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Antimicrobial mechanisms behind photodynamic effect in the presence of hydrogen peroxide

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Abstract

This study describes the use of methylene blue (MB) plus light (photodynamic inactivation, PDI) in the presence of hydrogen peroxide (H2O2) to kill Staphylococcus aureus, Escherichia coli, and Candida albicans. When H2O2 was added to MB plus light there was an increased antimicrobial effect, which could be due to a change in the type of ROS generated or increased microbial uptake of MB. To clarify the mechanism, the production of ROS was investigated in the presence and absence of H2O2. It was observed that ROS production was almost inhibited by the presence of H2O2 when cells were not present. In addition, experiments using different sequence combinations of MB and H2O2 were performed and MB optical properties inside the cell were analyzed. Spectroscopy experiments suggested that the amount of MB was higher inside the cells when H2O2 was used before or simultaneously with PDI, and ROS formation inside C. albicans cells confirmed that ROS production is higher in the presence of H2O2. Moreover enzymatic reduction of MB by E. coli during photosensitizer uptake to the photochemically inactive leucoMB could be reversed by the oxidative effects of hydrogen peroxide, increasing ROS formation inside the microorganism. Therefore, the combination of a photosensitizer such as MB and H2O2 is an interesting approach to improve PDI efficiency.

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Authors and Affiliations

  1. Centro de Pesquisa e Pós-Graduação São Leopoldo Mandic, Campinas, SP, Brazil

    Aguinaldo Silva Garcez

  2. Centro de Estudos, Treinamento e Aperfeiçoamento em Odontologia, São Paulo, SP, Brazil

    Silvia Cristina Núñez

  3. Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil

    Mauricio S. Baptista

  4. Universidade Federal do ABC, Santo André, SP, Brazil

    Nasser Ali Daghastanli

  5. Departamento de Física Aplicada, Instituto de Física, Universidade de São Paulo, São Paulo, SP, Brazil

    Rosangela Itri

  6. Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, MA, USA

    Michael R. Hamblin

  7. Department of Dermatology, Harvard Medical School, Boston, MA, USA

    Michael R. Hamblin

  8. Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA, USA

    Michael R. Hamblin

  9. Centro de Laser e Aplicaç ões, Instituto de Pesquisas Energéticas e Nucleares, IPEN-CNEN/SP, Av. Lineu Prestes, 2242, Cidade Universitária, 05508-000., São Paulo, SP, Brazil

    Martha Simões Ribeiro

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  1. Aguinaldo Silva Garcez
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  2. Silvia Cristina Núñez
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  3. Mauricio S. Baptista
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  4. Nasser Ali Daghastanli
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  5. Rosangela Itri
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  6. Michael R. Hamblin
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  7. Martha Simões Ribeiro
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Correspondence to Martha Simões Ribeiro.

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Garcez, A.S., Núñez, S.C., Baptista, M.S. et al. Antimicrobial mechanisms behind photodynamic effect in the presence of hydrogen peroxide. Photochem Photobiol Sci 10, 483–490 (2011). https://doi.org/10.1039/c0pp00082e

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  • DOI: https://doi.org/10.1039/c0pp00082e