An in vitro regenerated functional human endothelium on a nanofibrous electrospun scaffold
An in vitro regenerated functional human endothelium on a nanofibrous electrospun scaffold
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DOI：
10.1016/j.biomaterials.2010.02.017
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发表时间：
2010-05-01
期刊：
影响因子：
14
通讯作者：
Vohra, Yogesh K.
中科院分区：
文献类型：
--
作者：
Zhang, Xing;Thomas, Vinoy;Vohra, Yogesh K.
使用人主动脉内皮细胞（HAECs）研究了由明胶、弹性蛋白、聚己内酯（PCL）和聚卡普隆（PGC）组成的电纺双层支架的腔层促进内皮再生的能力。将不同密度的HAEC在安装在细胞冠上的支架的腔层的薄膜上培养期望的时间段。荧光图像显示HAECs在成功粘附到支架表面后24 h内形成单层。扫描电子显微镜（SEM）显示HAECs的覆盖令人满意。用荧光染色法测定HAECs的死亡率在最初的3天内低于5%，而用3-[4，5-二甲基（噻唑-2基）-3，5-二苯基]四唑溴化盐（MTT）法测定HAECs的增殖曲线在11天内呈指数增加。内皮单层的功能通过紧密连接形成的ZO-1染色、前列环素（PGI（2））分泌的6-酮-PGF（1 α）测定和人血小板的抗血栓形成能力进行探测。结果表明，再生内皮细胞具有天然内皮细胞的正常功能。这项研究表明，这种电纺双层支架是一个有前途的候选人，心血管移植的能力，促进功能性内皮细胞的再生，以防止小直径移植物中的血液凝固。(C)2010爱思唯尔有限公司版权所有。
The capacity of the luminal layer of an electrospun bi-layer scaffold composed of gelatin, elastin, polycaprolactone (PCL), and poliglecaprone (PGC) to promote endothelial regeneration was investigated using human aortic endothelial cells (HAECs). HAECs of different densities were cultured on a thin film of the luminal layer of the scaffold mounted on a cell crown for desired periods. Fluorescent images showed that HAECs formed a mono-layer within 24 h after having successfully adhered to the scaffold's surface. Scanning electron microscopy (SEM) revealed a satisfactory coverage by the HAECs. Death rates of HAECs populations determined by fluorescent staining were below 5% within the initial 3 days while the profile of proliferation exhibited an exponential increase within 11 days as determined by the 3-[4,5-dimethyl (thiazol-2y1)-3,5-diphery] tetrazolium bromide (MTT) assay. The functionalities of the endothelial monolayer were probed by ZO-1 staining for tight junction formation, by 6-keto-PGF(1 alpha), assay for prostacyclin (PGI(2)) secretion, and by human platelets for its anti-thrombotic capability. The results indicated that the regenerated endothelium possessed normal functions associated with native endothelium. This study suggests that this electrospun bi-layer scaffold is a promising candidate for cardiovascular grafting for its capability of promoting the regeneration of a functional endothelium to prevent blood clotting in small diameter grafts. (C) 2010 Elsevier Ltd. All rights reserved.