须癣毛癣菌的组织工程皮肤感染模型的构建

目的 体外构建须癣毛癣菌的组织工程皮肤感染模型。 方法 用牛Ⅰ型胶原溶液复合原代成纤维细胞培养3 d后，在其表面利用气液界面培养原代角质形成细胞和黑素细胞12 d得到组织工程皮肤，取新鲜的须癣毛癣菌悬液5 μl对其进行感染，分别在12、24、48、72 h进行HE染色和PAS染色观察。 结果 成纤维细胞复合胶原构建结构致密均匀的组织工程真皮，其上角质形成细胞和黑素细胞形成了层次清晰、结构良好的组织工程表皮。须癣毛癣菌感染该组织工程皮肤后，随时间延长，HE染色和PAS染色显示须癣毛癣菌对组织工程皮肤结构的破坏逐渐加重，至48 h，表皮破坏最明显，至72 h，组织工程皮肤的整体结构被菌丝和孢子所替代。 结论 初步建立了须癣毛癣菌的组织工程皮肤感染模型。

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

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 tissueengineered dermis.Then,primary keratinocytes and melanocytes were co-cultured 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.