模拟微重力动态培养构建组织工程化软骨修复猪关节软骨缺损研究  

Repair of Pig Cartilage Defects with Tissue-engineered Cartilage Constructed under Simulated Microgravity

作　　者：林威[1,2,3] 陈洁琳[2,3] 刘威[2,3] 杨坤[2,3] 王大明[2,3] 段莉[2,3] 朱伟民[2,3] 王大平[2,3] 

机构地区：[1]广州医科大学,广东广州510182 [2]深圳市运动医学技术工程研究中心,广东深圳518035 [3]深圳市第二人民医院组织工程实验室,广东深圳518035

出　　处：《中国运动医学杂志》2016年第5期432-437,455,共7页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金项目(No.81572198);广东省科技计划项目(2015A020212001);广东省自然科学基金(2015A030313772);深圳市科技计划项目(JCYJ20140414170821164;JSGG20140519105550503;JCYJ20140414170821200;JCYJ201404141708-21160;CXZZ20140 813160132596)

摘　　要：目的：通过大型动物实验探讨模拟微重力动态培养系统构建组织工程化软骨修复关节软骨缺损的效果。方法：10~12月龄的五指山猪8头,分别在每头猪髌骨切迹上建立4个软骨缺损模型,左右膝关节各2个缺损,共32个,取缺损位置的软骨组织进行细胞分离培养扩增后复合三维Ⅰ型胶原上,分别置于旋壁式生物反应器与培养板中培养7 d,一方面取适量的样本进行HE染色、DAPI染色检测;另一方面将复合物回植修复猪关节缺损。每头猪关节缺损随机分为4组：微重力动态培养组（A）、普通静态培养组（B）、单纯支架组（C）、空白对照组（D）,每个膝关节随机分配两组。术后12周处死实验动物,取实验标本进行核磁共振（MRI）、大体观察及组织学检测。结果：MOCART核磁共振评分：A组为74.37±3.90、B组为66.87±4.96、C组为58.75±3.30、D组为43.12±3.47,差异具有统计学意义（P〈0.05）;O’Driscoll组织学评分：A组为16.00±1.41、B组为12.37±1.69、C组为8.37±0.83、D组为3.25±1.26,各组之间差异具有统计学意义（P〈0.05）。结论：模拟微重力动态培养模式构建的组织工程化软骨较普通培养更有利于软骨缺损的修复。Objective Exploring the role of simulated microgravity dynamic system in the construction of tissue-engineered cartilage of pigs. Methods Cartilage defect model was established on patellar incisures of both knees in eight 10 ~12 months old Wuzhishan pigs（two defects in each knee）,and thus a total of 32 cartilage defects were investigated. The chondrocytes of defect tissue were separated,cultivated and amplified,then compound with the collagen type I. Thereafter,they were cultivated for 7 days in a rotating wall bioreactor to simulate microgravity dynamic cultivating chondrocytes, and an adequate amount of them were undertaken HE staining and immunofluorescence staining for histological observation and the remaining were randomly implanted back to the defected knee of some pigs（group A）. some of the knees of pigs were randomly implanted with regular static culture chondrocytes（group B）and pure scaffold（group C）. The remaining knees of pigs without any implantation were served as blank controls（group D）. The pigs were scarified 12 months later,and their defected cartilages were determined by MRI,gross observation,and histological examination. Results There were significant differences in MOCART MRI rating in group A（74.37 ±3.90）,group B（66.87 ± 4.96）,group C（58.75 ± 3.30）and group D（43.12 ± 3.47）（P 0.05）,as well as in O＇Driscoll histologic grades in group A（16.00 ± 1.41）,group B（12.37 ± 1.69）,group C（8.37 ± 0.83）and group D（3.25 ± 1.26）（P 0.05）. Conclusion The simulated microgravity dynamic cultivation of chondrocytes in the construction of engineered cartilage is more profitable than by the regular static culture.

关 键 词：微重力 组织工程 软骨损伤 生物反应器 

分 类 号：R68[医药卫生—骨科学]