Study on the rule of Ad-BMP-2 to promote the osteogenic potential of cultured rabbit mesenchymal stem cells

To study the influence of Ad-BMP-2 on osteogenic potential of cultured MSCs. MSCs were obtained from dual femoral greater trochanter of twenty Japanese Big Ear rabbits. The experimental group cells were collected from the left trochanters and the control group cells from the right ones. After the MSCs were transfected with Ad-BMP-2, the methods of RT-PCR and immunohistochemical staining were used to test the expression of BMP-2 protein. Gomori stain and ALP testing kit were used to measure ALP activity and BGP immumoradiatory testing kit were used to measure OCN.In addition, the collagen typeⅠ were tested with the methods of RT-PCR and Western blotting.At last, transmission electron microscopy was used to observe the ultrastructures of MSCs before and after gene transfection. Result showed that MSCs transfected with Ad-BMP-2 expressed BMP-2 protein effectively. After the gene transfection, the MSCs gained the structural and morphological characters which were similar to those of osteoblasts. The ALP activity was (7.01 0.59)U/L in the Ad-BMP-2 transfected group as compared with (5.23 0.55)U/L in the control group. Analysis revealed that collagen type Ⅰexpression remarkably increased after transfection (1.35±0.12 in the experimental group versus 3.65±0.37 in the control group).The OCN was 24.50±0.93ng/mL in the experimental group and 15.45±1.81ng/mL in the control group. It was concluded that the osteogenic potential of MSCs can be greatly promoted by gene transfection.