CAJ | 학술논문

目的利用人皮肤来源的细胞体外构建含色素组织工程皮肤白念珠菌感染模型.方法利用人皮肤原代培养成纤维细胞、角质形成细胞、黑素细胞,成纤维细胞与胶原孵育3d后得到组织工程真皮,在其表面接种角质形成细胞和黑素细胞培养12 d,获得组织工程皮肤,再在其表面滴加1个麦氏浓度的白念珠菌悬液5μl,分别在6、12、24、48和72 h进行HE染色和PAS染色观察.结果利用成纤维细胞、角质形成细胞、黑素细胞及胶原构建的组织工程皮肤结构良好,表皮分化层次分明.HE染色和PAS染色显示,感染白念珠菌后,随着时间的延长,白念珠菌对表皮结构的破坏逐渐加重,菌丝由浅表向真皮逐渐侵入.感染至72 h,表皮结构完全破坏,真皮内充满大量菌丝.结论利用人皮肤来源的细胞,培养出的含色素组织工程皮肤层次结构分明；初步建立了人皮肤来源的组织工程皮肤念珠菌感染模型.
목적 이용인피부래원적세포체외구건함색소조직공정피부백념주균감염모형.방법 이용인피부원대배양성섬유세포、각질형성세포、흑소세포,성섬유세포여효원부육3d후득도조직공정진피,재기표면접충각질형성세포화흑소세포배양12 d,획득조직공정피부,재재기표면적가1개맥씨농도적백념주균현액5μl,분별재6、12、24、48화72 h진행HE염색화PAS염색관찰.결과 이용성섬유세포、각질형성세포、흑소세포급효원구건적조직공정피부결구량호,표피분화층차분명.HE염색화PAS염색현시,감염백념주균후,수착시간적연장,백념주균대표피결구적파배축점가중,균사유천표향진피축점침입.감염지72 h,표피결구완전파배,진피내충만대량균사.결론 이용인피부래원적세포,배양출적함색소조직공정피부층차결구분명；초보건립료인피부래원적조직공정피부념주균감염모형.
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.