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JAC Advance Access published online on August 17, 2007
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkm304
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© The Author 2007. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Investigation of conditions involved in the susceptibility of the dermatophyte Trichophyton rubrum to photodynamic treatment

Threes G. M. Smijs1,*, Joke A. Bouwstra2, Mojgan Talebi1 and Stan Pavel1

1 Leiden University Medical Centre, Leiden, The Netherlands 2 University of Leiden, Leiden/Amsterdam Centre for Drugs Research, Leiden, The Netherlands

Received 8 May 2007; returned 12 June 2007; revised 10 July 2007; accepted 13 July 2007

* Correspondence address. Leiden University Medical Centre, Skin Research Laboratory, PO Box 9600, 2300 RC Leiden, The Netherlands. Tel: +31-71-5269376/60; Fax: +31-71-5248106; E-mail: G.M.T.Smijs{at}LUMC.nl

Background: Photodynamic treatment (PDT) refers to a treatment with light-activated agents (photosensitizers) in combination with visible light and molecular oxygen. Recently, we have demonstrated that the porphyrins, 5,10,15-tris(4-methylpyridinium)-20-phenyl-[21H,23H]-porphine trichloride (Sylsens B) and deuteroporphyrin monomethylester (DP mme) are excellent photosensitizers to be used against Trichophyton rubrum both in vitro and ex vivo.

Objectives and methods: The objective of this study was to investigate the key factors involved in PDT efficacy of both photosensitizers in an ex vivo situation during different fungal growth stages using a recently developed ex vivo model. The study focused on the influence of pH and ion strength of incubation media, photochemical properties of the photosensitizers (spectra and singlet oxygen production), and the effect of several scavengers of reactive oxygen species (sodium azide, histidine, mannitol) and phenylmethylsulphonylfluoride (keratinase inhibitor) on the PDT efficacy.

Results and conclusions: The results show that an optimal pH and low concentrations of calcium are crucial for a selective binding of Sylsens B to the fungus, leading to an increased PDT efficacy. This selective binding to T. rubrum cannot be accomplished for DP mme. It can be concluded that the prerequisite for successful treatment is a use of a low molarity solution of pH 5, supplemented with a chelating agent and a keratinase activity-repressing agent. Under these conditions, PDT with Sylsens B inactivates, initially via singlet oxygen, effectively the fungus in different fungal growth stages.

Key Words: ex vivo model , stratum corneum , pH , porphyrins , tinea infections , microconidia , surface potential


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