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.
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.