Synthesis of silver and zinc oxide nanoparticles using stingless bee honey and their biocompatibility in wound gauze application

This study investigates the synthesis of silver nanoparticles (Ag-NPs), zinc oxide nanoparticles (ZnO-NPs), and bimetallic Ag/ZnO-NPs using stingless bee honey (SBH) as a natural reducing and stabilising agent. Bioactive compounds in SBH, including flavonoids, phenolics, and organic acids, facilitated nanoparticles (NPs) formation. Physicochemical characterisation showed average particle sizes of 30.09 ± 12.36 nm (Ag-NPs), 19.33 ± 4.21 nm (ZnO-NPs), and 33.05 ± 10.63 nm (bimetallic Ag/ZnO-NPs), with spherical morphology. Statistical analysis using one-way ANOVA followed by Tukey’s post-hoc test confirmed significant differences (p < 0.05) in the antimicrobial efficacy of the NPs. Ag-NPs exhibited the highest antibacterial activity, with inhibition zones of 16.91 ± 0.40 mm against Staphylococcus aureus (S. aureus) and 17.43 ± 0.26 mm against Escherichia coli (E. coli), which were significantly greater than those of ZnO-NPs (13.50 ± 0.73 mm and 14.01 ± 0.95 mm) and bimetallic Ag/ZnO-NPs (13.02 ± 0.45 mm and 14.06 ± 0.54 mm). Cytotoxicity assays revealed that Ag-NPs and ZnO-NPs were toxic to HaCaT cells at IC50 values of 10 μg/ml and 85 μg/ml, respectively, whereas bimetallic Ag/ZnO-NPs showed no significant toxicity (p > 0.05). No cytotoxic effects were observed on 3T3 and MRC5 cells up to 100 μg/ml. In vivo wound healing analysis demonstrated that ZnO-NP-treated gauze significantly accelerated wound closure by day 4, achieving a closure rate of up to 51% (p < 0.05). Bimetallic Ag/ZnO-NPs showed consistent healing effects by day 8, with no statistical difference compared to ZnO-NPs at later stages (p > 0.05). Histopathological examination confirmed well-organised skin regeneration, including an intact dermis, keratinocyte proliferation, and adnexal structures, with minimal inflammation. Biochemical and hematological assessments showed no statistically significant differences among rats treated with synthesised NPs, commercial Durafiber Ag, and untreated gauze (p > 0.05). These findings highlight the potential of SBH’s synthesised NPs as innovative wound healing agents, offering statistically significant antimicrobial properties, enhanced wound closure, and biocompatibility. The accelerated healing effects of ZnO-NPs and the sustained performance of bimetallic Ag/ZnO-NPs support their application for various wound types. This study underscores the need for further research to optimise formulations, explore novel biomaterials, and conduct clinical evaluations for practical translation into wound care applications.

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