自组装多肽纳米支架与骨髓间充质干细胞的生物相容性研究△  

Biocompatibility of IKVAV peptide self-assembly nano-fibers with mesenchymal stem cells

作　　者：张伟[1,2] 岳靓[2] 易红蕾[1] 王传峰[1] 陈家瑜[1] 杨宗德[1] 魏显招[1] 张涤清[1] 李明[1] 傅强[1] 

机构地区：[1]第二军医大学长海医院骨科,上海200433 [2]解放军第401医院,山东青岛266071

出　　处：《中国矫形外科杂志》2011年第22期1906-1909,共4页Orthopedic Journal of China

基　　金：国家自然科学基金面上项目(编号:30872629);全军医学科学技术基金十二五计划重点项目(编号:08G073)

摘　　要：[目的]合成神经活性多肽IKVAV(isoleucine-lysine-valine-alanine-valine,异亮氨酸-赖氨酸-缬氨酸-丙氨酸-缬氨酸)两亲性分子,在体外进行自组装纳米纤维支架,并检测其与骨髓间充质干细胞(mesenchymal stemcells,MSCs)的生物相容性。[方法]IKVAV多肽两亲性分子委托上海波泰生物科技公司合成,并用高效液相色谱仪和质谱仪进行纯化和分析。将IKVAV多肽两亲性分子溶于0.1 M NaOH溶液中,加入稀盐酸降低pH值引发自组装,并利用透射电镜进行观察。分离兔骨髓间充质干细胞,流式分析其表面抗原标志。将MSCs与自组装支架体外共培养12 d,测定细胞粘附率,MTT方法检测IKVAV自组装材料对细胞生长增殖的影响,观察其生物相容性。[结果]IKVAV-PA可在一定条件下自组装形成凝胶,透射电镜可见其显示为纳米纤维,其直径为7.0～8.0 nm。分离的MSCs细胞表面标志CD105、CD44、CD34和CD45的阳性率分别为99.3%、99.5%、0.1%、0.4%。共培养过程后,随着时间延长,MSCs在多肽凝胶支架上黏附率上升,吸光度增加,细胞活性增强。[结论]IKVAV-PA能自组装形成纳米纤维凝胶,具有良好的生物相容性和生物活性。[Objective]To synthesize bioactive IKVAV（isoleucine-lysine-valine-alanine-valine） peptide-amphiphile（IKVAV-PA） and self-assemble it in vitro and investigate its biocompatibility to mesenchymal stem cells（MSCs）.[Method]IKVAV-PA was synthesized by the Shanghai Botai Biological Technology Company,which was then purified and tested with high-performance liquid chromatography and mass spectrometry respectively.The peptide was dissolved in NaOH solution,the diluted hydrochloric acid was added into IKVAV-PA solution to reduce the pH value and accordingly induce self-assembly.The morphological features of the assembled material were studied by transmission electron microscope（TEM）.The MSCs were separated and identified by flow cytometry for the cell surface markers.The MSCs were cultured with the assembled IKVAV-PA for 12 days.The number of the adhered cells was tested,MTT kit was used to determine the effect of IKVAV-PA assembled material on proliferation of MSCs.[Result]IKVAV-PA could be self-assembled to form a gel in certain condition.TEM showed that the assembled material was a kind of nanofiber with diameter of 7.0-8.0 nm.The surface markers of CD105,CD44,CD34 and CD45 on MSCs were 99.3 %,99.5%,0.1% and 0.4% respectively.The adhered cells on the assembled material were increased for more days.MTT assay showed that IKVAV-PA had promoted the proliferation of MSCs.[Conclusion]IKVAV-PA can be self-assembled into nano-fiber gel,which has good biocompatibility and neural bioactivity.

关 键 词：自组装多肽 纳米纤维 骨髓间充质干细胞 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]