采用酸溶、酶溶相结合的方法,从兔皮中提取了胶原蛋白.通过SDS-PAGE对其进行定性分析,利用FT-IR及TG/DTG表征其理化性能.结果表明,所提胶原为I型胶原,保持了完整的三股螺旋结构;热处理过程中,在100℃之前,200~450℃之间和450℃以后有3次明显失重.经戊二醛交联改性后,该胶原的含水量为(78.7±1.3)%,酶降解率从(88.65±0.11)%降为(6.03±0.38)%(p〈0.01).采用细胞生长抑制法测定了改性胶原对小鼠胚胎成纤维细胞(MEF-WT)的细胞毒性.结果表明,改性胶原对MEF-WT细胞无明显毒性,毒性评价为0级或1级.该胶原可用于组织工程.
The extraction and characterization modified of type I collagen from rabbit-skins are investigated in this paper.Collagen was extracted from rabbit-skins by the combination of acid solubilization and pepsin solubilization.A qualitative analysis was carried out by SDS-PAGE.Both FT-IR and TG/DTG were employed to characterize the physical and chemical properties of the collagen.The results show that the extracted collagen is referred as type I collagen with a complete three-helix structure.In the process of thermal treatment,three stages of weight loss are observed at the temperatures of below 100℃,between 200℃and 450℃,and after 450℃,respectively.After being cross-linked by glutaraldehyde,the collagen was submitted to water content and enzyme degradation with I-collagenase in vitro.It is indicated that,after modification,the water content was(78.7±1.3)%.The degradation rate decreases from(88.65±0.11)%to(6.03±0.38)%(p〈0.01).Cytotoxicity of the modified collagen was evaluated by growth suppression technique(MTT chromatometry)against wild type mouse embryonic fibroblasts(MEF-WT).Its result shows that the cytotoxicity of the collagen is of grade 1or grade 0,nearly no cytotoxicity.This collagen has a potential use in tissue engineering.