Abstract

This study reports the development and characterization of hierarchical electrospun scaffolds based on poly (L-lactic acid) (PLA) and polyacrylonitrile (PAN) reinforced with calcium oxide (CaO) nanoparticles (18.5 +/- 4.7 nm) for skin regeneration. Six configurations, including two five-layer multilayer systems (PLA/PAN/CaO and PAN/PLA/CaO), were evaluated to determine how composition and deposition sequence influence physicochemical, mechanical, and biological performance. FT-IR, XRD and DSC confirmed the successful integration of CaO, while thermal analysis evidenced an effect of chain mobility and interfacial interactions within multilayer systems. Cross-sectional SEM revealed the presence of both fibers with continuous interfaces. Nitrogen adsorption showed that CaO significantly increased the specific surface area (e.g., from 4.6 m(2)/g in neat PLA to 21.65 m(2)/g in PLA/CaO), with type IV isotherms indicating mesoporosity. Wettability assays demonstrated reduced contact angle in PLA (from 126.3 degrees to 91.8 degrees) and sequence-dependent surface properties in multilayers. Tensile testing confirmed that the multilayer architecture bridged the mechanical gap between compliant PLA and high-strength PAN, yielding intermediate moduli (similar to 10-11 MPa) and balanced toughness. Antibacterial assays against S. aureus and E. coli showed that CaO significantly reduced bacterial viability, with PLA/PAN/CaO achieving the highest inhibition (up to 37.1%). In vitro HaCaT assays and in vivo implantation in BALB/c mice confirmed high cytocompatibility and biocompatibility. These findings demonstrate that multilayer electrospinning of PLA/PAN/CaO enables the design of structurally integrated, bioactive, and mechanically balanced scaffolds for advanced wound healing and dermal repair.