Title:

Comparative Evaluation of Chitosan, Probiotic, and Herbal Mouthwashes in Terms of Dental Plaque Levels and Antibacterial Effect in Children with High Caries Risk – A Randomized Controlled Trial

Introduction and Aim:

Dental plaque is a major contributor to dental caries, especially in children at high caries risk due to poor oral hygiene, frequent sugar intake, and limited access to care. Mechanical plaque control techniques like brushing and flossing are often inadequately practiced in such populations. Chemical agents such as chlorhexidine are commonly prescribed for plaque control, but their side effects—like staining, altered taste, and mucosal irritation—have led to the search for safer alternatives. Natural agents like chitosan, probiotics, and herbal formulations are gaining popularity for their biocompatibility, antimicrobial effects, and minimal side effects. Chitosan, derived from marine shells, exhibits broad-spectrum antimicrobial action. Probiotics help by maintaining a healthy oral microbiome and reducing acidity, while herbal extracts possess proven antibacterial and anti-inflammatory benefits. This study aims to assess and compare the clinical and antibacterial effects of chitosan, probiotic, and herbal mouthwashes in children with high caries risk.

Methodology and Study Groups:

This double-blinded randomized controlled trial will be conducted in the Departments of Pediatric and Preventive Dentistry, Microbiology, and Pharmacology. Eighty children aged 7 to 12 years, screened from a residential school and identified as high caries risk using a standardized assessment tool, will be included based on specific inclusion and exclusion criteria. Participants will be divided into four equal groups: Group A – no mouthwash (control), Group B – chitosan mouthwash, Group C – probiotic mouthwash, and Group D – herbal mouthwash. The chitosan formulation will consist of 0.5% chitosan, ethanol, glycerine, sodium saccharine, castor oil, and flavoring in deionized water. All mouthwashes will be packed in 100 ml opaque bottles coded for blinding. Participants will rinse twice daily—once in the morning after brushing and once before bedtime—for one month, avoiding food or drink for 30–45 minutes afterward.

Assessment Procedures:

Oral hygiene will be clinically evaluated using the plaque index (Silness and Löe) and gingival index (Löe and Silness) at baseline, 15 days, and 1 month. Each tooth surface (distofacial, facial, mesiofacial, and lingual) will be scored, and mean values will be calculated per individual. For microbiological analysis, plaque samples will be collected at the same time points and transported to the Department of Microbiology within 2 hours. Samples will undergo serial dilution and be inoculated on mitis salivarius-bacitracin agar (MSB) for selective growth of Streptococcus mutans and S. salivarius. Plates will be incubated aerobically at 37°C for 48 hours, after which colony-forming units (CFUs) will be counted using a digital colony counter and expressed as CFUs per agar surface.

Statistical Analysis:

Data will be analyzed using SPSS version 21. Descriptive statistics will include mean and standard deviation. ANOVA followed by Tukey’s post hoc test will be used for intergroup comparisons, with significance set at p<0.05.