The antibacterial effect of photodynamic therapy in dental plaque-derived biofilms
C. R. Fontana
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Department of Diagnostic and Surgery, Araraquara Dental School, Sao Paulo State University, Sao Paulo, Brazil
Search for more papers by this authorA. D. Abernethy
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorS. Som
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorK. Ruggiero
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorS. Doucette
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorR. C. Marcantonio
Department of Diagnostic and Surgery, Araraquara Dental School, Sao Paulo State University, Sao Paulo, Brazil
Search for more papers by this authorC. I. Boussios
Laboratory for Information and Decisions Systems, Massachusetts Institute of Technology, Boston, MA, USA
Search for more papers by this authorA. C. R. Tanner
Department of Molecular Genetics, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorN. S. Soukos
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorC. R. Fontana
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Department of Diagnostic and Surgery, Araraquara Dental School, Sao Paulo State University, Sao Paulo, Brazil
Search for more papers by this authorA. D. Abernethy
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorS. Som
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorK. Ruggiero
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorS. Doucette
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorR. C. Marcantonio
Department of Diagnostic and Surgery, Araraquara Dental School, Sao Paulo State University, Sao Paulo, Brazil
Search for more papers by this authorC. I. Boussios
Laboratory for Information and Decisions Systems, Massachusetts Institute of Technology, Boston, MA, USA
Search for more papers by this authorA. C. R. Tanner
Department of Molecular Genetics, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorN. S. Soukos
Applied Molecular Photomedicine Laboratory, The Forsyth Institute, Boston, MA, USA
Search for more papers by this authorAbstract
Background and Objective: Photodynamic therapy has been advocated as an alternative to antimicrobial agents to suppress subgingival species and to treat periodontitis. Bacteria located within dense biofilms, such as those encountered in dental plaque, have been found to be relatively resistant to antimicrobial therapy. In the present study, we investigated the ability of photodynamic therapy to reduce the number of bacteria in biofilms by comparing the photodynamic effects of methylene blue on human dental plaque microorganisms in the planktonic phase and in biofilms.
Material and Methods: Dental plaque samples were obtained from 10 subjects with chronic periodontitis. Suspensions of plaque microorganisms from five subjects were sensitized with methylene blue (25 μg/mL) for 5 min then exposed to red light. Multispecies microbial biofilms developed from the same plaque samples were also exposed to methylene blue (25 μg/mL) and the same light conditions as their planktonic counterparts. In a second set of experiments, biofilms were developed with plaque bacteria from five subjects, sensitized with 25 or 50 μg/mL of methylene blue and then exposed to red light. After photodynamic therapy, survival fractions were calculated by counting the number of colony-forming units.
Results: Photodynamic therapy killed approximately 63% of bacteria present in suspension. By contrast, in biofilms, photodynamic therapy had much less of an effect on the viability of bacteria (32% maximal killing).
Conclusion: Oral bacteria in biofilms are affected less by photodynamic therapy than bacteria in the planktonic phase. The antibacterial effect of photodynamic therapy is reduced in biofilm bacteria but not to the same degree as has been reported for treatment with antibiotics under similar conditions.
References
- 1 Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284: 1318–1322.
- 2 Kroes I, Lepp P, Relman DA. Bacterial diversity within the human subgingival crevice. Proc Natl Acad Sci USA 1999; 96: 14547–14552.
- 3 Kumar PS, Griffen AL, Moeschberger ML, Leys EJ. Identification of candidate periodontal pathogens and beneficial species by quantitative 16S clonal analysis. J Clin Microbiol 2005; 43: 3944–3955.
- 4 Sakamoto M, Umeda M, Benno Y. Molecular analysis of human oral microbiota. J Periodont Res 2005; 40: 277–285.
- 5 Anderson GG, O’Toole GA. Innate and induced resistance mechanisms of bacterial biofilms. Curr Top Microbiol Immunol 2008; 322: 85–105.
- 6 Del Pozo JL, Patel R. The challenge of treating biofilm-associated bacterial infections. Clin Pharmacol Ther 2007; 82: 204–209.
- 7 Fux CA, Costerton JW, Stewart PS, Stoodley P. Survival strategies of infectious biofilms. Trends Microbiol 2005; 13: 34–40.
- 8 Wilson M. Lethal photosensitization of oral bacteria and its potential application in the photodynamic therapy of oral infections. Photochem Photobiol Sci 2004; 3: 412–418.
- 9 Wilson M. Photolysis of oral bacteria and its potential use in the treatment of caries and periodontal disease: a review. J Appl Bacteriol 1993; 75: 299–306.
- 10 Dougherty TJ, Gomer CJ, Henderson BW et al. Photodynamic therapy. J Natl Cancer Inst 1998; 90: 889–905.
- 11 Wilson M, Dobson J, Sarkar S. Sensitisation of periodontopathogenic bacteria to killing by light from a low-power laser. Oral Microbiol Immunol 1993; 8: 182–187.
- 12 Soukos NS, Ximenez-Fyvie LA, Hamblin MR, Socransky SS, Hasan T. Targeted antimicrobial photochemotherapy. Antimicrob Agents Chemother 1998; 42: 2595–2601.
- 13 Williams JA, Pearson GJ, Colles MJ, Wilson M. The effect of variable energy input from a novel light source on the photoactivated bactericidal action of toluidine blue O on Streptococcus mutans. Caries Res 2003; 37: 190–193.
- 14 Sarkar S, Wilson M. Lethal photosensitization of bacteria in subgingival plaque samples from patients with chronic periodontitis. J Periodont Res 1993; 28: 204–210.
- 15 Metcalf D, Robinson C, Devine D, Wood S. Enhancement of erythrosine-mediated photodynamic therapy of Streptococcus mutans biofilms by light fractionation. J Antimicrob Chemother 2006; 58: 190–192.
- 16 Wood S, Metcalf D, Devine D, Robinson C. Erythrosine is a potential photosensitizer for the photodynamic therapy of oral plaque biofilms. J Antimicrob Chemother 2006; 57: 680–684.
- 17 Zanin IC, Lobo MM, Rodrigues LK, Pimenta LA, Hofling JF, Goncalves RB. Photosensitization of in vitro biofilms by toluidine blue O combined with a light-emitting diode. Eur J Oral Sci 2006; 114: 64–69.
- 18 Zanin IC, Goncalves RB, Junior AB, Hope CK, Pratten J. Susceptibility of Streptococcus mutans biofilms to photodynamic therapy: an in vitro study. J Antimicrob Chemother 2005; 56: 324–330.
- 19 Soukos NS, Socransky SS, Mulholland SE, Lee S, Doukas AG. Photomechanical drug delivery into bacterial biofilms. Pharm Res 2000; 17: 405–409.
- 20 Soukos NS, Mulholland SE, Socransky SS, Doukas AG. Photodestruction of human dental plaque bacteria: enhancement of the photodynamic effect by photomechanical waves in an oral biofilm model. Lasers Surg Med 2003; 33: 161–168.
- 21 Ogura M, Abernethy AD, Blissett RD et al. Photomechanical wave-assisted molecular delivery in oral biofilms. World J Microbiol Biotechnol 2007; 23: 1637–1646.
- 22 Socransky SS, Smith C, Martin L, Paster BJ, Dewhirst FE, Levin AE. “Checkerboard” DNA-DNA hybridization. Biotechniques 1994; 17: 788–792.
- 23 Wood S, Nattress B, Kirkham J et al. An in vitro study of the use of photodynamic therapy for the treatment of natural oral plaque biofilms formed in vivo. J Photochem Photobiol B 1999; 50: 1–7.
- 24 Qin Y, Luan X, Bi L et al. Toluidine blue-mediated photoinactivation of periodontal pathogens from supragingival plaques. Lasers Med Sci 2008; 23: 49–54.
- 25 De Rosa MC, Crutchley RJ. Photosensitized singlet oxygen and its applications. Coord Chem Rev 2002; 233: 351–371.
- 26 Harris F, Chatfield LK, Phoenix DA. Phenothiazinium based photosensitizers - photodynamic agents with a multiplicity of cellular targets and clinical applications. Curr Drug Targets 2005; 6: 615–627.
- 27 Wainwright M, Phoenix DA, Gaskell M, Marshall B. Photobactericidal activity of methylene blue derivatives against vancomycin-resistant Enterococcus spp. J Antimicrobial Chemother 1999; 44: 823–825.
- 28 Usacheva MN, Teichert MC, Biel MA. The interaction of lipopolysaccharides with phenothiazine dyes. Lasers Surg Med 2003; 33: 311–319.
- 29 Bartlett JA, Indig GL. Effect of self-association and protein binding on the photochemical reactivity of triarylmethans. Implication of noncovalent interactions on the competition between photosensitization mechanisms Type I and Type II. Photochem Photobiol 1999; 70: 490–498.
- 30 Sedlacek MJ, Walker C. Antibiotic resistance in an in vitro subgingival biofilm model. Oral Microbiol Immunol 2007; 22: 333–339.
- 31 Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP. The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 1998; 280: 295–298.
- 32 Whiteley M, Gita Bangera M, Bumgarner RE et al. Gene expression in Pseudomonas aeruginosa biofilms. Nature 2001; 413: 860–864.
- 33 Bhatti M, MacRobert A, Meghji S, Henderson B, Wilson M. Effect of dosimetric and physiological factors on the lethal photosensitization of Porphyromonas gingivalis in vitro. Photochem Photobiol 1997; 65: 1026–1031.
- 34 Kömerik N, Wilson M. Factors influencing the susceptibility of gram-negative bacteria to toluidine blue-mediated lethal photosensitisation. J Appl Microbiol 2002; 92: 618–623.
- 35 Tegos GP, Hamblin MR. Phenothiazinium antimicrobial photosensitizers are substrates of bacterial multidrug resistance pumps. Antimicrob Agents Chemother 2006; 50: 196–203.
- 36 De Jong MH, Van der Hoeven JS, Van Os JH. Growth of micro-organisms from supragingival dental plaque on saliva agar. J Dent Res 1986; 65: 85–88.
- 37 Feres M, Haffajee AD, Allard K, Som S, Goodson JM, Socransky SS. Antibiotic resistance of subgingival species during and after antibiotic therapy. J Clin Periodontol 2002; 29: 724–735.
- 38 Sissons CH. Artificial dental plaque biofilm model systems. Adv Dent Res 1997; 11: 110–126.
- 39 Som S, Goodson JM, Abernethy A et al. Characterization and validation of a dental plaque microcosm laboratory biofilm. J Dent Res 2004; 83: (Spec. Iss. A):Abstr. 1408.
- 40 Müller P, Guggenheim B, Schmidlin PR. Efficacy of gasiform ozone and photodynamic therapy on a multispecies oral biofilm in vitro. Eur J Oral Sci 2007; 115: 77–80.
- 41 Stewart PS, Grab L, Diemer JA. Analysis of biocide transport limitation in an artificial biofilm system. J Appl Microbiol 1998; 85: 495–500.
- 42 O’Neill JF, Hope CK, Wilson M. Oral bacteria in multi-species biofilms can be killed by red light in the presence of toluidine blue. Lasers Surg Med 2002; 31: 86–90.
- 43 Stewart PS. Diffusion in biofilms. J Bacteriol 2003; 185: 1485–1491.
- 44 Rani SA, Pitts B, Stewart PS. Rapid diffusion of fluorescent tracers into Staphylococcus epidermidis biofilms visualized by time lapse microscopy. Antimicrob Agents Chemother 2005; 9: 728–732.
- 45 Foley I, Gilbert P. Antibiotic resistance of biofilms. Biofouling 1996; 10: 331–346.
- 46 Brown SM, Allison DG, Gilbert P. Resistance of bacterial biofilms to antibiotics: a growth-rate related effect? J Antimicrob Chemother 1988; 22: 777–783.
- 47 Fontana CR, Ruggiero K, Doucette S et al. Phototargeting of Dental Plaque Bacteria in Planktonic Phase Versus Biofilms. J Dent Res 2007; 86 (Spec. Iss. A): Abstr. 2960).
- 48 Andersen R, Loebel N, Hammond D, Wilson M. Treatment of periodontal disease by photodisinfection compared to scaling and root planning. J Clin Dent 2007; 18: 34–38.