Adenovirus-mediated TGF- β1 gene transfer to human degenerative lumbar int ervertebral disc cells

Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β(1) (TGF- β 1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerat ive lumbar disc disease. The cells were cultured and directly infected by two a denoviral constructs, Ad/CMV- EGFP containing the enhanced green fluorecence pro tein (EGFP) gene (marker gene) and Ad/CMV- TGF- β(1) containing the potentially therapeutic TGF- β(1) gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV- TGF- β(1)). Results Culture cells transducted by Ad/CMV- EGFP showed specific green fluorescence und er the fluoroscope and expression sustained for at least 4 weeks. When infe cted by Ad/CMV- TGF- β(1), approximally 30% of cultured cells were staind brown (+) with TGF- β(1) staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene , TGF- β(1), by using an adenoviral vector to human degenerative lumbar interve rtebral disc cells.

Adenovirus-mediated TGF-beta(1) gene transfer to human degenerative lumbar intervertebral disc cells

OBJECTIVE: To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor beta(1) (TGF-beta(1)) cDNA, and the expression of the encoded protein. METHODS: Nucleus pulposus samples were surgically obtained from 8 patients with degenerative lumbar disc disease. The cells were cultured and directly infected by two adenoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence protein (EGFP) gene (marker gene) and Ad/CMV-TGF-beta(1) containing the potentially therapeutic TGF-beta(1) gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-beta(1)). RESULTS: Culture cells transduced by Ad/CMV-EGFP showed specific green fluorescence under the fluoroscope and expression sustained for at least 4 weeks. When infected by Ad/CMV-TGF-beta(1), approximately 30% of cultured cells were stained brown (+) with TGF-beta(1) staining. CONCLUSION: This study established the strategy of delivering a potentially therapeutic gene, TGF-beta(1), by using an adenoviral vector to human degenerative lumbar intervertebral disc cells.