Transfection of vascular endothelial growth factor gene promotes osteogenesis activities of osteoblasts

Background: Ectogenesis vascular endothelial growth factor (VEGF) could enhance the activity of alkaline phosphatase (AKP) and concentration of cycli adenosine monophosphate (cAMP) fivefolds in cultured osteoblast cell. What's the effect of ectogenesis VEGF gene transfection on osteoblasts is still by no means clear. Objective: To investigate the effect of gene transfection and expression of ectogenesis VEGF on the osteogenesis activities of osteoblast cell. Dedign: A completely randomized controlled study. Setting: Laboratory of Transplantation and Immunity, West China Hospital, Sichuan University. Materials: Cranial osteoblasts of newborn two or three-day male SD rat. Methods: The experiment was conducted in the Laboratory of Transplantation and Immunology of Huaxi Hospital, Sichuan University from April to December 2003 The cranial osteolasts of newborn rat were separated and cultured with enzyme digestion method then were identified by AK-Pstaining. 1 pBLAST49-mVEGF gene was transferred into the osteoblasts cultured in vitro with cation liposomes transfection as gene transfection group. 2 Basticidin antibiotic was added into untransfection group as control 3 Culture the transfected and untransfected osteoblasts for 5 generations, immunohistochemical staining was performed on VEGF and collagen type I and osteocalcium were detected. Main outcome measures: 1 The expression of ectogenesis VEGF in the osteoblasts, 2 the influence of ectogenesis VEGF on synthesis of collagen I and secretion of osteocalcium of osteoblasts. Results: The concentration of osteocalcium and expression of type I collagen of the 1-5 generation osteoblast cell in pBLAST49-mVEGF gene transfer group were significantly higher than that in the control group (P < 0.05). Conclusion: It is found in this experiment that the synthesis of collagen I was enhanced obviously after sussceful transfection of pBLAST49-mVEGF plasmid. Compared with the control group, the diffence of intergrated optical density gained by Mias image analysis system was significant(P < 0.05), indicating that pBLAST49-mVEGF plasmid transfection can improve the synthesis of type I collagen and secretion of osteocalcium of osteoblasts.