改良Videman法复制兔膝骨关节炎的实验研究

Experimental Study of Modified Videman Method to Replicate Knee Osteoarthritis in Rabbits

目的:建立便捷、稳定、高效的膝骨关节炎(KOA)兔模型。方法:将60只健康雄性新西兰大白兔按照随机数字表法分为正常组、传统Videman法组(简称传统组)和改良Videman法组(简称改良组),每组20只。传统组采用普通管型石膏固定兔左膝关节伸直位6周进行造模;改良组依据传统组的造模原理,在膝前侧放置石膏托,予高分子石膏单层环形缠绕,固定兔左膝关节于伸直位6周进行造模。记录造模过程中兔石膏脱落、左后肢血运、死亡等一般情况。采用Lequesne MG膝关节级别评分法评估造模后兔左膝关节运动功能,X线和MRI影像学检测,MOAKS评分观察兔左膝关节间隙、关节腔积液、软骨光滑度,软骨及滑膜肉眼观、HE染色光镜观察及Mankins’s评分评价软骨组织形态。并比较传统组与改良组石膏固定、石膏拆卸平均耗时。结果:①造模一般情况比较,正常组无死亡情况;传统组石膏脱落重新固定6只,因会阴部感染死亡4只,断食死亡1只,下肢肿胀坏死3只,最后造模成功12只;改良组石膏脱落重新固定2只,因会阴部感染死亡1只,下肢肿胀坏死1只,最后造模成功18只。改良组造模成功率高于传统组(P<0.05)。②行为学结果比较,与正常组相比,改良组与传统组Lequesne MG评分均升高(P<0.05);改良组与传统组Lequesne MG评分比较,差异无统计学意义(P>0.05)。③影像学结果比较,与正常组相比,X线示改良组与传统组关节间隙均变窄(P<0.05),MRI示关节腔内积液增多、股骨髁软骨面欠光滑,MOAKS评分显著升高(P<0.05);改良组与传统组影像学结果比较,差异无统计学意义(P>0.05)。④形态学方面比较,与正常组相比,软骨及滑膜肉眼观示改良组与传统组软骨面无光泽,关节面变粗糙,部分有缺损;髌前滑膜增厚,色泽发黄,软骨肉眼观评分均升高(P<0.05);软骨HE染色光镜观察示软骨表层不平整,软骨细胞排列欠规整,偶见软骨细胞簇集,Mankin’s评分均升高(P<0.05);改良组与传统组形态学比较,差异无统计学意义(P>0.05)。⑤石膏固定、石膏拆卸平均耗时比较,改良组平均耗时明显短于传统组(P<0.05)。结论:改良Videman造模法建立KOA兔模型,具有操作简便、固定牢固、拆卸便利、造模成功率高等优点,有利于KOA机制的研究。

Objective:To establish a convenient,stable and efficient rabbit model of knee osteoarthritis(KOA).Methods:Sixty male New Zealand rabbits were selected as the normal group,traditional Videman method group(traditional group)and modified Videman method group(improved group)according to the random number table method,with twenty cases in each group.In the traditional group,the left knee joint was straightened for 6 weeks with ordinary tubular plaster.In the improved group,a plaster support was placed on the front of the knee and wrapped a single layer of polymer plaster in a ring.The left knee joint of the rabbit was fixed in a straight position for 6 weeks for modeling,according to the modeling principle of the traditional group.During the process of modeling,the general conditions of the falling off of the plaster,blood supply to the left hind limb and death were recorded.Lequesne MG knee grade score method was used to evaluate the left knee joint motor function of rabbits after modeling.The joint space,effusion,cartilage surface smoothness were evaluated by X-ray and MRI.The macropathology of cartilage and synovium,hematoxylin-eosin staining(HE staining)were observed by light microscope and Mankin's scores were applied to investigate morphological changes.The average time taken to fix and disassemble the plaster was compared with that of the traditional and improved groups.Results:1)Comparison of the general conditions of modeling:there was no death in the normal group.In the traditional group,there were 6 cases which plaster were slipped and re-fixed,4 cases died due to perineal infection and 1 case died from dstarving themselves,3 cases that had ischemic necrosis of lower limbs,12 cases were successfully established finally.In the improved group,the plaster was slipped and refixed in 2 cases,1 case died of perineal infection and 1 case died of lower limb swelling and necrosis,and 18 cases were successfully modeled.The success rate of the improved group was higher than that of the traditional group(P<0.05).2)Comparison of behavioral outcomes:in the comparison with the normal group,Lequesne MG knee joint evaluation method in the traditional group and the improved group were risen(P<0.05).There was no significant difference in Lequesne MG scores between the traditional group and the improved group(P>0.05).3)Comparison of imaging results:compared with the normal group,X-rays showed a narrowing of joint space between the modified group and the traditional group(P<0.05).MRI showed increased fluid accumulation in the articular cavity,less smooth femoral condyle cartilage,and MOAKS score increased significantly(P<0.05).There was no significant difference in imaging results between the traditional group and the improved group(P>0.05).4)Comparison of morphology:compared with the normal group,the cartilage and synovium of the improved group and the traditional group were observed to have dull cartilage surfaces,rough articular surfaces and some defects.The anterior patellar synovium was thickened and yellow,and the macropathology score of the cartilage was increased(P<0.05).HE staining of cartilage was observed by light microscopy to show that the surface of the cartilage was uneven and the chondrocytes were not arranged in an orderly manner.Chondrocyte clusters were occasionally observed,and the Mankin's scores were increased(P<0.05).There was no significant difference in morphology between the traditional group and the improved group(P>0.05).5)Comparison of the average time taken to fix and disassemble the plaster:the average time of the improved group was significantly shorter than that of the traditional group(P<0.05).Conclusion:The KOA rabbit model established by modified Videman method has the advantages of simple operation,firm fixation,convenient disassembly and high success rate of modeling,which is conducive to the study of KOA mechanism.