The photodynamic therapy on Streptococcus mutans biofilms using erythrosine and dental halogen curing unit

Young-Ho Lee, Ho-Won Park, Ju-Hyun Lee, Hyun-Woo Seo, Si-Young Lee, Young-Ho Lee, Ho-Won Park, Ju-Hyun Lee, Hyun-Woo Seo, Si-Young Lee

Abstract

The purpose of our study was to evaluate the effect of photodynamic therapy (PDT), using erythrosine as a photosensitizing agent and a dental halogen curing unit as a light source, on Streptococcus mutans in a biofilm phase. The S. mutans biofilms were formed in a 24-well cell culture cluster. Test groups consisted of biofilms divided into four groups: group 1: no photosensitizer or light irradiation treatment (control group); group 2: photosensitizer treatment alone; group 3: light irradiation alone; group 4: photosensitizer treatment and light irradiation. After treatments, the numbers of colony-forming unit (CFU) were counted and samples were examined by confocal laser scanning fluorescence microscopy (CLSM). Only group 4 (combined treatment) resulted in significant increases in cell death, with rates of 75% and 55% after 8 h of incubation, and 74% and 42% at 12 h, for biofilms formed in brain-heart infusion (BHI) broth supplemented with 0% or 0.1% sucrose, respectively. Therefore, PDT of S. mutans biofilms using a combination of erythrosine and a dental halogen curing unit, both widely used in dental clinics, resulted in a significant increase in cell death. The PDT effects are decreased in biofilms that form in the presence of sucrose.

Figures

Figure 1
Figure 1
Time based CFU analysis. The ability of Streptococcus mutans to colonize was increased in the presence of sucrose. The quantity of biofilms formation was increased up to 12 h incubation time. Error bars present standard deviations. Comparison of 0% sucrose (*), 0.1% sucrose (†), 1% sucrose (‡), statistically significant with P<0.05. CFU, colony-forming unit.
Figure 2
Figure 2
The PDT effect at 8 h incubation time. Cell death was significantly increased in only group 4 at both sucrose concentrations. *Compared with other groups, statistically significant with P<0.005. Error bars represent standard deviations. PDT, photodynamic therapy.
Figure 3
Figure 3
The PDT effect at 12 h incubation time. Cell death was significantly increased in only group 4 regardless of two concentrations of sucrose. *Compared with other groups, statistically significant with P<0.005. PDT, photodynamic therapy.
Figure 4
Figure 4
CLSM images of biofilms at 12 h incubation time in 0.1% sucrose. Green fluorescence represents viable bacteria and red fluorescence represents affected bacteria. (a) Group 1: non-treated biofilms with PDT. (b) Group 4: treated biofilms with PDT. The scale bar for (a) and (b) is 50 µm. CLSM, confocal laser scanning fluorescence microscopy; PDT, photodynamic therapy.

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