Subinhibitory Concentrations of Triclosan Promote Streptococcus mutans Biofilm Formation and Adherence to Oral Epithelial Cells

Triclosan is a general membrane-active agent with a broad-spectrum antimicrobial activity that is commonly used in oral care products. In this study, we investigated the effect of sub-minimum inhibitory concentrations (MICs) of triclosan on the capacity of the cariogenic bacterium Streptococcus mutans to form biofilm and adhere to oral epithelial cells. As quantified by crystal violet staining, biofilm formation by two reference strains of S. mutans was dose-dependently promoted, in the range of 2.2- to 6.2-fold, by 1/2 and 1/4 MIC of triclosan. Observations by scanning electron microscopy revealed the presence of a dense biofilm attached to the polystyrene surface. Growth of S. mutans in the presence of triclosan at sub-MICs also increased its capacity to adhere to a monolayer of gingival epithelial cells. The expression of several genes involved in adherence and biofilm formation in S. mutans was investigated by quantitative RT-PCR. It was found that sub-MICs of triclosan significantly increased the expression of comD, gtfC, and luxS, and to a lesser extent of gtfB and atlA genes. These findings stress the importance of maintaining effective bactericidal concentrations of therapeutic triclosan since sub-MICs may promote colonization of the oral cavity by S. mutans.

[1]  B. Nyvad,et al.  The Role of Bacteria in the Caries Process , 2011, Journal of dental research.

[2]  Y. Shimabukuro,et al.  Human gingival epithelial cells produce chemotactic factors interleukin-8 and monocyte chemoattractant protein-1 after stimulation with Porphyromonas gingivalis via toll-like receptor 2. , 2004, Journal of periodontology.

[3]  E. Mylonakis,et al.  Infective endocarditis in adults. , 2001, The New England journal of medicine.

[4]  B. Wondimu,et al.  Effect of triclosan on interleukin-1 beta production in human gingival fibroblasts challenged with tumor necrosis factor alpha. , 1998, European journal of oral sciences.

[5]  H. Schweizer Triclosan: a widely used biocide and its link to antibiotics. , 2001, FEMS microbiology letters.

[6]  T. Phan,et al.  Triclosan inhibition of membrane enzymes and glycolysis of Streptococcus mutans in suspensions and biofilms. , 2006, Canadian journal of microbiology.

[7]  S. Jankowski,et al.  Effects of subinhibitory concentrations of amikacin and ciprofloxacin on the hydrophobicity and adherence to epithelial cells of uropathogenic Escherichia coli strains. , 2007, International journal of antimicrobial agents.

[8]  Mingyun Li,et al.  Effect of nicotine on growth and metabolism of Streptococcus mutans. , 2012, European journal of oral sciences.

[9]  C. Mateo,et al.  Membranotropic effects of the antibacterial agent Triclosan. , 2001, Archives of biochemistry and biophysics.

[10]  R. Utsumi Bacterial signal transduction : networks and drug targets , 2008 .

[11]  L. Ni,et al.  Effects of sub-minimum inhibitory concentrations of antimicrobial agents on Streptococcus mutans biofilm formation. , 2012, International journal of antimicrobial agents.

[12]  S. Genovese,et al.  The plant coumarins auraptene and lacinartin as potential multifunctional therapeutic agents for treating periodontal disease , 2012, BMC Complementary and Alternative Medicine.

[13]  R. Davies,et al.  The effectiveness of a toothpaste containing triclosan and polyvinyl-methyl ether maleic acid copolymer in improving plaque control and gingival health: a systematic review. , 2004, Journal of clinical periodontology.

[14]  H. Koo,et al.  Extracellular Polysaccharides Matrix — An Often Forgotten Virulence Factor in Oral Biofilm Research , 2009, International Journal of Oral Science.

[15]  S. Ahn,et al.  A Hypothetical Protein of Streptococcus mutans Is Critical for Biofilm Formation , 2005, Infection and Immunity.

[16]  R. Carriveau,et al.  Effects of subinhibitory concentrations of ciprofloxacin on Staphylococcus saprophyticus adherence and virulence in urinary tract infections. , 2012, Journal of endourology.

[17]  A. Volpe,et al.  The comparative anticaries efficacy of a dentifrice containing 0.3% triclosan and 2.0% copolymer in a 0.243% sodium fluoride/silica base and a dentifrice containing 0.243% sodium fluoride/silica base: a two-year coronal caries clinical trial on adults in Israel. , 2001, The Journal of Clinical Dentistry.

[18]  R. Lamont,et al.  Streptococcus Adherence and Colonization , 2009, Microbiology and Molecular Biology Reviews.

[19]  Eugene Rosenberg,et al.  Adherence of bacteria to hydrocarbons: A simple method for measuring cell‐surface hydrophobicity , 1980 .

[20]  M. Parsek,et al.  Subinhibitory Concentrations of Azithromycin Decrease Nontypeable Haemophilus influenzae Biofilm Formation and Diminish Established Biofilms , 2007, Antimicrobial Agents and Chemotherapy.

[21]  W. Wade,et al.  Antibacterial activity of some triclosan-containing toothpastes and their ingredients. , 1992, Journal of periodontology.

[22]  Michael J. MacCoss,et al.  Aminoglycoside antibiotics induce bacterial biofilm formation , 2005, Nature.

[23]  Mel Rosenberg,et al.  Microbial cell surface hydrophobicity , 1990 .

[24]  R. Doyle,et al.  Subinhibitory concentrations of antibiotics affect cell surface properties of Streptococcus sobrinus , 1995, Journal of bacteriology.

[25]  J. Hacker,et al.  Effect of Subinhibitory Antibiotic Concentrations on Polysaccharide Intercellular Adhesin Expression in Biofilm-Forming Staphylococcus epidermidis , 2000, Antimicrobial Agents and Chemotherapy.

[26]  T. Modéer,et al.  Triclosan reduces prostaglandin biosynthesis in human gingival fibroblasts challenged with interleukin-1 in vitro. , 1996, Journal of clinical periodontology.

[27]  B. Nyvad,et al.  Caries Ecology Revisited: Microbial Dynamics and the Caries Process , 2008, Caries Research.

[28]  L. Majtánová,et al.  In vitro effect of subinhibitory concentrations of antibiotics on biofilm formation by clinical strains of Salmonella enterica serovar Typhimurium isolated in Slovakia , 2008, Journal of applied microbiology.

[29]  D. Senadheera,et al.  Quorum sensing and biofilm formation by Streptococcus mutans. , 2008, Advances in experimental medicine and biology.

[30]  Ya-Hsiung Shieh,et al.  Streptococcus mutans autolysin AtlA is a fibronectin‐binding protein and contributes to bacterial survival in the bloodstream and virulence for infective endocarditis , 2009, Molecular microbiology.