Gene Expression Of Extracellular Matrix Factors In Scratch-wound Model Of Articular Chondrocyte In Rabbit

Objective: To collect and culture cells from articular chondrocyte of rabbit and establish the injury model. To explore the expressions of extracellular matrix type II collagen(Col-II), matrix metalloproteinase13(MMP13), tissue inhibitor of metaloproteinases 1(TIMP 1) and hyaluronic acid(HA) genes factor genes in articular chondrocyte of rabbit at different time-points after injury. Methods: The healthy male or female 4 weeks old New Zealand White rabbits were chose. The knee joint cartilage chondrocytes were harvested in a sterile environment after anesthesia the rabbit. The cell were cultured and continued to train the next generation. Then good condition cells were cultured in six orifice. The scratch-wound model of chondrocyte was established by mechanical injury of normal and post-injury 1, 3, 5 and 7 days. The morphological characteristics of chondrocytes were observed and take pictures. The mRNA expressions of Col-II, MMP13, TIMP 1 and HA genes in articular chondrocytes were investigated by real-time quantitative polymerase chain reaction(qPCR) in normal and post-injury 1, 3, 5 and 7 day. Results: The injury model of rabbit articular chondrocyte was successfully established. Compared with the normal cell, the mRNA expression level of Col-II was lower than normal cells(P<0.05), the 3 and 7 day was higher than 1 day, 7 day was almost to be normal. The level of MMP13 mRNA decreased after 1 day of injury, 1 day about 1/47 of the normal(P<0.05), 3 day and 5 day was rised than 1 day, 5 days was 5 times to 3 day(P < 0.05), more than 37 times of the 7 day(P < 0.05). The TIMP1 mRNA expression increased during 1 day after injury, 3 day was lower than 1 and 5 day,1 day was higher than 3 day 2 times, 5 day was 2 times to 3 day(P<0.05), the mRNA expressions levels of TIMP1 raised after 3 day, was normal in 7 day. The 1 and 5 day expressions levels of HA mRNA was lower to normal, 5 day was the highest, 3 times higher than normal, 13 times of 1 day, 6 times of 3 day and 10 timesof 7 day(P<0.05). Conclusions: The expression patterns of MMP13, TIMP1, HA and type II collagen m RNA are different after articular chondrocyte injury, studying the expressions of extracellular matrix in injury model cartilage cells, which can provide an appropriate window period in gene therapy of cartilage damage in the future.