Sauerbier S., Stubbe K., Maglione M. Mesenchymal stem cells and bovine bone mineral in sinus lift procedures-- an experimental study in sheep. Tissue Engineering Part C Methods. 2010;16:1033-1039.
17. Seo B. M., Miura M., Gronthos S. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004;364:149-155.
18. Sirak S. V., Sletov A. A., Ibragimov I. M. Vliyanie poristogo titana na osteogennyi potential kletok kostnogo mozga in vitro. (Impact of porous titanium on osteogenic potential of bone marrow cells in vitro.) Medical News of the North Caucasus. 2012;27(3):22-25. 19. Sirak S. V., Sletov A. A., Pereverzev R. V. Eksperimentalnoe primenenie poristogo titana pri otkrytom sinus liftinge. (Experimental use of porous titanium at open sinus elevation). Palliativnaya meditsina i reabilitatsyia – Palliative medicine and rehabilitation. 2012;1:55-57.
20. Sirak S. V., Shchetinin E. V., Sletov A. A. Subantralnaya augmentaciya poristim titanom v eksperimente i klinike (Subantral augmentation with porous titanium in theexperiment and clinic). Stomatologiya – Stomatology. 2016;95(1):55-58.
21. Widera D., Zander C., Heidbreder M. et al. Adult palatum as a novel source of neural crest-related stem cells. Stem Cells 2009;27:1899-1910.
22. Widera D., Grimm W.-D., Moebius J. M. Highly efficient neural differentiation of human somatic stem cells, isolated by minimally invasive periodontal surgery. Stem Cells Dev. 2007;16:447-460.
Keywords: NCSC, osteoporotic sheep model, CT, 3D-reconstruction, density measurements, histological analysis, stem cell