丝素蛋白基人工角膜多孔支架的细胞相容性研究  

Study of cyto-compatibility in silk fibroin porous scaffold

作　　者：宋大伟[1] 马义宾[1] 聂蓉晖[1] 

机构地区：[1]泰安市中心医院眼科,271000

出　　处：《中华移植杂志（电子版）》2015年第2期36-39,共4页Chinese Journal of Transplantation(Electronic Edition)

摘　　要：目的研究不同丝素蛋白多孔支架的细胞相容性。方法将兔角膜成纤维细胞分别在盐析法制备的丝素蛋白支架、明胶含量为20%和30%的丝素蛋白-明胶多孔支架上进行培养,分别进行细胞活性和形貌分析,以评价不同丝素蛋白多孔支架的细胞相容性。结果所有支架体外培养2周后,细胞数目均显著增加。体外培养1 d后,细胞在所有支架上都表现出良好的黏附性,有大量伪足形成。2周后,细胞有显著增殖,并有大量细胞外基质形成。表明盐析法制备的丝素蛋白支架和丝素蛋白-明胶多孔支架都能够很好地促进细胞生长。丝素蛋白支架上细胞数目在体外培养10 d时达到最大值,然后细胞数目开始降低。丝素蛋白-明胶多孔支架上细胞数目在2周的培养过程中一直增加。14 d时,丝素蛋白-明胶多孔支架上的细胞数目高于在盐析法制备的丝素蛋白多孔支架上的细胞数目,明胶含量为30%的丝素蛋白-明胶支架培养细胞数目及活细胞数目明显多于20%者。激光共聚焦显微镜发现,14 d时盐析法制备的丝素蛋白多孔支架上有大量死细胞,而在丝素蛋白-明胶多孔支架上则基本没有死细胞,30%丝素蛋白-明胶多孔支架上的细胞数量及活细胞数量高于20%者。结论通过将明胶添加到丝素蛋白溶液,可提高支架的细胞相容性,使其能更好地满足角膜组织再生的要求。Objective To explore the cyto-compatibilitylity of different silk fibroin porous scaffolds. Methods The rabbit cornea fibroblasts were respectively cultured in silk fibroin scaffolds produced by salting out method,silk fibroin porous scaffolds with gelation content 20% and silk fibroin porous scaffolds with gelation content 30%,then analyzed their cell activities and compositions to evaluate the cyto-compatibility of different silk fibroin porous scaffolds. Result After all the scaffolds were cultured for 2 weeks,the number of cells increased significantly. After one day,all the cells on the scaffolds exhibited good adhesiveness and formed a number of pseudopodia. Two weeks later,the cells proliferated distinctively and formed massive extracellular matrixes. It proved that both silk fibroin scaffolds produced by salting out method and silk fibroin-gelatin porous scaffolds can promote the cell growth. The number of cells reached the maximum after being cultured on the silk fibroin scaffolds for10 days and after that it began to drop down. On the contrary,the number of cells cultured on the silk fibroin-gelatin porous scaffolds kept increasing during the whole 2 weeks. On the 14 th day,the number of cells on the silk fibroin-gelatin porous scaffolds was far more than that on silk fibroin porous scaffolds produced by salting out method. An amount of dead cells were found on the silk fibroin porous scaffolds produced by salting out method with the help of confocal laser scanning microscope,but few were found on the silk fibroin-gelatin porous scaffolds. Conclusion Putting gelation into the silk fibroin solution can improve the cyto-compatibility of the scaffolds and meet the needs of cornea tissue regeneration better.

关 键 词：丝素蛋白多孔支架 明胶 人工角膜 细胞相容性 

分 类 号：R318.08[医药卫生—生物医学工程]