Effect of Sclerostin on BMP-2-induced osteogenesis of human mesenchymal stem cells

ZHANG Yao; HE Yao-hua

doi:10.3969/j.issn.1002-266X.2015.28.002

Abstract：Objective To investigate the influence of Sclerostin on bone morphogenetic protein 2 ( BMP-2 )-induced osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs).Methods The bone marrow tissues taken from two males receiving amputation because of traffic accident were used to culture the human BMSCs in vitro.The cultured cells were randomly divided into three groups:groups 1, 2 and 3.The cells in the group 1 did not re-ceive transfection ( Mock) , the cells in the group 2 were transfected with control siRNA, and the cells in the group 3 were transfected with Sclerostin siRNA (BMPs inhibitor).Meanwhile, the osteoblasts were cultured with 0.1 μg/mL BMP-2. CCK-8 assay was used to detect the activities of the cells on the 0, 3rd and 7th day of induction, and the content of DNA was detected by CyQUANT cell proliferation assay kit.On the 14th day of culture, the expression levels of osteocalcin ( OCN) , osteocalcin ( OC) and osteopontin ( OPN) were detected by using RT-PCR, the alkaline phosphatase ( ALP) a-nalysis kit was used to analyze the activity of ALP, and the BCA protein assay reagent kit was used to analyze the cell pro-tein content.Results On the 3rd and 7th day of culture, the proliferation activity and total DNA content of group 3 were higher than those of the groups 1 and 2 (all P<0.05).On the 14th day of culture, the relative expression levels of OCN, OC and OPN mRNA and total protein content were higher than those of the groups 1 and 2 (all P<0.05).The activities of ALP in the groups 1, 2 and 3 were respectively 1.00%±0.08%, 1.12%±0.09% and 2.50%±0.03%, and group 3 was higher than group 1 and group 2 (all P<0.05).Conclusion Inhibiting the Sclerostin expression may increase the cell viability of human BMSCs and osteogenic differentiation.