小分子鹰嘴豆芽素A对人牙髓干细胞成骨分化的影响

目的:探讨不同浓度鹰嘴豆芽素A (biochanin A,BCA)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)成骨分化的影响及相关分子机制.方法:通过组织块法分离培养原代人牙髓干细胞,流式细胞术鉴定其细胞表型.通过CCK-8法检测不同浓度BCA对hDPSCs增殖活性的影响,通...

Effects of small molecule biochanin A on osteogenic differentiation of human dental pulp stem cells

PURPOSE: To study the effects of different concentrations of biochanin A (BCA) on osteogenic differentiation of human dental pulp stem cells (hDPSCs) and its related molecular mechanisms. METHODS: Primary human dental pulp stem cells were isolated and cultured by tissue explant method, cell phenotypes were identified by flow cytometry. Effects of different concentrations of BCA on proliferation of hDPSCs were detected by CCK-8 method, and effects of different concentrations of BCA on osteogenic differentiation of hDPSCs were analyzed by q-PCR, alkaline phosphatase (ALP) staining, and alizarin red staining. Finally, the influence of BCA on MAPK signaling pathway during osteogenic differentiation of hDPSCs was investigated through Western blot. SPSS 16.0 software package was used for statistical analysis of the data. RESULTS: hDPSCs obtained in the experiment were conformed to have the characteristics of mesenchymal stem cells by flow cytometry. CCK-8 results showed that BCA below 25 μmol/L had no effect on cell activity of hDPSCs, while over 30 μmol/L BCA inhibited the proliferation of hDPSCs. 10 μmol/L BCA significantly increased the expression of osteogenic-related genes ALP, COL-1, OPN and OCN. The staining of ALP and alizarin red was also more obvious than that of the control group and the other concentration groups. Western blot displayed that BCA activated P38/MAPK signaling pathway, but had no obvious effect on ERK and JNK pathway. CONCLUSIONS: Tissue explant method can obtain hDPSCs with higher purity. BCA may promote osteogenic differentiation of hDPSCs by activating P38/MAPK signaling pathway.