三种方法制备的猪小肠黏膜下层支架的生物相容性和免疫原性的对比研究  

A Comparative Study of Three Ways of Acellular Process on Small Intestinal Submucosa's Biocompatibility and Immunogenicity

作　　者：黄伟锋[1] 程鹏[1] 姜平[1] 

机构地区：[1]南方医科大学南方医院整形外科,广州510515

出　　处：《中国生物工程杂志》2015年第6期54-60,共7页China Biotechnology

基　　金：广东省自然科学基金(S2011010003864)资助项目

摘　　要：目的:采用三种不同的脱细胞方法制备小肠黏膜下层(small intestinal submucosa,SIS)支架,研究其生物相容性、免疫原性以及构建真皮替代物等方面,为组织工程化皮肤的细胞载体支架提供优选方案。方法:将新鲜猪小肠分别采用单纯机械法、机械-化学法、机械-酶消化法制备三种不同的SIS,分别作为A、B、C组;通过复合成纤维细胞构建真皮替代物,利用HE染色观察支架组织学形态以及细胞粘附情况、MTT法检测细胞增殖情况,同时观察支架移植SD大鼠皮下1、2、4周后炎症反应及血管化程度。结果:组织学观察A组有细胞残留,B、C组未见细胞残留。MTT结果显示细胞在支架上生长旺盛增殖能力强,其中A组优于B、C组;皮下移植后的HE结果表明B组引起炎症反应较A、C组弱,而C组血管化程度较A、B组更明显。结论:机械-化学法以及机械-酶消化法制备的SIS具有良好的生物相容性以及免疫原性,可作为构建组织工程化皮肤细胞载体的选择。Objective： To compare the effect of three ways of acellular process on biocompatibility and immunogenicity of the small intestinal submucosa,preparing for the scaffold of tissue engineering skin. Methods：Fresh jejunum of pig was prepared by mechanical method, mechanical-chemical method and machanicalenzymicmethod into SIS,kept as groups A,B,C,respectively. Fibroblasts were seeded on the SIS scaffolds to construct the derm substitute. The comparative examinations were performed by histological observation,MTT assay to observe the structure of the scaffolds,proliferation and adhesion of the cell on the scaffolds. The inflammation cause by the scaffolds after subcutaneous implantation for 1,2,4 weeks were also analysed. Results：Histological observation shows that there were no residual cells in groups B and C,but residual cells in group A.The proliferation and adhesion test indicated that group A was better than the other two（ P〈0. 05）. The subcutaneous implantation analysis showed that group B caused a less serious inflammation, and the vascularization capacity of group C was greater than groups A and B. Conclusion： Mechanical-chemical method and machanical-enzymic method were better ways to prepare SIS as a scaffold for tissue engineering skin.

关 键 词：小肠黏膜下层 脱细胞处理 成纤维细胞 真皮替代物细胞 载体支架 

分 类 号：Q813.1[生物学—生物工程]