<p>Delayed and impaired wound healing is a persistent issue, these wounds are prone to infections and suffer from poor cell re-epithelization. Nanoparticles can be utilized to aid in the wound healing process, they are able to carry and deliver biomolecules and drugs into wounds. Due to their small sizes, they can deeply penetrate tissues. This study reports the development of a dual-delivery system of chitosan nanoparticles (CSNPs) co-encapsulating epidermal growth factor (EGF) and vancomycin (VCM) for potential application in wound healing. The nanoparticles were fabricated using the ionotropic gelation method with tripolyphosphate anions (TPP) and characterized for their physicochemical properties including the size, shape, polydispersity index, and zeta potential of the nanoparticles. High-performance liquid chromatography (HPLC) was utilized to measure the encapsulation efficiency of the nanoparticles and to evaluate the release kinetics. The dual-loaded CSNPs exhibited a mean particle size of 458.39 nm and a high positive zeta potential. The encapsulation efficiency and in vitro release kinetics were quantified by HPLC. The formulation demonstrated a sustained release profile for both EGF and VCM. The nanoparticles showed no significant cytotoxicity against human dermal fibroblasts (HDFs) at concentrations up to 4 mg/ml and exhibited a minimum inhibitory concentration (MIC) of 2 mg/ml against <i>Staphylococcus aureus</i>. Furthermore, the dual-loaded nanoparticles significantly enhanced cell migration in an in vitro scratch assay, indicating their potential to accelerate wound closure. This work highlights the promise of dual-loaded chitosan nanoparticles as a novel therapeutic strategy to simultaneously promote tissue regeneration and combat bacterial infections in wounds.</p> Graphical abstract <p></p>