The Expressions Of Molecules In Joint Cartilage And Their Relevance To Osteoarthristis

Part one : The Gene Expressions in Joint Cartilage and Their Relevance to OsteoarthristisObjective:To investigate molecules involved in the development of OA through study of the gene expression in OA and femoral neck fracture, and provide a new treatment strategy for OA.Materials and methods: Deep arthritic joint cartilage tissue was obtained from forty-six patients undergoing hemiarthroplasty and total hip or knee arthroplasty because of osteoarthritis femerol neck fracture. 1.2g of deep cartilage sample was obtained as surgical waste at the time of hip or knee replacement surgery for clinical OA and for femoral neck fracture without clinical OA. The samples were immediatelly stored in RNAlater (Ambion) for RNA extraction(stored less than one week) . Total RNA was extracted from frozen biopsies (1.2g/sample) by using Rneasy Fibrous Tissue Midi Kit (Qiagen,guangDong,China) according to the manufacturer's recommendations.Total RNA was revers-transcripted to cDNA according to manufacturer's recommendations at a final volume of 10μl. The cDNA was stored at ?20°C until the samples were processed. Quantitative real-time polymerase chain reaction (RT-PCR) was used to determine the expression of IL-1β,NFAT1,NFAT1B,TNFα,COL2A1,COL10A1 and Aggrecan.The comparation was made between the femoral neck fracture and OA patients,and further between the knee OA and hip OA patients. The relation between gene expression and the knee OA severity was also studied.Result:The expressions of IL-1β,NFAT1,NFAT1B,TNFα,COL2A1,COL10A1 were significantly higher in the OA group, respetively. There was no significant difference in Aggrecan expression between the two groups(p>0.05).The expression of IL-1β,NFAT1,NFAT1B,TNFα,COL2A1,COL10A1 and Aggrecan had no significant difference between the knee OA and hip OA patients(p>0.05). The gene expressions was not relevant to the OA severity.Conclusion:The increased expressions of gene IL-1β,NFAT1,NFAT1B,TNFα,COL2A1, COL10A1 in joint cartilage contribute to the development of OA。Part two: The Expressions of Col2A1 and TNFαin Joint Cartilage and Their Relevance to OsteoarthristisObjective: To investigate the difference of Col2A1 and TNFαexpression invoved in the development of OA through comparation of the ones in hip OA and femoral neck fracture,and provide a theory foundation for treatment of OA.Materials and methods:Deep Arthritic joint cartilage tissue was obtained from eighteen patients undergoing hemiarthroplasty or total hip arthroplasty because of osteoarthritis or femerol neck fracture.The deep cartilage was stored at -80℃for western-blot. Antibodies specific to Col2A1 and TNFαwere used . Protein expression of the Col2a1 and TNFαof specimens were semi-quantitated by western-blot, and the quantitation of each protein was compared between OA and femerol neck group.Result:Protein expressions of the Col2A1 and TNFαof OA group are higher than fracture group ,which have significant difference.Conclusion:The increased expression of protein TNFαand Col2a1contribute to the development of OA。...