Design, Characterization, and Enhanced Performance of Electrospun Chitosan-Based Nanocomposites Reinforced with Halloysite Nanotubes and Cerium Oxide Nanoparticles for Wound Healing Applications.

The development of advanced wound dressings that integrate favorable physico-mechanical properties with the ability to support physiological healing processes remains a critical challenge in biomaterials science. An ideal dressing should modulate the wound microenvironment, prevent infection, maintain hydration, and possess adequate strength and elasticity. This study aimed to fabricate and characterize electrospun chitosan (CS)-based 3D scaffolds dual-reinforced with halloysite nanotubes (HNTs) and cerium oxide nanoparticles (CeONPs) to enhance material properties and biological performance. HNTs were incorporated to improve electrospinnability and provide mechanical reinforcement, while CeONPs were added for their redox-modulating and anti-inflammatory activities. Composite mats were fabricated via non-capillary electrospinning. The individual and synergistic effects of HNTs and CeONPs were systematically evaluated using physico-chemical methods (SEM, EDX, WAXS, TGA, mechanical testing) and biological assays (in vitro cytocompatibility with mesenchymal stem cells, in vivo biocompatibility, and wound healing efficacy in a rat model). Scaffolds containing only HNTs exhibited defect-free nanofibers with an average diameter of 151 nm, whereas the dual-filler (CS-PEO-HNT-CeONP) composites showed less uniform fibers with a rough surface and a larger average diameter of 233 nm. The dual-filler system demonstrated significantly enhanced mechanical properties, with a Young's modulus nearly double that of pure CS mats (881 MPa vs. 455 MPa), attributed to strong interfacial interactions. In vivo, the CS-PEO-HNT-CeONP scaffolds degraded more slowly, promoted earlier formation of a connective tissue capsule, and elicited a reduced inflammatory response compared to single-filler systems. Although epithelialization was temporarily delayed, the dual-filler composite ultimately facilitated superior tissue regeneration, characterized by a more organized, native-like collagen architecture. The synergistic combination of HNTs and CeONPs within a CS matrix yields a highly promising scaffold for wound management, offering a unique blend of tailored biodegradability, enhanced mechanical strength, and the ability to guide healing towards a regenerative rather than a fibrotic outcome, particularly for burns and traumatic injuries.