纳米PLLA膜复合内皮生长晕细胞的相容性研究  

Experimental Study of Compatibility of Endothelial Outgrowth Cells Cultured with Nanofibers PLLA Scaffold

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,南京210008 [2]东南大学生物电子学国家重点实验室,南京210096

出　　处：《中国普外基础与临床杂志》2009年第4期269-275,共7页Chinese Journal of Bases and Clinics In General Surgery

基　　金：江苏省卫生厅重大科研资助项目(项目编号:K200609)~~

摘　　要：目的通过观察内皮生长晕细胞（EOCs）与纳米聚-L-NL酸（PLLA）有序纤维膜体外复合培养的生物相容性及黏附、增殖情况，为构建组织修复材料提供理论依据。方法通过静电纺丝技术制备的纳米PLLA纤维支架，行低温等离子体技术改性及I型胶原表面涂覆，与EOCs复合培养，测定细胞黏附率及增殖率，观察细胞生长曲线，荧光显微镜及扫描电镜下观察支架材料与种子细胞EOCs形态特征和生物相容性。结果制备的纳米PLLA膜孔径在300-400nm之间，孔隙率〉90％。各组（单纯细胞组、无序膜组、有序膜组及超级有序膜组）吸光度（A）值均随共培养时间的增加而逐步升高；从第5天起，有序膜和超级有序膜组A值与无序膜和单纯细胞组比较，差异有统计学意义（P〈0．05）；单纯细胞组和无序膜组各检测时间点A值差异无统计学意义（P〉0．05）。复合培养12h及24h后有序膜组和超级有序膜组黏附率则明显高于无序膜组，差异有统计学意义（P〈0．01）。复合培养1d、3d及7d后，有序膜组和超级有序膜组的增殖率明显高于无序膜组（P〈0．05，P〈0．01）。细胞在支架膜上生长良好，纳米无序膜的细胞生长较散在、杂乱；有良好空间定向效果的有序膜及超级有序膜支架有利于细胞沿纤维定向附着、伸展、增殖，以超级有序膜更有利于保持其结构。结论EOCs是理想的组织工程种子来源细胞；纳米PLLA有序及超级有序膜支架能促进种子细胞在材料表面的黏附、增殖，并能较好地保持细胞的形态，是一种理想的组织修复材料。Objective To study the cellular biocompatibility, adhesion and proliferation of endothelial outgrowth cells （EOCs） isolated and expanded from rabbit peripheral blood cultured with aligned poly-L-lactic acid （PLLA） nanofibrous scaffolds in vitro so as to provide a basis for the applications of scaffolds biomaterials in tissue repair. Methods Nanofibrous scaffolds of PLLA by electrostatic spinning were modified by hypothermal plasmas body and type I collagen was coated onto the materials physically. In vitro, EOCs were cultured on the modified PLLA scaffold. Adhesion and proliferation were surveyed and morphological changes and biocompatibility of seeding cells on PLLA scaffold were observed by growth curves of the cells, fluorescent microscope and scanning electron microscope respectively. Results Fibers with diameters ranging from 300 nm to 400 nm were included in the nanofibrous scaffolds, whose porosities were more than 90 %. Absorbance （A） of each scaffold increased gradually after EOCs grew in the absence or presence of random, aligned, or super-aligned PLLA nanofibrous scaffold. Although there was no detectable effect of the random PLLA scaffold on the growth EOCs （P〉0.05）, both aligned and superaligned PLLA nanofibrous scaffold had significantly enhanced their growth since the 5th day （P〈0.05）. The rates of adhesion in both aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA scaffold after 12 h and 24 h incubation （P〈0.01）. The rates of proliferation after 1 d, 3 d and 7 d incubation in aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA nanofibrous scaffold （P〈0.05, P〈0.01）. EOCs grew well with PLLA scaffold, yet confused and disorderly in random nanofibers. EOCs could attach, extend and proliferate following fibrous orientation in aligned and super-aligned PLLA nanofibrous scaffold, in majority of the fibers were oriented along the longitudinal axis so that a unique

关 键 词：支架 聚-L-乳酸 纳米超级有序膜 内皮生长晕细胞 组织修复 

分 类 号：R318.08[医药卫生—生物医学工程]