Assement of a radiosterilized polycaprolactone-gelatin-alginate membrane as a scaffold for dermal fibroblast.

Abstract

Introduction. The development of temporary skin substitutes has become a topic of great interest due to their potential to improve burn wound healing, as well as to reduce inflammation, promote cell proliferation, and tissue regeneration. Consequently, various synthesis methods have been developed, with electrospinning being a novel technique for developing interconnected, nanofibrous, and porous scaffolds. It has been reported that combining two or more polymers can improve cell biocompatibility and mechanical properties to mimic the native extracellular matrix of the skin. Objective. We propose to evaluate a radiosterilized polycaprolactone-gelatin-alginate membrane as a scaffold for dermal fibroblasts. Methodology. The electrospinning solutions were prepared individually: PCL 60% w/v dissolved in acetic acid, GEL 25% w/v in formic acid, and ALG 15% w/v in DSMO and formic acid. The fibrillar membrane was obtained by electrospinning at 20 kV and 0.5 mL/h. Once obtained, the membranes were radio-sterilized. The morphology of the electrospun fibers was observed by Scanning Electron Microscopy (SEM). The chemical structure of the membranes was determined by Fourier Transform Infrared Spectroscopy (FTIR). To determine the biological response, dermal fibroblasts were seeded onto the three electrospun membranes (PCL, PCL-GEL, and PCL-GEL-ALG), respectively, and analyzed using the Calcein Am assay (Invitrogen®). Cell proliferation was determined using an immunofluorescence assay for Ki67. Results. We standardized the synthesis of an electrospun PCL-GEL-ALG membrane, which was radio-sterilized. This process did not alter the membrane's characteristics. The membranes were white, flexible, resistant to manipulation, and smooth to the touch, with thicknesses of approximately 0.8 mm (PCL), 0.86 mm (PCL-GEL), and 0.89 mm (PCL-GEL-ALG), respectively. SEM micrographs revealed a homogeneous fibrillar morphology with random orientation and interconnected porosity. We also determined that there were no chemical changes in the overall structure of the membranes. The cell viability assay showed that dermal fibroblasts seeded onto the membranes remained viable for up to seven days. The immunofluorescence assay confirmed that dermal fibroblasts seeded onto the PCL-GEL-ALG membrane were able to support cell proliferation for seven days. In conclusion, we can state that radiosterilization did not alter the characteristics of the electrospun membrane, keeping dermal fibroblasts viable and proliferating.