重组胶原蛋白海绵的制备及性状表征  

Preparation and properties of recombinant collagen sponge

作　　者：何越 侯增淼 李晓颖 高恩 刘建利[2] 赵金礼 He Yue;Hou Zengmiao;Li Xiaoying;Gao En;Liu Jianli;Zhao Jinli(Pharmaceutical Research and Development Center, Huikang Biotechnology Co., Ltd., Xi’an 710054, Shaanxi Province, China;College of Life Sciences, Northwest University, Xi’an 710069, Shaanxi Province, China)

机构地区：[1]陕西慧康生物科技有限责任公司医药研发中心,陕西省西安市710054 [2]西北大学生命科学学院,陕西省西安市710069

出　　处：《中国组织工程研究》2019年第6期912-916,共5页Chinese Journal of Tissue Engineering Research

基　　金：西安市科技计划项目(CX13120);项目负责人:赵金礼~~

摘　　要：背景:重组胶原蛋白具有良好的亲水性及生物相容性,但其存在溶解性强及机械强度不足等缺点。化学交联可显著提高材料的韧性、力学强度及耐降解性。目的:制备重组胶原蛋白海绵,并对其理化性质及安全性进行评价。方法:采用微生物发酵法获得重组胶原蛋白,经戊二醛交联后,将其置于模具中冷冻干燥,获得多孔重组胶原蛋白海绵,检测交联后重组胶原海绵的吸水率与孔隙率,采用扫描电镜观察交联前后重组胶原海绵的表面形态,红外光谱仪对比交联前后重组胶原海绵结构的变化。采用交联重组胶原海绵浸提液培养L929细胞,培养第68小时,采用MTT法检测细胞相对增殖率,评估交联重组胶原海绵的细胞毒性。结果与结论:(1)交联后重组胶原海绵的吸水率为(2 903.83±47.90)%,平均孔隙率在85%以上;(2)扫描电镜显示,交联前,重组胶原海绵的孔隙呈蜂窝状,致密;交联后,重组胶原海绵的孔隙较大,呈板层结构,可看到板层间桥键连接,纵向可看到较大褶皱;(3)红外光谱显示,交联后的重组胶原海绵特征性化学结构特征无明显变化;(4)交联重组胶原海绵浸提液培养L929细胞的相对增殖率为84%,细胞毒性为1级,生物安全性良好;(5)结果表明采用戊二醛交联制备的重组胶原海绵,理化性质稳定,生物相容性良好。BACKGROUND:Recombinant collagen has good hydrophilicity and biocompatibility,but it has the disadvantages of strong solubility and low mechanical strength.Chemical crosslinking can significantly improve the toughness,mechanical strength and degradation resistance of the material.OBJECTIVE:To prepare recombinant collagen sponge and to evaluate its physicochemical properties and safety.METHODS:The recombinant collagen was obtained via microbial fermentation and cross-linked with glutaraldehyde.The porous recombinant collagen sponge was finally obtained by freeze-drying in the mold.The water absorption,porosity and crosslinking agent residual of the recombinant collagen sponge were detected.The surface morphology of the recombinant collagen sponge before and after cross-linking was observed by scanning electron microscope.The changes in recombinant collagen structures before and after cross-linking were compared by infrared spectrometer.L929cells were cultured with cross-linked recombinant collagen sponge extract.At68hours after culture,the relative proliferation rate of the cells was detected by MTT,and the cytotoxicity of cross-linked recombinant collagen sponge was evaluated.RESULTS AND CONCLUSION:(1)The water absorption of the cross-linked collagen sponge was(2903.83±47.90)%and the average porosity was over85%.(2)Under the scanning electron microscope,the pores of the collagen sponge were honeycomb and dense before cross-linking.After cross-linking,the pores of the collagen sponge were large and bridged bond linkage of laminar layer structure could be seen,and also the large folds could be seen in the longitudinal direction.(3)There was no obvious change in the characteristic chemical structure of the recombinant collagen sponge after cross-linking.(4)The relative proliferation rate of L929cells was84%after culture in cross-linked recombinant collagen sponge extract,and the cytotoxicity was level1,indicating the biological safety was good.To conclude,the recombinant collagen sponge with glutaraldehyde cross-lin

关 键 词：重组胶原蛋白 交联 创面止血 胶原蛋白海绵 基因工程 宿主菌 基因工程菌 胶原 止血 交联试剂 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]