Bacterial cellulose (BC), a natural polymer with unique physical and mechanical properties, has several applications in the biomedical field, including drug loading and controlled drug delivery. For this study, a Box-Behnken experimental design was employed as a statistical tool to optimize the release of a model drug, amoxicillin, from BC membranes. Independent variables studied were the concentration of the drug (X1), the concentration of glycerol (X2) and the concentration of a permeation enhancer (X3). From the variables studied, drug concentration had the highest effect on drug release. Among the other independent variables, th linear and quadratic X2 terms, the linear X3 term and the interaction term X2X3 were found to affect the release of amoxicillin from bacterial cellulose membranes.