J Bacteriol Virol.  2016 Mar;46(1):27-35. 10.4167/jbv.2016.46.1.27.

Effect of Methylene Blue-mediated Photodynamic Therapy on Wild-type and Ciprofloxacin-resistant Mycobacterium smegmatis

• Shim I
• Choi M
• Min Y
• Seok KH
• Kim JK
• Jeong JY
• Oak CH
• Park I

• KMID: 2168740
• DOI: http://doi.org/10.4167/jbv.2016.46.1.27

Abstract

Tuberculosis (TB) patients are normally treated with a combination of antibiotics. However, with improper or incomplete treatment of antibiotics, the disease may progress to multidrug-resistant TB (MDR-TB). The treatment of MDR-TB is very costly and inefficient. Therefore, there is a great demand of new therapeutic approaches for MDR-TB such as photodynamic therapy. In this study, we tried to optimize the conditions for photodynamic inactivation of TB using methylene blue as a photosensitizer. Different combinations of methylene blue concentrations and light doses were tested for their photodynamic effects to A549 cells or Mycobacterium smegmatis (M. smegmatis). We also tested the effect of photodynamic therapy on ciprofloxacin-resistant M. smegmatis. Methylene blue treatment alone did not affect the survival rates of A549 cells or bacteria up to 5 µg/ml. When the A549 and M. smegmatis cells treated with methylene blue were irradiated with laser light (wavelength, 630 nm), photodynamic inactivation of cells was increased in methylene blue concentration- and light dose-dependent manners. Interestingly, the ciprofloxacin-resistant M. smegmatis exhibited higher level of susceptibility to methylene blue-mediated photodynamic inactivation. This study suggests that photodynamic therapy at 3.6 J/cm2 in the presence of 5 µg/ml methylene blue may be an appropriate range for therapy due to the high bactericidal activity against high level of ciprofloxacin-resistant M. smegmatis and the low damaging effect to mammalian cells. This study demonstrates that photodynamic therapy could be a potential alternative for MDR-TB treatment.

Keyword

Photodynamic therapy; Methylene blue; Mycobacterium smegmatis; Ciprofloxacin; Tuberculosis

MeSH Terms

Anti-Bacterial Agents
Bacteria
Ciprofloxacin
Humans
Methylene Blue
Mycobacterium smegmatis*
Mycobacterium*
Photochemotherapy*
Survival Rate
Tuberculosis
Anti-Bacterial Agents
Ciprofloxacin
Methylene Blue

Figure

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  Figure 1. Effects of photodynamic therapy with methylene blue on A549 human lung cell. Different concentrations of methylene blue (MB) were incubated with A549 cells and irradiated with different light doses. Means are shown as points.

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  Figure 2. Effects of photodynamic therapy with methylene blue on Mycobacterium smegmatis. Different concentrations of methylene blue (MB) were incubated with Mycobacterium smegmatis and irradiated with different light doses. Data were presented as Mean ± SD of three independent experiments. Asterisks indicate statistically significant differences with respect to the corresponding controls (student t-test: ∗, p < 0.05; ∗∗, p < 0.01).

• 

  Figure 3. Effects of photodynamic therapy with methylene blue on ciprofloxacin resistant Mycobacterium smegmatis. Different concentrations of methylene blue (MB) were incubated with Mycobacterium smegmatis and irradiated with different light doses. Data were presented as Mean ± SD of three independent experiments. Asterisks indicate statistically significant differences with respect to the corresponding controls (student t-test: ∗∗, p < 0.01).

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