脱细胞猪角膜基质支架的制备及其生物相容性  

Preparation of an acellular porcine corneal stroma scaffold and its biocompatibility

作　　者：霍银平[1] 周利晓[1] 孙成林[1] 

机构地区：[1]郑州大学第五附属医院眼科,河南省郑州市450052

出　　处：《中国组织工程研究》2016年第30期4449-4455,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(201503131)~~

摘　　要：背景:研究发现多种生物材料可制备角膜基质支架且具有良好的生物相容性,而关于脱细胞猪角膜基质支架的制备和生物相容性研究不多。目的:制备脱细胞猪角膜基质支架,探讨其与人角膜基质细胞的生物相容性。方法:制备脱细胞猪角膜基质支架及其浸提液。取生长状态良好的人角膜基质细胞,分别以脱细胞猪角膜基质支架浸提液(实验组)、完全培养基培养(对照组)培养,培养后1,2,3 d,采用MTT法检测人角膜基质细胞的增殖;将人角膜基质细胞直接接种于脱细胞猪角膜基质支架上,接种1,3,7 d,采用免疫化学法测定人角膜基质细胞在支架内的生长情况。结果与结论:(1)随着时间的增加,实验组与对照组人角膜基质细胞数量不断增加,两组不同时间点的吸光度值比较差异无显著性意义;(2)人角膜基质细胞在脱细胞猪角膜基质支架上生长良好,细胞整联蛋白β1、波形蛋白、乙醛脱氢酶3A1呈阳性表达,CD34、CDK2和K-Ras蛋白也呈阳性表达;(3)结果表明,脱细胞猪角膜基质支架无细胞毒性,与人角膜基质细胞具有良好的生物相容性。BACKGROUND： Studies have found that a variety of biological materials can be used for preparing corneal stroma scaffolds that have good biocompatibility, but research on preparation and biocompatibility of the acellular porcine corneal stroma scaffold is little. OBJECTIVE： To explore the preparation and biocompatibility of the acellular porcine corneal stroma scaffold. METHODS： Acellular porcine corneal stroma scaffold and its extract were prepared. Well-grown human corneal stromal cells were selected and cultured in the extract of acellular porcine comeal stroma scaffold （experimental group） or in the complete medium （control group）, respectively. After 1,2 and 3 days of culture, the proliferation ability of human corneal stromal cells was detected by MTT assay. In the meanwhile, human corneal cells were directly seeded onto the acellular porcine corneal stroma scaffold, and then the cell growth on the scaffold was detected using immunochemical method. RESULTS AND CONCLUSION： The number of human corneal stromal cells was in a rise with time in the two groups, and absorbance values had no significant difference between two groups at different time points of culture. Human corneal stromal cells grew well on the scaffold, and were positive for cell integrin 131, vimentin, aldehyde dehydrogenase 3A1, as well as CD34, CDK2 and K-Ras. These results show that the acellular porcine corneal stroma scaffold has no cytotoxicity, and has good biocompatibility.

关 键 词：生物相容性材料 角膜基质 组织工程 生物材料 材料相容性 脱细胞猪角膜基质支架 生物相容性 人角膜基质细胞 整联蛋白β1 波形蛋白 乙醛脱氢酶3A1 

分 类 号：R318[医药卫生—生物医学工程]