This study reports the development and characterization of chitosan–poly(vinyl alcohol) (CH/PVA) nanofibers (NFs), functionalized with bioactive compounds (ACs) relevant for wound healing and tissue regeneration. CH/PVA NFs loaded with L-arginine (ARG), allantoin (ALA), royal jelly (RJ) and curcumin (CUR), either as single or co-loaded systems, were prepared by electrospinning. The polymer solutions were characterized in terms of key physicochemical properties relevant to electrospinning. The CH/PVA@ACs NFs were characterized morphologically and structurally through scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Additionally, surface-related, physical, and functional properties such as wettability, swelling behavior, and in vitro release profiles were examined. The NFs were successfully produced in a uniform and continuous manner, with the fiber diameter and morphology being influenced by the type of ACs. FTIR analysis validated the characteristic functional groups linked to both the polymeric matrix and ACs. The nanofibrous systems demonstrated a high swelling capacity and a release behavior that is dependent on pH. Analyses of surface free energy and wettability revealed favorable interfacial interactions between solid and liquid, indicating compatibility with aqueous biological environments. In summary, the developed CH/PVA@ACs NFs exhibited appropriate morphological, structural, surface, and functional properties, underscoring their potential as effective materials for wound dressings.

Background: Wound healing remains a major clinical challenge, often impaired by persistent inflammation, oxidative stress, and abnormal extracellular matrix remodeling. Electrospun nanofibers (NFs) have emerged as promising wound dressing platforms due to their biomimetic structure and capacity to incorporate multiple bioactive compounds (ACs) with synergistic therapeutic effects. Objectives: This study aimed to biologically assess novel chitosan/poly(vinyl alcohol) (CH/PVA) NFs functionalized with natural active compounds (L-arginine—ARG, allantoin—ALA, royal jelly—RJ, and curcumin—CUR) as multifunctional systems for wound healing and tissue remodeling. Methods: The nanofibrous systems performed the in vitro evaluation of antioxidant activity (DPPH, ABTS, FRAP, PRAP), anti-inflammatory potential (protein denaturation test), hemocompatibility, and cytocompatibility using dermal fibroblasts. In vivo healing performance was evaluated in an excisional wound model using macroscopic wound contraction analysis, histopathology, and immunohistochemical staining (MMP-9, CD31, VEGF-A, α-SMA). Results: The bioactive-enriched CH/PVA NFs exhibited strong antioxidant and anti-inflammatory activity, excellent hemocompatibility (hemolysis < 5%), and excellent cytocompatibility, with promoting fibroblast proliferation. In vivo experiments revealed that the treated groups exhibited accelerated wound closure, improved re-epithelialization, increased angiogenesis, and showed more efficient tissue remodeling compared to the controls, as validated by histological and immunohistochemical studies. Conclusions: The findings indicate that bioactive-enriched CH/PVA NFs serve as effective, biocompatible, and multifunctional matrices for wound healing, hence endorsing their potential for further translational advancement in skin regeneration applications.