电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜预防腱周粘连的相关研究  

The related research of the PLGA/type-I collagen-PCL double-layer electrospun membrane's ability of preventing peritendinous adhesion

作　　者：李鹏昊[1] 刘舒云[2] 王亚洁[1] 全琦[2] 王玉[2] 汪爱媛[2] 彭江[2] 许文静[2] 卢世璧[2] 郭全义[2] 

机构地区：[1]南开大学医学院,天津300071 [2]中国人民解放军总医院骨科研究所骨科再生医学北京市重点实验室/全军骨科战创伤重点实验室,北京100853

出　　处：《中国医药生物技术》2017年第4期289-296,共8页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(21134004);国家高科技研究发展计划(863计划)(2012AA020502;2015AA020303);骨科再生医学北京市重点实验室2016年度科技创新基地培育和发展项目子专项(Z161100005016059)

摘　　要：目的探讨聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜的结构、成分、亲水性、组织相容性和细胞毒性,以及该膜对鼠L929成纤维细胞黏附和活性的影响,从而明确该电纺膜在预防肌腱断裂后腱周粘连的可行性。方法制备单纯电纺聚乳酸-羟基乙酸共聚物膜和电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜。通过扫描电镜、红外光谱、接触角检测分析电纺膜的超微结构、成分以及亲水性能;通过溶血试验、热原试验、致敏试验和MTT法验证电纺膜的组织相容性和细胞毒性;将鼠L929细胞种植在单纯电纺聚乳酸-羟基乙酸共聚物膜和电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜上,体外培养4 d后进行死/活细胞染色,通过共聚焦显微镜观察比较L929细胞在两组膜上的黏附和活性情况。结果两种电纺膜由纳米级-微米级的纤维丝有序排列,形成致密多孔的网状结构。且溶血试验、热原试验、致敏试验结果均呈阴性。MTT检测发现,L929细胞在电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜的DMEM浸提液组和正常培养液组中的OD570值的差异没有统计学意义(P>0.05);死/活细胞染色提示,鼠L929成纤维细胞在电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜的Ⅰ型胶原-聚己内酯层上第4天的黏附率和生存率显著高于对照组(P<0.05),而L929细胞在单纯电纺聚乳酸-羟基乙酸共聚物膜上的黏附率和生存率显著低于对照组(P<0.05)。结论电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜是由纳米级-微米级有序排列的纤维丝构成的多孔膜片,而且组织相容性良好,无明显细胞毒性。其聚乳酸-羟基乙酸共聚物层可以有效抑制成纤维细胞的黏附和生存,Ⅰ型胶原-聚己内酯层可以促进细胞的黏附和存活。电纺聚乳酸-羟基乙酸共聚物/Ⅰ型胶原-聚己内酯双层膜对�Objective To study the structure, composition, hydrophile, histocompatibility and cytotoxicity of the electrospun PLGA/type-I collagen-PCL double-layer membrane, and evaulate the effect of the membrane in mouse＇s L929 fibroblasts＇ adhesion and activity. Methods Pure PLGA electrosptm membrane and PLGA/type-I collagen-PCL double-layer electrospun membrane were produced via electrospinning. The membranes＇ ultrastructure, ingredients and hydrophile were analyzed through scanning electron microscope, fourier transform infrared spectroscopy and water contact angle testing. The histocompatibility and cytotoxicity of the electrospinning membranes were verified through hemolysis test, pyrogen test, sensitization test and MTT test. In addition, mouse L929 fibroblasts were cultured on pure PLGA electrospun membrane and PLGA/type-I collagen-PCL double-layer electrospun membrane for four days, and then stained with FDA/PI, and the condition of the cells＇ adhesion and activity were observed and compared using confocal microscopy. Results Well-ordered nano-scale to micron-scale fibers formed dense porous mesh structure in two kinds of electrospun membranes. And the results of hemolysis test, pyrogen test and sensitization test were negative. The values of ODsTo had no significant statistical difference between the PLGA/type-I collagen-PCL double-layer electrospun membrane＇s DMEM leaching solution ＋L929 cells group and the normal culture solution ＋L929 cells group （P 〉 0.05）, determined by MTT test. In addition, on the fourth day, the mouse＇s L929 fibroblasts＇ proliferation and activity on the PLGA/type-I collagen-PCL double-layer electrospun membrane was significantly higher than that of control group （P 〈 0.05）, and the mouse＇s L929 fibroblasts＇ proliferation and activity on the pure PLGA membrane was significantly lower than that of control group （P 〈 0.05）. Conclusion The PLGA/type-I collagen-PCL double-layer electrospun membrane is made into dense porous mesh structure with abundant w

关 键 词：组织粘连 生物相容性材料 胶原I型 肌腱断裂 静电纺丝 

分 类 号：R318.08[医药卫生—生物医学工程] R68[医药卫生—基础医学]