Experimental Studies on the effects of BMP-2 Expressing Adenoviral Vector to the Distraction Osteogenesis

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1.To examine the effectiveness of a gene transfer procedure for the delivery of BMP-2 to the distraction osteogenesis, we constructed a human BMP-2--expressing replication-deficient adenoviral vector, AxCAOBMP-2, and evaluated its osteoinductive activity in vitro and in vivo.2.C2C12 myoblasts were infected in vitro with this viral vector or an Escherichia coil LacZ gene-expressing control adenovirus vector (AxCALacZ). Twenty-four hours after the infection, indirect immunofluorescence was performed. On day 5 after the infection, alkaline phosphatase (ALP) in the cells and osteocalcin in the culture medium were measured. Furthermore, to examine the effectiveness of gene transfer of BMP-2 in vivo, we evaluated osteoinduction by AxCAOBMP-2, under transient immunosuppression with cyclophosphamide, given at a dose of 125 mg/kg intraperitoneally the day before injection of the adenoviral vector. Twenty-five microliters of AxCAOBMP-2 (8.75 x 10^8 plaque-forming units [pfu],'Group I) and AxCALa … More cZ (1.75 x 10^8 pfu, control group) and 5 microl of AxCAOBMP-2 (1.75x 10^8 pfu, Group II) were injected into a right calf muscle of Wistar rats. On day 21, bone formation in each group was investigated radiologically and histologically.3.Abundant BMP-2 expression in C2C12 cells infected with this viral vector was confirmed by immunofluorescence. C2C12 cellstransferred with the BMP-2 gene by this vector produced ALP in the cells and also produced and secreted osteocalcin in the culture medium. Osteoinduction was found only in the AxCAOBMP-2 treated groups with immunosuppression. Osteoinduction activity was higher in Group I than in Group II.3.Twelve animals, performed distraction, were treated with AxCAOBMP-2 (8.75 x 10^8 pfu) 25μg in each, during distraction osteogenesis at 7 days and 14 days. In results, Bone formation and ALP activity were increased significantly in comparison with non-treatment groups. It was suggested that Gene transfer procedure for the delivery of BMP-2 effected to the distraction osteogenesis and had a potential shorting distraction period. Less