软骨组织工程支架材料的胶原结构分析

Analysis of collagenous structures for cartilage tissue engineered scaffolds

背景:在软骨组织工程化组织培养过程中发现不同类型的胶原材料影响软骨细胞的分化与表达,胶原材料的来源、结构及性质的稳定性决定软骨组织工程研究的可靠性。目的:表征Ⅰ型、Ⅱ型和Ⅲ型胶原的结构,阐明用于软骨组织修复的不同类型的胶原材料的结构特点,指导软骨组织工程支架材料的选取。方法:取来自新生小牛皮的Ⅰ型胶原、来自成猪关节软骨的Ⅱ型胶原及来自新生小牛皮的Ⅲ型胶原,分别将3种胶原材料进行傅里叶变换红外光谱、示差扫描量热法、SDS-PAGE和苦味酸-天狼腥红染色分析,比较结构上的差异。结果与结论:新生小牛皮Ⅰ型和Ⅲ型胶原与成猪关节软骨Ⅱ型胶原具有相似的官能团,热变性温度分别是104.2℃、99.7℃和92.5℃。SDS-PAGE分析Ⅰ型胶原中Ⅲ型胶原的百分含量为(11.29±0.91)%。苦味酸-天狼腥红染色显示Ⅰ型为红色和橙色的紧密排列的粗纤维,有很强的双折光;Ⅱ型胶原为红色的疏松的网状纤维;Ⅲ型胶原为绿色的疏松的细纤维。3种胶原的官能团结构和重要特征十分相似,但其高级结构有较大的差异,其相互关系的分析对软骨组织工程支架材料的选择和制备有十分重要的意义。

BACKGROUND：Various types of collagen affect differentiation and expression of chondrocytes in the process of tissue engineered culture for cartilage.The reliability of cartilage tissue engineering is determined by resource,structure and property of collagen.OBJECTIVE：To characterizing the structure of collagen type Ⅰ,Ⅱ and Ⅲ,to clarify the structural characteristics of collagen for cartilage tissue repairing and provide guidance for choosing cartilage tissue engineering scaffolds.METHODS：The type Ⅰ and Ⅲ collagen was obtained from neonatal calf skins,and type Ⅱ collagen was obtained from porcine articular cartilage.The differences of structures of collagen type Ⅰ,Ⅱ and Ⅲ were analyzed and compared by FTIR,DSC,SDS-PAGE and picrosirius-red staining.RESULTS AND CONCLUSION：The neonatal calf collagen typeⅠ,Ⅲ and porcine collagen type Ⅱ had the similar functional group.The thermal denaturation temperature of the three type collagen was 104.2℃,99.7℃ and 92.5℃.SDS-PAGE analysis showed the collagen type Ⅲ percentage in collagen type Ⅰ was（11.29±0.91）%.Picrosirius-red staining exhibited that the collagen type Ⅰ was comprised red and orange raw fibers,with strongly birefringent.Collagen type Ⅲ was lax and thin,greenish fibers,but collagen type Ⅱ was red,loosely cancellated fibers.The functional groups and important characteristics of the three of collagen are similar.However,there are some differences in advanced structure.To analyze the relationships of collagen type Ⅰ,Ⅱ and Ⅲ is important to choose and prepare scaffolds for cartilage tissue engineering.