Effects of alpha-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells

Zou Bin; Zong Shao-hui; Zeng Gao-feng; Fang Ye; Gao Tai-hang

doi:10.3969/j.issn.2095-4344.2014.06.003

Abstract：BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts, andα-zearalanol can promote the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. <br> OBJECTIVE:To observe the effects ofα-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. <br> METHODS:Mouse bone marrow mesenchymal stem cells were isolated through whole bone marrow adherence method. Passage 3 cells were divided into non-induced group (cultured by the Iscove’s modified Dulbecco’s medium containing 10%fetal bovine serum and 1%penicil in and streptomycin), blank induced group (induced by the classic osteoblast-induced system with 0.1μmol/L dexamethasone, 10 mmol/Lβ-glycerine sodium, and 50μmol/L vitamin C), positive control induced group (induced by the classic osteoblast-induced system and 10-8 mol/L oestrogen), and low-, median-, high-concentrationα-zearalanol groups (induced by the classic osteoblast-induced system and 10-7, 10-6, 10-5 mol/Lα-zearalanol). The morphological changes of induced cells were observed by phase contrast microscopy, and the growth curves of cultured cells were plotted based on MTT results. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. <br> RESULTS AND CONCLUSION:In the non-induced group, cells showed long fusiform shape mostly, and contact inhibition was observed while the cells were close-packed. After induced, cells proliferated effectively, showed triangle, astero-form, polygon or irregular shape, and clumps and multilayer of cells became evident. Compared to both the blank induced group, the alkaline phosphatase activity and osteocalcin expression were significantly lower in the non-induced group, but higher in the positive control induced group and low concentrationα-zearalanol group (P<0.01). These findings indicate thatα-zearalanol can promote the proliferation of bone marrow mesenchymal stem cells, and enhance the expression of alkaline phosphatase and osteocalcin expression, especial y in the low concentration group. This demonstrates that low concentrationα-zearalanol obviously enhances the osteogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro. Up-regulating the release of alkaline phosphatase and osteocalcin may be one of the mechanisms by whichα-zearalanol promotes osteogenic differentiation of bone marrow mesenchymal stem cells.