| Background: With the aging process and the popularity of sports, osteoarthritis, articular chondrocytes in cartilage injury and other related diseases every year. A large number of studies have shown that the cytoskeleton in OA cartilage degeneration, and significant change has occurred in the process. Purpose of this project is to investigate the cytoskeleton of the three components of cartilage extracellular matrix metabolism in the role, then revealed osteoarthritic cartilage degeneration pathogenesis. Also can be applied to regulate the cytoskeleton components in some means to interfere with the degradation of osteoarthritis occur, for the treatment of osteoarthritis open new avenues.Objective: Using laser scanning confocal microscope observe the morphology of cartilage cells in cytoskeleton, and cytoskeleton proteins chondrocytes quantitative determination of fluorescence intensity to observe the cytoskeleton of normal and added three agents damaged cartilage morphology and cytoskeleton destruction of protein to determine the validity of cytoskeleton. Actin cytoskeleton in the clear, microtubules and intermediate filaments are the corresponding three components of destructive agent destruction of cytoskeletal filaments later analysis, three types of microtubule and intermediate filament cytoskeleton component of articular cartilage damage in vitro cellular matrix metabolism (collagenâ…¡and GAG) in.Methods: 10 New Zealand white rabbits aged 2 months, we take the knee cartilage chondrocytes cultured until the cells adhered, were randomly divided into four groups, namely normal control group, microfilament destruction group (adding 0.01mg / L cellscytochalasin-D destruction of actin), microtubule damage group (adding 0.4mg / L colchicine destruction of tubulin), intermediate filament destruction group (adding 175mg / L acrylamide destruction of vimentin). Cultured for 3 days, some cells from each group were digested and climbing films, line by immunofluorescence staining and confocal fluorescence microscope, semi-quantitative detection. The cells were observed actin protein tubulin and vimentin form and content. 3,6,9 in the first days of culture supernatant every group, with the ELISA method and Alcian blue were detected in the supernatant of each group typeâ…¡collagen and glycosaminoglycans content.Results: Cartilage cell filamentous actin was distributed in the inner plasma membrane along the cell periphery contour arranged actin filaments can be seen after the destruction of cells in the periphery of the solid ring highlighted weakened. Radial microtubules scattered in the nucleus to the cell, microtubule damage were observed in microtubules, protein structure sparse. Vimentin filaments were formed interwoven network structure throughout the cytoplasm, intermediate filaments and round cells were observed in damage, protein fluorescence reduced. Cytoskeletal protein composition of the average of three fluorescence values corresponding analysis shows that the value of the corresponding fluorescence CSK damage group compared with the control group were significantly lower (p <0.05). Meanwhile, the destruction of microtubules and intermediate filaments damage group group at 3,6,9 days in the supernatant of typeâ…¡collagen and GAG were significantly lower than the control group (p <0.05), while the destruction of actin filaments in the control group 3,6,9 days in the supernatant of the first typeâ…¡collagen and GAG content were not statistically different.Conclusion: The concentration of cytoskeleton default agent can destroy cartilage damage corresponding cytoskeleton components, so that the specific fluorescence cytoskeleton component was more reduced, and the destruction of microtubules and intermediate filament group of cartilage cells in monolayer culture supernatants of day 3,6,9 Solution typeâ…¡collagen and GAG content was significantly decreased, while the destruction of actin filaments 3,6,9 days in monolayer culture group on chondrocytes in the supernatant of collagen and GAG typeâ…¡showed no significant effect. |
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