General Study and Gene Expression Profiling of Endotheliocytes Cultivated on Electrospun Materials.

Alena O Stepanova,Vera S Chernonosova,Georgiy Yu Shevelev,Alexander M Karaskov,Anna V Cherepanova,Petr P Laktionov,Pavel P Laktionov,Ivan A Zaporozhchenko

doi:10.3390/ma12244082

Abstract

Endothelization of the luminal surface of vascular grafts is required for their long-term functioning. Here, we have cultivated human endothelial cells (HUVEC) on different 3D matrices to assess cell proliferation, gene expression and select the best substrate for endothelization. 3D matrices were produced by electrospinning from solutions of poly(D,L-lactide-co-glycolide) (PLGA), polycaprolactone (PCL), and blends of PCL with gelatin (Gl) in hexafluoroisopropanol. Structure and surface properties of 3D matrices were characterized by SEM, AFM, and sessile drop analysis. Cell adhesion, viability, and proliferation were studied by SEM, Alamar Blue staining, and 5-ethynyl-2’-deoxyuridine (EdU) assay. Gene expression profiling was done on an Illumina HiSeq 2500 platform. Obtained data indicated that 3D matrices produced from PCL with Gl and treated with glutaraldehyde provide the most suitable support for HUVEC adhesion and proliferation. Transcriptome sequencing has demonstrated a minimal difference of gene expression profile in HUVEC cultivated on the surface of these matrices as compared to tissue culture plastic, thus confirming these matrices as the best support for endothelization.

Highlights

Endothelization of the surfaces of cardio-vascular implants is necessary for their long-term functioning
The matrices were prepared from PCL, PCL with Gl and PLGA by ES in conditions listed in PCL blended with Gl (PCL-Gl) 3D matrices positively correlated with Gl concentration up to 10%, but further increase had no significant effect on cell adhesion
XPS demonstrated that the concentration of Gl on the surface of the fibers in these matrices was no less than 21%, and most of the surface-exposed Gl was tightly bound to the fibers [44]

Summary

Introduction

Endothelization of the surfaces of cardio-vascular implants is necessary for their long-term functioning. Seeding of the surfaces with endothelium is as important as mechanical compliance and the capability for long-term functioning without loss of stiffness [1]. Ineffective endothelization is considered as the primary cause of low long-term patency of small diameter vascular grafts (VG) and Materials 2019, 12, 4082; doi:10.3390/ma12244082 www.mdpi.com/journal/materials. A number of studies have demonstrated that the absence of functional endothelial layer can reduce hemocompatibility, induce blood clotting and intensive neointima growth in the lumen of small diameter VG [4]. Malformation or malfunctioning of the endothelial layer can induce systemic or local inflammation with subsequent fibrosis or calcinosis of the damaged area [5]. Endothelization is affected not just by the surface and the structural properties of the material

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