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Evaluation of an Explanted Porcine Skin Model to Investigate Infection with the Dermatophyte Trichophyton rubrum

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Abstract

Dermatophytosis is a fungal infection of skin, hair and nails, and the most frequently found causative agent is Trichophyton rubrum. The disease is very common and often recurring, and it is therefore difficult to eradicate. To develop and test novel treatments, infection models that are representative of the infection process are desirable. Several infection models have been developed, including the use of cultured cells, isolated corneocytes, explanted human skin or reconstituted human epidermis. However, these have various disadvantages, ranging from not being an accurate reflection of the site of infection, as is the case with, for example, cultured cells, to being difficult to scale up or having ethical issues (e.g., explanted human skin). We therefore sought to develop an infection model using explanted porcine skin, which is low cost and ethically neutral. We show that in our model, fungal growth is dependent on the presence of skin, and adherence of conidia is time-dependent with maximum adherence observed after ~ 2 h. Scanning electron microscopy suggested the production of fibril-like material that links conidia to each other and to skin. Prolonged incubation of infected skin leads to luxurious growth and invasion of the dermis, which is not surprising as the skin is not maintained in conditions to keep the tissue alive, and therefore is likely to lack an active immune system that would limit fungal growth. Therefore, the model developed seems useful to study the early stages of infection. Furthermore, we demonstrate that the model can be used to test novel treatment regimens for tinea infections.

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Authors and Affiliations

  1. Department of Pharmacy and Pharmacology, University of Bath, Bath, BA2 7AY, UK

    Fritz Ka-Ho Ho, M. Begoña Delgado-Charro & Albert Bolhuis

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  1. Fritz Ka-Ho Ho
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  2. M. Begoña Delgado-Charro
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  3. Albert Bolhuis
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Correspondence to Albert Bolhuis.

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The authors declare that they have no conflict of interest.

Ethical Approval

Ethical approval for the study was obtained from the relevant ethics committees. Porcine skin was collected from a local slaughterhouse, and no experimental procedures were performed on live animals.

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Supplementary Fig.

 1. Proteolytic activity in the culture supernatant.T. rubrum was grown for 10 days in rich medium (PDB) or minimal salts medium (MS) in the presence or absence of 0.2% keratin (± ker). Proteolytic activity, which is expressed in arbitrary units (AU), was then determined using azocasein. Error bars indicate standard deviation of the mean from three independent experiments. (TIFF 3460 kb)

Supplementary Fig.

 2. Skin samples were treated on each side with chlorine gas with the times indicated. Specimens were then fixed, cryoprotected, and stained with haematoxylin/eosin, and imaged using light microscopy. The scale bars indicated are 50 µm. (TIFF 18199 kb)

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Ho, F.KH., Delgado-Charro, M.B. & Bolhuis, A. Evaluation of an Explanted Porcine Skin Model to Investigate Infection with the Dermatophyte Trichophyton rubrum. Mycopathologia 185, 233–243 (2020). https://doi.org/10.1007/s11046-020-00438-9

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