Construction of a Trichophyton mentagrophytes-infected tissue-engineered skin model

Abstract

Objective To establish a Trichophyton mentagrophytes-infected tissue-engineered skin model in vitro. Methods Primary fibroblasts were cultured with the presence of type Ⅰ bovine collagen for 3 days to form tissue-engineered dermis. Then, primary keratinocytes and melanocytes were cocultured on the surface (namely air-liquid interface) of the artificial dermis for another 12 days to form a tissue-engineered skin model. Five microliters of fresh Trichophyton mentagrophytes suspensions were used to infect the tissue-engineered skin model. Hematoxylin and eosin(HE) staining and periodic acid-Schiff (PAS) staining were conducted to observe the structure of the cultures at 12, 24, 48 and 72 hours separately after the infection. Results A tissue-engineered dermal equivalent with a compact and uniform structure was formed by the culture of fibroblasts with the presence of type Ⅰ bovine collagen, and a well-stratified and -structured epidermis was formed by cocultured keratinocytes and melanocytes on the surface of the dermis. After infection with Trichophyton mentagrophytes, the structure of tissue-engineered skin was gradually destroyed over time, and epidermal destruction was severest at 48 hours; the entire structure of the tissue-engineered skin was replaced by hyphae and spores at 72 hours. Conclusion A Trichophyton mentagrophytes-infected tissue-engineered skin model has been preliminarily established. Key words: Tissue engineering; Skin; Animal testing alternatives; Infection; Trichophyton mentagrophytes