Electrospun Multilayer Scaffolds Based on Poly (L-Lactic Acid) and Poly (Acrylonitrile) Reinforced with CaO Nanoparticles for Enhanced Skin Regeneration and Wound Healing

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 m2/g in neat PLA to 21.65 m2/g in PLA/CaO), with type IV isotherms indicating mesoporosity. Wettability assays demonstrated reduced contact angle in PLA (from 126.3° to 91.8°) 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 (~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.