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 48h 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.
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 48h 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.
Authors: Dongli Li; Antonio Pellegrino; Andre Hallack; Nik Petrinic; Antoine Jérusalem; Robin O Cleveland Journal: Biophys J Date: 2018-03-27 Impact factor: 4.033