Tabart Jeremy, Baldo Aline, Vermout Sandy, Losson Bertrand, Mignon Bernard
Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
Vet Dermatol. 2008 Jun;19(3):130-3. doi: 10.1111/j.1365-3164.2008.00661.x.
A fully differentiated reconstructed interfollicular feline epidermis (RFE) was recently developed in vitro. It was shown to be relevant for the study of Microsporum canis-epidermal interactions. In this study, RFE was evaluated as a potential model for the in vitro screening of drugs against M. canis. As a preliminary step, the minimum inhibitory concentration of miconazole nitrate against M. canis IHEM 21239 grown on Sabouraud's dextrose agar was determined to be 0.3 microg mL(-1). RFE grown at the air-liquid interface was cultured for 24 h in RFE culture medium, supplemented with either miconazole (range 0.1-1 microg mL(-1)) or its solvent (dimethylsulfoxide). Then, RFE was inoculated in triplicate with 1 x 10(5 )M. canis arthroconidia and incubated for five additional days. To evaluate fungal growth, RFE was processed for routine histopathology, three serial sections being performed across the block at 100 microm intervals. No fungal growth was detected invading or on the surface of infected RFE in the presence of miconazole concentrations equal to or higher than 0.3 microg mL (final concentration in the culture medium). This study demonstrates that RFE is an adequate model for the in vitro screening of drugs against M. canis and potentially against other skin pathogens.
最近在体外构建了一种完全分化的毛囊间猫表皮重建模型(RFE)。研究表明,该模型与犬小孢子菌 - 表皮相互作用的研究相关。在本研究中,评估了RFE作为体外筛选抗犬小孢子菌药物潜在模型的可能性。作为初步步骤,测定了硝酸咪康唑对在沙氏葡萄糖琼脂上生长的犬小孢子菌IHEM 21239的最低抑菌浓度为0.3μg/mL(-1)。将在气液界面生长的RFE在补充有咪康唑(浓度范围为0.1 - 1μg/mL(-1))或其溶剂(二甲基亚砜)的RFE培养基中培养24小时。然后,用1×10(5)个犬小孢子菌关节孢子一式三份接种RFE,并再孵育五天。为了评估真菌生长情况,对RFE进行常规组织病理学处理,以100μm的间隔在整个组织块上制作三个连续切片。在咪康唑浓度等于或高于0.3μg/mL(培养基中的最终浓度)的情况下,在受感染的RFE表面或侵入其中均未检测到真菌生长。本研究表明,RFE是体外筛选抗犬小孢子菌以及潜在抗其他皮肤病原体药物的合适模型。