The present study was done in order to examine the durability of the tetracycline-induced antimicrobial capacity, and also to assess the reproducibility of the bacterial growth-inhibitory assay used. Standardized enamel and dentin specimens were impregnated in aqueous solutions of tetracycline HCl, oxytetracycline HCl or doxycycline HCl, rinsed in water, and stored dry for 200 days. Another series of specimens was impregnated in solutions of doxycycline HCl and then rinsed in tap water for varying periods up to 35 days. In addition, drug-impregnated specimens were used for reproducibility tests without storage or prolonged rinsing. Impregnated specimens were tested for antimicrobial capacity on agar plates seeded with S. sanguis. After 24 h aerobic incubation in 10% CO2 atmosphere, the plates were inspected and the diameter of the bacterial growth inhibition zones measured. The drug-impregnated enamel and dentin specimens consistently demonstrated growth-inhibitory capacity. The results of the reproducibility tests showed moderate intrasample and day-to-day variation. Two hundred days of dry storage or 35 days soaking in water reduced, but did not eliminate, the bacterial growth-inhibitory capacity of the impregnated dental specimens. The results show that a short-term exposure of dental hard tissues to tetracyclines may result in a long-lasting antibacterial capacity.