纳米化左旋聚乳酸有序膜的细胞亲和性研究  

Experimental study of affinity of endothelial outgrowth cells cultured on aligned plla nanofibrous scaffolds

作　　者：卢辉俊[1] 冯章启[2] 顾忠泽[2] 刘长建[1] 

机构地区：[1]南京大学医学院附属鼓楼医院血管外科,210008 [2]东南大学生物电子学国家重点实验室

出　　处：《中华胸心血管外科杂志》2009年第4期263-266,共4页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：本课题受江苏省卫生厅重大科研项目资助（K200609）

摘　　要：目的制备具有较好细胞亲和性、利于内皮生长晕细胞（EOCs））黏附增殖的纳米左旋聚乳酸有序膜，为构建组织工程血管材料提供理论依据。方法改性后纳米纤维膜与细胞复合培养，观察细胞与材料生物亲和性。结果纳米纤维孔径在300—400nm ，孔隙率〉90％。有序和超级有序膜组吸光度A值与无序膜、单纯细胞组差异有显著统计学意义（P〈0．05）。无序膜细胞生长较散在、杂乱；有序及超级有序纤维膜有利于细胞沿纤维定向附着、增殖。结论EOCs是理想的组织工程血管种子细胞来源。有序及超级有序膜是一种理想的组织工程血管材料。Objective Tissue engineering holds great promisc in providing vascular grafts as substitutes for damaged small-diameter blood vessels. Two of the key factors in vascular tissue engineering are bieeompatible scaffolds that mimic the effects of extracellular matrix and the source of seeding cells. To study the cellular affinity and adhesion and proliferation of endothelial outgrowth cells （EOCs） obtained from rabbit peripheral blood cultured with the aligned synthetic poly-L-lactic acid（PLLA） nanofibers in vitro so as to provide a solid foundation for future vascular tissue engineering involving both PLLA and EOCs. Methods The random, aligned and super-aligned PLLA nanofibers were prepared by an insulated high-speed roller collector covered with parallel steel sticks rotating at 0 rpm, 1000 rpm and 2500 rpm, respectively. Finally, the eleetorspun PLLA nanofibers were modified by hypothermy plasma and type I collagen was coated onto the materials physically. The resulting rabbit peripheral blood mononuclear cell （PBMC） were collected and resuspended in endothelial basal medium （EBM-2） supplemented with EBM-2-SingleQuots, which were continually cuhttred with complete EBM-2 medium for 3 weeks. To confirm EOC generation during culturing, morphological changes of adherent cells were visualized with Olympus phase-contrast microscopy. The attached Cells were either incubated with Ac-LDL that was labeled with the fluorochrome DiI or incubated with FITC- conjugated UEA-I. Incorporation of DiI-labelled Ac-LDL and binding of F1TC-UEA-I was determined by fluorescent microscopy. First passage EOCs were seeded onto sterilized PLLA nanofther scaffolds in 96-well tissue culture plates. To determine the growth curves, cells were taken from 4 randomly chosen wells for growth each condition every other day from day 3 until day 17. Cells were incubated in WTT （Thiazolyl blue ） and subsequently resuspended in DMSO（ Dimethyl sulfoxide ） with shaking. The absorbance A representing cell density in each well was me

关 键 词：人工血管 组织工程 支架 纳米超级有序膜 内皮生长细胞 

分 类 号：R686[医药卫生—骨科学]