Ectopic bone formation using bone morphogenetic protein 2 adenoviral vector / 吉林大学学报(医学版)

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 immunohistochemical analysis,and ALP was determined 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 ?L AdBMP-2 solution(2?10~(12)pfu?mL~(-1)) was injected into the muscle of the left hind limb of the nude rat,and 30 ?L Ad? gal solution(2?10~(12)pfu?mL~(-1)) was 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 immunohistochemistry 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 stained 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.