MoSAIC/Biomedical NMR Unit, Department of Medicine, Catholic University of Leuven, B3000 Leuven, Belgium.
Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, MA 01003, United States.
Biomaterials. 2014 Dec;35(37):9941-9950. doi: 10.1016/j.biomaterials.2014.08.031. Epub 2014 Sep 15.
The effects of nanoparticle (NP)-related parameters on cellular interactions are currently uncertain as analysis is complicated by the combinatorial diversity arising from the array of size, shape and surface properties. Here, we present a validated multiparametric high-content imaging method, with the utility of this approach demonstrated by in-depth analysis of the role of hydrophobicity on the interaction of Au NPs with cultured cells. In this methodology, we evaluate cell viability, membrane damage, induction of reactive oxygen species, mitochondrial health, cell area, skewness and induction of autophagy. High-content cell cycle phase studies and in-depth gene expression studies then serve to elucidate the underlying mechanisms. The data reveal a clear influence of the degree of NP surface hydrophobicity with membrane damage and autophagy induction, which is stronger than the effect of surface charge, for charges ranging between -50 and +20 mV. All labeling experiments occur in the same format, and can be further supplemented with additional parameters providing a broadly accessible format for studying cell-NP interactions under highly reproducible conditions.
纳米颗粒（NP）相关参数对细胞相互作用的影响目前尚不确定，因为分析受到尺寸、形状和表面特性组合多样性的影响而变得复杂。在这里，我们提出了一种经过验证的多参数高内涵成像方法，通过深入分析疏水性对 Au NPs 与培养细胞相互作用的影响，展示了该方法的实用性。在该方法中，我们评估细胞活力、膜损伤、活性氧的诱导、线粒体健康、细胞面积、偏度和自噬的诱导。高内涵细胞周期相研究和深入的基因表达研究随后用于阐明潜在的机制。数据显示，NP 表面疏水性对膜损伤和自噬诱导的影响明显大于表面电荷的影响，对于-50 至+20 mV 之间的电荷。所有标记实验都以相同的格式进行，并且可以进一步补充其他参数，为在高度可重复的条件下研究细胞-NP 相互作用提供广泛适用的格式。
