Development of a model for Candida albicans infection using tissue-engineered human skin substitutes

Abstract

Objective To develop a model for Candida albicans infection using a pigmented tissueengineered skin substitute derived from human skin cells.Methods Fibroblasts,keratinocytes and melanocytes were isolated from the foreskin of Chinese adults,and subjected to serial passages.Fibroblasts were incubated in collagen for three days to construct tissue-engineered dermis.Then,keratinocytes and melanocytes were cocultured above the dermis for 12 days to construct epidermis.Subsequently,the constructed tissue-engineered skin was inoculated with 5 μl of C.albicans suspension (106 spores/ml) followed by culture for 6,12,24,48 and 72 hours.Hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining were carried out to observe the infected skin constitute.Results The tissue-engineered skin constructed by fibroblasts,keratinocytes,melanocytes and collagen showed a well-organized structure and highly differentiated epidermis.After infection with C.albicans,the epidermal structure was gradually destructed with the extension of hyphae from the superficial epidermis to dermis,and was completely destroyed with the dermis filled with hyphae at 72 hours.Conclusions Pigmented tissue-engineered skin is successfully constructed using human skin cells with a well-organized structure and highly differentiated epidermis,and a model for C.albicans infection has also been preliminarily established on the pigmented tissue-engineered skin. Key words: Tissue engineering; Skin, artificial; Candida albicans; Models, biological