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

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.

Activated Carbon-Enriched Electrospun-Produced Scaffolds for Drug Delivery/Release in Biological Systems

“…The viability of human vascular endothelial cells cultured on biodegradable polymer poly(l-lactic acid) scaffolds was 50% lower compared to cells cultivated in TCPS [ 33 ]. The adhesion and proliferation of HUVEC cells on PLGA scaffolds was 30–40% [ 34 ]. According to our data, PCL-based ACN-enriched scaffolds have demonstrated good biocompatibility in comparison with others materials.…”

“…Gingival fibroblasts (GFs) obtained from human gingiva, as described in [ 37 ], were kindly gifted by Dr. Cherepanova. Human umbilical vein endothelial cells (HUVECs) were isolated and cultivated, as described previously [ 34 , 38 ]. Cells were cultured in IMDM (Invitrogen, Carlsbad, CA, USA) medium supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and 100 units of streptavidin/penicillin in a CO 2 incubator at 37 °C and 5% CO 2 .…”

Electrospun Scaffolds Enriched with Nanoparticle-Associated DNA: General Properties, DNA Release and Cell Transfection

“…Since silicates readily bind to DNA and are widely used for DNA isolation [39][40][41], we used silica NPs as pDNA carriers for the production of long-term pDNA-releasing scaffolds. To produce these scaffolds, we used polycaprolactone as a basic polymer combined with 10% gelatin, in order to increase cell adhesion [56]. Previously, we demonstrated that drug-enriched PCL-based scaffolds are well suited for vascular stent coatings, demonstrating satisfactory mechanical properties and biphasic long-term drug delivery [57,58].…”

Electrospun Nano-Fibers for Biomedical and Tissue Engineering Applications: A Comprehensive Review

“…Nowadays, nano-fiber technology advances permit the fabrication of nano-fibers from a wide range of materials including natural and synthetic polymers [200][201][202][203][204][205][206][207][208][209][210][211][212][213], ceramics, metals, as well as inorganic/inorganic or organic/inorganic composite systems [214][215][216][217][218][219][220]. A unique combination of properties such as the conductivity of metallic dopants accompanied by the flexibility of polymers can be obtained by a combination of various materials.…”