Interdisciplinary Program of Nano-Medical Device Engineering, Graduate School, Hallym University, Chuncheon 24252, Korea.
Integrative Materials Research Institute, Hallym University, Chuncheon 24252, Korea.
Int J Mol Sci. 2021 Feb 22;22(4):2160. doi: 10.3390/ijms22042160.
Owing to the prohibition of cosmetic animal testing, various attempts have recently been made using skin-on-a-chip (SOC) technology as a replacement for animal testing. Previously, we reported the development of a pumpless SOC capable of drug testing with a simple drive using the principle that the medium flows along the channel by gravity when the chip is tilted using a microfluidic channel. In this study, using pumpless SOC, instead of drug testing at the single-cell level, we evaluated the efficacy of α-lipoic acid (ALA), which is known as an anti-aging substance in skin equivalents, for skin tissue and epidermal structure formation. The expression of proteins and changes in genotyping were compared and evaluated. Hematoxylin and eosin staining for histological analysis showed a difference in the activity of fibroblasts in the dermis layer with respect to the presence or absence of ALA. We observed that the epidermis layer became increasingly prominent as the culture period was extended by treatment with 10 μM ALA. The expression of epidermal structural proteins of filaggrin, involucrin, keratin 10, and collagen IV increased because of the effect of ALA. Changes in the epidermis layer were noticeable after the ALA treatment. As a result of aging, damage to the skin-barrier function and structural integrity is reduced, indicating that ALA has an anti-aging effect. We performed a gene analysis of filaggrin, involucrin, keratin 10, integrin, and collagen I genes in ALA-treated human skin equivalents, which indicated an increase in filaggrin gene expression after ALA treatment. These results indicate that pumpless SOC can be used as an in vitro skin model similar to human skin, protein and gene expression can be analyzed, and it can be used for functional drug tests of cosmetic materials in the future. This technology is expected to contribute to the development of skin disease models.
由于化妆品动物测试的禁令，最近人们尝试使用皮肤芯片（SOC）技术替代动物测试。之前，我们曾报道过一种无需泵的 SOC 的开发，该 SOC 可使用微流控通道倾斜芯片时，通过重力使介质沿通道流动的原理，在简单的驱动下进行药物测试。在这项研究中，我们使用无需泵的 SOC，而不是在单细胞水平上进行药物测试，评估了 α-硫辛酸（ALA）在皮肤等效物中作为抗衰老物质的功效，以评估其对皮肤组织和表皮结构形成的效果。比较并评估了蛋白质的表达和基因分型的变化。用于组织学分析的苏木精和伊红染色显示，真皮层中成纤维细胞的活性存在差异，这与是否存在 ALA 有关。我们观察到，随着培养时间的延长，10 μM ALA 处理使表皮层变得更加明显。由于 ALA 的作用，表皮结构蛋白丝聚合蛋白、兜甲蛋白、角蛋白 10 和胶原蛋白 IV 的表达增加。ALA 处理后，表皮层的变化明显。随着衰老，皮肤屏障功能和结构完整性的损伤减少，表明 ALA 具有抗衰老作用。我们对 ALA 处理的人皮肤等效物中的丝聚合蛋白、兜甲蛋白、角蛋白 10、整合素和胶原蛋白 I 基因进行了基因分析，结果表明 ALA 处理后丝聚合蛋白基因表达增加。这些结果表明，无需泵的 SOC 可作为类似人体皮肤的体外皮肤模型，可分析蛋白质和基因表达，并可用于未来化妆品材料的功能药物测试。这项技术有望为皮肤疾病模型的开发做出贡献。
