Biofabrication of Leucas aspera-Mediated Chitosan–Zinc Oxide Nanocomposites for In Vitro Antioxidant, Antibacterial, Anti-Inflammatory and Wound-Healing Properties

<b>Background/Objectives</b>: Nanostructured biomaterials based on natural polymers have gained increasing attention in pharmaceutics due to their biocompatibility, multifunctionality, and diverse biomedical applications. This novel study aimed to biofabricate chitosan-doped zinc oxide nanocomposites (CS-ZnONCs) using <i>Leucas aspera</i> leaf extract and to evaluate their physicochemical properties and in vitro biomedical performance. <b>Methods</b>: CS-ZnONCs were synthesized using <i>L. aspera</i> leaf extract through a green precipitation approach, and the resulting nanocomposites were characterized by various spectroscopic techniques. The in vitro antioxidant, antibacterial, and anti-inflammatory activities were evaluated, while wound-healing potential was assessed using L929 fibroblast cell migration assays. <b>Results</b>: UV-visible analysis confirmed the formation of CS-ZnONCs, with a characteristic absorption peak at 362 nm, and FTIR spectra indicated the presence of various important functional groups. XRD results demonstrated the crystalline nature of ZnO within the chitosan matrix. Well-dispersed, quasi-spherical nanoparticles with an average size of 44 ± 3.1 nm were identified by HR-TEM, and a positive zeta potential (+9 mV) suggested considerable colloidal stability. CS-ZnONCs showed a high swelling capacity (88 ± 2.75% for 2%) and significant phytocompound release (65.38 ± 2.79% at pH 7.4). The CS-ZnONCs showed significant antioxidant activity (ABTS of 88.19 ± 1.59%), notable antibacterial efficacy against <i>Staphylococcus aureus</i> (18.78 ± 0.98 mm) and <i>Escherichia coli</i> (17.14 ± 0.96 mm), and significant anti-inflammatory activity (82.12 ± 1.47% membrane stabilization). In vitro biocompatibility and wound-healing assays revealed significant cytocompatibility in Vero cells, with 98.75 ± 1.17% cell viability observed, whereas the fibroblast migration assay demonstrated near-complete wound closure (96.55 ± 6.46%). <b>Conclusions</b>: The green-synthesized CS-ZnONCs exhibit favorable physicochemical properties, biocompatibility, and multifunctional biological activities, supporting their potential as a promising sustainable biomaterial nanomedicine for pharmaceutical formulations, wound healing, and regenerative medicine applications.