碱性成纤维细胞生长因子与胶原对组织工程软骨体外构建的影响  

Effects of bFGF and Collagen on the Construction of Tissue-engineered Cartilage in vitro

作　　者：吴凤麟[1] 宇丽[2] 

机构地区：[1]广东药学院生命科学与生物制药学院,广州510006 [2]暨南大学医学院生物化学教研室,广州510632

出　　处：《中国生物工程杂志》2007年第7期45-49,共5页China Biotechnology

基　　金：教育部科学技术研究重点项目(02190);广州市科技计划重大科技攻关重点基础项目(200221-E0032)

摘　　要：目的:以三维成团培养为培养系统,探讨bFGF与胶原对组织工程软骨体外构建的影响。方法:成团培养兔生长板软骨细胞,设bFGF、胶原及联合作用组。HE染色观察新生组织形态;免疫组化检测Ⅰ、Ⅱ型胶原表达以观察细胞表型;Hoechst33258法检测细胞DNA含量;羟脯氨酸法与阿新蓝法测定基质中胶原与蛋白多糖的合成。结果:新生软骨的组织学形态近似自然软骨;各实验组软骨细胞DNA含量明显上升;胶原可以显著促进基质的合成;各实验组Ⅰ型胶原的表达少于对照组,Ⅱ型胶原的表达则高于对照组;联合作用组效果更加明显。结论:三维的成团培养可以促进基质合成,有效维持软骨细胞表型;bFGF与胶原有利于工程化软骨构建,其效果具有协同效应,两者联合应用可进一步促进软骨再生。Objective： To investigate effects of bFGF and collagen on tissue-engineered cartilage in pellet culture. Methods： Growth-plate chondrocytes were incubated in pellet culture, bFGF and Collagen was applied alone or together based on the results of previous research. The histological structure of neocartilage was observed by stained with H＆E and Immunohistochemical analysis. The DNA quantity of neocartilage was measured by hoechst33258 dye. The production of matrix was estimated based on hydroxyproline quantity and Alcian Blue stain method. Results： The histological structure of in vitro -formed tissue was similar to growth-plate cartilage. The DNA quantity of neocarfilage increased by the supplement of bFGF and collagen. The production of matrix was promoted by collagen, bFGF and collagen suppressed the expression of collagen Ⅰ ,and decelerated the decrease of collagen Ⅱ. The combined usage of bFGF and collagen had synergistic effects. Conclusion： Pellet culture maintained the phenotype of chondrocyte and promoted matrix production, bFGF and Collagen improved the construction of engineered cartilage. The combined usage of bFGF and collagen had synergistic effects on the regeneration of cartilage.

关 键 词：BFGF 胶原 软骨 构建 

分 类 号：Q813.1[生物学—生物工程]