Calcitonin On The Rat Knee OA Model Of Cartilage Degeneration And Subchondral Bone Resorption

Objective:Modified Hulth mode method the formation of the rat knee osteoarthritismodel to evaluate the effectiveness of the model; effective model to analyze the changes incharacteristics of the articular cartilage and synovial matrix metalloproteinase; researchcalcitonin (CT) on the knee cartilage, synovium and subchondral bone changes, theprotective effect of CT on the articular cartilage and subchondral bone resorptioninhibition.Methods:105SD rats were randomly divided into control group (ACLT+MMX),calcitonin (CT) in the intervention group and sham operation group (Sham),35in eachgroup.(1) Modified Hulth mode method of osteoarthritis (OA) model, the anterior cruciateligament transection and medial meniscectomy (ACLT+MMX). The Sham group onlyopen joint cavity, without cutting off the posterior cruciate ligament nor meniscectomy.CT group: calcitonin15IU/kg the next day, after modeling, the next day before asubcutaneous injection, the control group and sham group injected the same amount ofsaline. Modeling,1w,2w,4w,6w,8w,12w rats were killed. We observed the jointspecimens, took the medial femoral condyles, the medial tibial platform subchondral boneand joint synovial tissue HE staining, toluidine blue staining, Masson staining. Mankinscore increase Masson staining resulted score on the course of OA stages.(2) Took the cartilage and synovial specimens, the cartilage from the medial femoralcondyle. Specimens of each time point after treatment, stored in liquid nitrogen, theenzyme-linked immunosorbent assay (ELISA) in Transcription of Sox9, type II collagen, MMP-1,-3,-13protein content, real time quantitative PCR (Realtime PCR), detectionSox9, type II collagen, MMP-1,-3,-13changed in gene expression analysis of early OAand changed characteristics of the middle and late three kinds of matrix metalloproteinases.Compared the differences of the three large mouse cytokines, analysis of the protectiveeffect of calcitonin intervention for osteoarthritis cartilage.(3) Took the medial tibial plateau specimens, slashed the surface cartilage andsurrounding soft tissue, RNA was extracted after grinding, real-time quantitative PCR(Realtime PCR), detection of RANK and cathepsin K (CK) expression changes, analysis ofcalcitonin hormone on bone resorption in the cartilage of early OA stage. Read by HEstaining of the subchondral bone, trabecular bone volume, number, separation, compareOA2w and12w modeling of trabecular bone differences.Result(1) After the model2w articular cartilage lose their luster, dark color, mild hyperplasiaof the number of HE staining chondrocytes; the4w cartilage destruction, increase in cellnumber, Masson staining red dye, the emergence of the little blue; the6w cartilage ulcerdestruction significantly, osteophytosis, cell derangement, Masson staining red and blue;8w cartilage defects, osteophytosis, cartilage necrosis, most of toluidine blue level issignificantly lighter to reduce Masson stain blue;12w cartilage, exposed bone, the cellssignificantly reduced the number loss of dye, Masson staining showed a large red dyetoluidine blue. Sham group is generally close to normal and the tissue sections.(2) ELISA and Realtime PCRControl group and sham group: cartilage MMP-1: the control group and sham groupcompared to a significant increase (P <0.01); MMP-3: control group and sham groupcompared to a significant increase (P <0.01), disease progression in up trend (P <0.05);compared to MMP-13: the control group and sham group, was significantly upregulated (P<0.01), disease progression in up trend (P <0.01). Synovium of MMP-1,-3,-13expressionis higher than the sham group, the upward trend (P <0.01).CT group compared with control group: CT group and control group, Transcription of Sox9expression increase (P <0.05); type II collagen in OA modeling4w, downregulated,upregulated6w from, changes significant difference (P <0.01); of MMP-1,-13expressionwas significantly lowered (P <<0.01); of MMP-3in OA early (2w) is to increaseperformance (<0.01), in the late stage is lowered (P <0.01); of CK downregulated (P<0.01); RANK expression in OA early reduced (P <0.01).(3) bone histomorphometry parameters changeControl group and sham group:2w, trabecular bone volume and trabecular numberwas significantly less than the sham group, the trabecular separation is greater than thesham group (P <0.01), subchondral bone volume and reduce the number of spacingincreases.12w when trabecular bone volume, trabecular number were higher than the shamgroup, the trabecular separation is less than the sham group (P <0.01).CT group compared with control group:2w, trabecular bone volume and trabecularnumber were higher than control group, trabecular separation is less than control group (P<0.01);12w when there is no significant difference (P>0.05). Sham and Sham+CTgroup there was no difference (P>0.05).Conclusion1.A modified Hulth rat OA modeling method, the transection of the anterior cruciateligament and medial meniscus, is a more practical and effective knee OA modelingmethod.The mean Mankin 's score can basically reflect the OA as early, middle and lateinstallments, Masson staining can be used as the complement of the Mankin score.2.Cause sometimes the phase of the cartilage extracellular matrix degradation MMPssecretion. Chondrocyte MMP-1,-13early in OA is high expression of MMP-3expressionis low expression of the early, and then gradually increased.3.Synovium of OA during the early stage of the degradation of cartilage, not just thesecondary synovitis. Early cartilage damage, synovial play an important role.4.CT promote articular cartilage formation and inhibit the degradation of cartilagematrix.5.OA early cartilage, bone resorption activity. Trabecular bone volume and reduce the number of intervals widened, osteoclast markers change. Calcitonin inhibits boneresorption early in OA, the treatment of OA in the early application of calcitonin.