title: Efficient gene delivery to articular cartilage using electroporation.
abstract: 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.
authors.affiliation: Department of Orthopaedic Surgery, Faculty of Medicine, Toyama Medical and Pharmaceutical University , 2630 Sugitani, Toyama 930-0194 , Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Katayama
authors.forename: Tomoatsu
authors.initials: T
authors.lastname: Kimura
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Tomita
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Matsuno
authors.forename: Yuji
authors.initials: Y
authors.lastname: Morita
authors.forename: Isao
authors.initials: I
authors.lastname: Matsushita
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Gejo