纤维蛋白凝胶基质材料用于组织工程修复兔关节软骨缺损与Ⅰ型胶原凝胶和混合凝胶性能特征的比较  

Different fibrin gel matrix for tissue engineering to repair cartilage defect in rabbits: A comparison with collagen Ⅰ gel and mixed gel in performance characteristics

作　　者：王立春[1] 张旗涛[1] 张宏颖 

机构地区：[1]哈尔滨医科大学附属第二临床医学院骨外科,黑龙江省哈尔滨市150086 [2]哈尔滨市穆斯林医院内科,黑龙江省哈尔滨市150042

出　　处：《中国临床康复》2005年第26期116-118,共3页Chinese Journal of Clinical Rehabilitation

基　　金：哈尔滨市青年科学研究基金资助项目(2004AFQXJ006)~~

摘　　要：目的:观察纤维蛋白凝胶、Ⅰ型胶原凝胶、混合凝胶3种不同生物型细胞外基质用于组织工程修复兔关节软骨缺损形态学及性能特征比较。方法:实验于2002-01/2003-12在哈尔滨医科大学附属二院实验中心进行。6月龄日本大耳白兔48只。①人工软骨制备:传代培养兔骨髓基质干细胞,与纤维蛋白原凝胶、Ⅰ型胶原凝胶、纤维蛋白-Ⅰ型胶原混合凝胶混合制成3种人工软骨。②模型制备与动物分组:兔麻醉后取膝关节外侧切口,显露髌股关节面,作直径4.5mm,深3mm转孔获得关节软骨缺损模型。48只兔分为4组,每组12只。纤维蛋白凝胶组,缺损区充填骨髓基质细胞纤维蛋白凝胶;Ⅰ型胶原凝胶组,充填骨髓基质细胞Ⅰ型胶原凝胶;混合凝胶组,充填骨髓基质细胞纤维蛋白-Ⅰ型胶原混合凝胶;空白对照组,不充填材料。术后各组动物膝关节不予外固定,自由活动。③标本观察:分别于4,8,12周每组取3只(6侧膝关节)麻醉后处死,行膝关节大体观察,选择损伤区股骨干切片作苯胺蓝、苏木精-伊红染色、Ⅱ型胶原免疫组织化学染色采用光学显微镜观察,并按改良的Seller评分(0分为正常关节,分数越高表示关节退行性变越严重)进行量化评分。结果:①各组兔膝关节4周及12周时大体形态学观察:4周时:纤维蛋白凝胶组、混合凝胶组为淡粉色半透明的软骨样结构,Ⅰ型胶原凝胶组、空白对照组多数缺损区仍呈暗红色,表面不规则。12周时:纤维蛋白凝胶组、混合凝胶组修复组织与周围软骨平滑过渡高度一致、光泽度较正常软骨略差;Ⅰ型胶原凝胶组、空白对照组修复区多数不平整,中央凹陷且存在龟裂。②各组兔股骨组织切片观察结果:纤维蛋白原凝胶、混合凝胶组各时间点组织化学染色均显著优于Ⅰ型胶原凝胶组和空白对照组;纤维蛋白原凝胶、混合凝胶组无显著差异。③组织切片的Seller评分结果:�AIM： To compare the morphological and performance characteristic difference among three different biologic extracellular matrix, fibrin gel, collagen Ⅰ gel and mixed gel applied in tissue engineering to repair rabbit articular cartilage defect. METHODS： This experiment was completed in the experiment center of the Second Hospital Affiliated to Harbin Medical University from January 2002 to December 2003. Forty-eight 6-month Japanese flap-eared rabbits were used as the studying material. ①Preparation of artificial cartilage： The subcuhured marrow stromal cells of rabbits were mixed with fibrinogen gel, collagen Ⅰ gel and fibrin-collagen Ⅰ mixed gel to make 3 types of artificial cartilage. ②Preparation of models and grouping of animals： After the rabbits were anesthetized, the rabbit patellofemoral joint surface was exposed through lateral incision of knee joint, and a hole 4.5 mm in diameter and 3 mm in depth was made to get articular cartilage defect model. Forty-eight rabbits were randomly divided into 4 groups with 12 in each： fibrin gel group, filled with bone marrow stromal cells-fibrin gel in the defect area; collagen Ⅰ gel group, filled with bone marrow stromal cells.collagen Ⅰ gel; mixed gel group, filled with bone marrow stromal cells-fibrin.collagen Ⅰ mixed gel; blank group, filled with nothing. After the operation, all the animals received no external fixation and could move freely. ③Observation on the specimens： At week 4, 8, 12, 3 rabbits（6 knees） of each group were anesthetized and put to death. The knee joint was observed grossly. The shaft of femur in defect area was chosen to make sections which were then stained with aniline blue and hematine-eosine, processed by collagen Ⅱ immunohistochemical staining. The sections were observed by light microscopy and scored according to improved Seller＇s score method（0 for normal joint and higher marks for more serious degeneration of joint）. RESULTS： ①Morphological and gross observation of rabbit knee joint in eac

关 键 词：纤维蛋白凝胶基质材料 组织工程修复 兔 关节软骨缺损 Ⅰ型胶原凝胶 混合凝胶 性能特征 

分 类 号：R684[医药卫生—骨科学] R318.08[医药卫生—外科学]