Cytokine co-stimulation effect on odontogenic differentiation of stem cells.

ABSTRACT

OBJECTIVE: The study aims to evaluate the effect of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-beta 1 (TGF-ß1) co-stimulation on odontogenic differentiation of human dental pulp stem cells (hDPSCs). MATERIALS AND METHODS: The viability/proliferation of hDPSCs treated with BMP-2 (group B), TGF-ß1 (group T), or BMP-2/TGF-ß1 (group BT) were evaluated. The experiments on odontogenic differentiation were done for 14 days. The following subgroups were added to investigate the effect of co-stimulation with different timing subgroup B1, TGF-ß1 co-stimulation in the first week; subgroup B2, TGF-ß1 co-stimulation in the second week; subgroup T1, BMP-2 co-stimulation in the first week; and subgroup T2, BMP-2 co-stimulation in the second week. The mineralization was assessed using alizarin red staining. The expression of following genes was assessed using quantitative real-time polymerase chain reaction dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP1), osteopontin (OPN), and alkaline phosphatase. RESULTS: All groups showed viability similar to the control group (P > .05). The greater mineralization was detected in B groups on day 14. The expressions of DSPP, DMP-1, and OPN increased on day 14 (P < .05). In the combination groups, the higher expressions of DSPP and DMP-1 were observed in subgroups B1 and B2 than groups B and T (P < .05). CONCLUSIONS: BMP-2 was the key in odontogenic differentiation of hDPSCs, which was further enhanced by co-stimulation with TGF-ß1. Continuous stimulation with TGFß-1 did not improve the differentiation of hDPSCs. CLINICAL RELEVANCE Combined use of the BMP-2 and TGFß-1 at the specific sequence can provide a tissue engineering approach for the future guided dentin regeneration.