Study Of Cytoskeleton In Chondrocytes From A Rabbit Osteoarthritis(OA)-induced Model

Objective:To observe the morphology of cytoskeleton(CSK) of control and osteoarthritis(OA) chondrocytes and estimated the fluorescence intensity of CSK proteins.And explore the effect of cytoskeleton in pathogenesy of OA.Materials and methods:Eight New Zealand white rabbits were divided into two groups randomly,each group 4.The animal model of OA was induced by anterior cruciate ligament transaction(ACLT) in animals of experiment group,and the same process of operation was performed on ones of control group,but no ACLT.After 20 weeks,rabbits were sacrificed and evaluated at two levels:macroscopic and histologic (HE) in all left knees.The chondrocytes of their right knee joints were taken out under aseptic conditions and cultured in the plates.The chondrocytes of control and experiment were inoculated into 24 well plates which round cover slips were put at the bottoms.Cell climbing slices were fixed when 80%confluence was reached in the inverted microscope.The CSK proteins:actin,vimentin,tubulin and vinculin were stained by immunofluorescence antibody on control and cell climbing slices respectively.The morphology of CSKs were Observed by LCSM and the fluorescence intensity of CSK proteins were detected quantitatively by the fluorescence intensity software.Fluorescein staining and laser scanning confocal microscope(LSCM) was used to observe the change of morphology of chondrocytes isolated from cartilage of right knees in two groups.Results:The knee joints of experiment group had obvious degeneration of articular cartilage compared with control ones.The score of macroscopic of experiment group is significantly higher than control one(P<0.05).Intermediate filament,microtubule and vinculin in experiment group is aggregation and disrupt, and is significantly lower than control one by fluorescent quantitation assay (P<0.001).Conclusion:The morphology and protein quantity of OA chondrocytes CSK changed accompanied with control ones,which suggested that the change of CSK maight correlate with the pathogenesy of OA.