Antimicrobial Efficacy of Calcium Hydroxide with Different Herbal Additives: An In-vitro Study

Abstract

Introduction: Microorganisms such as Enterococcus faecalis, Enterobacteriaceae, and Candida albicans are associated with persisting infections and can cause endodontic failure. The addition of herbal additives such as Triphala, Clove, and Activated charcoal to Calcium hydroxide enhances the microbial efficacy within the root canal. Aim: To evaluate and compare the anti-microbial efficacy of calcium hydroxide with various herbal additives against E. faecalis and C. albicans using Agar Diffusion Test (ADT) and Direct Contact Test (DCT). Materials and Methods: The present in-vitro study was conducted in Bangalore, Karnataka, between July 2023 and September 2023. Ethical clearance was obtained from the institution. Sabouraud Dextrose Agar (SDA) and Mueller-Hinton Agar were prepared and used to culture C. albicans and E. faecalis, respectively. In each plate, four wells were filled with groups (n=10): Group-I: Nanocalcium hydroxide (NCH)+sterile distilled water, Group-II: NCH+Nanoparticle Clove, Group-III: NCH+Nanoparticle Triphala, Group-IV: NCH+nanoparticle activated charcoal. The plates were incubated at 37°C, and zones of inhibition around the wells were measured on the first, third, and seventh day. DCT was evaluated in Colony Forming Units (CFUs)/ml by adding each bacterial suspension to four groups and inoculating on blood agar plates. One-way ANOVA test followed by Tukey’s post-hoc analysis was used to compare the mean Zone Of Inhibition (ZOI). Kruskal-Wallis Test followed by Mann-Whitney post-hoc test was used to compare the mean CFUs. Results: One-way ANOVA test showed that there was a significant difference in the mean ZOI between the four groups at p=0.005. The maximum mean ZOI against C. albicans was achieved by the combination of calcium hydroxide and clove with an average mean of 22.20±1.92 mm, 20±1.87 mm, and 17.20±2.28 mm on the 1st, 3rd, and 7th day, respectively. Calcium hydroxide with distilled water combination showed the highest mean ZOI against E. faecalis with an average mean of 18.20±0.84 mm on day 1, 17±1 mm on the 3rd and 7th day. The mean CFUs of E. faecalis and C. albicans showed a significant difference between the four groups at p<0.001. Conclusion: The addition of herbals to Calcium hydroxide increased anti-microbial activity against Candida albicans and Enterococcus faecalis.

Keywords

• activated charcoal
• candida albicans
• enterococcus faecalis
• triphala