大鼠软骨非全层损伤模型的建立及活化细胞与整合素β_1表达关系的研究  

MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β_1 IN A RAT MODEL

作　　者：张凯彬[1] 石晶[1] 李杨[1] 蒋逸秋[1] 丁朋[1] 杨晓霏[1] 桂鉴超[1] 

机构地区：[1]南京医科大学附属南京医院骨科,南京210000

出　　处：《中国修复重建外科杂志》2016年第4期432-439,共8页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的探讨大鼠软骨非全层损伤模型的制备及术后细胞变化及细胞活化与整合素β_1表达的关系。方法 10周龄SD雄性大鼠45只,体质量300~400 g,随机分为手术组、假手术组和对照组,每组15只。手术组采用划痕法制造软骨非全层损伤模型;假手术组打开膝关节囊后直接缝合;对照组不行手术。术后1、7、14 d各组分别处死5只大鼠,取材行大体观察;并行HE、番红O组织学染色,评估并比较各组HE染色改良组织学评分及番红O染色灰度值;行Brd U、CD105免疫荧光染色和CD105/整合素β_1双标记染色,采用标准双盲法计数并比较各组各时间点CD105阳性细胞数。结果大体观察示手术组划痕周围软骨欠光滑,非透明,有软骨软化、纤维化改变,且随造模时间延长逐渐加重;假手术组和对照组软骨呈白色透明状,无软化、纤维化改变。HE染色示手术组在划痕周围细胞数增多,大小不等,排列分布不均;假手术组和对照组细胞大小均一,染色均匀,排列有序。手术组术后各时间点HE染色改良组织学评分均显著高于假手术组及对照组（P〈0.05）,各组组内各时间点间比较差异均无统计学意义（P〉0.05）。番红O染色示手术组各时间点染色欠均匀,划痕周围区域着色较浅;假手术组和对照组各时间点染色均匀,无失染。术后各时间点各组间比较以及各组内各时间点间比较番红O染色灰度值,差异均无统计学意义（P〉0.05）。Brd U免疫荧光染色示手术组划痕周围细胞排列紊乱,分布不均;假手术组和对照组细胞排列分布均匀,无聚集现象。CD105免疫荧光染色示手术组划痕周围有较多阳性细胞聚集,细胞大小不一、分布不均;假手术组和对照组软骨层见少数阳性细胞,分布均匀。手术组术后各时间点CD105阳性细胞数均显著多于假手术组和对照组（P〈0.05）;手术组内随时间延长CD105阳性细胞数逐渐增多,各时间点间比较差�Objective To investigate the relationships between the expression of integrin β_1 and activated cells in a partial-thickness articular cartilage injury model of adult rats. Methods Forty-five male Sprague Dawley rats（aged 10 weeks and weighing 300-400 g） were randomly divided into operated group（n=15）, sham-operated group（n=15）, and control group（n=15）. Partial-thickness articular cartilage injury model was made by scarification in operated group, direct suture after opening of the knee joint was performed in sham-operated group, and no operation was done in control group. Five rats were sacrificed at 1, 7, and 14 days after operation respectively for macroscopic evaluation, HE staining, Safranin O staining, CD105, Brd U, CD105/integrin β_1 immunofluorescence and double labeling staining. The histological score of HE staining, gray value of Safranin O staining and CD105-positive cells count were compared among groups at each time point. Results Macroscopic evaluation showed chondromalacia and cartilage fibrosis around the linear injury with aggravating tendency with time in operated group, but no chondromalacia and cartilage fibrosis in sham-operated and control groups. HE staining demonstrated a number of activated cells accumulating around the linear injury with nonuniform distribution in operated group, and uniform size and distribution in sham-operated and control groups. The histological scores at each time point in operated group were significantly higher than those in sham-operated group and control group（P〈0.05）, but no significant difference was found between different time points in 3 groups（P〉0.05）. Safranin O staining was nonuniform with hypochromasia around linear injury in operated group, but the staining was uniform in sham-operated group and control group. Gray value of Safranin O staining had no significant difference among groups and among different time points in the same group（P〉0.05）. Brd U-positive and CD105-positive cells distributed unevenly around the l

关 键 词：软骨非全层损伤 软骨前体细胞 CD105 BRDU 整合素Β1 大鼠 

分 类 号：R687[医药卫生—骨科学] R-332[医药卫生—外科学]