Recently the practical applications of mussel inspired catechol-containing polymers have been greatly expanded due to their fascinating chemical properties. In this study, we present a facile one-step electrospinning process to synthesize electrospun mussel-inspired copolymer, poly (dopamine methacrylamide-co- methyl methacrylate) (MADO)) with enhanced antibacterial activities after surface functionalization with silver nanoparticles (AgNPs). The assembly of silver nanostructures was driven by the catecholic moiety. The morphologies and structures of the nanofibers were analyzed by field emission scaning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. The nanofibers had a smooth surface with an average diameter of 800 nm. The diameters of the AgNPs were in the range of 15–22 nm. We also evaluated the biocompatibility of the nanofibers, particularly their cytotoxicity to human skin fibroblasts and potential to cause primary skin irritation. The in vitro antibacterial activity and in vivo wound healing capacity of the nanofibers were also investigated. The nanofibers significantly inhibited growth of Escherichia coli bacteria. The nanocomposite nanofibers accelerated the rate of wound healing over that of the control. The results of our in vitro and in vivo animal experiments suggest that catecholic nanofibers show potential to be used as a bioactive wound dressing.