Antibacterial Effectiveness of Biofilm-Degrading Enzyme, Nano-Silver–Copper Solution, and Quaternary Ammonium Silane as a Three-Step Irrigation Protocol on Enterococcus faecalis: An Ex Vivo Study

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Objective: To compare the three-step irrigation protocol containing extracellular polymeric substance (EPS)-degrading enzyme, nano-silver–copper solution, and quaternary ammonium silane (QAS) versus single-step conventional irrigation against Enterococcus faecalis. Methods: Thirty-six single-rooted teeth were randomly selected in the experimental study, which was conducted between 2022 and 2024. The sample size of 36 was calculated based on G* effect size calculation. The inclusion criteria were intact non-carious teeth with a fully formed apex. Teeth with resorption, gross decay, or an open apex were excluded from this study. After access cavity preparation and root canal preparation, E. faecalis bacteria inoculation and incubation were performed for 1 week. The teeth were allocated into groups: Group 1: normal saline, Group 2: 3% sodium hypochlorite, and Group 3: a three-step irrigation protocol of EPS-degrading enzyme, nano-silver–copper solution, and QAS used separately. After irrigation, paper points were used to collect the sample from the teeth, and quantification was performed using colony-forming units. To compare the study parameters, the tests used were paired t test and one-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. Results: Comparing the three groups, ANOVA and the Tukey post hoc test showed a statistically significant difference between the saline, hypo, and three-step irrigation groups, while there was no statistically significant difference between the hypo and three-step irrigation groups. Conclusion: The results of this study showed that the three-step irrigation protocol containing an EPS-degrading enzyme, nano-silver–copper solution, and QAS was effective as a single-step conventional irrigation against E. faecalis .