须癣毛癣菌的组织工程皮肤感染模型的构建  

Construction of a Trlchophyton mentagrophytes.infected tissue-engineered skin model

作　　者：曹玉萍[1] 沈永年[2] 吕桂霞[2] 王乐[3] 曾荣[3] 李玲珺[4] 马鹏程[4] 刘维达[2] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所性病科,南京210042 [2]中国医学科学院北京协和医学院皮肤病研究所真菌科,南京210042 [3]中国医学科学院北京协和医学院皮肤病研究所医务处,南京210042 [4]中国医学科学院北京协和医学院皮肤病研究所药物研究室,南京210042

出　　处：《中华皮肤科杂志》2016年第1期32-35,共4页Chinese Journal of Dermatology

基　　金：国家科技重大专项（2009ZX09303-005）;江苏省自然科学基金（BK2008092）;江苏省临床医学科技专项（BL2012003）;北京协和医学院协和青年基金（33320140051）

摘　　要：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。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 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 typeⅠbovine 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.

关 键 词：组织工程 皮肤 动物替代试验 感染 须癣毛癣菌 

分 类 号：R756[医药卫生—皮肤病学与性病学]