Department of Mechanical Engineering, KAIST, 291 Daehakro, Yuseong-gu, Daejeon 34034, Republic of Korea.
School of Environmental Engineering, University of Seoul, 163 Seoulsiripdae-ro, Dongdaemun-gu, Seoul 02504, Republic of Korea.
Toxicol Appl Pharmacol. 2021 Aug 1;424:115589. doi: 10.1016/j.taap.2021.115589. Epub 2021 May 23.
Changes in the physical state of the cells can serve as important indicators of stress responses because they are closely linked with the changes in the pathophysiological functions of the cells. Physical traits can be conveniently assessed by analyzing the morphological features and the stresses at the cell-matrix and cell-cell adhesions in both single-cell and monolayer model systems in 2D. In this study, we investigated the mechano-stress responses of human bronchial epithelial cells, BEAS-2B, to two functionally distinct groups of biocides identified during the humidifier disinfectant accident, namely, guanidine (PHMG) and isothiazolinone (CMIT/MIT). We analyzed the physical traits, including cell area, nuclear area, and nuclear shape. While the results showed inconsistent average responses to the biocides, the degree of dispersion in the data set, measured by standard deviation, was remarkably higher in CMIT/MIT treated cells for all traits. As mechano-stress endpoints, traction and intercellular stresses were also measured, and the cytoskeletal actin structures were analyzed using immunofluorescence. This study demonstrates the versatility of the real-time imaging-based biomechanical analysis, which will contribute to identifying the temporally sensitive cellular behaviors as well as the emergence of heterogeneity in response to exogenously imposed stress factors. This study will also shed light on a comparative understanding of less studied substance, CMIT/MIT, in relation to a more studied substance, PHMG, which will further contribute to more strategic planning for proper risk management of the ingredients involved in toxicological accidents.
细胞物理状态的变化可以作为应激反应的重要指标，因为它们与细胞的病理生理功能变化密切相关。通过分析单细胞和单层模型系统中细胞-基质和细胞-细胞黏附处的形态特征和应力，可以方便地评估物理特征。在这项研究中，我们研究了人支气管上皮细胞 BEAS-2B 对两种功能不同的杀菌剂的机械应激反应，这两种杀菌剂是在加湿器消毒剂事故中发现的，分别是胍（PHMG）和异噻唑啉酮（CMIT/MIT）。我们分析了物理特征，包括细胞面积、核面积和核形状。虽然结果显示细胞对杀菌剂的平均反应不一致，但用标准差衡量的数据集中的离散度在 CMIT/MIT 处理的细胞中所有特征都显著更高。作为机械应激终点，还测量了牵引力和细胞间应力，并使用免疫荧光法分析了细胞骨架肌动蛋白结构。这项研究展示了基于实时成像的生物力学分析的多功能性，这将有助于识别对外源性施加的应激因素的时间敏感的细胞行为以及异质性的出现。这项研究还将有助于比较研究不太受关注的物质 CMIT/MIT 与更受关注的物质 PHMG，这将有助于更有策略地规划与毒理学事故相关的成分的适当风险管理。
