325 288

Cited 59 times in

Biologic response of human intervertebral disc cells to gene therapy cocktail

Authors
 Seong-Hwan Moon  ;  Kotaro Nishida  ;  Lars G. Gilbertson  ;  Hwan-Mo Lee  ;  Hyang Kim  ;  Ronald A. Hall  ;  Paul D. Robbins  ;  James D. Kang 
Citation
 SPINE, Vol.33(17) : 1850-1855, 2008 
Journal Title
 SPINE 
ISSN
 0362-2436 
Issue Date
2008
MeSH
Adenoviridae/genetics ; Adult ; Cells, Cultured ; Gene Transfer Techniques ; Genetic Therapy/methods* ; Genetic Vectors/administration & dosage ; Genetic Vectors/genetics ; Humans ; Intervertebral Disc/cytology* ; Intervertebral Disc/physiology* ; Middle Aged ; Proteoglycans/biosynthesis ; Proteoglycans/genetics
Keywords
gene therapy ; intervertebral disc ; proteoglycan
Abstract
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 transduction. IVD cultures with Ad/TGF-beta1, Ad/IGF-1, Ad/BMP-2 (MOI of 75) showed 2.9, 1.8, and 1.9 fold increases, respectively, in proteoglycan synthesis compared to control. Human IVD cultures with double gene combination (MOI of 75) showed 3.2 to 3.9 fold increases of proteoglycan synthesis. Lastly, Human IVD cultures with triple gene combination (TGF-beta1+IGF-1+BMP-2 genes with an MOI of 75) transfer demonstrated 4.7 fold increase in proteoglycan synthesis compared control. CONCLUSION: Combination or "cocktail" gene therapy offers a promising mechanism for maximizing matrixsynthesis with low dose of adenoviral mixtures, circumventing systemic, local toxic effect, and immune response.
Files in This Item:
T200800109.pdf Download
DOI
10.1097/BRS.0b013e31817e1cd7
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Orthopedic Surgery (정형외과학교실) > 1. Journal Papers
Yonsei Authors
Moon, Seong Hwan(문성환)
Lee, Hwan Mo(이환모) ORCID logo https://orcid.org/0000-0002-5405-3832
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/106234
사서에게 알리기
  feedback

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse

Links