Effect of Herbal Toothpaste Without Fluoride Against Candida Albicans and Streptococcus Mutans: An In Vitro Study
Abstract
Objective: This study aimed to evaluate the antimicrobial effects of herbal toothpaste without fluoride on Candida albicans ATCC 90028 (C. albicans ATCC 90028) and Streptococcus mutans ATCC 25175 (S. mutans ATCC 25175). Despite limitations, such as the discrepancy between in vitro conditions and the human oral environment, the findings have provided valuable insights for consumers and dental professionals, guiding informed decisions in toothpaste selection.
Material and Methods: C. albicans ATCC 90028 was cultured in Sabouraud’s dextrose agar (SDA), and S. mutans ATCC 25175 was cultured in brain heart infusion broth (BHI) at 37 °C with 5% CO₂ for 18 hours. The bacteria suspension was adjusted to the turbidity standard according to the 0.5 McFarland standard (1.5×10⁸ colony forming unit (CFU)/ml). Twenty-seven commercially available herbal toothpaste formulations were diluted with 2% Dimethyl Sulfoxide at a 1:1 ratio and mixed using a vortex mixer. Antimicrobial sensitivity testing was performed using SDA for C. albicans and BHI for S. mutans. The results were 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 colony forming units. Biofilm formation was analyzed using crystal violet staining and the optical density measurement at 560 nm. All tests were performed in triplicate.
Results: All 27 herbal toothpaste formulations tested at 1 g/ml effectively inhibited and killed C. albicans ATCC 90028 and S. mutans ATCC 25175 in the MIC and MBC tests. In the antimicrobial sensitivity testing, 5 formulations (A005, A018, A025, A004, and A023) showed notable inhibition zones against C. albicans, with A005, A018, and A025 demonstrating superior antifungal activity. The biofilm formation inhibition assay indicated that 15 formulations completely inhibited biofilm formation. All formulations successfully inhibited acid production by S. mutans. In patient saliva tests, most herbal Fluoride-free formulations effectively inhibited S. mutans growth, while some demonstrated reduced antibacterial and antifungal activity. Formulations A018 and A025 showed the highest overall efficacy.
Conclusion: The study confirms the antifungal potential of herbal toothpaste formulations against C. albicans ATCC 90028 and S. mutans ATCC 25175. Some formulations were more effective than others, highlighting the need for further investigation into their active ingredients and mechanisms of action. The ability to inhibit biofilm formation is particularly valuable in preventing persistent infections. These findings support the potential use of herbal toothpaste as a preventive measure against C. albicans and S. mutans-related oral infections.
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