Effect of Herbal Dentifrice Without Fluoride Against Oral Microbial : An in vitro Study

Abstract

Objective: This study aims to evaluate the antimicrobial effects of fluoride-free herbal toothpaste on Candida albicans (C. albicans) and Streptococcus mutans (S. mutans). Despite its limitations, the findings offer valuable insights for both consumers and dental professionals, helping to guide informed decisions in toothpaste selection.Material and Methods: C. albicans ATCC 90028 was cultured in Sabouraud’s dextrose agar, and S. mutans ATCC 25175 was cultured at 37°C with 5% CO₂ for 18 hours. Twenty-seven commercially available herbal toothpaste formulations were diluted with 2% Dimethyl Sulfoxide (DMSO) at a 1:1 ratio and mixed using a vortex mixer. Antibiotic sensitivity testing was performed using SDA for C. albicans and BHI for S. mutans, with results recorded by measuring the diameter of the clear zone. The minimum inhibitory concentration (MIC) was determined using a 96-well plate with resazurin, while the minimum bactericidal concentration (MBC) was assessed by culturing on agar and counting CFUs. Biofilm formation was analyzed using crystal violet staining and OD measurement at 560 nm. All tests were performed in triplicate.Results: The MIC and MBC tests showed no microbial growth, confirming that all 27 herbal toothpaste formulations at a concentration of 1 g/ml effectively inhibited and killed C. albicans. Five formulations (A005, A018, A025, A004, and A023) demonstrated notable antifungal activity. In the biofilm formation assay, all 27 formulations exhibited biofilm formation, while 15 formulations completely inhibited C. albicans and S. mutans biofilm formation.Conclusion: The study confirms the antifungal and antibacterial potential of herbal toothpaste against C. albicans and S. mutans. Variations in effectiveness highlight the need for further research on the active ingredients. Inhibiting biofilm formation is crucial for preventing persistent infections. These findings support the use of herbal toothpaste as a preventive measure for C. albicans and S. mutans-related oral infections.