Yamaguchi Fumiko, Watanabe Shin-Ichi, Harada Fusae, Miyake Miyuki, Yoshida Masaki, Okano Tomomichi
Human & Environment Safety Evaluation Center, Lion Corporation.
J Oleo Sci. 2014;63(10):995-1004. doi: 10.5650/jos.ess13154. Epub 2014 Sep 10.
We investigated the effect of the alkyl-chain length of anionic surfactants on the skin using an in vitro model. The evaluated anionic surfactants were sodium alkyl sulfate (AS) and sodium fatty acid methyl ester sulfonate (MES), which had different alkyl-chain lengths (C8-C14). Skin tissue damage and permeability were examined using a reconstructed human epidermal model, LabCyte EPI-MODEL24. Skin tissue damage was examined by measuring cytotoxicity with an MTT assay. Liquid chromatography/tandem mass spectrometry (LC/MS-MS) and liquid chromatography/mass spectrometry (LC/MS) were used to detect surfactants that permeated into the assay medium through an epidermal model. To assess the permeation mechanism and cell damage caused by the surfactants through the epidermis, we evaluated the structural changes of Bovine Serum Albumin (BSA), used as a simple model protein, and the fluidity of 1,2-dipalmitoyl-sn-glycero-3-phosphpcholine (DPPC) liposome, which serves as one of the most abundant phospholipid models of living cell membranes in the epidermis. The effects of the surfactants on the proteins were measured using Circular Dichroism (CD) spectroscopy, while the effects on membrane fluidity were investigated by electron spin resonance (ESR) spectroscopy. ET50 (the 50% median effective time) increased as follows: C10 < C12 < C8 < C14 in AS and C8, C10 < C12 < C14 in MES. The order of permeation through the LabCyte EPI-MODEL24 was C10 > C12 > C14, for both AS and MES. For both AS and MES, the order parameter, which is the criteria for the microscopic viscosity of lipid bilayers, increased as follows: C10 < C12 < C14, which means the membrane fluidity is C10 > C12 > C14. It was determined that the difference in skin tissue damage in the LabCyte EPI-MODEL24 with C10 to C14 AS and MES was caused by the difference in permeation and cell membrane fluidity through the lipid bilayer path in the epidermis.
我们使用体外模型研究了阴离子表面活性剂的烷基链长度对皮肤的影响。所评估的阴离子表面活性剂为烷基硫酸钠（AS）和脂肪酸甲酯磺酸钠（MES），它们具有不同的烷基链长度（C8 - C14）。使用重组人表皮模型LabCyte EPI - MODEL24检查皮肤组织损伤和通透性。通过MTT法测定细胞毒性来检查皮肤组织损伤。液相色谱/串联质谱（LC/MS - MS）和液相色谱/质谱（LC/MS）用于检测通过表皮模型渗透到测定培养基中的表面活性剂。为了评估表面活性剂通过表皮引起的渗透机制和细胞损伤，我们评估了用作简单模型蛋白的牛血清白蛋白（BSA）的结构变化，以及1,2 - 二棕榈酰 - sn - 甘油 - 3 - 磷酸胆碱（DPPC）脂质体的流动性，DPPC脂质体是表皮中活细胞膜最丰富的磷脂模型之一。使用圆二色性（CD）光谱测量表面活性剂对蛋白质的影响，同时通过电子自旋共振（ESR）光谱研究对膜流动性的影响。ET50（50％中位有效时间）的增加顺序如下：AS中C10 < C12 < C8 < C14，MES中C8、C10 < C12 < C14。对于AS和MES，通过LabCyte EPI - MODEL24的渗透顺序均为C10 > C12 > C14。对于AS和MES，作为脂质双层微观粘度标准的序参数增加顺序如下：C10 < C12 < C14，这意味着膜流动性为C10 > C12 > C14。已确定，在LabCyte EPI - MODEL24中，C10至C14的AS和MES对皮肤组织损伤的差异是由通过表皮脂质双层路径的渗透和细胞膜流动性差异引起的。
