猪耳廓及关节来源软骨细胞构建组织工程软骨的实验研究  

Comparison study of tissue engineered cartilage constructed with chondrocytes derived from porcine auricular and articular cartilage

作　　者：康宁 刘霞 曹谊林 肖苒 

机构地区：[1]中国医学科学院整形外科医院研究中心北京100144

出　　处：《中华整形外科杂志》2014年第1期33-40,共8页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金（31300801）;北京市科技计划项目（D090800046609003）;卫生部临床学科重点项目

摘　　要：【摘要】目的探讨耳廓及关节来源软骨细胞构建组织工程软骨的差异。方法分离培养健康长枫杂交猪耳廓、关节来源软骨细胞及骨髓间充质干细胞（bonemarrowmesenchymalstemcells，BMSCs），分别以软骨细胞单独接种或与BMSCs1：1混合接种聚羟基乙酸／聚乳酸（Polyglycolicacid／Polylacticacid，PGA／PLA）支架2种模式构建组织工程软骨。针对每种构建模式按软骨细胞来源随机分为4组，每组4例样本，分别为耳廓软骨细胞组、关节软骨细胞组、耳廓软骨细胞＋BMSCs1：1共培养组、关节软骨细胞＋BMSCs1：1共培养组。于体外培养8周、裸鼠皮下植入6周后分别取材进行大体观察，比较单独培养或共培养构建过程中4组样本的组织厚度和湿重，HE、番红O、lI型胶原免疫组化和丽春红／维多利亚蓝染色比较软骨组织形成情况，Real—timePCR检测软骨相关基因的表达，采用两组独立样本f检验进行统计学分析。结果体外培养8周，各组均能形成软骨样结构，只有耳廓软骨细胞＋BMSCs组可见少量弹性纤维分泌；单独应用关节软骨细胞构建的组织工程软骨厚度为（3．12±0．24）mm，较耳廓软骨细胞组平均增厚38．1％（P〈0．05），关节软骨细胞＋BMSCs1：1共培养构建的组织工程软骨厚度为（2．41±0．21）mm，较耳廓软骨细胞＋BMSCs组平均增厚19．3％（P〈0．05）；体内重塑6周后，耳廓软骨细胞组及耳廓软骨细胞＋BMSCs组均可见大量弹性纤维，关节软骨细胞组及关节软骨细胞＋BMSCs组均形成了典型的透明软骨结构并发生了骨化，X型胶原和MMPl3的表达水平均显著上调（P〈0．05）。因此体内重塑6周后，耳廓软骨细胞单独培养或与BMSCs共培养均可形成弹性软骨结构，关节软骨细胞单独培养或与BMSCs共培养可形成透明软骨结构。结论应用不同组织来源软骨细胞构建的组织工程软骨，回植体内环�Objective To compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage. Methods Chondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1 ： 1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage（ n = 4）. The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including

关 键 词：【关键词】软骨细胞 骨髓间质干细胞 组织工程 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]