Background:Enterococcus faecalis is a persistent organism that plays a major role in the etiology of persistent periradicular lesions after root canal treatment has been associated with different forms of periradicular disease including primary endodontic infections and persistent infections. The present study compares the antibacterial activities of calcium hydroxide, calcium hydroxide mixed with silver zeolite, and calcium hydroxide mixed with 2% chlorhexidine against E. faecalis using direct contact test.Materials and Methods:The test materials of the in vitro experimental study were grouped as group 1—calcium hydroxide mixed with sterile water, group 2—2% silver zeolite added in calcium hydroxide mixed with sterile water, and group 3—calcium hydroxide mixed with 2% chlorhexidine. The bottom of microtiter plate were coated with freshly mixed tested material and a 10 μL of bacterial suspension was placed. After 1 h of incubation at 37°C, brain–heart infusion (BHI) broth (245 μL) was added and mixed for 2 min. These were designated as “subgroup 1” wells. A volume of 15 μL of broth then transferred from subgroup 1 wells to an adjacent set of four wells containing fresh BHI medium (215 μL); these wells were designated as “subgroup 2”’ wells. The optical density was measured by a spectrophotometer after the first day, third day, and seventh day. One-way analysis of variance (ANOVA) and Tukey tests were performed for the analysis.Results:Calcium hydroxide mixed with silver zeolite showed maximum antibacterial activity.Conclusion:Silver zeolite can be added in calcium hydroxide to enhance the latter's antibacterial activity against E. faecalis.