Effects Of Alpha-zearalanol On The Osteogenic Differentiation Of Murine Bone Marrow Mesenchymal Stem Cells

Objective:To observe the effects of a-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells.Methods:Mouse bone marrow mesenchymal stem cells were isolated through whole bone marrow adherence method. Passage3cells were divided into non-induced group (cultured by the Iscove’s modified Dulbecco’s medium (IMDM) containing10%fetal bovine serum and1%penicillin and streptomycin), blank induced group (induced by the classic osteoblast-induced system with0.1μmol/L dexamethasone,10mmol/L P-glycerine sodium, and50μmol/L vitamin C), positive control induced group (induced by the classic osteoblast-induced system and10"8mol/L oestrogen), and low, median, high concentration a-zearalanol group (induced by the classic osteoblast-induced system and10-7,10-6,10-5mol/L a-zearalanol). The morphological changes of induced cells were observed by phase contrast microscopy, and the growth curves of cultured cells were drawn via MTT results. The expression of alkaline phosphatase and osteocalcin were detected by ELISA.Results: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 concentration a-zearalanol group (P<0.01). These findings indicate that a-zearalanol can promote the proliferation of bone marrow mesenchymal stem cells, and enhance the expression of alkaline phosphatase and osteocalcin expression, especially in the low concentration group.Conclusions:This study was demonstrates that low concentration a-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 which a-zearalanol promotes osteogenic differentiation of bone marrow mesenchymal stem cells.