In this study, a series of nanofibrous membranes were prepared from cellulose acetate (CA) and polyester urethane (PEU) using coelectrospinning or blend-electrospinning. The drug release, in vitro antimicrobial activity and in vivo wound healing performance of the nanofiber membranes were evaluated for use as wound dressings. To prevent common clinical infections, an antimicrobial agent, polyhexamethylene biguanide (PHMB) was incorporated into the electrospun fibers. The presence of CA in the nanofiber membrane improved its hydrophilicity and permeability to air and moisture. CA fibers became slightly swollen upon contacting with liquid phase. CA not only increased the liquid uptake but also created a moist environment for the wound, which accelerated wound recovery. PHMB release dynamics of the membranes was controlled by the structure and component ratios of the membranes. The lower ratio of CA: PEU helped to preserve the physical and thermal properties of the membranes, and also reduced the burst release effectively and slowed down diffusion of PHMB during in vitro tests. The controlled-diffusion membranes exerted long-term antimicrobial effect for wound healing. (c) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 100B: 15561565, 2012.