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Objective: This study was designed to research the contributions of synovial cells whose Wnt/β-catenin signal pathway are activated to the cartilage cells, to discuss the ways and molecular mechanism of Osteoarthritis (OA). Methods: β-catenin over expressing lentivirus vector was constructed to transfect synovial cells of Human and fluorescence microscope was used to observe viral transfection efficiency. Expression of β-catenin protein of the synovial cells was detected by Western blot. Synovial cells and chondrocytes were cultured in transwell chamber and the influences of synovial cells to chondrocytes were observed under optical inverted microscope. Changes of MMP-7 in culture supernatant of chondrocytes was tested by Enzyme linked immunosorbent assay (ELISA) method at different time points to find the action time that synovial cells influence the chondrocytes best. Moreover, ELISA method was used to test CTX-II, COMP levels in the culture supernatant of chondrocytes at optimal time point. Results: After been transfected by lentivirus vector, fluorescent microscopy showed that most of the synovial cells were arise fluorescence. Expression of β-catenin increased significantly in the cytoplasm and nucleus of synovial cells (P<0.01). The growth morphology of chondrocytes did not change significantly under light microscope after co-cultured with synovial cells which Wnt/β-catenin signal pathway are activated. But chondrocytes' growth and adhesion force was reduced with the prolongation of the culture time, this may be related to the degradation of extracellular matrix. ELISA results showed that MMP-7 expression of chondrocytes supernatant was significantly increased, there was a large difference compared with the normal group (P<0.01). MMP-7 expression of chondrocytes culture supernatant were time-dependent. It increased gradually with the time prolonging at 2 h, 6 h and 24 h, but decreased slightly at 24 h. The expression of MMP-7 changed significantly after 6 hours'co-culture. ELISA results showed that the level of COMP, CTX-II in the supernatant of chondrocytes within singal pathway activated group was significantly higher than that in normal control group at 6 hours'co-culture (P<0. 01). Conclusions: β-catenin over expressing lentivirus can activate Wnt/β-catenin signaling pathway. The grouth of chondrocytes can be affected by synovial cells whose cause the abnormal of internal environment within synovial- cartilage system,promote cartilage matrix degradation at same time. This may be exist molecular interactions between two cells as a possible mechanism of synovitis promoting OA. [ABSTRACT FROM AUTHOR]

Objective: This study was designed to research the contributions of synovial cells whose Wnt/β-catenin signal pathway are activated to the cartilage cells, to discuss the ways and molecular mechanism of Osteoarthritis (OA). Methods: β-catenin over expressing lentivirus vector was constructed to transfect synovial cells of Human and fluorescence microscope was used to observe viral transfection efficiency. Expression of β-catenin protein of the synovial cells was detected by Western blot. Synovial cells and chondrocytes were cultured in transwell chamber and the influences of synovial cells to chondrocytes were observed under optical inverted microscope. Changes of MMP-7 in culture supernatant of chondrocytes was tested by Enzyme linked immunosorbent assay (ELISA) method at different time points to find the action time that synovial cells influence the chondrocytes best. Moreover, ELISA method was used to test CTX-Ⅱ, COMP levels in the culture supernatant of chondrocytes at optimal time point. Results: After been transfected by lentivirus vector, fluorescent microscopy showed that most of the synovial cells were arise fluorescence. Expression of β-catenin increased significantly in the cytoplasm and nucleus of synovial cells (P<0.01). The growth morphology of chondrocytes did not change significantly under light microscope after co-cultured with synovial cells which Wnt/β-catenin signal pathway are activated. But chondrocytes' growth and adhesion force was reduced with the prolongation of the culture time, this may be related to the degradation of extracellular matrix. ELISA results showed that MMP-7 expression of chondrocytes supernatant was significantly increased, there was a large difference compared with the normal group (P<0.01). MMP-7 expression of chondrocytes culture supernatant were time-dependent. It increased gradually with the time prolonging at 2 h, 6 h and 24 h, but decreased slightly at 24 h. The expression of MMP-7 changed significantly after 6 hours' co-culture. ELISA results showed that the level of COMP, CTX-Ⅱ in the supernatant of chondrocytes within singal pathway activated group was significantly higher than that in normal control group at 6 hours' co-culture(P<0.01). Conclusions: β-catenin over expressing lentivirus can activate Wnt/β-catenin signaling pathway. The growth of chondrocytes can be affected by synovial cells whose cause the abnormal of internal environment within synovial-cartilage system,promote cartilage matrix degradation at same time.This may be exist molecular interactions between two cells as a possible mechanism of synovitis promoting OA. [ABSTRACT FROM AUTHOR]