纳米PLLA有序膜制备及其对内皮生长晕细胞的促进作用  

Fabrication of Alignate Nanofibers PLLA Scaffold to Enhance the Growth of Endothelial Outgrowth Cells

作　　者：卢辉俊[1] 冯章启[2] 刘晨[1] 乔彤[1] 黄佃[1] 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：《中国生物医学工程学报》2009年第3期449-455,共7页Chinese Journal of Biomedical Engineering

基　　金：江苏省卫生厅重大科研项目资助项目(K200609)

摘　　要：通过观察内皮生长晕细胞（EOCs）在纳米PLLA有序膜表面黏附、增殖的情况,为优化组织工程材料提供一种新途径。通过静电纺丝技术制备的纳米PLLA纤维支架,进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。采用细胞生长曲线和光镜、荧光显微镜及扫描电镜观察支架材料对种子细胞黏附、增殖、形态特征等方面的影响。结果显示：制得的纳米PLLA纤维孔径为300~400 nm,孔隙率〉90%;有序膜和超级有序膜组吸光度A值与无序膜、单纯细胞组有显著性差异（P〈0.05）;细胞在支架膜上生长良好,纳米无序膜细胞生长较散在、杂乱;有良好空间定向效果的有序纤维及超级有序纤维支架有利于细胞沿纤维定向附着、伸展、增殖,分泌胞外基质,而超级有序膜更有利于保持其结构。内皮生长晕细胞是理想的血管组织工程种子细胞来源;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,是一种理想的血管组织工程支架材料。The purpose of this study is to observe the adhesion, extension and proliferation of endothelial outgrowth cells （EOCs） isolated and expanded from rabbit peripheral blood cultured with alignate nanofibers PLLA scaffold in vitro for vascular tissue engineering. Nanofibrous scaffolds of PLLA by electrospinning were modified by hypothermy plasma treatment and type I collagen coating. EOCs were cultured on the modified PLLA scaffold. The cells attachment, proliferation and morphological change of EOCs in the scaffold were examined by growth curves of the cells, contrast phase microscope, fluorescent microscope and scanning electron microscope. The nanofibrous scaffolds comprised of fibers with diameters of 300 - 400 nm, porosity was more than 90 % . There were significant difference of absorbance between aligned nanofibers group, super-aligned nanofibers group, random nanofibers group, and control group （ P 〈 0.05 ）. EOCs exhibited seperate and disorder morphology in the random nanofibrous scaffold, while attached, extended, proliferated and secreted extracellular matrix on the aligned nanofibrous scaffold, as well as oriented along the direction of the nanofibers. The super-aligned nanofibrous scaffold displayed the strongest function of keeping morphology of the cells. EOCs can adhere to aligned nanofibrous scaffold, super-aligned nanofibrous scaffold of PLLA. The cells proliferate and keep morphology well on the scaffolds. Thus the two types of scallfolds are candidates for EOCs transplantation in vascular tissue engineering.

关 键 词：支架 PLLA 纳米超级有序膜 内皮生长晕细胞 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程] R322[医药卫生—基础医学]