Eskandari Ali, Yim Evelyn K F, Glerum D Moira, Tsui Ting Y
Department of Chemical Engineering, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
Materials (Basel). 2023 May 18;16(10):3814. doi: 10.3390/ma16103814.
Tissues and organs consist of cells organized in specified patterns that support their function, as exemplified by tissues such as skin, muscle, and cornea. It is, therefore, important to understand how external cues, such as engineered surfaces or chemical contaminants, can influence the organization and morphology of cells. In this work, we studied the impact of indium sulfate on human dermal fibroblast (GM5565) viability, production of reactive oxygen species (ROS), morphology, and alignment behavior on tantalum/silicon oxide parallel line/trench surface structures. The viability of cells was measured using the alamarBlue™ Cell Viability Reagent probe, while the ROS levels in cells were quantified using cell-permeant 2',7'-dichlorodihydrofluorescein diacetate. Cell morphology and orientation on the engineered surfaces were characterized using fluorescence confocal and scanning electron microscopy. When cells were cultured in media containing indium (III) sulfate, the average cell viability decreased by as much as ~32% and the concentration of cellular ROS increased. Cell geometry became more circular and compact in the presence of indium sulfate. Even though actin microfilaments continue to preferentially adhere to tantalum-coated trenches in the presence of indium sulfate, the cells are less able to orient along the line axes of the chips. Interestingly, the indium sulfate-induced changes in cell alignment behavior are pattern dependent-a larger proportion of adherent cells on structures with line/trench widths in the range of 1 μm and 10 μm lose the ability to orient themselves, compared to those grown on structures with line widths smaller than 0.5 μm. Our results show that indium sulfate impacts the response of human fibroblasts to the surface structure to which they adhere and underscores the importance of evaluating cell behaviors on textured surfaces, especially in the presence of potential chemical contaminants.
组织和器官由以特定模式组织起来的细胞构成,这些模式支持其功能,皮肤、肌肉和角膜等组织就是例证。因此,了解诸如工程表面或化学污染物等外部线索如何影响细胞的组织和形态非常重要。在这项工作中,我们研究了硫酸铟对人皮肤成纤维细胞(GM5565)活力、活性氧(ROS)产生、形态以及在钽/氧化硅平行线/沟槽表面结构上的排列行为的影响。使用alamarBlue™细胞活力试剂探针测量细胞活力,同时使用细胞渗透性的2',7'-二氯二氢荧光素二乙酸酯对细胞中的ROS水平进行定量。使用荧光共聚焦显微镜和扫描电子显微镜对工程表面上的细胞形态和取向进行表征。当细胞在含有硫酸铟(III)的培养基中培养时,平均细胞活力下降多达约32%,细胞内ROS浓度增加。在硫酸铟存在下,细胞几何形状变得更加圆形和紧凑。尽管在硫酸铟存在下肌动蛋白微丝仍继续优先附着在钽涂层的沟槽上,但细胞沿芯片线轴定向的能力较弱。有趣的是,硫酸铟诱导的细胞排列行为变化取决于图案——与生长在线宽小于0.5μm的结构上的细胞相比,生长在1μm至10μm线/沟槽宽度范围内结构上的附着细胞中,有更大比例失去了自我定向的能力。我们的结果表明,硫酸铟会影响人成纤维细胞对其附着表面结构的反应,并强调了评估细胞在纹理表面上行为的重要性,尤其是在存在潜在化学污染物的情况下。