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MR Vol.13 No.3 indexに戻る
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MODERN RHEUMATOLOGY
Vol.13 No.3 |
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Efficient gene delivery to articular cartilage
using electroporation |
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| 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 |
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| Received: 22 November 2002 Accepted: 10 January
2003 |
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| 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. |
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| Key words |
| Articular cartilage - Electroporation (EP) -
Gene transfer |
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