The development of wound dressing materials for effortless and easy treatment is crucial to accelerate the wound healing process. It is particularly preferred that the material used for this purpose is biocompatible. In this study, controlled release of the most commonly used antibiotic, mupirocin, against Staphylococcus aureus (ATCC 25923), gram-positive bacteria species and one of the leading causes of severe human infections in both health and community settings, has been studied. Polyvinyl alcohol (PVA) and sodium alginate (NaAlg) biopolymers were used to prepare microspheres. Microspheres were prepared by emulsion crosslinking method and crosslinked with calcium chloride (CaCl2). The prepared microspheres were covered with ethyl cellulose (EC) to produce wound dressing materials. Antibacterial activities and swelling behaviors of prepared biomaterials were determined. The effects of PVA/NaAlg ratio, mupirocin amount and membrane thickness on mupirocin release were investigated. The release of mupirocin increased with the increasing PVA/NaAlg ratio from 1/2 to 2/1. At the end of 100 h, the highest release of mupirocin was found to be 86% for the PVA/NaAlg ratio 2/1, drug/polymer ratio 1/2 (C1 formulated) microspheres. The surface morphology and structure of the wound dressing material were investigated using Scanning Electron Microscopy and Fourier Transform Infrared Spectroscopy.