The process of wound healing is composed of coagulation, inflammation, fibroplasia, collagenation, epithelization, and wound contraction. The wound dressing should protect the wound from bacterial infection, maintain a moist healing environment, and promote cell migration to reconstruct damaged tissue, and be easy to apply and remove to improve patient comfort. The purpose of our study was to develop multifunctional hydrogels composed of genipin-crosslinked biodegradable biomaterials of poly(gamma-glutamic acid) and gelatin, encapsulating gentamicin to accelerate wound healing. The results of swelling ratio measurements clearly indicate that hydrogel composition of poly(gamma-glutamic acid) gelatin had a higher swelling ratio and lower peel adhesion properties than gelatin hydrogel alone. In an in vitro study, the gentamicin incorporated in prepared hydrogels effectively inhibited target microorganisms and produced a higher expression of Type I collagen in fibroblast cells. Confocal laser scanning microscopy revealed that the fibroblast cells cultured in the hydrogel membranes produced fibroblast cell migration and showed a continuous lined cytoskeletal distributing status. In the in vivo study, it was found that the gentamicin incorporated in genipin-crosslinked gamma-PGA gelatin wound dressing demonstrates the potential of such biologically functionalized dressing to accelerate wound closure and, hence, its potential clinical usefulness. (C) 2010 Wiley Periodicals, Inc. J Appl Polym Sci 120: 1057-1068, 2011