The impact of antimicrobial photodynamic therapy on Streptococcus mutans in an artificial biofilm model

Martin Schneider; Gregor Kirfel; Felix Krause; Michael Berthold; Olivier Brede; Matthias Frentzen; Andreas Braun

doi:10.1117/12.841809

5 March 2010 The impact of antimicrobial photodynamic therapy on Streptococcus mutans in an artificial biofilm model

Martin Schneider, Gregor Kirfel, Felix Krause, Michael Berthold, Olivier Brede, Matthias Frentzen, Andreas Braun

Proceedings Volume 7549, Lasers in Dentistry XVI; 75490A (2010) https://doi.org/10.1117/12.841809
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

The aim of the study was to assess the impact of laser induced antimicrobial photodynamic therapy on the viability of Streptococcus mutans cells employing an aritificial biofilm model. Employing sterile chambered coverglasses, a salivary pellicle layer formation was induced in 19 chambers. Streptococcus mutans cells were inoculated in a sterile culture medium. Using a live/dead bacterial viability kit, bacteria with intact cell membranes stain fluorescent green. Test chambers containing each the pellicle layer and 0.5 ml of the bacterial culture were analyzed using a confocal laser scan microscope within a layer of 10 μm at intervals of 1 μm from the pellicle layer. A photosensitizer was added to the test chambers and irradiated with a diode laser (wavelength: 660 nm, output power: 100 mW, Helbo) for 2 min each. Comparing the baseline fluorescence (median: 13.8 [U], min: 3.7, max: 26.2) with the values after adding the photosensitizer (median: 3.7, min: 1.1, max: 9), a dilution caused decrease of fluorescence could be observed (p<0.05). After irradiation of the samples with a diode laser, an additional 48 percent decrease of fluorescence became evident (median: 2.2, min: 0.4, max: 3.4) (p<0.05). Comparing the samples with added photosensitizer but without laser irradiation at different times, no decrease of fluorescence could be measured (p>0.05). The present study indicates that antimicrobial photodynamic therapy can reduce living bacteria within a layer of 10 μm in an artificial biofilm model. Further studies have to evaluate the maximum biofilm thickness that still allows a toxic effect on microorganisms.

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Martin Schneider, Gregor Kirfel, Felix Krause, Michael Berthold, Olivier Brede, Matthias Frentzen, and Andreas Braun "The impact of antimicrobial photodynamic therapy on Streptococcus mutans in an artificial biofilm model", Proc. SPIE 7549, Lasers in Dentistry XVI, 75490A (5 March 2010); https://doi.org/10.1117/12.841809

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KEYWORDS

Luminescence

Photodynamic therapy

Bacteria

Pellicles

Laser irradiation

Confocal microscopy

Microorganisms

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