壳聚糖温敏水凝胶复合rhBMP-14促兔急性肩袖损伤愈合的实验研究  

The experimental study of the promoting function of chitosan thermosensitive hydrogel-rhBMP-14 for rabbit acute rotator cuff tear

作　　者：邢春苗[1] 陈情忠[1] 龚炎培[1] XIN Chunmiao CHEN Qingzhong GONG Yanpei(Department of Hand Surgery, the Affiliated Hospital of Nantong University, Nantong 226001)

机构地区：[1]南通大学附属医院手外科,南通226001

出　　处：《南通大学学报（医学版）》2016年第5期425-429,共5页Journal of Nantong University(Medical sciences)

基　　金：南通市科技计划项目(HS2013035)

摘　　要：目的:研究重组人骨形态发生蛋白-14(recombinant human bone morphogenetic protein-14,rh BMP-14)在兔急性肩袖损伤后肌腱止点重建过程中的作用。方法:共取45只8月龄雄性新西兰大白兔,随机选取3只作为正常组,进行生物力学测试。其余42只随机分成3组,每组14只。采取双侧冈上肌腱完全离断模型作为肩袖急性损伤模型,并一期行肌腱止点重建术。实验组在腱-骨界面注射以壳聚糖温敏水凝胶(chitosan/glycerophosphate,CS/GP)为载体的rh BMP-14;实验对照组仅填充CS/GP;空白对照组不给予任何处理。分别于处理后第4周和第8周随机处死实验用兔7只,切取肩袖标本。随机选取各组4例肩袖标本进行HE染色观察腱-骨愈合情况,取各组4例标本进行苦味酸天狼星红染色进行Ⅰ型胶原和Ⅲ型胶原面积测定并进行比较。另取各组6例标本进行生物力学测试。结果:(1)组织学观察:HE染色显示术后4周实验组有大量软骨细胞增生,并伴有胶原纤维和软骨组织形成,胶原方向较为恒定;在实验对照组及空白对照组仅有少量胶原纤维及软骨形成,方向性差。实验组腱-骨界面组织于术后8周连接紧密,胶原纤维方向一致,并伴有矿化胶原纤维形成,腱-骨界面出现明显的Sharpey’s纤维连接,有类似直接止点形成;实验对照组和空白对照组有部分似直接止点的矿化胶原纤维形成,大部分由结缔组织充填。苦味酸天狼星红染色显示随着时间的增加,实验对照组及空白对照组Ⅰ型胶原面积和Ⅲ型胶原面积之比均有所提高,而实验组的面积比例在选取的相同时间段较对照组明显提高(P<0.01)。(2)生物力学测试显示:术后3组最大载荷及刚度都随时间的延长而增强。实验组在第4周、第8周的最大载荷与刚度都较实验对照组与空白对照组明显增高(P<0.01)。结论:rh BMP-14可促进肌腱止点的成熟,并可诱导直接止点形成,明显提高腱-骨界面的最�Objective： To study the promoting function of recombinant human bone morphogenetic protein-14（rh BMP-14） in the healing of tendon-bone of rabbit rotator cuff injury after reattachment. Methods： Forty-five adult male New Zealand white rabbits aged 8 months, randomly selected three as normal group for biomechanical testing. The other forty-two rabbits were randomized into three groups（fourteen animals/group）. Underwent bilateral detachment and repair of the supraspinatus tendon in the experiment. The animals received one of three treatments at the tendon-bone interface. The experimental group received a mixture of rh BMP-14 and chitosan thermosensitive hydrogel（CS/GP）. The CS/GP control group received only CS/GP. The blank control group were not handled. Seven animals in each group were sacrificed at 4 and 8 week. At each time point, four specimens were observed by H.E. stain. Collagens were observed by Picrate Sirius Red staining on four specimens.Measured the areas of type Ⅰ and type Ⅲ collagens and calculated their ratio. Six specimens were allocated for biomechanical testing. Results：（1）At 4-week, histological analysis of the tendon-bone interface showed an increasing collagen fibrils and cartilage tissue formed in the tendon-bone interface in the rh BMP-14 group, and it had more cartilage cells, the collagenous matrix was oriented in line. In the CS/GP control group and blank group, there were little collagen fibrils and cartilage tissue formed in the tendon-bone interface. At 8-week, the interface tissue in the rh BMP-14 group was denser and the collagenous matrix was more oriented in line with the tendon. Sharpey ＇s fibers were found in the interface at the 8-week of the rh BMP-14 group. In the CS/GP control grouprh and blank control group, there was little appearing fibrocartilage zone.Picrate Sirius red staining showed that the ratio of type Ⅰcollagens and type Ⅲ collagens raised with the increase of time for each group, and the difference was statistically significant（P�

关 键 词：骨形态发生蛋白 肩袖 创伤和损伤 生物力学 组织学 兔 

分 类 号：R658.2[医药卫生—外科学]