Effect Of Human Bone Morphogenetic Protein-7 Gene Transfection On Biological Activity Of Primary Cultured Rabbit Nucleus Pulposus Cells In Vitro

Objectives: To study the effect of human bone morphogenetic protein-7 (hBMP-7) gene transfection on biological activity of primary cultured rabbit nucleus pulposus cells in vitro. To determine the feasibility of hBMP-7 gene which will on gene therapy research of intervertebral disc degeneration (IDD) and to provide basis for further study. Method: hBMP-7 gene was integrated into an adenovirus vector to construct Ad-hBMP7. Then Ad-hBMP7 infected primary cultured rabbit nucleus pulposus cells. And the expression of hBMP-7 was determined by RT-PCR, Western blot and immunocytochemistry. The effect of hBMP-7 on cell proliferation was surveyed by MTT and flow cytometry. Furthermore, the effect of hBMP-7 gene on synthesis of proteoglycan and type II collagen was detected by modified dimethylmethylene blue method and ELISA, respectively. Result: The adenoviral vector was successfully constructed. After transduction into the nucleus pulposus cells, high level of hBMP-7 expression was observed and lasted over 3 weeks. Also hBMP-7 gene can promote cell proliferation and synthesis of proteoglycan and type II collagen with significant difference compared with control groups (P<0.05). Conclusion: hBMP-7 gene mediated by adenovirus can be the target gene for the further study on gene therapy of IDD.