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

SummaryObjectiveThe 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.MethodsChondrocytes 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.ResultsRadial 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.ConclusionIn 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

Extracorporeal shock wave therapy improves motor dysfunction and pain originating from knee osteoarthritis in rats

SummaryObjectiveAlthough there have been several reports on the use of extracorporeal shock wave therapy (ESWT), the efficacy of ESWT for knee osteoarthritis (OA) has not been clarified. The aim of this study is to investigate the effect of ESWT on OA in a rat knee model.MethodsThe rats were divided into three groups: (1) control, (2) OA, and (3) ESWT (knee OA+shock wave therapy). Behavioral analysis consisted of measuring the duration of walking on a treadmill. The expression of calcitonin gene-related peptide (CGRP) in dorsal root ganglion (DRG) neurons innervating the knee using immunohistochemistry was examined in the three groups at their peak time point on the treadmill.ResultsWalking duration was significantly extended 4, 7 and 14 days after ESWT in rats with knee OA (peak time point: 4 days), again decreasing by days 21 and 28. Immunohistochemical studies revealed that the OA group had significantly higher percentages of CGRP positive neurons in the DRG than were found in the control group. In addition, ESWT reduced the ratio of CGRP positive DRG neurons in the OA model.ConclusionThe improvement in walking ability and the reduction of CGRP positive neurons in DRG indicates that ESWT is a useful treatment for knee OA

Kinetic Modelling of Hydrothermal Lignin Depolymerisation

Although lignin is one of the most abundant renewable organic materials in the world, it is principally a waste product of the paper industry which is used for the production of heat and power. Hydrothermal lignin depolymerisation aids in facilitating the valorization of lignin in aqueous solutions or suspensions. For the recovery of valuable phenolic products from lignin it is crucial to understand the main reaction pathways of lignin degradation and the reaction kinetics. Batch experiments were carried out for studying the depolymerisation of an enzymatic hydrolysis lignin from spruce wood in near critical water. Phenolic products were extracted from the aqueous phase and quantified via gas chromatography. The main reaction products were grouped (lumped), the main reaction pathways of hydrothermal lignin depolymerisation were discovered and formal kinetic rate coefficients were determined. Optimization of these formal kinetic parameters yielded a satisfying approximation of the experimental yields of phenolic products and describes the most important tendencies over temperature and residence time of solid residue and gas. The model is validated by the comparison with other kinetic studies of the degradation of lignin as well as the decomposition of intermediate phenolics, such as catechols and methoxyphenols