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Adenovirus-mediated TGF-beta(1) gene transfer to human degenerative lumbar intervertebral disc cells
Jian Zhao1, Lianshun Jia, Deyu Chen
1Department of Orthopaedics, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China. drzhaojian@163.com
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.