An in vitro regenerated functional human endothelium on a nanofibrous electrospun scaffold.
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-2yl)-3,5-diphery] tetrazolium bromide (MTT) assay. The functionalities of the endothelial mono-layer were probed by ZO-1 staining for tight junction formation, by 6-keto-PGF(1alpha) 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.
Zhang
Xing
X
Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Thomas
Vinoy
V
Xu
Yuanyuan
Y
Bellis
Susan L
SL
Vohra
Yogesh K
YK
D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D004549:Elastin / Q000737:chemistry / Q000378:metabolism; D055664:Electrochemical Techniques; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005780:Gelatin / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008422:Materials Testing; D057139:Nanofibers / Q000737:chemistry*; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism; D012038:Regeneration / Q000502:physiology*; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
D001672:Biocompatible Materials; D011091:Polyesters; C016240:polycaprolactone; D005780:Gelatin; D004549:Elastin