Experiment of treatment of bone defect with adenovirus-mediated expression of bone morphogenetic protein-2 / 上海第二医科大学学报

ABSTRACT

Objective To explore the feasibility of mouse muscle-derived cells with adenovirus-mediated expression of BMP-2 in the treatment of mouse tibia bone defect via collagen sponge as scaffold. Methods Mouse adenovirus BMP-2 plasmids were prepared and identified by PCR.The adherent cells of muscle-derived stem cells cultured in vitro were passaged,cells of the third generation were selected,and recombinant Ad-BMP-2 plasmids were expressed and incubated in mouse myoblasts.The number of BMP-2 positive cells was measured using indirect immunofluorescence method,and alkaline phosphatase(ALP) method was adopted to determine the cell activity in each group.The mouse models of proximal tibial bone defect were established(n=80) and were divided into Ad-BMP-2 transfection group,Ad-LacZ transfection group,blank group and Ad-BMP-2 implantation group.The histological and radiological changes were observed 3 weeks after operation. Results It was revealed by indirect immunofluorescence that BMP-2 positive cells were frequently found in Ad-BMP-2 transfection group,while none were observed in Ad-LacZ transfection group and blank group.The ALP activity was higher in Ad-BMP-2 transfection group,while that was extremely low in Ad-LacZ transfection group and blank group.There was most typical imaging of repair and healing of bone defect in Ad-BMP-2 transfection group,followed by Ad-BMP-2 implantation group,while there was no typical imaging in Ad-LacZ transfection group and blank group. Conclusion The constructed Ad-BMP-2 plasmids can effectively transfect mouse muscle-derived cells,and the transfected cells can continuously express effective amount of BMP-2.The mouse muscle-derived cells with adenovirus-mediated expression of BMP-2 play a definite role in the treatment of bone defect,which provides an effective method and material for clinical application.