Published: 2019-10-22

Effectiveness of chitosan mouthwash on plaque formation and levels of salivary Streptococcus and Lactobacillus count: an in vivo study

Aparna Indu S. R., Sharan S. Sargod, Sham S. Bhat, Shamjith Manikkoth, N. Ramakrishnan


Background: The objectives of the present study were to clinically evaluate the effectiveness of chitosan mouthwash on Plaque formation and to evaluate the antimicrobial activity against salivary Streptococcus mutans (SM) and Lactobacillus (LB).

Methods: A total of 20 participants with decay-missing-filled index ≥4 and simplified oral hygiene index score >1.3 were recruited for the study after taking informed consent. All the participants were provided a bottle of chitosan mouthwash in addition to their usual oral hygiene procedure. Baseline salivary SM and LB levels were determined, using chair-side CRT® bacteria test prior giving chitosan mouthwash and after 14 days, during which children are advised to use 10 ml of mouthwash for 60 seconds two times daily. Plaque index score recorded using Silness and Loe plaque index. Paired t-test (or corresponding non-parametric) and percentage comparison method using cross tables were used for statistical analysis.

Results: The baseline plaque index score of 0.758 and after 14 days of mouthwash use, it reduced to 0.434. The mean difference in the plaque index showed significant reduction in the plaque score (i.e., with the difference of 0.324). The results showed the antiplaque effects with a short duration of 14 days use of chitosan (CH) mouthwash. The antibacterial activity demonstrated a range of inhibitory effect on salivary SM and LB.

Conclusions: CH showed an evident strong effect against salivary SM and LB levels and controlling the plaque biofilm formation. So water soluble CH can be used in new formulations for oral applications not only as antimicrobial agent but also for plaque biofilm control.


Chitosan, SM, LB, Mouthwash, Plaque, In vivo

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Archana V, Prabhuji ML, Karthikeyan BV, Selvan A. Control of Streptococcus sanguinis oral biofilm by novel chlorhexidine-chitosan mouthwash: an in vitro study. J Exp Integrative Med. 2013;3(2).

Sano H, Shibasaki Ki, Matsukubo T, Takaesu Y. Effect of chitosan rinsing on reduction of dental plaque formation. Bulletin Tokyo Dent Coll. 2003;44(1):9-16.

Chen CY, Chung YC. Antibacterial effect of water-soluble chitosan on representative dental pathogens Streptococcus mutans and Lactobacilli brevis. J Appl Oral Sci. 2012;20(6):620-7.

Gupta SC, Kapoor VK. Fundamentals of mathematical statistics. Delhi, India: Sulthan Chand and Sons Publications Pvt Ltd; 2002.

Nagappan N, John J. Antimicrobial efficacy of herbal and chlorhexidine mouth rinse: a systematic review. J Dent Med Sci. 2012;2(4):5-10.

Van Leeuwen MP, Slot DE, Van der Weijden GA. Essential oils compared to chlorhexidine with respect to plaque and parameters of gingival inflammation: a systematic review. J Periodontol. 2011;82(2):174-94.

Flotra L, Gjermo Pe, Rolla G, Waerhaug J. Side effects of chlorhexidine mouth washes. Europ J Oral Sci. 1971;79(2):119-25.

Kowitz GM, Lucatorto FM, Cherrick HM. Effects of mouthwashes on the oral soft tissues. J Oral Med. 1976;31(2):47-50.

Gagari E, Kabani S. Adverse effects of mouthwash use: a review. Oral Surg Oral Med Oral Pathol Oral Radiol Endodontics. 1995;80(4):432-9.

Aliasghari A, Khorasgani MR, Vaezifar S, Rahimi F, Younesi H, Khoroushi M. Evaluation of antibacterial efficiency of chitosan and chitosan nanoparticles on cariogenic streptococci: an in vitro study. Iran J Microbiol. 2016;8(2):93.

Stamford TC, Stamford-Arnaud TM, de Medeiros Cavalcante HM, Macedo RO, de Campos-Takaki GM. Microbiological chitosan: Potential application as anticariogenic agent. In Practical Applications Biomed Engineer; 2013.

Weaver A, Fleming SM, Smith DB. Mouthwash and oral cancer: carcinogen or coincidence?. J Oral Surg (Am Dent Assoc: 1965). 1979;37(4):250-3.

Wynder EL, Kabat GC. Oral cancer and mouthwash use: reply. J Natl Cancer Institute. 1983;71(6):1105.

Blot WJ, Winn DM, Fraumeni JF. Oral cancer and mouthwash. J Natl Cancer Institute. 1983;70(2):251-3.

Mashberg A, Barsa P, Grossman ML. A study of the relationship between mouthwash use and oral and pharyngeal cancer. J Am Dent Assoc (1939). 1985;110(5):731-4.

Kabat GC, Hebert JR, Wynder EL. Risk factors for oral cancer in women. Cancer Res. 1989;49(10):2803-6.

Winn DM, Blot WJ, McLaughlin JK, Austin DF, Greenberg RS, Preston-Martin S, et al. Mouthwash use and oral conditions in the risk of oral and pharyngeal cancer. Cancer Res. 1991;51(11):3044-7.

Lemos-Junior CA, Villoria GE. Reviewed evidence about the safety of the daily use of alcohol-based mouthrinses. Brazil Oral Res. 2008;22:24-31.

Massey CC, Shulman JD. Acute ethanol toxicity from ingesting mouthwash in children younger than age 6, 1989-2003. Pediatric Dentist. 2006;28(5):405-9.

Costa EM, Silva S, Madureira AR, Cardelle-Cobas A, Tavaria FK, Pintado MM. A comprehensive study into the impact of a chitosan mouthwash upon oral microorganism's biofilm formation in vitro. Carbohydrate Polymers. 2014;101:1081-6.

Rahmani F, Moghadamnia AA, Kazemi S, Shirzad A, Motallebnejad M. Effect of 0.5% Chitosan mouthwash on recurrent aphthous stomatitis: a randomized double-blind crossover clinical trial. Electronic physician. 2018;10(6):6912.

Shah HP, Bhavsar NV, Chavda MG. Comparative evaluation of clinical efficacy of chlorhexidine, phenolic and chitosan mouth rinses on plaque and gingivitis: single-centre, double blind, randomized controlled clinical study. Austin J Dent. 2017; 4(1):1064.

De Carvalho MM, Stamford T, Pereira E, Dos Santos P, Sampaio F. Chitosan as an oral antimicrobial agent. Formatex. 2011;2012(1):13.

Uraz A, Boynuegri D, Ozcan G, Karaduman B, Uc D, Senel S, et al. Two percent chitosan mouthwash: A microbiological and clinical comparative study. J Dent Sci. 2012;7(4):342-9.

Twetman L, Twetman S. Comparison of two chair-side tests for enumeration of Streptococci mutans in saliva. Oral Health Dent Manag. 2014;13(3):580-3.

Walsh LJ, Tsang AK. Chairside testing for cariogenic bacteria: current concepts and clinical strategies. Int Dentistr South Africa. 2008;10(2):50-65.

Dutta PK, Dutta J, Tripathi VS. Chitin and chitosan: chemistry, properties and applications. J Scientific Industrial Res. 2004;63:20-31.

Decker EM, Von Ohle C, Weiger R, Wiech I, Brecx M. A synergistic chlorhexidine/chitosan combination for improved antiplaque strategies. J Periodont Res. 2005;40(5):373-7.