Abstract:Objective To observe the effect of MMP-2 on the osteogenic differentiation of muscle-derived mesenchymal stem cells.Methods The rat thigh muscle tissue was taken for primary culture,and mesenchymal stem cells were isolated and identified by flow cytometry.The Plvx-IRES-ZsGreen1-MMP-2 overexpression vector and Plvx-shRNA2-sh-MMP-2 interference vector were constructed,and the mesenchymal stem cells were treated with MMP-2 overexpression and suppression.Western blotting was used to detect the expression of osteogenic related genes RUNX2,OCN and ALP in the muscle-derived mesenchymal stem cells on the first,7th,and 14th days after plasmid transfection.Alizarin red staining was further performed to detect the osteogenic differentiation of mesenchymal stem cells on the 7th and 14th days.Results The primary cultured muscle-derived mesenchymal stem cells were identified as rat muscle stem cells by flow cytometry;the construction of Plvx-IRES-ZsGreen1-MMP-2 overexpression vector and Plvx-shRNA2-sh-MMP-2 interference vector could effectively interfere the expression of MMP-2 in mesenchymal stem cells.Western blot detection results showed that the expression of osteogenic genes RUNX2,OCN and ALP in the MMP-2-upregulating group were significantly increased on the 7th and 14th days,and were significantly decreased on the 7th and 14th days in the MMP-2-downregulating group.However,on the 7th day of culture,there was little difference in Alizarin Red staining both in the MMP-2-upregulating group and MMP-2-downregulating group.On the 14th day of culture,the Alizarin Red staining of the MMP-2 overexpression group had the most calcium nodules.Conclusion MMP-2 can promote the osteogenic differentiation of myogenic mesenchymal stem cells and the formation of calcium nodules.
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