改良型人发角蛋白/丝素蛋白复合材料体外理化及生物学特性评估  

Evaluation of physicochemical and biological properties of modified keratin/silk fibroin compound biomaterial in vitro

作　　者：冯超[1] 李喆[2] 吕向国[1] 徐月敏[1] 傅强[1] 

机构地区：[1]上海交通大学附属第六人民医院泌尿外科,上海200233 [2]东华大学材料学院,上海201620

出　　处：《上海交通大学学报（医学版）》2015年第1期29-35,共7页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81100488)~~

摘　　要：目的探索人发角蛋白/丝素蛋白复合材料的改良技术并进行理化鉴定及生物学特性评估。方法自人发及蚕丝中分别提取人发角蛋白及丝素蛋白。采用SDS聚丙烯酰胺凝胶电泳(SDS-PAGE)技术对两种蛋白进行定性。按不同浓度比例将两种蛋白进行混合,并加入不同浓度的明胶。对最终制备完成的复合材料进行力学拉伸测试,筛选出最佳的蛋白与明胶浓度组合。采用红外光谱分析复合材料蛋白空间结构变化。兔皮下埋植复合材料,观察生物相容性,MTT法检测细胞毒性。结果经过提取、纯化,人发角蛋白与丝素蛋白浓度及纯度达到满足后续实验水平。力学拉伸测试结果显示:当明胶浓度为2%,丝素蛋白与人发角蛋白的比例为60∶40时,制备材料的力学特性最佳。红外光谱分析结果显示复合材料各蛋白组分的空间结构无明显变化。兔皮下埋植实验显示复合材料具有较为理想的生物相容性及降解速率;MTT检测结果提示复合材料无明显细胞毒性。结论利用人发角蛋白、丝素蛋白以及明胶在合适比例下,可对已报道的人发角蛋白/丝素蛋白复合材料进行进一步的理化特性改良,使其能够更适合后续组织工程修复重建的基础研究及临床应用。Objective To explore the feasibility of modified technique for the keratin/silk fibroin compound biomaterial, identify physicochemical properties, and evaluate biological properties. Methods The keratin and silk fibroin were extracted from human hair and worm silk, respectively. The SDS-PAGE was adopted to determine the properties of those two proteins. Two proteins were mixed at different concentrations and different concentrations of gelatin were added. The tensile mechanical tests were used to detect the keratin/silk fibroin compound biomaterial and then the optimal combination of protein concentration and gelatine concentration was screened. Variations of the spatial structure of proteins were analyzed by the Fourier transform infrared spectroscopy （FTIR）. The compound biomaterial was embedded beneath the rabbit skin to evaluate its biocompatibility and the cytotoxicity was detected by the MTT assay. Results The concentration and purity of keratin and silk fibroin met the requirements of subsequent tests after extraction and purifcation. Mechanical tests showed that best mechanical properties were achieved when the concentration of gelatin was 2% and the ratio of keratin and silk fibroin was 60：40. Results of the FTIR indicated that the spatial structure of each protein of the compound biomaterial did not vary significantly. Tests of embedding the compound biomaterial beneath the rabbit skin showed that the biocompatibility and degradation rate of the compound biomaterial were ideal. Results of the MTT indicated that the compound biomaterial has no significant cytotoxicity. Conclusion The physicochemical properties of reported keratin/silk fibroin compound biomaterial can be improved by proper ratio of keratin and silk fibroin and proper concentration of gelatine, which is more suitable for subsequent fundamental researches and clinical applications of repair and reconstruction of tissue engineering.

关 键 词：组织工程 人发角蛋白 丝素蛋白 

分 类 号：R318.08[医药卫生—生物医学工程]