PDF(Pubmed)
Abstract：
Two-dimensional (2D) cell culture is an important tool in the discovery of skin-active agents. Fibroblasts and keratinocytes, more rarely fibroblast-keratinocyte cocultures, are usually used for that purpose, where test compounds are added by mixing with the overlaying growth medium. However, such an approach is suboptimal because it lacks the stratum corneum component. The stratum corneum acts as a selective gatekeeper and opposes the intradermal permeation of many compounds that are bioactive when placed in direct contact with cells. One solution is to use reconstituted epidermis, but this approach is costly and time consuming. Here, a model is proposed, where the simplicity and convenience of the 2D cell culture is combined with the advantage of a hydrophobic barrier reminiscent of the skin horny layer. This model was tested with skin-relevant solvents, as well as with \"naked\" hydrophilic and encapsulated compounds. Cell viability and collagen stimulation were used as readouts. The results showed that the incorporation of a stratum corneum-substitute barrier on top of a 2D cell culture reduced the cytotoxicity of a common cosmetic solvent, dimethyl isosorbide (DMI), in cell culture and modified the bioactivity of the added actives (magnesium ascorbyl phosphate [MAP] and oligomeric proanthocyanidins [OPCs]/levan biopolymer), which became dependent on their ability to penetrate through a lipidic layer. Taken together, these results indicate a better physiological relevance of this cell culture model in workflows aimed at the discovery and analysis of skin-active compounds than conventional 2D systems.
摘要：
二维（2D）细胞培养是发现皮肤活性剂的重要工具。成纤维细胞和角质形成细胞，更罕见的成纤维细胞-角质形成细胞共培养，通常用于此目的，其中通过与覆盖的生长培养基混合来添加测试化合物。然而,这种方法是次优的，因为它缺乏角质层成分。角质层充当选择性看门人,并且对抗许多化合物的皮内渗透,所述化合物在与细胞直接接触时具有生物活性。一种解决方案是使用重组表皮，但是这种方法既昂贵又耗时。这里,提出了一个模型，其中2D细胞培养的简单和方便与让人联想到皮肤角质层的疏水屏障的优点相结合。这个模型是用皮肤相关溶剂测试的，以及“裸露的”亲水和包封的化合物。使用细胞活力和胶原刺激作为读数。结果表明，在2D细胞培养物顶部掺入角质层替代物屏障降低了常见化妆品溶剂的细胞毒性，二甲基异山梨醇（dmi），在细胞培养中，并修改了添加的活性物质（抗坏血酸磷酸镁[MAP]和低聚原花青素[OPCs]/果聚糖生物聚合物）的生物活性，这取决于它们穿透脂质层的能力。一起来看,这些结果表明，与传统的2D系统相比，该细胞培养模型在旨在发现和分析皮肤活性化合物的工作流程中具有更好的生理相关性。
