Aim: To develop a novel stabilizing agent for silver nanoparticles (AgNPs) with the aim of enhancing its antibacterial efficacy against wound associated pathogens while mitigating their cytotoxic effect on human cells. Materials & methods: In this study, monodispersed gelatin nanoparticles were synthesized to stabilize AgNPs. The stability, antibacterial activity and biocompatibility of the gelatin-stabilized AgNPs (Gel-AgNPs) were compared with citrate-stabilized AgNPs (citrate-AgNPs) or silver ions. Results & conclusion: Gelatin-stabilized AgNPs showed significantly better antibacterial activities compared with citrate-stabilized AgNPs against both Gram-positive and Gram-negative bacteria. These Gel-AgNPs showed significantly lower cytotoxicity to human dermal fibroblasts compared with Ag⁺. These findings provided the first evidence substantiating a novel functionality of gelatin nanoparticles in both stabilizing and enhancing the activity of AgNPs.