| |
|
 |
|
MODERN RHEUMATOLOGY Vol.13 No.3
>MR13-3
|
| Efficient gene delivery to articular cartilage using electroporation |
| |
| Rie Katayama1 , Tomoatsu Kimura1, Tetsuya Tomita2, Hiroaki Matsuno1, Yuji Morita1, Isao Matsushita1 and Ryuichi Gejo1 |
(1) Department of Orthopaedic Surgery, Faculty of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama, 930-0194, Japan
(2) Department of Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Japan |
| |
| Received: 22 November 2002 Accepted: 10 January 2003 |
| |
Full Text
> Click Here （member's only） |
| |
| Abstract |
| Effective in vivo gene transfer into articular cartilage has not yet been established. Since chondrocytes are embedded within a rich extracellular matrix, various gene transfer methods have failed to introduce genes into deeper layers of the articular cartilage. In this study, we developed new superfine pointed needle electrodes for in situ electroporation (EP), and investigated the efficiency of gene transfer into articular cartilage with different degrees of degeneration. Full-thickness articular cartilage slices were obtained from the knee joint of a 3?4-month-old rabbit. The cartilage tissues were treated briefly with trypsin to partly remove matrix proteoglycan. Human articular cartilage with different grades of degeneration was also used. For EP, the articular cartilage surface was soaked in a solution containing green fluorescent protein (GFP) plasmid. Then, the superfine pointed 7-needle electrodes were gently stabbed into the surface layer of the articular cartilage and the gene was transfected by an electroporator. GFP expression was examined by immunohistochemical analysis. Cartilage tissue was successfully transfected with the GFP gene by the electrodes and EP. Transfection efficiency was enhanced by depleting the matrix proteoglycan in rabbit articular cartilage. Chondrocytes in the deeper layer of the articular cartilage were also transfected and expressed GFP. In human osteoarthritic cartilage, ca. 30% of the cells in the deeper layer were transfected by selecting optimal EP conditions. No adverse effects of EP on damaged articular cartilage were obvious from histological analysis or TUNEL staining. The results indicated that EP-mediated in vivo gene transfer into articular cartilage may provide a useful therapeutic strategy to treat cartilage degeneration. |
| |
| Key words |
| Articular cartilage - Electroporation (EP) - Gene transfer |
|
| |
| 一覧に戻る |
|
|
