Comparative Evaluation Of Effect Of Three Different Mouthwashes On Salivary Streptococcus mutans And Streptococcus salivarius Count And Plaque Level In Children Between Age 8-10 Years: A Clinical Study

TITLE:-

COMPARATIVE EVALUATION OF EFFECT OF THREE DIFFERENT MOUTH WASHES ON SALIVARY STREPTOCOCCUS MUTANS AND STREPTOCOCCUS SALIVARIUS COUNT AND PLAQUE LEVEL IN CHILDREN BETWEEN AGE 8-10 YEARS: A CLINICAL STUDY

INTRODUCTION:-

Oral hygiene and it’s persistent maintenance by children has always been a challenge for the family and dental professionals. The dental caries and the plaque accumulation leads to imbalanced dental health. The main cause of dental caries is attributed to dental plaque. The oral flora consists more than 350 cultivable species. Historically, natural products have been used since ancient times and in the folklore for treatment of many diseases and illnesses. Various products such as turmeric, camphor, clove oil are sought to relieve pain caused due to odontogenic infections, whereas pomegranate, cocoa husk extract, ginger, neem, tulsi, amla, triphala and aloe-vera, are proven to reduce S. mutans(Streptococcus Mutans) counts when used as mouth rinses. Natural, organic, and herbal mouth rinses do not contain alcohol, artificial preservatives, or colours and flavours, and have unique therapeutic properties. Hence, they are attaining popularity among today’s relatively more aware consumers. S. salivarius(Streptococcus salivarius) is one of the first colonizers of the human oral cavity and gut after birth and therefore may contribute to the establishment of immune homeostasis and regulation of host inflammatory responses. Various agents have been used to control dental caries forming bacteria as well as for plaque reduction. These include dental varnishes, mouthwash, toothpaste, dentifrices and other mechanical agents too. Mouthwashes have been found to be one of the safe and effective delivery system as anti-microbial and antiplaque agents. These mouthwashes are capable of inhibiting bacterial adhesion, colonization and metabolic  activity which ultimately affects bacterial growth. Amongst all, chlorhexidine as been the gold standard mouthwash. It is a cationic bisguanide, with a broad-spectrum antibacterial effect by virtue of its high intra oral substantivity and bactericidal and bacteriostatic activity. However, prolonged use of chlorhexidine has led to tooth discolouration in many children so, various herbal extract mouthwashes has been attempted to help reduce the incidence of dental caries and plaque accumulation. One of the recent herbal products is Punica granatum, which literally translates to seeded (“granatus”) apple (“pomum”). Punica granatum Linn belongs to family Punicaceae, mostly known as “pomegranate.” It is a shrub native from Asia where several of its parts have been used as an astringent, haemostatic, and for diabetes control. The genus name, Punica, was the Roman name for Carthage, where the best pomegranates were known to grow. The incorporation of cocoa powder or chocolate into hamster diets was reported to reduce caries. Subsequently it was suggested that phenolic substances could be responsible for the observed anti-caries effect of cocoa powder. The cocoa bean husk is a by product material generated in the chocolate industry. It has been shown to possess two types of cariostatic sub stances, one showing anti-glycosyltranferase activity and the other antibacterial activity. In the literature there are no studies evaluating the comparative effects of cocoa bean husk, pomegranate and chlorhexidine mouthwash in reduction of S. mutans and S. salivarius in saliva and its clinical effect on dental plaque formation in young children. The purpose of this study is to compare the effects of cocoa bean husk, pomegranate and chlorhexidine mouthwash on levels of Streptococcus mutans and streptococcus salivarius in the saliva and its clinical effect on dental plaque formation of the children between age of 8 to 10 years old.

METHODOLOGY:-

 Study Design:

Type: Clinical observational study

Duration: 18 months (12 months data collection + analysis)

Setting: Departments of Pediatric Dentistry, Microbiology, and Pharmaceuticals

 Participants:

Age Group: 8–10 years

Sample Size: 30 children (10 per group)

Sampling: Random allocation using AI-generated table

Groups:

Group A: 0.2% Chlorhexidine (positive control)

Group B: 0.1% Cocoa Bean Husk Extract (CBHE)

Group C: 38% Hydroalcoholic Pomegranate Extract

 Eligibility Criteria:

Inclusion: Healthy children with cooperative behavior, no recent antibiotics, no active caries or oral appliances

Exclusion: Special needs, recent surgery or fluoride treatment, known allergies, or lack of consent

 Study Procedure:

Phase I – Microbial Evaluation:

Saliva Collection: Baseline & after 7 days in 3 stages (with 15-day washout)

Testing: Saliva cultured on Mitis Salivarius agar for S. mutans and S. salivarius CFU count

Tools: Sterile cups, culture plates, CFU counter

Phase II – Clinical Plaque Evaluation:

Index Used: Simplified Oral Hygiene Index (OHI-S)

Tools: Mouth mirror and explorer

Procedure: Plaque and calculus scored on 6 teeth surfaces; OHI-S calculated at same intervals as saliva

 Preparation of Mouthwashes:

CBHE: Extracted using ethanol and lyophilized; sweetened with saccharin

Pomegranate: Hydroalcoholic extract from crushed fruit, filtered and concentrated

 Mouthwash Use Instructions:

10 ml, twice daily for 7 days per stage

No food/drink for 30–45 minutes post-use

Blinded bottles coded for data masking

Latin square design used for sequence control

And then Streptococcus mutans count and Streptococcus salivarius count will be assessed and analysed at baseline and after intervention