Via the use of Mimedisc?as a scaffold,the normal human dermal fibroblasts were first seeding into the scaffold to form a dermal equivalent after 3 weeks;and then seeding normal human epidermal keratinocytes on it,followed by immersion and air-liquid inter-phase culture stage,to finally form a 3D sponge matrix skin model which has a very similar structure of normal human skin(NHS).The results of Masson staining,immuno-histochemical(IHC)and immuno -fluorescence(IF)staining show that this skin model displays a very similar structure and key protein expression of NHS.Meanwhile,this model was used as a tool to investigate the effect of Himalaya cedar extract on Dermal-Epidermal Junction(DEJ)in vitro.Results show that the Himalayan cedar extract can significantly increase the thickness of the epidermis,thus increase the barrier capacity of the skin to resist external environment.Further,the Himalaya cedar extract can promote the expression of Laminin 5 and Collagen VII of DEJ. The expression of the Laminin 5 and Collagen VII increases 20% and 47%respectively.So the Himalaya cedar extract displays the potential to be applied in development of anti-aging cosmetics.%使用Mimedisc作为支架,接种正常人真皮成纤维细胞,经过3周培养形成类似真皮结构,然后在其上接种正常人表皮角质细胞,依次经过浸没培养阶段和气-液相培养阶段,最终在体外形成3D皮肤结构模型.通过Masson染色及一系列免疫荧光染色显示此3D皮肤模型有近似真实皮肤的结构及关键蛋白表达.以此模型为工具研究了活性物喜马拉雅雪松精粹对皮肤真表皮连接处(DEJ)的作用.结果显示喜马拉雅雪松精粹可以明显提高表皮的厚度,增加皮肤抵抗外界环境的屏障能力,并且可以显著促进DEJ重要蛋白层粘连蛋白5(Laminin 5)和VII型胶原蛋白(Collagen VII,Col VII)的表达,使Laminin 5上调20%,Col VII上调47%,具有应用于抗衰老类化妆品的潜力.
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