Activity study of bone marrow mesenchymal stem cells-derived osteoblast transfected by vascular endothelial growth factor gene

Objective This experiment aims at exploring the best conditions of differentiation from bone marrow mesenchymal stem cells(MSCs) to osteoblast and observing the effects of extra vascular endothelial growth factor(VEGF) gene expression on the regeneration of osteoblast cells in rats.Methods In vitro,MSCs P2 cells were selected to be induced to osteoblasts.The cells were divided into two groups:control group and 1×10-8 mol/L DEX group.The expression of alkaline phosphatase(ALP) and type-Ⅰ collagen of the osteoblast cells were observed.Alizarin red staining was applied to observe the calcium nodules.The extra VEGF gene was transfered into the osteoblast cells in vitro by the type 5 adenovirus vector and analyzed by immunohistochemistry staining.Results ALP activity(A405):the ALP activity of two groups at the day 3,6,9,12 was measured.The results of 1×10-8 mol/L DEX group were 0.144±0.004,0.175±0.003,0.207±0.010,0.224±0.004.The results of control group were 0.101±0.003,0.124±0.016,0.133±0.005,0.139±0.005.Significant statistical difference of interblock,time dissimilarity and interaction of interblock with time dissimilarity existed between two groups(P0.05).The cells expressed positive staining of ALP when they were induced for 7-9 days and showed alizarin red positive staining of calcium nodules when they were induced for 20 days.The expression of extra VEGF in the osteoblast cells that derived from MSCs showed positive staining.Conclusion The ad5-h-VEGF gene could be integrated with the osteoblast cells that derived from MSCs and expressed in the cytoplasm.