Comparison Study On Two Experimental Diannan Mini Pig Models Of Osteoarthritis Made By Intra-articular Papain Injection And Hulth Procedure Surgery

[Objective]To compare two different experimental models with osteoarthritis in Diannan mini pig made by papain injection and Hulth procedure surgery and to provide the new OA model for study.[Methods]OA was induced in 10-15 kg Diannan mini pigs by surgical and chemical injection.27 pigs were divided into three groups.The surgical group(n=9),papain injection group(n=9),and blank control group(n=9).The surgical group and made by the medial meniscectomy(MSX),anterior cruciate ligament transection(ACLT),medial collateral ligament transection(MCLT),and posterior cruciate ligament transection(PCLT).Papain injected with on first,fourth,and seventh days.All pigs were sacrificed at 4,8,12 weeks post-surgery and post-injection,and the synovial fluid,membrane,and cartilage were collected for following test:Histologic examination with ?(H&E staining,Safranin-O staining,and Immunofluorescence);? ELISA was used to measure the level of SDF-1 in synovial mambrane;? Q-PCR was used to test the collagen type ?,MMP-3,aggrecan in cartilage;? Western blot was used to test the levels of collagen type ?,MMP-3,aggrecan in cartilage.SPSS 23 was used to analyses the data.The data will be expressed as mean �standard(X � S),P<0.05 was used as significant difference standard.[Results]1)Histologic examination(H&E staining,Safranin-O staining,and Immunofluorescence):O? The result of H&E staining,Safranin-O staining of surgery at 4 weeks showed articular cartilage surface damage;at 8 weeks showed articular cartilage damage more and tide line damage;at 12 weeks showed loss of articular cartilage,uneven of nucleus,bone trabecular damage and tide line discontinue.Chemistry(papain injection)at 4 weeks showed articular cartilage rough;at 8 weeks showed articular damage to subchondral bone and bone trabecular damage;at 12 weeks showed more severe damage of articular cartilage to subchondral bone.There were statistically significant(P<0.05).? Immunofluorescence test:The result of immunofluorescence IOD/area,at the same time among surgery group,chemical group,and control group showed that the levels of Coll ? a trend decrease tendency.In chemical group the levels of Coll ? showed more decrease tendency when compared with the control group and surgery group.There was statistically significant(P<0.05).2).ELISA test:SDF-1 in the membrane at the same time among surgery group,chemical group,and control group.Both of the surgery and chemical group showed increase tendency when compared with the control group.There was statistically significant(P<0.05).3).Q-PCR test:The result of Q-PCR test,at the same time compared with surgery group,chemical group,and control group,The levels of collagen ? and aggrecan showed surgery group decreases more than the control group,and chemistry group decreases more than surgery group.The levels of MMP-3 showed surgery group increases more than the control group and chemistry group increase more than surgery group.There were statistically significant(P<0.05).4).Western Blot test:The result of MMP-3,Collagen ?,and Aggrecan were tested by Western Blot;at the same time,the levels of MMP-3 showed increase tendency;at 4,8,12 weeks among surgery and chemistry collagen ?,and aggrecan showed decrease tendency.There were statistically significant(P<0.05).[Conclusion]1.Diannan mini pig could be induced OA with two methods.2.Papain injection was a better choice for the reason that easier,and less impact to soft tissue when compared-with Hulth procedure surgery.3.It was more severe in papain injection than surgical procedure.The time for OA in chemical group was shorter than surgery group.Papain injection was more effective than surgery for OA models of the Diannan mini pig.4.Papain articular damage existed in all compartment of the knee joint.Surgery articular damage existed in the medial compartment of the knee.5.SDF-1 was a key biomarker of OA in Diannan mini pig,Therefore,the SDF-1/CXCR4 signaling pathway may play a key role in the pathological process of OA in Yunnan province.It will lay a foundation for the development of high efficiency targeted therapy drugs.