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akermanite bioceramics for bone regeneration. Biomaterials , 2009; 30:5041-5048. 8. Qu T, Jing J, Jiang Y, Taylor RJ, Feng JQ, Geiger B, Liu X . Magnesium-containing nanostructured hybrid scaffolds for enhanced dentin regeneration. Tissue Eng , 2014; 20:2422-2433. 9. Saltman PD, Strause LG . The role of trace minerals in osteoporosis. J Am Coll Nutr , 1993; 12:384-389. 10. Beattie JH, Avenell A . Trace element nutrition and bone metabolism. Nutr Res Rev , 1992; 5(1):167-188. 11. Rodríguez JP, Ríos S, González M . Modulation of the proliferation and differentiation

Isolation of Dental Pulp Stem Cells and their In Vitro Differentiation into Odontoblast-like Cells

Background: Recently, tooth tissue engineering has attracted more and more attention. Stem cell based tissue engineering is thought to be a promising way to replace the missing tooth. Mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into a variety of cell types.

Material and Methods: We isolated stem cells from dental pulp and measured their self-renewal capacities. Adipogenic, chondrogenic as well as odontogenic differentiation potentials were investigated, using bone morphogenic protein 2 (BMP2) for the odontogenic differentiation.

Results: The cumulative number of the isolated cells was high. Polycomb ring finger oncogene (Bmi1) and Signal transducer and activator of transcription 3 (Stat3) were continuously expressed suggesting longer proliferative lifespan and self-renewal capacity of the isolated cells. Peroxisome proliferatoractivated receptor was expressed showing adipogenic conversion that was also confirmed by positive staining of cells with Oil red O stain and chondrogenic differentiation was confirmed by positive staining of cells with Alcian blue stain. BMP2 stimulated the expression of dentin sialophosphoprotein (DSPP) and enamelysin, indicating successful odontogenic differentiation that was also confirmed by the positive staining of the cells with Alizarin red stain.

Conclusion: Thus, adult pulp contains stem cells, which are useful for cell therapy with BMP2 for dentin regeneration.

REFERENCES 1. D’Souza R. Development of the pulpodentin complex. In Hargreaves KM, Goodis HE, editors. Seltzer and Bender’s dental pulp. Carol Stream, IL: Quintessence Publishing Co., 2002. 13-40. 2. Bhasker SN. Orban’s oral histology and embryology. St. Louis: Mosby, 1991. 3. Huang GT, Sonoyama W, Liu Y, et al. The Hidden Treasure in Apical Papilla: The Potential Role in Pulp/Dentin Regeneration and BioRoot Engineering. J Endod, 2008, 34, 645-651. 4. Chrepa V, Pitcher B, Henry M, et al. Survival of the Apical Papilla and Its Resident Stem Cells in a Case of

polyalkenoates. J Dent Res 2012;91:454-9. doi: 10.1177/0022034512443068 44. Grech L, Mallia B, Camilleri J. Characterization of set Intermediate Restorative Material, Biodentine, Bioaggregate and a prototype calcium silicate cement for use as root-end filling materials. Int Endod J 2013,46:632-41. doi: 10.1111/ iej.12039 45. About I. Dentin regeneration in vitro: the pivotal role of supportive cells. Adv Dent Res 2011;23:320-4. doi: 10.1177/0022034511405324 46. Novicka A, Lipski M, Parafiniuk M, Sporniak-Tutak K, Lichota D, Kosierkiewicz A, Kaczmarek W, Buczkowska- Radlińska J

treated teeth implanted subcutaneously in mice [ 101 ]. bFGF-releasing scaffolds promoted robust dentin formation in a rat model of molar defect [ 102 ]. Controlled bFGF release results in localized dentin formation in the defect area [ 102 , 103 ]. The dose administered is a critical factor for the dentin formation. A low dose (0.05 mg/ml) fails to promote dentin regeneration, while a high dose (5 mg/ml) results in scattered and incomplete dentin formation [ 103 ]. Correspondingly, bFGF (dose 30 ng) did not promote dentin bridge formation, but instead fibrous