Hemolytic, Biocompatible, and Functional Effect of Cellularized Polycaprolactone-Hydrolyzed Collagen Electrospun Membranes for Possible Application as Vascular Implants

In search of bioactive vascular prostheses that exhibit greater biocompatibility through the combination of natural and synthetic polymers, tissue engineering from a biomimetic perspective has proposed the development of three-dimensional structures as therapeutic strategies in the field of cardiovascular medicine. Techniques such as electrospinning allow obtaining of scaffolds that emulate the microarchitecture of the extracellular matrix of native vessels; thus, this study aimed to evaluate the biological influence of microarchitecture on polycaprolactone (PCL) and hydrolyzed collagen (H-Col) electrospun scaffolds, which have a homogeneous (microscale) or heterogeneous (micro-nanoscale) fibrillar structure. The hemolytic, biocompatible, and functional effect of the scaffolds in interaction with an in vitro fibroblast model was determined, in view of its potential use for vascular implants. Scaffolds were characterized by scanning electron microscopy and atomic force microscopy, Fourier transform infrared spectroscopy, wettability, static permeability, tensile test, and degradation. In addition, direct and indirect 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were used to identify the cell viability of fibroblasts, fluorescence assays were performed to establish morphological changes of the cell nuclei, and the hemolytic effect of the scaffolds was calculated. Results showed that ethanol-treated biocompositescaffolds exhibited mass losses lower than 6.65% and slow wettability and absorption, resulting from an increase in secondary structures that contribute to the crystalline phase of H-Col. The scaffolds demonstrated stable degradation in saline during the incubation period because of the availability of soluble structures in aqueous media, and the inclusion of H-Col increased the elastic properties of the scaffold. As regards hemocompatibility, the scaffolds had hemolysis levels lower than 1%; moreover, in terms of biocompatible characteristics, scaffolds exhibited good adhesion, proliferation, and cell viability and insignificant changes in the circularity of the cell nuclei. However, scaffolds with homogeneous fibers showed cell agglomerates after 48 h of interaction. By contrast, permeability decreased as the incubation period progressed, because of the cellularization of the three-dimensional structure. In conclusion, multiscale scaffolds could exhibit a suitable behavior as a bioactive small-diameter vascular implant.