Silver sulfadiazine (SSD) is a useful antimicrobial agent for wound treatment. However, recent findings indicate that conventional SSD cream has several drawbacks for use in treatments. Bacterial cellulose (BC) is a promising material for wound dressing due to its outstanding properties of holding water, strength and degradability. Unfortunately, BC itself exhibits no antimicrobial activity. A combination of SSD and BC is envisaged to form a new class of wound dressing with both antimicrobial activity and biocompatibility, which has not been reported to date. To achieve antimicrobial activity, SSD particles were impregnated into BC by immersing BC into SSD suspension after ultrasonication, namely SSD-BC. Parameters influencing SSD-BC impregnation were systematically studied. Optimized conditions of sonication time for no less than 90 min and the proper pH value between 6.6 and 9.0 were suggested. The absorption of SSD onto the BC nanofibrous network was revealed by XRD and SEM analyses. The SSD-BC membranes exhibited significant antimicrobial activities against Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus evaluated by the disc diffusion method. In addition, the favorable biocompatibility of SSD-BC was verified by MTT colorimetry, epidermal cell counting method and optical microscopy. The results demonstrate the potential of SSD-BC membranes as a new class of antimicrobial and biocompatible wound dressing.