内侧半月板失稳诱发C57BL/6小鼠骨关节炎模型中关节腔微量注射方法的建立

Standard operation procedures of joint cavity micro-aspiration in destabilization of medial meniscus-induced osteoarthritis model of C57BL/6 mice

目的建立C57BL/6小鼠骨关节炎(OA)模型关节腔微量注射的图文操作流程。方法 24只雄性小鼠分假手术组及内侧半月板失稳术(DMM)组,显微操作下离断右膝内侧半月板胫副韧带(MMTL)诱发关节力学失稳,在OA模型基础上通过甲苯胺蓝关节腔注射验证微量给药的可行性,采用OARSI评分系统评估OA小鼠膝关节病理等级。结果 DMM组小鼠关节软骨番红O着色缺失,按关节分区评分,股骨内侧髁评分为3.417 0±0.642 1(n=12),内侧胫骨平台评分为9.583 0±0.811 5(n=12);假手术组对应部位评分依次为0.416 7±0.148 6(n=12),0.583 3±0.148 6(n=12)。DMM组和假手术组OA病理评分相比较,股骨内侧髁(P=0.000 2)及胫骨平台(P<0.000 1)评分具有统计学差异。此外,区别于对照组关节腔注射生理盐水,微量注射甲苯胺蓝成功时可使透明软骨着色。结论内侧半月板失稳术可诱导C57BL/6小鼠产生典型OA病损,OASRI评分提示内侧股骨髁及胫骨平台OA进展严重;可利用甲苯胺蓝验证小鼠OA模型中关节腔给药的重复性和可靠性。

Objective To establish an operation procedure of joint cavity micro-aspiration in destabilization of medial meniscus(DMM)-induced osteoarthritis(OA)model in C57BL/6 mice.Methods We included 24 male mice in this study,and sectioning of medial meniscotibial ligament led to increased local joint mechanical instability.Then DMM-induced OA models were administered with normal saline and toluidine blue in each individual s right knee joint.Besides,histological assessment was made with OARSI score system after the knee joints were collected.Results DMM-induced sections exhibited irregular articular surface and reduced Safranin O fast green staining in weight-bearing area,compared with sham group.According to the site-specific OARSI score system,medial femoral condyle(MFC)was 3.417±0.642 1(n=12)and medial tibial condyle(MTP)was 9.583±0.811 5(n=12)in DMM group.In comparison with sham group(MFC:0.416 7±0.148 6,MTP:0.583 3±0.148 6,respectively),there was a significant difference(P=0.000 2,P<0.000 1,respectively).Additionally,with the imaging system we established previously,the femoral and tibial articular cartilages were found labeled with toluidine blue staining.Conclusion Typical OA progression was presented in DMM-induced OA model through OARSI system assessment.The procedure could improve the positive results when we performed micro-aspiration in OA model of C57BL/6 mice,indicating that micro-injection method in mouse knee joint cavity is repeatable and reliable in vivo.