Bacterial cellulose (BC) has attracted increasing attention as a novel wound dressing material, but its antimicrobial activity, which is one of the critical skin-barrier functions in wound healing, is not sufficient for use in practical applications. To overcome such a deficiency, silver nanoparticles were generated and self-assembled on the surface of BC nanofibers, forming a stable and evenly distributed Ag nanoparticle coated BC nanofiber (AgNP-BC). The performance of AgNP-BC was systematically studied in terms of antibacterial activities, cytocompatibility and effects on wound healing. The results showed that AgNP-BC exhibited significant antibacterial activity against Staphylococcus aureus. Moreover, AgNP-BC allowed attachment, and growth of rat fibroblasts with low cytotoxicity emerged. Based on these advantages, AgNP-BC samples were applied in a second-degree rat wound model. Wound flora showed a significant reduction during the healing. The fresh epidermal and dermis thicknesses with AgNP-BC samples were 111 and 855 µm respectively, higher than 74 and 619 µm for BC groups and 57 and 473 µm for untreated control wounds. The results demonstrated that AgNP-BC could reduce inflammation and promote scald wound healing.