Skip to main content

Advertisement

SpringerLink
Log in

Photomechanical wave-assisted molecular delivery in oral biofilms

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Photomechanical waves (PW), the product of an intense light beam interaction with a target material, enhance molecular delivery across biological membranes and skin. The ability to deliver methylene blue (MB), a fluorescent probe and photosensitizer, into bacterial biofilms was demonstrated by applying PW on saliva-derived multi-species biofilms that were developed on agar surfaces in 24-well plates. PW were generated with a Q-switched Nd:YAG laser and were directed into the biofilms in the presence of 25 μg/ml MB. The biofilms were then irradiated with red light at 665 nm. After illumination, adherent bacteria were scraped and spread over the surface of blood agar plates. Survival fractions were calculated by counting bacterial colonies. Microbial analysis was performed via a colony lift method and a DNA checkerboard assay using whole genomic probes to 40 oral microorganisms. Visual analysis by confocal scanning laser microscopy demonstrated that the application of PW enhanced the penetration depth of MB in biofilms. Exposure to MB, PW and light led to a significant reduction of the mean levels of log10 CFU counts compared with the group that received MB and light (P = 0.006). The DNA checkerboard assay showed some benefit from PW-assisted phototargeting in 25 biofilm microorganisms relative to phototreatment alone. Our data provide a basis for further exploration and optimization of PW parameters for complete eradication of microorganisms in oral microcosm biofilms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abernethy A, Som S, Ruggiero K, Doukas AG, Soukos NS (2005) Photodestruction of dental plaque bacteria in planktonic phase versus biofilms. J Dental Res 84 (abstract)

  • Bartlett JA, Indig GL (1999) 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 70:490–498

    Article  CAS  Google Scholar 

  • Brown SM, Allison DG, Gilbert P (1988) Resistance of bacterial biofilms to antibiotics: a growth-rate related effect? J Antimicrob Chemother 22:777–783

    Article  CAS  Google Scholar 

  • Carmen JC, Roeder BL, Nelson JL, Ogilvie RL, Robison RA, Schaalje GB, Pitt WG (2005) Treatment of biofilm infections on implants with low-frequency ultrasound and antibiotics. Am J Infect Control 33:78–82

    Article  Google Scholar 

  • Caubet R, Pedarros-Caubet F, Chu M, Freye E, de Belem Rodrigues M, Moreau JM, Ellison WJ (2004) A radio frequency electric current enhances antibiotic efficacy against bacterial biofilms. Antimicrob Agents Chemother 48:4662–4664

    Article  CAS  Google Scholar 

  • Cochran WL, McFeters GA, Stewart PS (2000) Reduced susceptibility of thin Pseudomonas aeruginosa biofilms to hydrogen peroxide and monochloramine. J Appl Microbiol 88:22–30

    Article  CAS  Google Scholar 

  • Costerton JW (1999) Introduction to biofilm. Int J Antimicrob Agents 11:217–221

    Article  CAS  Google Scholar 

  • Costerton JW, Ellis B, Lam K, Johnson F, Khoury AE (1994) Mechanism of electrical enhancement of efficacy of antibiotics in killing biofilm bacteria. Antimicrob Agents Chemother 38:2803–2809

    CAS  Google Scholar 

  • Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284:1318–1322

    Article  CAS  Google Scholar 

  • Davies D (2003) Understanding biofilm resistance to antibacterial agents. Nature Rev Drug Discovery 2:114–122

    Article  CAS  Google Scholar 

  • Dobson J, Wilson M (1992) Sensitization of oral bacteria in biofilms to killing by light from a low-power laser. Arch Oral Biol 37:883–887

    Article  CAS  Google Scholar 

  • Dougherty TJ, Gomer CJ, Henderson BW, Jori G, Kessel D, Korbelik M, Moan J, Peng Q (1998) Photodynamic therapy. J National Cancer Institute 90:889–905

    Article  CAS  Google Scholar 

  • Doukas AG, Flotte TJ (1996) Physical characteristics and biological effects of laser-induced stress waves. Ultrasound Medicine Biology 22:151–164

    Article  CAS  Google Scholar 

  • Doukas AG, Kollias N (2004) Transdermal drug delivery with a pressure wave. Adv Drug Delivery Rev 56:559–579

    Article  CAS  Google Scholar 

  • Foley I, Gilbert P (1996) Antibiotic resistance of biofilms. Biofouling 10:331–346

    Article  CAS  Google Scholar 

  • Harris F, Chatfield LK, Phoenix DA (2005) Phenothiazinium based photosensitizers – Photodynamic agents with a multiplicity of cellular targets and clinical applications. Current Drug Targets 6:615–627

    Article  CAS  Google Scholar 

  • Kinniment SL, Wimpenny JW, Adams D, Marsh PD (1996) The effect of chlorhexidine on defined, mixed culture oral biofilms grown in a novel model system. J Appl Bacteriol 81:120–125

    CAS  Google Scholar 

  • Khoury AE, Lam K, Ellis B, Costerton JW (1992) Prevention and control of bacterial infections associated with medical devices. Am Soc Arti Internal Organs J 38:M174–178

    CAS  Google Scholar 

  • Larsen T (2002) Susceptibility of Porphyromonas gingivalis in biofilms to amoxicillin, doxycycline and metronidazole. Oral Microbiol Immunol 17:267–271

    Article  CAS  Google Scholar 

  • Lee S, Anderson T, Zhang H, Flotte TJ, Doukas AG (1996) Alteration of cell membrane by stress waves in vitro. Ultrasound Med Biol 22:1285–1293

    Article  CAS  Google Scholar 

  • Lee S, McAuliffe DJ, Flotte TJ, Kollias N, Doukas AG (1998) Photomechanical transcutaneous delivery of macromolecules. J Investigative Dermatol 111:925–929

    Article  CAS  Google Scholar 

  • Lee S, McAuliffe DJ, Flotte TJ, Kollias N, Doukas AG (2001) Photomechanical transdermal delivery: the effect of laser confinement. Lasers Surgery Med 28:344–347

    Article  CAS  Google Scholar 

  • Lin TYD, Mcauliffe DJ, Michaud N, Zhang H, Lee S, Doukas AG, Flotte TJ (2003) Nuclear transport by laser-induced pressure transients. Pharmaceutical Res 20:879–883

    Article  CAS  Google Scholar 

  • Marsh PD, Bradshaw DJ (1993) Microbiological effects of new agents in dentifrices for plaque control. Int Dental J 43:399–406

    CAS  Google Scholar 

  • Marsh PD (2005) Dental plaque: biological significance of a biofilm and community life-style. J Clin Periodontol 32(Suppl 6):7–15

    Article  CAS  Google Scholar 

  • Metcalf D, Robinson C, Devine D, Wood S (2006) Enhancement of erythrosine-mediated photodynamic therapy of Streptococcus mutans biofilms by light fractionation. J Antimicrob Chemother 58:190–192

    Article  CAS  Google Scholar 

  • Mitragotri S, Kost J (2004) Low-frequency sonophoresis: a review. Adv Drug Delivery Rev 56:589–601

    Article  CAS  Google Scholar 

  • Mulholland SE, Lee S, McAuliffe DJ, Doukas AG (1999) Cell loading with laser-generated stress waves: the role of the stress gradient. Pharmaceutical Res 16:514–518

    Article  CAS  Google Scholar 

  • Noiri Y, Okami Y, Narimatsu M, Takahashi Y, Kawahara T, Ebisu S (2003) Effects of chlorhexidine, minocycline, and metronidazole on Porphyromonas gingivalis strain 381 in biofilms. J Periodontol 74:1647–1651

    Article  CAS  Google Scholar 

  • Ogura M, Sato S, Nakanishi K, Uenoyama M, Kiyozumi T, Saitoh D, Ikeda T, Ashida H, Obara M (2004) In vivo targeted gene transfer in skin by the use of laser-induced stress waves. Lasers Surgery Med 34:242–248

    Article  Google Scholar 

  • Pratten J, Wilson M (1999) Antimicrobial susceptibility and composition of microcosm dental plaques supplemented with sucrose. Antimicrob Agents Chemother 43:1595–1599

    CAS  Google Scholar 

  • Qian Z, Sagers RD, Pitt WG (1997) The effect of ultrasonic frequency upon enhanced killing of P. aeruginosa biofilms. Annals Biomed Eng 25:69–76

    CAS  Google Scholar 

  • Rediske AM, Roeder BL, Nelson JL, Robison RL, Schaalje GB, Robison RA, Pitt WG (2000) Pulsed ultrasound enhances the killing of Escherichia coli biofilms by aminoglycoside antibiotics in vivo. Antimicrob Agents Chemother 44:771–772

    Article  CAS  Google Scholar 

  • Sarkar S, Wilson M (1993) Lethal photosensitization of bacteria in subgingival plaque samples from patients with chronic periodontitis. J Periodontal Res 28:204–210

    Article  CAS  Google Scholar 

  • Shani S, Friedman M, Steinberg D (2000) The anticariogenic effect of amine fluorides on Streptococcus sobrinus and glucosyltransferase in biofilms. Caries Res 34:260–267

    Article  CAS  Google Scholar 

  • Socransky SS, Smith C, Martin L, Paster BJ, Dewhirst FE, Levin AE (1994) “Checkerboard” DNA–DNA hybridization. Biotechniques 17:788–792

    CAS  Google Scholar 

  • Socransky SS, Haffajee AD (2002) Dental biofilms: difficult therapeutic targets. Periodontology 2000 28:12–55

    Article  Google Scholar 

  • Som S, Goodson JM, Abernethy A, Ruggiero K, Dunham J, Skobe Z, Rogers RA, Tegos G, Hamblin MR, Doukas AG, Soukos N (2004) Characterization and validation of a dental plaque microcosm laboratory biofilm. J Dental Res 83 (abstract)

  • Soukos NS, Ximenez-Fyvie LA, Hamblin MR, Socransky SS, Hasan T (1998) Targeted antimicrobial photochemotherapy. Antimicrob Agents Chemother 42:2595–2601

    CAS  Google Scholar 

  • Soukos NS, Socransky SS, Mulholland SE, Lee S, Doukas AG (2000) Photomechanical drug delivery into bacterial biofilms. Pharmaceutical Res 17:405–409

    Article  CAS  Google Scholar 

  • Soukos NS, Mulholland SE, Socransky SS, Doukas AG (2003) Photodestruction of human dental plaque bacteria: enhancement of the photodynamic effect by photomechanical waves in an oral biofilm model. Lasers Surgery Med 33:161–168

    Article  Google Scholar 

  • Stewart PS, Grab L, Diemer JA (1998) Analysis of biocide transport limitation in an artificial biofilm system. J Appl Microbiol 85:495–500

    Article  CAS  Google Scholar 

  • Stewart PS, Wattanakaroon W, Goodrum L, Fortun SM, McLeod BR (1999) Electrolytic generation of oxygen partially explains electrical enhancement of tobramycin efficacy against Pseudomonas aeruginosa biofilm. Antimicrobial Agents and Chemotherapy 43:292–296

    CAS  Google Scholar 

  • Stoodley P, de Beer D, Lappin-Scott H (1997) Influence of electrical fields and pH on biofilm structure as related to the bioelectric effect. Antimicrob Agents Chemother 41:1876–1879

    CAS  Google Scholar 

  • Sutherland IW (2001) The biofilm matrix – an immobilized but dynamic microbial environment. Trends Microbiol 9:222–227

    Article  CAS  Google Scholar 

  • Terakawa M, Ogura M, Sato S, Wakisaka H, Ashida H, Uenoyama M, Masaki Y, Obara M (2004) Gene transfer into mammalian cells by use of a nanosecond pulsed laser-induced stress wave. Optics Lett 29:1227–1229

    Article  Google Scholar 

  • Usacheva MN, Teichert MC, Biel MA (2003) The interaction of lipopolysaccharides with phenothiazine dyes. Lasers Surgery Med 33:311–319

    Article  Google Scholar 

  • Wainwright M, Phoenix DA, Marland J, Wareing DRA, Bolton FJ (1997) A study of photobacterial activity in the phenothiazinium series. FEMS Immunol Med Microbiol 19:75–80

    Article  CAS  Google Scholar 

  • Wellman N, Fortun SM, McLeod BR (1996) Bacterial biofilms and the bioelectric effect. Antimicrob Agents Chemother 40:2012–2014

    CAS  Google Scholar 

  • Whiteley M, Gita Bangera M, Bumgarner RE, Parsek MR, Teitzel GM, Lory S, Greenberg EP (2001) Gene expression in Pseudomonas aeruginosa biofilms. Nature 413:860–864

    Article  CAS  Google Scholar 

  • Williams JA, Pearson GJ, Colles MJ, Wilson M (2003) 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 37:190–193

    Article  CAS  Google Scholar 

  • Wilson M (1993) Photolysis of oral bacteria and its potential use in the treatment of caries and periodontal disease: a review. J Appl Bacteriol 75:299–306

    CAS  Google Scholar 

  • Wilson M, Dobson J, Harvey W (1992) Sensitization of oral bacteria to killing by low-power laser radiation. Curr Microbiol 25:77–81

    Article  CAS  Google Scholar 

  • Wilson M, Dobson J, Sarkar S (1993) Sensitisation of periodontopathogenic bacteria to killing by light from a low-power laser. Oral Microbiol Immunol 8:182–187

    CAS  Google Scholar 

  • Wood S, Nattress B, Kirkham J, Shore R, Brookes S, Griffiths J, Robinson C (1999) 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: Biol 50:1–7

    Google Scholar 

  • Wood S, Metcalf D, Devine D, Robinson C (2006) Erythrosine is a potential photosensitizer for the photodynamic therapy of oral plaque biofilms. J Antimicrob Chemother 57:680–684

    Article  CAS  Google Scholar 

  • Zanin IC, Goncalves RB, Junior AB, Hope CK, Pratten J (2005) Susceptibility of Streptococcus mutans biofilms to photodynamic therapy: an in vitro study. J Antimicrob Chemother 56:324–330

    Article  CAS  Google Scholar 

  • Zanin IC, Lobo MM, Rodrigues LK, Pimenta LA, Hofling JF, Goncalves RB (2006) Photosensitization of in vitro biofilms by toluidine blue O combined with a light-emitting diode. Eur J Oral Sci 114:64–69

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the NIDCR grant RO1-DE-14360. Dr. Ogura acknowledges support by the Ministry of Education, Science, Sports and Culture, Grant-in Aid for JSPS Fellows, 7596, 2004. Dr. Doukas acknowledges support by the DoD Medical Free Electron Laser Program (FA9550-04-1-0079). We thank the Palomar Corp., Lexington, MA for the loan of the Nd:YAG laser. We thank Dr. Rick Rogers, Harvard School of Public Health, Boston for his help with the confocal scanning laser microscopy. We thank Dr. Phil Stewart, The Center for Biofilm Engineering, Montana State University-Bozeman, and Drs. Federico Foschi and Carla Fontana, Applied Molecular Photomedicine Laboratory, The Forsyth Institute for critically reading the manuscript. Dr. Ogura thanks Dr. Monoru Obara, Keio University, and Dr. Shunichi Sato, National Defense Medical College Research Institute, for helpful discussions.

Author information

Authors and Affiliations

  1. Applied Molecular Photomedicine Laboratory, The Forsyth Institute, 140 The Fenway, Boston, MA, 02115, USA

    Makoto Ogura, Abraham D. Abernethy, Ryan D. Blissett, Karriann Ruggiero, Sovanda Som & Nikolaos S. Soukos

  2. Department of Electronics and Electrical Engineering, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, 223-8522, Japan

    Makoto Ogura

  3. Harvard School of Dental Medicine, 188 Longwood Avenue, Boston, MA, 02115, USA

    Ryan D. Blissett

  4. Clinical Collaborative, The Forsyth Institute, 140 The Fenway, Boston, MA, 02115, USA

    J. Max Goodson

  5. Department of Biostatistics, The Forsyth Institute, 140 The Fenway, Boston, MA, 02115, USA

    Ralph Kent

  6. Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, 50 Blossom Street, Boston, MA, 02114, USA

    Apostolos G. Doukas

Authors
  1. Makoto Ogura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Abraham D. Abernethy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ryan D. Blissett
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Karriann Ruggiero
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sovanda Som
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J. Max Goodson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ralph Kent
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Apostolos G. Doukas
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nikolaos S. Soukos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nikolaos S. Soukos.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ogura, M., Abernethy, A.D., Blissett, R.D. et al. Photomechanical wave-assisted molecular delivery in oral biofilms. World J Microbiol Biotechnol 23, 1637–1646 (2007). https://doi.org/10.1007/s11274-007-9411-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-007-9411-x

Keywords

Search

Navigation