Cytotoxicity of Mouthrinses on Epithelial Cells by Micronucleus Test

ABSTRACT

Objectives: To determine the cytotoxicity of three commercial mouthrinses Klorhex, Andorex and Tanflex on buccal epithelial cells using micronucleus (MN) test.

Materials and Methods: 28 patients with aged 16-24 undergone three mouthrinses’ application were analyzed before and after one week exposure. Physiologic saline was used for the control group. The MN incidence was scored in the buccal epithelial of each participants. The difference in pre- and posttreatment after one week incidence of MN and plaque (PI) and gingival indices (GI) was compared by non-parametric statistical tests.

Results: The micronuclei incidence increased in Klorhex, Tanflex and Andorex groups after exposure to mouth rinses (P<.05). But when compared with the control group, there was not any difference between Andorex and control group (P>.05). In the other study groups, MN incidence was significantly increased after 7 days treatment (P<.05). GI scores of all groups were decreased significantly (P<.05). PI scores were decreased only in the Klorhex group (P<.05).

Conclusions: Our primary findings support the presence of possible cytotoxic effects of the mouthrinses on gingival epithelial cells. (Eur J Dent 2007;2:80-85)

Key words

Mouthrinses - Cytotoxicity - Micronucleus test

• References

• 1 Mandel ID. Chemotherapeutic agents for controlling plaque and gingivitis. J Clin Periodontol 1988; 15: 488-498
• 2 Brecx M. Strategies and agents in supragingival chemical plaque control. Periodontol 2000 1997; 15: 100-108
• 3 Hickel R. Wirkstoffe gegen Plaque und Bakterien. Quint Bonusausgabe 1977; 1: 045-057
• 4 Rosin M, Welk A, Kocher T, Majic-Todt A, Kramer A, Pitten FA. The effect of a polyhexamethylene biguanide mouthrinse compared to an essential oil rinse and a chlorhexidine rinse on bacterial counts and 4-day plaque regrowth. J Clin Periodontol 2002; 29: 392-399
• 5 Sreenivasan P, Gaffar A. Antiplaque biocides and bacterial resistance: a review. J Clin Periodontol 2002; 29: 965-974
• 6 Guggenheim B, Regolati B, Schmid R, Mühlemann RH. Effects of the topical application of mutanase on rat caries. Caries Res 1980; 14: 128-135
• 7 Lindquist B, Edward S, Torell P, Krasse B. Effect of different caries preventive measures in children highly infected with mutans streptococci. Scand J Dent Res 1989; 97: 330-337
• 8 Knuuttila M, Söderling E. Effect of chlorhexidine on the release of lysosomal enzymes from cultured macrophages. Acta Odont Scand 1981; 39: 285-289
• 9 Babich H, Wurzburger BJ, Rubin YL, Sinensky MC, Blau L. An in vitro study on the cytotoxicity of chlorhexidine digluconate to human gingival cells. Cell Biol Toxicol 1995; 11: 79-88
• 10 Goldschmidt P, Cogen R, Taubman S. Cytopathologic effects of chlorhexidine on human cells.. J Periodontol 1977; 48: 212-215
• 11 Purcher JJ, Daniel JC. The effects of chlorhexidine digluconate on human fibroblasts in vitro. J Periodontol 1992; 63: 526-532
• 12 Gendron R, Grenier D, Sorsa T, Mayrand D. Inhibition of the activities of matrix metalloproteinases 2, 8, and 9 by chlorhexidine. Clin Diagnos Lab Immunol 1999; 6: 437-439
• 13 Kidd EAM. Role of chlorhexidine in the management of dental caries.. Int Dent J 1991; 41: 279-286
• 14 Emilson CG. Potential efficacy of chlorhexidine against mutans streptococci and human dental caries. J Dent Res 1994; 73: 682-691
• 15 Van Rijkom HM, Truin GJ, Van’t Hof MA. A meta analysis of clinical studies on the caries-inhibiting effect of chlorhexidine treatment. J Dent Res 1996; 75: 790-795
• 16 Baca P, Munoz MJ, Bravo M, Junco P, Baca AP. Effectiveness of chlorhexidine–thymol varnish (Cervitec) for caries reduction in permanent first molars of 6–7-year-old children. 24-month clinical trial. Commune Dent Oral Epidemiol 2002; 30: 363-368
• 17 Matthews RW, Scully CM, Levers BG, Hislop WS. Clinical evaluation of benzydamine, chlorhexidine, and placebo mouthwashes in the management of recurrent aphthous stomatitis. Oral Surg Oral Med Oral Pathol 1987; 63: 189-191
• 18 Heddle JA, Hite M, Kirkhart B, Mavourinin K, Gregor JTM, Newell GW, Salamone MF. The indication of micronuclei as a measure of genotoxicity. Mutat Res 1983; 123: 61-118
• 19 Albertini RJ, Anderson GR, Douglas L, Hagmar K, Hemminki F, Merlo AT, Natarajan H, Norppa DEG, Shuker R, Tice MD, Waters Waters, Aitio A. IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans. Mutat Res 2000; 463: 111-172
• 20 Cairns J. Mutation selection and the natural history of cancer. Nature 1975; 255: 197-200
• 21 Burgaz S, Iscan A, Büyükbingöl ZK, Bozkurt A, Karakaya AE. Evaluation of micronuclei in exfoliated urothelial cells and urinary thioether excretion of smokers. Mutat Res 1995; 335: 163-169
• 22 Lehucher-Michel MP, Favre-Eloff G, Devictor B, Duffaud F, Botta A. Effect of smoking on micronucleated epithelial cells in smears from the uterine cervix.. Carcinogen 1997; 18: 749-753
• 23 Valverde M, del Carmen Lopez, Lopez I, Sanchez ITI, Fortoul P, Ostrosky-Wegman E. Rojas DNA damage in leukocytes and buccal and nasal epithelial cells of individual exposed to air pollution in Mexico City. Environ Mol Mutagen 1997; 30: 147-152
• 24 Salama SA, Serrana M, Au WW. Biomonitoring using accessible human cells for exposure and health risk assessment. Mutat Res 1999; 436: 99-112
• 25 Burgaz S, Çakmak G, Erdem O, Yilmaz M, Karakaya AE. Micronuclei frequencies in exfoliated nasal mucosa cells from pathology and anatomy laboratory workers exposed to formaldehyde. Neoplasma 2001; 48: 144-147
• 26 Burgaz S, Demircigil GC, Yilmazer M, Ertas N, Kemaloglu Y, Burgaz Y. Assessment of cytogenetic damage in lymphocytes and in exfoliated nasal cells of dental laboratory technicians exposed to chromium, cobalt and nickel. Mutat Res 2002; 521: 47-56
• 27 Silness J, Löe H. Periodontal disease in pregnancy. II. Correlation between oral hygiene and periodontal condition. Acta Odontol Scand 1964; 22: 121-135
• 28 Löe H, Silness J. Periodontal disease in pregnancy. I. Prevalence and severity. Acta Odontol Scand 1963; 21: 533-551
• 29 Countryman PI, Heddle JA. The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes.. Mutat Res 1976; 41: 321-332
• 30 Eren K, Ozmeric N, Sardas S. Monitoring of buccal epithelial cells by alkaline comet assay (single cell gel electrophoresis technique) in cytogenetic evaluation of chlorhexidine.. Clin Oral Invest 2002; 6: 150-154
• 31 Wilken R, Botha SJ, Grobler A, Germishuys PJ. In vitro cytotoxicity of chlorhexidine gluconate, benzydamine-HCl and povidone iodine mouthrinses on human gingival fibroblasts. SADJ 2001; 56: 455-460
• 32 Zeiger E. Identification of rodent carcinogens and noncarcinogens using genetic toxicity tests: premises, promises and performance. Regul Toxicol Pharmacol 1988; 28: 85-95
• 33 Kirsch-Volders M, Elhajouji A, Cundari E, Van Hummelen JM. The in vitro micronucleus test: a multi-endpoint assay to detect simultaneously mitotic delay, apoptosis, chromosome breakage, chromosome loss and non-disjunction.. Mutat Res 1977; 392: 19-30
• 34 Miller B, Pötter-Locher F, Seelbach A, Stoper H, Utesch D, Madle S. Evaluation of in vitro micronucleus test as an alternative to the in vitro chromosomal aberration assay: position of the GUM Working Group on the in vitro micronucleus test. Mutat Res 1998; 410: 81-116
• 35 Addy M, Moran J, Wade J. Chemical plaque control in the prevention of gingivitis and periodontitis.. In: Lang NP, Karring T, eds. Proceedings of the 1st European Workshop on Periodontology; London: Quintessence; 1994: 244-257
• 36 Bollen CML, Quirynen M. Microbiological response to mechanical treatment in combination with adjunctive therapy: a review of the literature. J Periodontol 1996; 67: 1143-1150
• 37 Quirynen M, Avondroot P, Peeters W, Pauwels M, Coucke W, Van Steenberghe D. Effect of different chlorhexidine formulations in mouthrinses on de novo plaque formation.. J Clin Periodontol 2001; 28: 1127-1136
• 38 Hidalgo E, Dominguez C. Mechanisms underlying chlorhexidine induced cytotoxicity. Toxicol in Vitro 2001; 15: 271-276
• 39 Waaler SM, Rolla G, Skorland KK, Ogaard B. Effects of oral rinsing with triclosan and sodium lauryl sulphate on dental plaque formation: a pilot study. Scand J Dent Res 1993; 101: 192-195
• 40 Barkvoll P, Rolla G. Triclosan protects the skin against dermatitis caused by sodium lauryl sulphate exposure. J Clin Periodontol 1994; 21: 717-719
• 41 Bonassi M, Fenech C, Lando Y. et al Human micronucleus project: international database comparison for results with the cytokinesis-block micronucleus assay in human lymphocytes. 1. Effect of laboratory protocol, scoring criteria, and host factors on the frequency of micronuclei. Environ Mol Mutagen 2001; 37: 31-45