Ludwig-Maximilians-University Munich, Department of Chemistry and Center for NanoScience, Butenandtstr.11, Gerhard-Ertl-Gebäude, 81377 Munich, Germany.
Small. 2013 Dec 9;9(23):3970-80, 3906. doi: 10.1002/smll.201301004. Epub 2013 May 17.
In this study, it is shown that the cytotoxic response of cells as well as the uptake kinetics of nanoparticles (NPs) is cell type dependent. We use silica NPs with a diameter of 310 nm labeled with perylene dye and 304 nm unlabeled particles to evaluate cell type-dependent uptake and cytotoxicity on human vascular endothelial cells (HUVEC) and cancer cells derived from the cervix carcinoma (HeLa). Besides their size, the particles are characterized concerning homogeneity of the labeling and their zeta potential. The cellular uptake of the labeled NPs is quantified by imaging the cells via confocal microscopy in a time-dependent manner, with subsequent image analysis via a custom-made and freely available digital method, Particle_in_Cell-3D. We find that within the first 4 h of interaction, the uptake of silica NPs into the cytoplasm is up to 10 times more efficient in HUVEC than in HeLa cells. Interestingly, after 10 or 24 h of interaction, the number of intracellular particles for HeLa cells by far surpasses the one for HUVEC. Inhibitor studies show that these endothelial cells internalize 310 nm SiO₂ NPs via the clathrin-dependent pathway. Remarkably, the differences in the amount of taken up NPs are not directly reflected by the metabolic activity and membrane integrity of the individual cell types. Interaction with NPs leads to a concentration-dependent decrease in mitochondrial activity and an increase in membrane leakage for HUVEC, whereas HeLa cells show only a reduced mitochondrial activity and no membrane leakage. In addition, silica NPs lead to HUVEC cell death while HeLa cells survive. These findings indicate that HUVEC are more sensitive than HeLa cells upon silica NP exposure.
在这项研究中，我们表明细胞的细胞毒性反应以及纳米颗粒（NPs）的摄取动力学是依赖于细胞类型的。我们使用直径为 310nm 的带有苝染料标记的二氧化硅 NPs 和 304nm 的未标记颗粒来评估人血管内皮细胞（HUVEC）和源自宫颈癌（HeLa）的癌细胞上的细胞类型依赖性摄取和细胞毒性。除了它们的尺寸外，这些颗粒的特征还包括标记的均匀性和它们的 Zeta 电位。通过共焦显微镜以时间依赖性方式对标记的 NPs 的细胞摄取进行定量成像，随后通过定制的免费数字方法 Particle_in_Cell-3D 进行图像分析。我们发现，在相互作用的前 4 小时内，HUVEC 中二氧化硅 NPs 进入细胞质的摄取效率比 HeLa 细胞高 10 倍。有趣的是，在 10 或 24 小时的相互作用后，HeLa 细胞内的颗粒数量远远超过 HUVEC 细胞内的颗粒数量。抑制剂研究表明，这些内皮细胞通过网格蛋白依赖性途径内化 310nm SiO₂ NPs。值得注意的是，摄取的 NPs 数量的差异并没有直接反映在单个细胞类型的代谢活性和膜完整性上。与 NPs 的相互作用导致 HUVEC 的线粒体活性呈浓度依赖性下降和膜渗漏增加，而 HeLa 细胞仅显示线粒体活性降低而无膜渗漏。此外，二氧化硅 NPs 导致 HUVEC 细胞死亡，而 HeLa 细胞存活。这些发现表明，在暴露于二氧化硅 NPs 时，HUVEC 比 HeLa 细胞更敏感。
