Related Articles Biologic Response of Human Intervertebral Disc Cells to Gene Therapy Cocktail. Spine. 2008 Jul 10; Authors: Moon SH, Nishida K, Gilbertson LG, Lee HM, Kim H, Hall RA, Robbins PD, Kang JD STUDY DESIGN.: In vitro experiment using human intervertebral disc (IVD) cells and adenovirus-therapeutic gene constructs. OBJECTIVE.: To examine the biologic effect of "cocktail" therapeutic gene transfer to human IVD cells in three-dimensional cultures. SUMMARY OF BACKGROUND DATA.: Gene therapy is regarded as a potential option for the treatment of degenerative disc disease. Although various anabolic genes have previously been introduced for this purpose, cocktail gene transfer of anabolic genes to IVD cells has never been attempted. METHODS.: Human IVDs were harvested during surgical disc procedures and cultured. We prepared recombinant adenovirus constructs bearing the TGF-beta1 gene (Ad/TGF-beta1), the IGF-1 gene (Ad/IGF-1), and the BMP-2 gene (Ad/BMP-2). Transgene expression was detected by luciferase assays, enzyme linked immunosorbent assays, and Western blot analysis. Newly synthesized proteoglycan was measured by S-sulfate incorporation on Sephadex G-25 M in PD 10 columns. Human IVD cells were transduced by single, double, and triple combination of Ad/TGF-beta1, Ad/IGF-1, Ad/BMP-2 with an MOI of 75, then cultured three-dimensionally in alginate beads. RESULTS.: Transgene expression was detected at 18 hours after viral transd ...
Anytime
