Bacterial cellulose (BC) and bacterial cellulose-vaccarin (BC-Vac) membranes were successfully produced in large scale. BC was synthesized by Gluconacetobacter xylinum. BC-Vac membranes were prepared by immersing BC in vaccarin solution. The surface morphologies of BC and BC-Vac membranes were examined by a scanning electron microscope (SEM) and an atomic force microscopy (AFM). The images showed that BC-Vac exhibited the characteristic 3D nanofibrillar network of BC matrix but there was adhesion between fibers. The mechanical properties of BC and BC-Vac membranes were evaluated and the results indicated that the adding of drug vaccarin into the BC membranes increased the malleability indicated by the increment in elongation at break compared with BC. Fourier transform infrared spectroscopy (FTIR) analysis was conducted to confirm the incorporation of vaccarin in BC-Vac and investigate the hydroxyl interactions between BC and drug vaccarin. Cell viability and cell attachment studies demonstrated that BC and BC-Vac membranes had no cytotoxicity and could be a good carrier for cell growth. The wound healing performance was examined in vivo by rat skin models. Histological observations revealed that wounds treated with BC-Vac epithelialized and regenerated faster than treated with BC. Therefore, BC-Vac was considered as a potential candidate for wound dressing materials.