JAC Advance Access originally published online on January 9, 2007
Journal of Antimicrobial Chemotherapy 2007 59(3):433-440; doi:10.1093/jac/dkl490
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A novel ex vivo skin model to study the susceptibility of the dermatophyte Trichophyton rubrum to photodynamic treatment in different growth phases
1 Leiden University Medical Centre, Skin Research Laboratory, PO Box 9600, 2300 RC Leiden, The Netherlands 2 University of Leiden, Leiden/Amsterdam Centre for Drugs Research, Leiden, The Netherlands 3 PhotoBioChem NV, Leiden, The Netherlands
Received 19 July 2006; returned 14 October 2006; revised 20 October 2006; accepted 2 November 2006
* Corresponding author. Tel: +31-71-5269376 / +31-71-5269360; Fax: +31-71-5268286; E-mail: G.M.T.Smijs{at}LUMC.nl
Background: Dermatophytes are fungi that can cause infections of skin, hair and nails because of their ability to feed on keratin. Superficial mycoses are among the most prevalent infectious diseases worldwide. Two important restrictions of current therapeutic options are the recurrence of the infection and prolonged treatment. This is especially true for infections caused by Trichophyton rubrum, a widely distributed dermatophyte. The application of photosensitizers for treatment of fungal infections is, within the field of photodynamic treatment (PDT), relatively new. Recently, we demonstrated that the porphyrins 5,10,15-tris(4-methylpyridinium)-20-phenyl-[21H,23H]-porphine trichloride (Sylsens B) and deuteroporphyrin monomethylester (DP mme) were excellent photosensitizers towards T. rubrum when using red light.
Objectives and methods: To evaluate the photodynamic effectiveness of the porphyrins in a situation that mimics the clinical situation, we developed an ex vivo model using human stratum corneum. This model offers the possibility of applying PDT at different time points during the germination and subsequent development of T. rubrum microconidia. The model was used for two different incubation media, Dulbecco's modified Eagle medium (DMEM) and distilled water.
Results and conclusions: We demonstrated that the PDT susceptibility of T. rubrum depended on the time of PDT application after spore inoculation. A decrease in susceptibility was observed with increasing time of PDT application for both photosensitizers in DMEM. Changing the incubation medium to distilled water resulted in an increased fungicidal effect for Sylsens B and in a decreased effect for DP mme. We conclude that T. rubrum is susceptible to PDT in a situation that mimics the clinical situation. The fungicidal effect of PDT on fungal spores is of particular importance.
Keywords: ex vivo model , stratum corneum , porphyrins , tinea , microconidia
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