Intradiscal Gene Therapy: Therapeutic Implications in Degenerative Disc Disease.
- Author:
Seong Hwan MOON
1
;
Moon Soo PARK
;
Yong Ho KANG
;
Hyung Jung KIM
;
Hwan Mo LEE
;
Nam Hyun KIM
;
Lars G GILBERTSON
;
James D KANG
Author Information
1. Department of Orthopedic Surgery, Yonsei University College of Medicine, Seoul, Korea. shmoon@yumc.yonsei.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Gene Therapy;
Proteoglycan;
Disc Degeneration;
TGF-beta1
- MeSH:
Anabolic Agents;
Chromatography;
Culture Media, Conditioned;
Enzyme-Linked Immunosorbent Assay;
Genetic Therapy*;
Humans;
Intercellular Signaling Peptides and Proteins;
Intervertebral Disc;
Intervertebral Disc Degeneration;
New Zealand;
Proteoglycans;
Rabbits;
Transforming Growth Factor beta1;
Up-Regulation
- From:Journal of Korean Society of Spine Surgery
2000;7(4):507-513
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
STUDY DESIGN: In vitro and in vivo studies to determine the anabolic effects of intervertebral disc (IVD) to adenovirus-mediated therapeutic gene transfer. OBJECTIVES: To quantify the anabolic effect of human IVD cells in vitro and rabbit IVD in vivo to therapeutic gene transfer. SUMMARY OF LITERATURE REVIEW: An alternative possibility to delivery of growth factors, in continuous manner, is the genetic modification of disc cells through gene transfer. Contemplating to extend this approach to treatment of disc degeneration, it is necessary to demonstrate anabolic effect of human IVD cells and rabbit disc to therapeutic gene transfer. MATERIALS AND METHODS: In vitro: IVD tissue was obtained from twelve patients. IVD cells were then isolated, cultured, and transduced with Ad/TGF-beta1. Genetically modified disc cells were incorporated into alginate beads and cultured. In vivo: Fifteen skeletally mature New Zealand white rabbit were used. 15ul of saline containing Ad/TGF-beta1 were injected into the nucleus pulposus of the disc in six rabbits. All rabbits were sacrificed 6 weeks after surgery. Nucleus pulposus tissues were harvested, weighted, and cultured. Conditioned medium of alginate bead and rabbit disc tissue cultures were subjected to ELISA to detect TGF-beta1 production. Newly synthesized proteoglycan were analyzed using chromatography on Sephadex G-25 in PD-10 columns after S35-sulfate incorporation. RESULTS: Concentration of TGF-beta1 increased over time in alginate beads cultures transduced with Ad/TGF- beta1. At 6 weeks nucleus pulposus tissue from the disc injected with Ad/TGF-beta1 exhibited 200% (p<0.05) increase in TGF- beta1 production. There was statistically significant 290% increase in newly synthesized proteoglycan in alginate cultures transduced with Ad/TGF- beta1 (p<0.05) compared to control. At 6 weeks nucleus pulposus tissue from the disc injected with Ad/TGF- beta1 exhibited 85% increase in proteoglycan synthesis (p<0.05) over that of intact control. CONCLUSION: In this study, we observed the robust upregulation of proteoglycan synthesis in gene transferred disc cells in vitro and in vivo - indicating good prospects for biologic effects of therapeutic gene therapy in the disc using adenovirus-mediated approach.
