아데노바이러스를 이용한 치료적 유전자 전달 : 인간 추간판 세포에서의 기질 생성

Abstract

Study Design : In vitro experiment to determine the matrix synthesis of human intervertebral disc (IVD) cell to adenovirus-mediated therapeutic gene transfer. Objectives : To elucidate proteogycan and collagen synthesis of human IVD cells in vitro to adenovirus-mediated transfer of cDNA of transforming growth factor-beta 1(TGF-βl). Summary of Literature Review : Sophisticated method to delivery of growth factors, in continuous manner, is the genetic modification of disc cells through gene transfer. Confirming susceptibility of human IVD cell to adenovirus, anabolic response of human IVD cells to therapeutic gene transfer should be next step. Materials and Methods: IVD tissue was obtained from fourteen patients with grade III, IV degeneration. Isolation and culture of disc cells were performed. Disc cells were treated with either Ad/TGF-βl exogenous TGF-βl. Control cultures were treated with either saline or Ad/luciferase. Newly synthesized proteoglycans were assessed by 35S-sulfate incorporation using chro-matography on Sepadex G-25 in PD-10 columns. Uptake of 3H proline was used to measure synthesis of collagen and noncol-lagen protein. Results: Culture treated with Ad/TGF-βl showed 3 fold increase in proteoglycan synthesis (p<0.05), culture with exogenous TGF-β1 failed to demonstrate increase in proteoglycan synthesis. In collagen and noncollagen synthesis, cultures with Ad/TGF-β1 and exogenous TGF-βl showed similar 3.7 fold increase in collagen and 2.7 fold increase in noncollagen synthesis comparing control (p<0.05). Conclusion: Adenovirus-mediated gene transfer appears to be an efficient technique for modulating biologic activity of human intervertebral disc cells in terms of matrix synthesis.