Effects Of Ginsenoside On Proliferation And Osteoblast Differentiation Of Murine MSC In Vitro

Background:The depression of MSCs proliferation and osteogenic differentiation is an important part of pathogenesis of osteoporosis. It has been shown that ginsenoside can prevent and cure this disease. Recently, the authors report that ginsenoside can promote the proliferation of rat bone marrow-derived mesenchymal stem cell in vitro. Little has been known about the effects of ginsenoside on the differentiation of murine bone marrow-derived mesenchymal stem cells into osteoblasts.Purpose:This study was performed to examine the effect of ginsenoside on proliferation and osteogenic differentiation of murine bone marrow-derived mesenchymal stem cell and reveal its mechanism. A profound understanding of pharmacological effects of ginsenoside on MSCs was expected.Methods:The MSC colony-forming assays, subculture culture and osteoinductive culture were performed, concentrations of ginsenoside were added to these cultures respectively (final concentration 25～100mg/L). The clonogenic capacity of MSCs was evaluated by conoly-forming assays. Cells were counted in the MSC subcultures. MTT test was used to assess cell viability. After osteoinductive cultures, osteoblasts were stained by Alizarin red to test the forming of calcium nodules, ALP test was used to assess osteoblasts viability, the mRNA level for the gene ALP and BMP-2 in MSCs subcultures were determined by real-time fluorescent quantitative PCR.Results:The experiment results showed that the conoly-forming efficiency of MSCs was markedly increased, the number of cells in MSC subculture was dramatically higher than control group at concentration of 100mg/L of GS. However, there are no distinctive differences in the conoly-forming efficiency of MSCs and the number of cells in MSC subculture in range of 25 to 50mg/L of GS. MTT absorbance value(A490) were remarkably higer than control group at 100mg/L of GS for 24h; while at 50mg/L and 100mg/L for 48h, A490 values were obviously higher than control. Another results showed that the number of calcium nodules in the group of osteoinductive medium added 100mg/L GS were more than osteoinductive control group(P<0.05); other experimental groups have no significant difference compared with osteoinductive control groupCP>0.05). At 100mg/L of GS, compared with Omg/L GS, the level ofALP mRNA expression were increased, at 50mg/L and 100mg/L of GS, the level ofBMP-2 mRNA expression were increased (P<0.05).Conclusion:These results indicate that ginsenosides are able to enhance the conoly-forming capacity of MSCs and the proliferation in the MSC subcultures. Ginsenosides are able to induce osteoblasts-forming and have a up-regulating effect on mRNA expression of ALP and BMP-2 in the MSC subcultures. These effects are dose-dependent and time-dependent.