Microstructural Effects of Melt Electrowritten-Reinforced Hydrogel Scaffolds for Engineering Thick Skin Substitutes.

Engineering thick skin tissue substitutes resembling the physiochemical and mechanical properties of native tissue is a significant challenge. Melt electrowriting (MEW) is a powerful technique with the capability of fabricating highly ordered structures with fine fiber diameters, closely replicating the native extracellular matrix (ECM). In this study, we constructed melt electrowritten porous polycaprolactone (PCL) scaffolds with three different geometries by depositing fibers at 0-90 and 60-120° in a mesh structure and in a honeycomb-like orientation to assess the effects of the microstructure on the mechanical strength of the scaffold and cellular behavior. These scaffolds were subsequently infilled with gelatin hydrogel, encapsulating human skin dermal fibroblasts (HSFs) and human umbilical vein endothelial cells (HUVECs). Mechanical tensile tests revealed that the honeycomb microstructure of the hybrid PCL/gelatin scaffold exhibited greater elongation at failure, along with an acceptable elastic modulus suitable for skin tissue applications. All scaffolds provided a cytocompatible microenvironment that maintained over 90% cell viability and preserved typical cell morphology. HSFs were guided through the PCL fibers to the apical surface, while HUVECs were distributed within the gelatin hydrogel within the hybrid structure. Additionally, HSFs' alignment was regulated by the scaffold geometry. Notably, the expression of CD31 in HUVECs─a key transmembrane protein for capillary formation─increased significantly over a 14 day incubation period. Among those, 0-90° mesh and honeycomb geometries showed the greatest effects on the upregulation of CD31. These findings demonstrate that the microstructural guidance of HSFs and their interaction with HUVECs in hybrid structures play a crucial role in promoting vascularization. In conclusion, the honeycomb MEW-gelatin hybrid scaffold demonstrates significant potential for effectively replicating both the mechanical and physicochemical properties essential for full-thickness skin tissue substitutes.