Influence of Sub-Lethal and Lethal Concentrations of Chlorhexidine on Morphology and Glucosyltransferase Genes Expression in Streptococcus mutans UA159
Andréa Cristina Barbosa da Silva1*, Rafael Nóbrega Stipp2, Renata de Oliveira Mattos-Graner2, Fábio Correia Sampaio3, Demetrius Antônio Machado de Araújo4
1Graduate Program in Dentistry, Center of Sciences, Technology and Health, State University of Paraiba, UEPB, Campina Grande, Brazil.
2Department of Oral Diagnosis, Piracicaba Dental School, University of Campinas, UNICAMP, Sao Paulo, Brazil.
3Department of Clinical and Social Dentistry, Health Science Center, Brazilian Northeast Network on Biotechnology, Federal University of Paraiba, Joao Pessoa, Brazil.
4Department of Biotechnology, Center of Biotechnology, Brazilian Northeast Network on Biotechnology, Federal University of Paraiba, Joao Pessoa, Brazil.
DOI: 10.4236/aim.2014.413105   PDF   HTML   XML   3,621 Downloads   4,240 Views   Citations


Chlorhexidine (CHX) is regarded as one of the most successful antiplaque agents in controlling the formation of dental biofilm. Nevertheless, molecular mechanisms of their effects in Streptococcus mutans are largely unknown. In this work, the effects of sub-lethal and lethal concentrations of chlorhexidine (CHX) on planktonic or biofilm-organized Streptococcus mutans cells were investigated in dose- and time-dependent manner. The Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) for planktonic cells and biofilm conditions were determined by standard methods. Quantitative PCR (qPCR) was used to quantify the relative levels of glucosyltransferase B (gtfB), gtfC and gtfD transcription of S. mutans in the presence of CHX. The CHX activity in the initial biofilm structure and morphological alterations in planktonic cells were examined by Scanning Electron Microscopy (SEM). The results indicate that CHX increased expression of gtfC and gtfD in planktonic S. mutans cells and CHX reduced the expression of gtfB, gtfC, and gtfD in biofilms. High concentrations of CHX resulted in several wilted S. mutans planktonic cells with spilled intracellular material, while decreased cells’ chain length and matrix was found when the initial biofilm was exposed to increasing concentrations of CHX. CHX’s effects against bacteria depend on the type of growth organization and the concentration and time of exposure to the drug. At sub-lethal concentrations, CHX affects the expression of glucosyltransferases, which may have anticariogenic effect.

Share and Cite:

Silva, A. , Stipp, R. , Mattos-Graner, R. , Sampaio, F. and Araújo, D. (2014) Influence of Sub-Lethal and Lethal Concentrations of Chlorhexidine on Morphology and Glucosyltransferase Genes Expression in Streptococcus mutans UA159. Advances in Microbiology, 4, 945-954. doi: 10.4236/aim.2014.413105.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Lang, N.P. and Brecx, M.C. (1986) Chlorhexidine Gluconate—An Agent for Chemical Plaque Control and Prevention of Gingival Inflammation. Journal of Periodontal Research, 21, 74-89.
[2] Kidd, E.A.M. (1991) Role of Chlorhexidine in the Management of Dental Caries. International Dental Journal, 41, 279-286.
[3] Zickert, I., Emilson, C.G. and Krasse, B. (1982) Effect of Caries Preventive Measures in Children Highly Infected with the Bacterium S. mutans. Archives of Oral Biology, 27, 861-868.
[4] Loesche, W.J. (1986) Role of S. mutans in Human Dental Decay. Microbiological Reviews, 50, 2118-2135.
[5] Silva, A.C.B., Cruz, J.S., Sampaio, F.C. and Araújo, D.A.M. (2008) Detection of Oral Streptococci in Dental Biofilm from Caries-Active and Caries-Free Children. Brazilian Journal of Microbiology, 39, 648-651.
[6] Kuramitsu, H.K. (1993) Virulence Factors of Mutans Streptococci: Role of Molecular Genetics. Critical Reviews in Oral Biology & Medicine, 4, 159-176.
[7] Hamada, S. and Slade, H.D. (1980) Biology, Immunology, and Cariogenicity of S. mutans. Microbiological Reviews, 44, 331-384.
[8] Zanatta, F.B., Antoniazzi, R.P. and Rosing, C.K. (2007) The Effect of 0.12% Chlorhexidine Rinsing in Previously Plaque-Free and Plaque-Covered Surfaces. A Randomized Controlled Clinical Trial. Journal of Periodontology, 78, 2127-2134.
[9] Gjermo, P. and Eriksen, H.M. (1974) Unchanged Plaque Inhibiting Effect of Chlorhexidine in Human Subjects after Two Years of Continuous Use. Archives of Oral Biology, 19, 317-319.
[10] Baehni, P.C. and Takeuchi, Y. (2003) Anti-Plaque Agents in the Prevention of Biofilm-Associated Oral Diseases. Oral Diseases, 9, 23-29.
[11] Gunsolley, J.C. (2006) A Meta-Analysis of Six-Month Studies of Antiplaque and Antigingivitis Agents. Journal of the American Dental Association, 137, 1649-1657.
[12] Hope, C.K. and Wilson, M. (2004) Analysis of the Effects of Chlorhexidine on Oral Biofilm Vitality and Structure Based on Viability Profiling and an Indicator of Membrane Integrity. Antimicrobial Agents and Chemotherapy, 48, 1461-1468.
[13] Tattawasart, U., Hann, A.C., Maillard, J.Y., Furr, J.R. and Russel, A.D. (2000) Cytological Changes in Chlorhexidine-Resistent Isolates of Pseudomonas stutzeri. Journal of Antimicrobial Chemotherapy, 45, 145-152.
[14] Ajdic, D., Mcshan, W.M., Mclaughlin, R.E., Savic, G., Carson, M.B., Primeaux, C., Tian, R., Kenton, S., Jia, H., Lin, S., Qian, Y., Li, S., Zhu, H., Najar, F., Lai, H., White, J., Roe, B.A. and Ferretti, J. (2002) Genome Sequence of Streptococcus mutans UA159, a Cariogenic Dental Pathogen. Proceedings of the National Academy of Sciences of the United States of America, 99, 14434-14439.
[15] Koo, H., Rosalen, P.L., Cury, J.A., Park, Y.K. and Bowen, W.H. (2002) Effects of Compounds Found in Propolis on Streptococcus mutans Growth and on Glucosyltransferase Activity. Antimicrobial Agents and Chemotherapy, 46, 1302-1309.
[16] Stipp, R.N., Goncalves, R.B., Hofling, J.F., Smith, D.J. and Mattos-Graner, R.O. (2008) Transcriptional Analysis of gtfB, gtfC, and gbpB and Their Putative Response Regulators in Several Isolates of Streptococcus mutans. Oral Microbiology and Immunology, 23, 466-473.
[17] Pfaffl, M.W. (2001) A New Mathematical Model for Relative Quantification in Real-Time RT-PCR. Nucleic Acids Research, 29, e45.
[18] Socransky, S.S. and Haffajee, A.D. (2002) Dental Biofilms: Difficult Therapeutic Targets. Periodontology 2000, 28, 12-55.
[19] Davies, D. (2003) Understanding Biofilm Resistance to Antibacterial Agents. Nature Reviews Drug Discovery, 2, 114-122.
[20] Shemesh, M., Tam, A. and Steinberg, D. (2007) Differential Gene Expression Profiling of Streptococcus mutans Cultured under Biofilm and Planktonic Conditions. Microbiology, 153, 1307-1317.
[21] Cury, J. and Koo, H. (2007) Extraction and Purification of Total RNA from Sreptococcus mutans Biofilms. Analytical Biochemistry, 365, 208-214.
[22] Cury, J.A., Seils, J. and Koo, H. (2008) Isolation and Purification of Total RNA from Streptococcus mutans in Suspension Cultures and Biofilms. Brazilian Oral Research, 22, 216-222.
[23] Shemesh, M., Tam, A., Feldman, M. and Steinberg, D. (2006) Differential Expression Profiles of Streptococcus mutans ftf, gtf and vicR Genes in the Presence of Dietary Carbohydrates at Early and Late Exponential Growth Phases. Carbohydrate Research, 341, 2090-2097.
[24] Tam, A., Shemesh, M., Wormser, U., Sintov, A. and Steinberg, D. (2006) Effect of Different Iodine Formulations on the Expression and Activity of Streptococcus mutans Glucosyltransferase and Frutosyltransferase in Biofilm and Planktonic Environments. Journal of Antimicrobial Chemotherapy, 57, 865-871.
[25] Koo, H., Seils, J., Abranches, J., Burne, R.A., Bowen, W.H. and Quivey, R.G. (2006) Influence of Apigenin on gtf Gene Expression in Streptococcus mutans UA159. Antimicrobial Agents and Chemotherapy, 50, 542-546.
[26] Fujiwara, T., Hoshino, T., Ooshima, T. and Hamada, S. (2002) Differential and Quantitative Analysis of mRNA Expression of Glucosyltransferases from Streptococcus mutans MT8148. Journal of Dental Research, 81, 109-113.
[27] Li, Y. and Burne, R.A. (2001) Regulation of the gtfBC and ftf Genes of Streptococcus mutans in Biofilms in Response to pH and Carbohydrate. Microbiology, 147, 2841-2848.
[28] Klein, M.I., De Baz, L., Agidi, S., Lee, H., Xie, G., Lin, A.H.M., et al. (2010) Dynamics of Streptococcus mutans Transcriptome in Response to Starch and Sucrose during Biofilm Development. PLoS ONE, 5, e13478.
[29] Wade, W.G. (2010) New Aspects and New Concepts of Maintaining “Microbiological” Health. Journal of Dentistry, 38, S21-S25.
[30] Marsh, P.D. (2010) Controlling the Oral Biofilm with Antimicrobials. Journal of Dentistry, 38, S11-S15.
[31] Ciancio, S.G. (1995) Chemical Agents: Plaque Control, Calculus Reduction and Treatment of Dentinal Hypersensitivity. Periodontology 2000, 8, 75-85.
[32] Radford, J.R., Homer, K.A., Naylor, M.N. and Beighton, D. (1992) Inhibition of Human Subgingival Plaque Protease Activity by Chlorhexidine. Archives of Oral Biology, 37, 245-248.
[33] Dunne Jr., W.M. (2002) Bacterial Adhesion: Sean Any Good Biofilms Lately? Clinical Microbiology Reviews, 15, 155-166.
[34] Vitkov, L., Hermann, A., Krautgartner, W.D., Herrmann, M., Fuchs, K., Klappacher, M. and Hannig, M. (2005) Chlorhexidine-Induced Ultrastructural Alterations in Oral Biofilm. Microscopy Research and Technique, 68, 85-89.
[35] Hugo, W.B. and Longworth, A.R. (1966) The Effect of Chlorhexidine on the Electrophoretic Mobility, Cytoplasmatic Constituents, Dehydrogenase Activity and Cell Walls of Escherichia coli and Staphylococcus aureus. Journal of Pharmacy and Pharmacology, 18, 569-578.

Copyright © 2021 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.