一种含BMP3软骨钉制备及其对兔股骨软骨缺损修复的研究  

Preparation of a BMP3-containing cartilage nail and its repair of femoral cartilage defects in rabbits

作　　者：刘伟伟[1] 丁力 刘大卫[1] 崔晓雪[1] 史梦柔 陈斌[1] 李昕[1] 唐瑞辰 王长江 LIU Wei-wei;DING Li;LIU Da-wei;CUI Xiao-xue;SHI Meng-rou;CHEN Bin;LI Xin;TANG Rui-chen;WANG Chang-jiang(Tianjin Institute of Medical and Pharmaceutical Sciences,Tianjin 300020,China;Tianjin Zhongjin Biological Development Co.,LTD.,Tianjin 300380,China)

机构地区：[1]天津市医药科学研究所,天津300020 [2]天津中津生物发展有限公司,天津300380

出　　处：《生物医学工程与临床》2025年第2期142-149,共8页Biomedical Engineering and Clinical Medicine

基　　金：天津市科技计划项目——科技重大专项与工程(21ZXSYSY00020)。

摘　　要：目的制备一种以牛心包和牛松质骨为原料且含BMP3的软骨钉,并观察其对兔软骨缺损修复的影响。方法牛心包和牛松质骨经处理后复合加入提纯的BMP3牛皮质骨粗提取物制成软骨钉。选择雄性日本大耳白兔22只,兔龄120~150 d;体质量2.5~3.0 kg。在其股骨关节面内侧髁钻直径4 mm、深度8 mm孔,左侧植入软骨钉(样品组,22个植入点),右侧仅钻孔操作(模型组,12个植入点)或不处理(空白对照组,10个植入点)。植入后8周、12周分别处死半数动物,用苏木精-伊红染色,观察植入部位的局部组织反应和新生软骨情况,番红O-固绿染色观察软骨形成,免疫组织化学染色观察Runt相关转录因子2(RUNX2)和Ⅱ型胶原蛋白(COLⅡ)表达。结果①苏木精-伊红染色:植入后8周模型组关节局部缺损明显,未见明显软骨组织形成;样品组植入材料周围可见少量软骨组织形成,软骨细胞排列较为紊乱。植入后12周模型组关节缺损部位表面少量软骨组织形成,软骨细胞排列不规则;样品组软骨组织形成较8周增多,软骨细胞排列较规则。②番红O-固绿染色:8周模型组未见红染的软骨组织;样品组少量散在红染的软骨组织。12周模型组关节面少量薄层红染的软骨组织,样品组关节可见较大量深红色软骨组织。③免疫组织化学染色:空白对照组各时间点(灰度值:8周83.15±6.28,12周86.76±8.20)和COLⅡ(灰度值:8周78.15±4.79,12周82.10±4.31)高表达,模型组表达明显降低(RUNX2灰度值:8周125.48±5.91,12周117.53±2.59;COLⅡ灰度值:8周129.05±2.18,12周122.03±7.27。P<0.01),样品组表达量较模型组增加(RUNX2灰度值:8周100.62±7.04,12周90.48±8.15;COLⅡ灰度值:8周98.89±5.55,12周86.19±8.28。P<0.01);随着时间点延长,模型组和样品组12周表达较8周有所增加,且样品组增加明显(P<0.01)。结论以牛心包和牛松质骨为原料,添加含BMP3牛皮质骨粗提取物制备的软骨钉可以促进兔股骨�Objective To prepare a cartilage nail containing BMP3 with bovine pericardium and bovine cancellous bone as raw materials,and observe its effect on the repair of rabbit femoral cartilage defect.Methods After treatment of bovine pericardium and bovine cancellous bone,the purified BMP3 bovine cortical bone crude extract was added to make cartilage nails.A total of 21 male Japanese white rabbits were sacrificed,which aged 120-150 days with body mass of 2.5-3.0 kg.The medial condyle of femoral articular surface was drilled a hole with diameter of 4 mm and depth of 8 mm.The cartilage nails were implanted on the left side(sample group,22 implantation sites),the right side was treated with only drilling(model group,12 implantation sites)or non-treatment(blank control group,10 implantation sites).Half of rabbits were sacrificed respectively at 8 and 12 weeks after implantation.Hematoxylin-eosin staining was used to observe local tissue reaction and novel cartilage at implantation site.The safranin O-fast green staining was used to observe cartilage formation,and immunohistochemical staining was used to observe the expression of Runt-related transcription factor 2(RUNX2)and typeⅡcollagen(COLⅡ).Results①The hematoxylin-eosin staining showed that after implantation for 8 weeks,the local defects of the joints were obvious,and no obvious cartilage tissue was formed in model group,a small amount of cartilage tissue was formed around implantation material,and the arrangement of chondrocytes was disordered in sample group.After implantation for 12 weeks,a small amount of cartilage tissue was formed on the surface of joint defect,and the chondrocytes were irregularly arranged in model group,while formation of cartilage tissue was more than 8 weeks,and the arrangement of chondrocytes was more regular in sample group.②The saffranine O-fast green staining showed after implantation for 8 weeks,no red-stained cartilage tissue was observed in model group,a small amount of red-stained cartilage tissue was scattered in sample grou

关 键 词：骨软骨缺损 股骨 牛心包 牛松质骨 BMP3 RUNX2 Ⅱ型胶原蛋白 兔 

分 类 号：R687[医药卫生—骨科学] R-332[医药卫生—外科学] R318.08[医药卫生—临床医学]