晚期内皮祖细胞在纳米聚-L-乳酸支架膜表面的粘附和增殖  

Experimental Study of Adherency and Proliferation of Late Endothelial Progenitor Cells on Nanofibers Poly-L-Lactic Acid 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,江苏省南京市210008 [2]东南大学生物电子学国家重点实验室,江苏省南京市210096

出　　处：《中国动脉硬化杂志》2008年第10期824-828,共5页Chinese Journal of Arteriosclerosis

摘　　要：目的通过比较晚期内皮祖细胞在纳米无序及有序聚-L-乳酸支架膜表面粘附、增殖情况,为优化组织工程材料提供一种新途径。方法通过静电纺丝技术制备纳米聚-L-乳酸纤维支架,行低温等离子体技术改性及I型胶原表面涂覆,与晚期内皮祖细胞复合培养,并进行细胞双荧光染色鉴定,通过不同时间点两组材料晚期内皮祖细胞粘附和增殖率的测定比较及光镜、荧光显微镜和扫描电镜观察支架材料对种子细胞形态特征等方面的影响。结果制得的纳米聚-L-乳酸纤维孔径在300～400nm之间,孔隙率>90%;细胞粘附12h基本完成;各时间点有序膜组增殖率明显高于无序膜组(P<0.05)。细胞在支架膜上生长良好,无序膜组细胞生长较散在、杂乱;有良好空间定向效果的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。结论晚期内皮祖细胞是理想的组织工程种子细胞来源;纳米聚-L-乳酸有序膜支架能促进种子细胞在材料表面的粘附、增殖,并能较好地保持细胞形态,是一种理想的血管组织工程支架材料。Aim To compare the adhesion and proliferation of late endothelial progenitor cells （EPC） isolated and expanded from rabbit peripheral blood cultured with nanofibers and alignate nanofibers synthetic poly-L-lactic acid （PL- LA） scaffold in vitro for applications in vascular tissue engineering, thus to optimize PLLA scaffold materials. Methotis Nanofibrous scaffolds of PLLA by electrospinrfing were modified by hypothermy plasma and type I collagen was coated onto the materials physically. In vitro, late EPC were cultured on the PLLA scaffold. Furthermore, cell incorporation of DiI-labelled ac-LDL and binding of FITC-UEA-I was determined by fluorescent microscopy. Adherency and proliferation of EPC in two groups at different time points were surveyed and to observe morphological change of late EPC on PLLA scaffold was observed by contrast phase microscope, fluorescent microscope and scanning electron microscope. Results The nanofibrous scaffolds consists of fibers with diameters ranging from 300 nm to 400 nm, whose porosities were more than 90%. The adherency of late EPC on scaffold were completed in 12 hours. The proliferation of late EPC on aliglnate nanofibers scaffold was markedly increased compared to control groups in every time （ P 〈 0.05 ）. Late EPC grew well with PLLA scaffold, yet confused and disorderly in control groups. Late EPC could attach, extend, proliferate and secrete extracellular matrix following fibrous orientation in alignate nanofibers groups of PLLA, which majority of the fibers were oriented along the longitudinal axis which form a unique aligned topography. Conclusions Late EPC are ideal seeding cells for tissue engineering. Late EPC can adhere well to alignate nanofibers of PLLA and proliferate, keep well on cell morphology. This type of PLLA scallfold is a optimal candidate material for late EPC transplantation in vascular tissue engineering.

关 键 词：外科学 聚-L-乳酸 纳米有序纤维 晚期内皮祖细胞 组织工程 

分 类 号：R6[医药卫生—外科学]