26RFa effects on osteogenic differentiation of human bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism. OBJECTIVE:To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells. METHODS:In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. cells were inoculated into 6-wel plates and then divided into two groupsexperimental group treated with 10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay. RESULTS AND CONCLUSION:After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group (P<0.05, P<0.01, P<0.05). After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group (P<0.05). These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions.