Responsiveness of Human Intervertebral Disc Cells in Matrix Synthesis To Adenovirus-Mediated Gene Transfer(IGF-1, TGF-beta1 Encoding Genes).
10.4184/jkss.2001.8.3.195
- Author:
Hwan Mo LEE
1
;
Moon Soo PARK
;
Hyang KIM
;
Hak Sun KIM
;
Eung Shick KANG
;
Nam Hyun KIM
;
Seong Hwan MOON
Author Information
1. Department of Orthopedic Surgery, Yonsei University College of Medicine, Seoul, Korea. shmoon@yumc.yonsei.ac.kr
- Publication Type:Original Article
- Keywords:
Combination gene therapy;
Intervertebral disc;
Proteoglycan;
TGF-beta1;
IGF-1
- MeSH:
Anabolic Agents;
Cells, Cultured;
Chromatography;
Genetic Therapy;
Humans*;
Insulin-Like Growth Factor I;
Intercellular Signaling Peptides and Proteins;
Intervertebral Disc*;
Logic;
Musculoskeletal Diseases;
Proteoglycans;
Regeneration;
Signal Transduction;
Transforming Growth Factor beta1*
- From:Journal of Korean Society of Spine Surgery
2001;8(3):195-201
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Genetic modification of cells through gene transfer gains popularity as a sophisticated delivery system in the management of musculoskeletal disease. Anabolic growth factors like IGF-1 BMP-2 OP-1, and TGF-beta1 were candidate for therapeutic purposus for regenerating matrix of intervertebral disc. TGF-beta1, OP-1 and BMP-2 share same pathway i.e. Smad while IGF-1 utilize P13K pathway. Accordingly it is logical to use two cytokine encoding genes which using different signal transduction pathway to upregulate matrix synthesis. PURPOSE: To elucidate the anabolic effect of the combination gene transfer(IGF-1 and TGF-beta1 encoding gene) to human disc cells, cultured in alginate beads. MATERIALS AND METHODS: Lumbar and cervical intervertebral disc tissue was obtained from surgical disc procedure from fifteen patients. After isolation and culture of the cells, cultures were transduced with first, Adenovirus-TGFbeta1 construct(Ad/TGF-beta1) and Ad/IGF-1 respectively, second, with combination of two viral constructs(Ad/TGF-beta1+Ad/IGF-1). Cultures treated with saline and Ad/luciferase served as control. Then cultures were incorporated into alginate beads. Exogenous TGF-beta1(2ng/ml) and IGF-1(100ng/ml) were administered also. Newly synthesized proteoglycan was assessed using S35 incorporation using chromatography on Sephadex G-25 in PD-10 column. RESULTS: In cultures transduced with single therapeutic gene construct, there were statistically significant 2.9 fold(Ad/TGF-beta1 and 1.8 fold(Ad/IGF-1) increase in newly synthesized proteoglycan comparing control(p<0.05). In culture transduced with double combination of therapeutic gene construct, there were 3.9 fold(Ad/TGF-beta1+Ad/IGF-1) increase in newly synthesized proteo-glycan comparing control(p<0.05). Culture treated with TGF-beta1(2ng/ml) showed 3.9 fold increase and IGF-1(100ng/ml) 2.9 fold increase, TGF-beta1+IGF-1 4.2 fold increase in proteoglycan synthesis compared to control. CONCLUSIONS: Combination gene therapy provided efficient mechanism of upregulating matrix regeneration of the human inter-vertebral disc cells.
