Cellulose acetate nanofiber mats containing tetracycline hydrochloride were prepared by electrospinning. Incorporation of tetracycline hydrochloride (20wt% based on the weight of cellulose acetate) in the cellulose acetate solution (17% wt/vol in 2:1vol/vol ethanol-dimethyl sulfoxide) did not affect the morphology of the obtained fibers, as tetracycline hydrochloride-loaded cellulose acetate fibers were smooth. The average diameters of these fibers ranged between 50 and 90nm. Determination of the release characteristics of tetracycline hydrochloride from the cellulose acetate nanofiber mats was carried out by total immersion in phosphate buffer solution, that is, in the total immersion method, the maximum amounts of the tetracycline hydrochloride released from the cellulose acetate nanofiber into the medium were 10 and 25%mu g/100ml. Antibacterial activity of prepared tetracycline hydrochloride-loaded as-spun cellulose acetate fiber mate was achieved using Escherichia coli and Staphylococcus aureus as a representative each of Gram-negative and Gram-positive bacteria, respectively. Antibacterial activity of tetracycline hydrochloride-loaded as-spun cellulose acetate fiber mat showed effectiveness with 77-88% bacteria reduction after 10min-1h for Escherichia coli and approximately 83-85% reduction after 10min-1h for Staphylococcus aureus.