关键词: 骨形态发生蛋白类, 基因, 遗传载体

Abstract: Objective To evaluate the osteoinductive activity of recombinant adenoviral vector carrying the human BMP-2 gene (AdBMP-2) and investigate the mechanism of bone formation induced by the bone morphogenetic protein 2 (BMP-2). Methods AdBMP-2 infected rabbit bone messenchymal stem cells (BMSC) were cultivated in vitro. The expression of hBMP-2 was determined by in situ hybridization and im munohistochemical analysis, and ALP wasdetermined by enzyme cytochemistry 7 d after infection. At 21th day, Alizarin red S staining was used to evaluate calcium-rich deposits by cells transfected in culture. 30 μL AdBMP-2 solution (2×10¹pfu•mL^-1) was injected into the muscle of the left hind limb of the nude rat, and 30 μL Adβ gal solution (2×10^1＊pfu•mL^-1) w as injected into the muscle of the right hind limb as control. Animals were sacrificed at 20th day following injection to investigate the bone formation histologically and the expression of BMP-2 in the specimens of the injection sites by immunohistoch emistry and in situ hybridization method. Results The expressions of BMP-2 and ALP in the transduced BMSC were confirmed at 7 th day after infection. A deeply stain ed mineralized layer was evident at 21th day in transduced cells. In vivo experiment, BMP-2 gene therapy induced mesenchymal monocytes chemotaxis, proliferation,differentiation into chondrocytes, with subsequent endochondral bone formation. At the same time, hBMP-2 expression was observed in the chondrocytes, osteoblasts and mesenchymal monocytes. Conclusion AdBMP-2 posseses the osteoinductive activity in vitro and in vivo.

Key words: human, bone morphogenetic proteins, gene, genetic vectors