| Osteoarthritis(OA) is a chronic progressive disease of bone and joint which usually occurs in middle-aged. It is a cartilage degenerative changes to the core, involving bone, including synovium, joint capsule and other structures outside the joint capsule of the different levels of inflammatory lesions.With the aging of our population and the trend of younger onset of OA, OA has become hazardous to human health and quality of life of major diseases.The research of occurrence and development mechanism of the OA has become the focus of orthopedic research in the field.The Studies have shown that OA is usually accompanied by an imbalance of intra-articular microenvironment,and synovial cells is the main factor.Object ives:Establishment the co-culture system of synovial cells and cartilage cell and build a class of OA microenvironment,to study the role of synovial cells in the process of OA and provide a basis for the pathogenesis of OA.Methods:1) We use the modified Hulth method to copy knee osteoarthritis model of rabbit.We removal of the medial meniscus, cut off the anterior cruciate ligament and medial collateral ligament.The joint are swelling after 4 weeks of surgery and get a lot of synovial tissue.2) To obtain synovial tissue after 4 weeks,mechanically isolat synovial tissues and enzyme-digested by Trypsin-Ⅱtype collagenase.To observe the morphology and activity of Synovial cells,and identifie by CD68 immunohistochemistry.3) To Isolate and culture rabbit's chondrocytes and observe the growth of cartilage cells.4) The original and passage synovial cells were in different proportions (1:1,1:1/2, 1:1/4,1:1/8), respectively, to co-culthur with cartilage cells (cell number of 4x104). The control group is normal cartilage cells. To observe morphology and biochemistry activity of chondrocytes which are effected by synovial cells of primary and passage.5) To observe the gene expression of cartilages'Ⅱcollagen and proteoglycan in RNA level after co-cultured with synovial cells of primary and passage.Results:1) The synovial tissues are hyperplasia obviously by the methode of Modified Hulth:the lining cell layer of synovial were increased,interstitial fibrosis and vascular were proliferation.2) The methed by Trypsin-Ⅱtype collagenase can obtain synovial cells which are great activity above 98%.Macrophage-like synovial cells are positive by CD68 immunohistochemical staining.3) Trypsin-I type collagenase-Ⅱtype collagenase enzyme-digestion could be successfully isolated and obtained a large number of highly active chondrocytes.4) To establish the co-culture system of synovial cells and cartilage cells.The quantity and culturing time of synovial cells have a closely related to the morphology and proliferation of cartilage cells.The more the number of synovial cells, the role of the longer, the proliferation of cartilage cells lower.5) In the co-culture system of synovial cells and cartilage cells,the quantity and passages of synovial cells have a closely related to the gene expression of cartilages'II collagen and proteoglycan.The gene expression of cartilages of co-cultured has decrease significantly.The role of primary synovial cells for cartilages is stronger than passage synovial cells. The higher the number of synovial cells, the lower gene expression of cartilage cells.Conclusions:1) Hulth modeling method can be improved quickly formed osteoarthritis and solve the difficult problem of synovial drawn.In the method,synovial tissue hyperplasia in a short time, lining cell layers overlying increased and interstitial fibrosis. The method is suitable for extraction of synovial cells and study the pathogenesis of OA.2) Separable enzymatic digestion can obtain synovial cells and cartilage cells.This methods is proved to be simple and feasible and can obtain synovial cells which are great activity in a short time.3) The co-culture system of synovial cells and cartilage cells is for the study of micrenvironment of OA.The co-culture system can simulate the process of articular cartilage injury of OA.4) The change of synovial and synovial cells has a closely relate to the vivo environment of articular.So,the change in the environment can be an important factor in the pathogenesis of OA... |
PDF Full Text Request