Institute of Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Warszawa, Poland.
Skin Res Technol. 2013 Feb;19(1):e200-8. doi: 10.1111/j.1600-0846.2012.00628.x. Epub 2012 Jun 7.
BACKGROUND/PURPOSE: The knowledge how surfactants and hydrolyzed proteins influence the elastic properties of living epidermal keratinocytes is sparse. We demonstrate that the stiffness of cells measured by atomic force microscope (AFM) can be correlated with viability test.
The effects of sodium lauryl sulphate (SLS) and hydrolyzed collagen (HK) of molecular weight 9 kDa were examined with respect to human keratinocytes viability and elasticity. MTT assay was applied to determine the survival fraction of keratinocytes treated with SLS and HK solutions of various molar ratios. The AFM measurements of the keratinocytes stiffness were carried out immediately after the exposure of cells to the SLS and HK, respectively.
The increase of the SLS concentration resulted in the decrease of cells proliferation and this effect was inhibited by addition of HK. The strongest inhibition was observed for the SLS:HK molar ratio equals to 2:1. AFM study shows decrease in the cell stiffness for cells treated with SLS. Fluorescence microscopy reveals remodeling of actin filaments of SLS-treated cells. SLS:HK mixture treatment results in mechanical stiffness close to untreated cells.
These results provide possible correlations between mechanical properties and viability of keratinocytes when the chemical stress occurs.
背景/目的:关于表面活性剂和水解蛋白如何影响活表皮角质形成细胞的弹性特性,目前知之甚少。我们证明了原子力显微镜(AFM)测量的细胞硬度可以与生存能力测试相关联。
研究了十二烷基硫酸钠(SLS)和分子量为 9 kDa 的水解胶原蛋白(HK)对人角质形成细胞活力和弹性的影响。MTT 测定法用于确定用 SLS 和 HK 溶液处理的角质形成细胞的存活率,其中 SLS 和 HK 溶液的摩尔比不同。在将细胞分别暴露于 SLS 和 HK 后,立即进行 AFM 测量以确定角质形成细胞的硬度。
随着 SLS 浓度的增加,细胞增殖减少,而添加 HK 则抑制了这种作用。当 SLS:HK 的摩尔比等于 2:1 时,观察到最强的抑制作用。AFM 研究表明,用 SLS 处理的细胞的硬度降低。荧光显微镜显示 SLS 处理的细胞中的肌动蛋白丝重塑。SLS:HK 混合物处理导致细胞的力学硬度接近未处理的细胞。
当发生化学应激时,这些结果提供了角质形成细胞的机械性能和活力之间可能的相关性。