A Pilot Study on the Evaluation of Hetafu Mouthwash in Oral Biofilm of Orthodontic Patients: A Clinical Trial

Introduction Orthodontic patients are at a high risk of plaque buildup and alterations in oral microbiota due to appliance‑related retention sites. Antimicrobial mouth rinses containing bioactive components have been proposed as adjuncts to support oral microbial balance during treatment. This pilot study evaluated the effect of Hetafu mouthwash on selected beneficial and cariogenic bacteria in patients undergoing fixed orthodontic therapy. Methods A prospective clinical trial was conducted on patients undergoing orthodontic treatment at Adhiparasakthi Dental College and Hospital who met the inclusion criteria and provided informed consent. Ethical approval was obtained from the Institutional Ethical Committee of Adhiparasakthi Dental College and Hospital, Melmaruvathur (ethical approval number: ECR/1742/APDCH/ORTHO/FM 05/TN2025). The study included a total of 20 patients. The participants were instructed to use 10 mL of Hetafu mouthwash, rinsing for 30 seconds, two times daily for 240 days without altering their routine oral hygiene practices. Plaque samples were collected at the start of the study, at 60 days, and at 240 days. The samples were cultured on selective media to quantify four target organisms: Streptococcus mutans, Streptococcus oralis (cariogenic bacteria), Lactobacillus sporogenes, and Streptococcus salivarius (beneficial commensals). Colony‑forming units were recorded at each time point, and the data were subjected to statistical analysis. Results Statistical analysis was conducted using SPSS software version 26.0 (IBM Corp., Armonk, NY), with changes in bacterial counts over time analyzed through the Friedman test, followed by post hoc pairwise comparisons performed using Dunn’s test with Bonferroni modification, and significance set at p < 0.05. A significant and progressive reduction was observed in Streptococcus mutans counts across all three time points (p = 0.001), with each interval showing a statistically meaningful decline. Streptococcus oralis also demonstrated a marked decrease over time (p = 0.001), with significant differences between all follow‑ups. In contrast, Streptococcus salivarius exhibited a steady and significant increase, with the highest levels recorded at 240 days (p = 0.001). Lactobacillus sporogenes showed a modest rise, reaching statistical significance only between baseline and the final follow‑up (p = 0.001). Conclusion The use of Hetafu mouthwash for 240 days resulted in a significant reduction in Streptococcus mutans and a marked increase in Streptococcus salivarius, reflecting a shift toward a more favorable oral microbial profile in orthodontic patients. These preliminary findings indicate that the Hetafu mouthwash may serve as a simple adjunct to support oral hygiene during fixed appliance therapy. Future studies with larger sample sizes are necessary to confirm these findings.