中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (1): 32-35.

• 论著 • 上一篇    下一篇

须癣毛癣菌的组织工程皮肤感染模型的构建

曹玉萍1,沈永年2,吕桂霞2,王乐1,曾荣1,李玲珺1,马鹏程2,刘维达2   

  1. 1. 中国医学科学院皮肤病研究所
    2. 南京 中国医学科学院北京协和医学院皮肤病研究所
  • 收稿日期:2015-06-04 修回日期:2015-06-28 出版日期:2016-01-15 发布日期:2015-12-31
  • 通讯作者: 刘维达 E-mail:liumyco@hotmail.com
  • 基金资助:

    “重大新药创制”科技重大专项;江苏省自然科学基金项目;江苏省临床医学科技专项;2014年度北京协和医学院协和青年基金

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

  • Received:2015-06-04 Revised:2015-06-28 Online:2016-01-15 Published:2015-12-31

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。

Abstract:

Cao Yuping, Shen Yongnian, Lyu Guixia, Wang Le, Zeng Rong, Li Lingjun, Ma Pengcheng, Liu Weida Department of Sexually Transmitted Disease, Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, Nanjing 210042, China (Cao YP); Department of Mycology, Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, Nanjing 210042, China (Shen YN, Lyu GX, Liu WD); Division of Medical Affairs, Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, Nanjing 210042, China (Wang L, Zeng R); Pharmacal Research Laboratory, Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, Nanjing 210042, China (Li LJ, Ma PC) Corresponding authors: Liu Weida, Email: liumyco@hotmail.com; Ma Pengcheng, Email: mpc815@163.com 【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 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.

中图分类号: 

  • R751