The effects of radial shock waves on gene transfer in rabbit chondrocytes in vitro

• Published in: Osteoarthritis and cartilage Vol. 15; no. 11; pp. 1275 - 1282
• Main Authors: Murata, R., M.D, Nakagawa, K., M.D., Ph.D, Ohtori, S., M.D., Ph.D, Ochiai, N., M.D., Ph.D, Arai, M., M.E, Saisu, T., M.D., Ph.D, Sasho, T., M.D., Ph.D, Takahashi, K., M.D., Ph.D, Moriya, H., M.D., Ph.D
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.11.2007
• Subjects: Animals; Cartilage, Articular - metabolism; Cell viability; Chondrocyte; Chondrocytes - enzymology; Chondrocytes - metabolism; Gene therapy; Gene transfer; Knee Joint; Luciferases - genetics; Luciferases - metabolism; Rabbits; Radial shock waves; Rheumatology; Transfection - methods; Transfection efficiency; Ultrasonics; Cell viability; Radial shock waves; Gene transfer; Chondrocyte; Gene therapy; Transfection efficiency
• ISSN: 1063-4584; 1522-9653
• DOI: 10.1016/j.joca.2007.04.001

Summary Objective The purpose of this study was to develop a new technique of gene transfer utilizing radial shock waves. The effects of radial shock waves on gene transfer in rabbit chondrocytes were examined by varying the parameters of exposure conditions in vitro. Methods Chondrocytes were obtained from New Zealand white rabbits and cultured in a monolayer. A luciferase-encoding gene expression vector, or vector alone, was added to chondrocyte cell suspensions, and the cells were then exposed to radial shock waves. Parameters such as pressure amplitude, number of pulses, frequency, and DNA concentration were varied, and luciferase activity was measured 48 h after transfection. Transfection efficiency of radial shock waves was compared with the FuGENE6 transfection method using a green fluorescence protein (GFP)-encoding gene vector by fluorescent-activated cell sorter (FACS) analysis. Results Radial shock wave exposure significantly increased luciferase activity over 140-fold as compared to the control under the optimal exposure conditions. Both pressure amplitude and number of pulses were relevant to transfection efficiency and cell viability, but frequency was not. Transfection efficiency increased in a dose-dependent manner with DNA concentration. FACS analysis showed 4.74% of GFP-encoding gene using radial shock waves. FuGENE6 transfection was almost similar in transfection efficiency to radial shock wave. Conclusion In spite of certain degree of cell disruption, radial shock waves significantly augmented reporter gene transfection in rabbit chondrocytes in vitro . Radial shock waves may potentially contribute to the treatment of the cartilage morbidities by enhancing the potency of tissue healing and gene transfection of growth factors.