Lasers in Medical Science

, Volume 33, Issue 5, pp 983–990 | Cite as

Photodynamic antimicrobial chemotherapy (PACT) using toluidine blue inhibits both growth and biofilm formation by Candida krusei

  • Bruna Graziele Marques da Silva
  • Moisés Lopes Carvalho
  • Isabela Bueno Rosseti
  • Stella Zamuner
  • Maricilia Silva CostaEmail author
Original Article


Among non-albicans Candida species, the opportunistic pathogen Candida krusei emerges because of the high mortality related to infections produced by this yeast. The Candida krusei is an opportunistic pathogen presenting an intrinsic resistance to fluconazol. In spite of the reduced number of infections produced by C. krusei, its occurrence is increasing in some groups of patients submitted to the use of fluconazol for prophylaxis. Photodynamic antimicrobial chemotherapy (PACT) is a potential antimicrobial therapy that combines visible light and a nontoxic dye, known as a photosensitizer, producing reactive oxygen species (ROS) that can kill the treated cells. The objective of this study was to investigate the effects of PACT, using toluidine blue, as a photosensitizer on both growth and biofilm formation by Candida krusei. In this work, we studied the effect of the PACT, using TB on both cell growth and biofilm formation by C. krusei. PACT was performed using a light source with output power of 0.068 W and peak wavelength of 630 nm, resulting in a fluence of 20, 30, or 40 J/cm2. In addition, ROS production was determined after PACT. The number of samples used in this study varied from 6 to 8. Statistical differences were evaluated by analysis of variance (ANOVA) and post hoc comparison with Tukey-Kramer test. PACT inhibited both growth and biofilm formation by C. krusei. It was also observed that PACT stimulated ROS production. Comparing to cells not irradiated, irradiation was able to increase ROS production in 11.43, 6.27, and 4.37 times, in the presence of TB 0.01, 0.02, and 0.05 mg/mL, respectively. These results suggest that the inhibition observed in the cell growth after PACT could be related to the ROS production, promoting cellular damage. Taken together, these results demonstrated the ability of PACT reducing both cell growth and biofilm formation by C. krusei.


Candida krusei Photodynamic antimicrobial chemotherapy PACT Toluidine blue Biofilm formation 



This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

Funding information

The authors would like to thank FAPESP and CNPq for the financial support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

In this study, all experiments were performed using cultures of Candida krusei (ATCC 6258); therefore, there was no need for approval by local authorities.

Informed consent

We have obtained permission from all the authors; we declare that the material has not been published in whole or in part elsewhere and the paper is not currently being considered for publication elsewhere.


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Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2018

Authors and Affiliations

  • Bruna Graziele Marques da Silva
    • 1
  • Moisés Lopes Carvalho
    • 1
  • Isabela Bueno Rosseti
    • 2
  • Stella Zamuner
    • 3
  • Maricilia Silva Costa
    • 1
    Email author
  1. 1.Instituto de Pesquisa e Desenvolvimento (IP&D)Universidade do Vale do Paraíba (UNIVAP)São José dos CamposBrazil
  2. 2.Anhanguera EducacionalSão José dos CamposBrazil
  3. 3.Posgraduated Program in MedicineUniversidade Nove de Julho (UNINOVE)São PauloBrazil

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