title: Morphological changes of the dermatophyte Trichophyton rubrum after photodynamic treatment: a scanning electron microscopy study.
abstract: Treatment strategies for superficial mycosis caused by the dermatophyte Trichophyton rubrum consist of the use of topical or oral antifungal preparations. We have recently discovered that T. rubrum is susceptible to photodynamic treatment (PDT), with 5,10,15-tris(4-methylpyridinium)-20-phenyl-[21H,23H]-porphine trichloride (Sylsens B) as a photosensitizer. The susceptibility appeared to depend on the fungal growth stage, with PDT efficacy higher with microconidia when compared to mycelia. The aim of this study was to investigate, with the use of scanning electron microscopy, the morphological changes caused by a lethal PDT dose to T. rubrum when grown on isolated human stratum corneum. Corresponding dark treatment and light treatment without photosensitizer were used as controls. A sub-lethal PDT dose was also included in this investigation The morphologic changes were followed at various time points after the treatment of different fungal growth stages. Normal fungal growth was characterized by a fiber-like appearance of the surface of the hyphae and microconidia with the exception of the hyphal tips in full mycelia and the microconidia shortly after attachment to the stratum corneum. Here, densely packed globular structures were observed. The light dose (108 J/cm2) in the absence of Sylsens B, or the application of the photosensitizer in the absence of light, caused reversible fungal wall deformations and bulge formation. However, after a lethal PDT, a sequence of severe disruptions and deformations of both microconidia and the mycelium were observed leading to extrusion of cell material and emptied fungal elements. In case of a non-lethal PDT, fungal re-growth started on the remnants of the treated mycelium.
authors.affiliation: Leiden University Medical Centre, Leiden, The Netherlands. G.M.T.Smijs@LUMC.nl
authors.forename: Threes G M
authors.initials: TG
authors.lastname: Smijs
authors.forename: Aat A
authors.initials: AA
authors.lastname: Mulder
authors.forename: Stan
authors.initials: S
authors.lastname: Pavel
authors.forename: Jos J M
authors.initials: JJ
authors.lastname: Onderwater
authors.forename: Henk K
authors.initials: HK
authors.lastname: Koerten
authors.forename: Joke A
authors.initials: JA
authors.lastname: Bouwstra
mesh_terms: D002473:Cell Wall / Q000187:drug effects / Q000648:ultrastructure; D003624:Darkness; D004817:Epidermis / Q000382:microbiology; D006801:Humans; D025301:Hyphae / Q000187:drug effects / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000494:pharmacology; D011166:Porphyrins / Q000494:pharmacology; D011726:Pyridinium Compounds / Q000494:pharmacology; D013172:Spores, Fungal / Q000187:drug effects / Q000648:ultrastructure; D013995:Time; D014249:Trichophyton / Q000187:drug effects / Q000648:ultrastructure*
chemical_list: C509254:5,10,15-tris(4-methylpyridinium)-20-phenyl-(21H,23H)-porphine; D017319:Photosensitizing Agents; D011166:Porphyrins; D011726:Pyridinium Compounds