Evaluating the Effectiveness of Punica granatum as a Natural Dental Plaque Disclosing Agent Against Streptococcus mutans, Lactobacillus and Enterococcus faecalis: An In-vitro Study

Background: The potential health risks associated with synthetic and inorganic dyes have become a growing concern in recent years. As a safer, eco-friendly alternative, Punica granatum (pomegranate) has demonstrated promising antibacterial and antioxidant properties. This study aims to evaluate the effectiveness of Punica granatum as a natural dental plaque-disclosing agent compared to conventional synthetic dyes in staining Streptococcus mutans, Lactobacillus and Enterococcus faecalis. The study also investigates its color spreadability, absorption properties and cytotoxicity levels. Methods: Cultures of Streptococcus mutans, Lactobacillus and E. faecalis were reactivated and maintained for testing. Samples were divided into three groups: Group I (Control-Eosin dye), Group II (Pomegranate extract) and Group III (Pomegranate extract with ethanol). Color spreadability, UV spectroscopy and cytotoxicity using nauplii fish were analyzed. Potential variables such as pH differences, extraction quality and solvent interactions were carefully monitored to ensure result accuracy. Results: UV spectroscopy revealed that Group I exhibited peak absorption at 550 nm (orange-red region), while Groups II and III peaked at 350 nm (UV region). Cytotoxicity assessment showed significantly reduced mortality rates in Groups II (10 deceased nauplii fish) and III (20 deceased nauplii fish) compared to Group I at 40 µg/mL. This suggests a notable reduction in cytotoxicity with the pomegranate-based formulations. Additionally, Group III (pomegranate extract with ethanol) achieved superior staining efficiency, comparable to the control group. Conclusion: Within the study's limitations, Punica granatum demonstrated significant potential as a natural and effective dental plaque-disclosing agent with reduced cytotoxicity. Its eco-friendly profile makes it a viable alternative to conventional dyes. Further clinical studies are recommended to explore its long-term stability, cost-effectiveness and practical application in dental hygiene practices.