Nanobiotechnology could aid in developing honey-based natural wound healing agents. In the present work, stingless bee honey (SBH) loaded alginate nanoparticles (H-ALG NPs) were formulated with an ionic crosslinking method and thoroughly characterized. The average size of the H-ALG NPs was observed to be 312 ± 4.32 nm, with 0.112 ± 0.04 PDI and - 21.2 ± 0.29 mV Zeta potential. TEM and SEM imaging confirmed the presence of spherical-shaped NPs. Encapsulation efficiency and loading capacity were observed to be 84.74% and 23.12%, respectively. Further, H-ALG NPs showed dose-dependent radical scavenging activity against DPPH with IC50 23.20 compared to 35.99 mg/mL for SBH, suggesting antioxidant potential. The H-ALG NPs were also tested for cytotoxicity on a human dermal fibroblast cell line and exhibited low toxicity with a higher IC50 than SBH (970.07 vs. 755.67 μg/mL; P < 0.001). H-ALG NPs also significantly promoted the migration of fibroblasts as indicated by scratch assay. The qPCR analysis for gene expression demonstrated that H-ALG NPs potentially promote the production of proinflammatory mediators (i.e., TNF-α, IL-6, IL-1β and IL-8) in fibroblasts, which play a crucial role in wound healing, albeit at lower levels compared to SBH. The mouse wound model confirmed that H-ALG NPs accelerated wound healing (P < 0.05) with improved histological characteristics. These findings highlight H-ALG NPs as a potential natural and effective therapeutic agent for wound management and advocate for further exploration of the antioxidant-based medicinal properties of SBH.​