细胞共培养人体皮肤类似物超微结构观察及其渗透性测试

发布时间：2018-12-15 18:52

【摘要】：目的观察人体皮肤类似物模型形态结构,对其屏障功能进行初步评价。方法通过胶原蛋白包被成纤维细胞,制备真皮层,真皮层表面接种角质细胞,浸没培养1周后,气-液界面培养以促进角质细胞的分化形成角质层,3周后收获皮肤模型;通过肉眼观察、电镜观察皮肤类似物的宏观结构和超微结构;以吲哚美辛作为模型药物,进行体外透皮实验考察细胞共培养皮肤类似物模型的屏障功能。结果皮肤类似物形成与人类皮肤相似的真皮层和表皮层,表皮层表面形成完整的角质层,具有与人类皮肤类似的超微结构;24 h内渗透性实验结果显示皮肤类似物屏障功能略低于大鼠腹部皮肤,但22~24 h,皮肤类似物对模型药的透过速率远小于大鼠腹部皮肤(约为其28%),且2种皮肤模型对吲哚美辛经皮渗透的稳态流速没有显著性差异(P0.05)。结论皮肤类似物有与人体皮肤相类似的形态结构特征,与大鼠腹部皮肤相比对吲哚美辛有相似的屏障作用,有望替代动物皮肤成为经皮给药系统评价的新型皮肤模型。
[Abstract]:Objective to observe the morphologic structure of human skin analogue model and evaluate its barrier function. Methods the fibroblasts were coated with collagen protein to prepare the dermis. The keratinocytes were inoculated on the surface of the dermis. After 1 week of immersion and culture, the air-liquid interface was cultured to promote the differentiation of the keratinocytes into the keratinocytes, and the skin model was harvested after 3 weeks. The macrostructure and ultrastructure of the skin analogue were observed by naked eye and electron microscope, and the barrier function of the skin analogue model co-cultured by cell culture was investigated by transdermal experiments in vitro with indomethacin as the model drug. Results the skin analogue formed the dermis and epidermis similar to the human skin, and the surface of the epidermis formed a complete cuticle, which had the ultrastructure similar to that of the human skin. The results of permeability test within 24 h showed that the barrier function of skin analogue was slightly lower than that of abdominal skin of rats, but the permeation rate of skin analogue to the model drug was much lower than that of abdominal skin (about 28%) at 22 ~ 24 h. There was no significant difference in the steady flow rate of indomethacin permeation between the two skin models (P0.05). Conclusion the skin analogue has similar morphological and structural characteristics to human skin and has similar barrier effect to indomethacin compared with the abdominal skin of rats. It is expected to replace animal skin as a new skin model for the evaluation of drug delivery system.
【作者单位】：北京中医药大学中药学院;北京中医药大学科研实验中心;
【基金】：国家自然科学基金项目(No.81473365) 北京市自然科学基金项目(No.7132127) 北京中医药大学创新团队发展计划项目(No.2011-CXTD-13);北京中医药大学自主选题项目(No.2015-JYB-XS095)
【分类号】：R329.2

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