Effect Of Transfected Bone Morphogenetic Protein-2 Gene And Insulin-like Growth Factor Gene On Gene Expression Of Wistar Rat Mesenchymal Stem Cells

Background Fracture healing process consisted of two parts, the regulation of cell factor on bone marrow mesenchymal stem cell differentiation and proliferation and cytokine on bone induction. Bone morphogenetic protein-2 (BMP-2) and insulin-like growth factor-1 (IGF-1) were important cytokines of fracture healing. BMP-2 could induce bone marrow mesenchymal stem cells (BMSCs) to be differentiated into osteocytes, which was the strongest ossifying factor; IGF-1 could motivate the formation and differentiation of osteocytes, and there was insulin-like fuction on it. The fractures in patients with diabetes was difficult to heal, which was the result of many factors. For the experiment confirmed, the levels of BMP-2 and IGF-1 in diabetes patients' body were decreased in the early fracture and serum. The decreased expression of IGF-Ⅰand BMP-2 and the decreased level of BMP-2 and IGF-Ⅰin early serum were the important reasons of healing-difficulity in the Wistar rats with diabete。But there was no systematic research on how to get exogenous BMP-2 and IGF-Ⅰinto BMSCs to enhance the expression for healing the fracture patients with diabetes.Objective To investigate whether bone morphogenetic protein -2 and insulin-like growth factor -1 in high glucose are able to transfect Wistar rat mesenchymal stem cells, and the effect of the gene expression.Methods The Wistar male rats BMSCs on the third generation after generation were chose to be the transfecting target cell of adenovirus, which were divided into the control group, the blank-carrier group, the BMP-2 group, the IGF-1 group, the BMP-2+IGF-1 group. There were 3 samples in every group. The control group was the cell without being transfected by virus, the blank-carrier group was the cell with being transfected by virus without target gene, the other groups were the cell with being transfected by respective target gene. After transfection, real-time PCR and Enzyme linked immunosorbent assay method (ELISA) were used to measure the gene production. The experimental data was expressed with x±s.The SPSS 17.0 was used to deal with all of the data. The was significant difference when P<0.05 and highly significant difference when P<0.01.Results After the succession of Adenovirus mediated BMP-2 and IGF-1 gene being transfected the cells separately and jointly in high glucose, all of bone marrow stromal cells were survived; and bone marrow stromal cells can express the target gene product, BMP-2 gene product expression peaked in 72 hours after transfection, the grey degree of BMP-2 group was (1.86±0.06), the concentration of BMP-2 group gene product was (58.6±1.61)μg/L, the grey degree of BMP-2+IGF-1 group was (2.37±0.34), the concentration of BMP-2+IGF-1 group gene product was (67.61±1.91)μg/L, there were significant statistical differences between them; the grey degree of BMP-2 group was (1.48±0.03), the concentration of BMP-2 group gene product was (65.17±3.64)μg/L, the grey degree of BMP-2+IGF-1 group was (1.93±0.17), the concentration of BMP-2+IGF-1 group gene product was (73.94±4.30)μg/L, there were significant statistical differences between them.Conclusion Adenovirus mediated BMP-2 and IGF-1 gene were able to transfect the Wistar rat bone marrow stromal cells in high glucose, the gene products after transfection were increased significantly, there is a synergistic effect on gene expression between BMP-2 and IGF-1.