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Quartz crystal microbalances (QCM) are suitable for real-time dosimetry in nanotoxicological studies using VITROCELL®Cloud cell exposure systems.
Particle and Fibre Toxicology ( IF 10 ) Pub Date : 2020-09-16 , DOI: 10.1186/s12989-020-00376-w
Yaobo Ding 1, 2 , Patrick Weindl 1, 2, 3 , Anke-Gabriele Lenz 1, 2 , Paula Mayer 1, 2 , Tobias Krebs 3 , Otmar Schmid 1, 2
Affiliation  

Accurate knowledge of cell−/tissue-delivered dose plays a pivotal role in inhalation toxicology studies, since it is the key parameter for hazard assessment and translation of in vitro to in vivo dose-response. Traditionally, (nano-)particle toxicological studies with in vivo and in vitro models of the lung rely on in silio computational or off-line analytical methods for dosimetry. In contrast to traditional in vitro testing under submerged cell culture conditions, the more physiologic air-liquid interface (ALI) conditions offer the possibility for real-time dosimetry using quartz crystal microbalances (QCMs). However, it is unclear, if QCMs are sensitive enough for nanotoxicological studies. We investigated this issue for two commercially available VITROCELL®Cloud ALI exposure systems. Quantitative fluorescence spectroscopy of fluorescein-spiked saline aerosol was used to determine detection limit, precision and accuracy of the QCMs implemented in a VITROCELL®Cloud 6 and Cloud 12 system for dose-controlled ALI aerosol-cell exposure experiments. Both QCMs performed linearly over the entire investigated dose range (200 to 12,000 ng/cm2) with an accuracy of 3.4% (Cloud 6) and 3.8% (Cloud 12). Their precision (repeatability) decreased from 2.5% for large doses (> 9500 ng/cm2) to values of 10% and even 25% for doses of 1000 ng/cm2 and 200 ng/cm2, respectively. Their lower detection limit was 170 ng/cm2 and 169 ng/cm2 for the Cloud 6 and Cloud 12, respectively. Dose-response measurements with (NM110) ZnO nanoparticles revealed an onset dose of 3.3 μg/cm2 (or 0.39 cm2/cm2) for both cell viability (WST-1) and cytotoxicity (LDH) of A549 lung epithelial cells. The QCMs of the Cloud 6 and Cloud 12 systems show similar performance and are highly sensitive, accurate devices for (quasi-) real-time dosimetry of the cell-delivered particle dose in ALI cell exposure experiments, if operated according to manufacturer specifications. Comparison with in vitro onset doses from this and previously published ALI studies revealed that the detection limit of 170 ng/cm2 is sufficient for determination of toxicological onset doses for all particle types with low (e.g. polystyrene) or high mass-specific toxicity (e.g. ZnO and Ag) investigated here. Hence, in principle QCMs are suitable for in vitro nanotoxciological studies, but this should be investigated for each QCM and ALI exposure system under the specific exposure conditions as described in the present study.

中文翻译:

石英晶体微量天平(QCM)适用于使用VITROCELL®Cloud细胞暴露系统进行纳米毒理学研究的实时剂量测定。

对细胞/组织递送剂量的准确了解在吸入毒理学研究中起着关键作用,因为它是危险性评估以及体外到体内剂量反应转化的关键参数。传统上,使用体内和体外肺部模型进行的(纳米)颗粒毒理学研究都依靠剂量计算的计算机内计算或离线分析方法。与在浸没细胞培养条件下进行的传统体外测试相比,更具生理性的气液界面(ALI)条件为使用石英晶体微量天平(QCM)进行实时剂量测定提供了可能性。但是,目前尚不清楚QCM是否足够敏感以进行纳米毒理学研究。我们针对两个市售的VITROCELL®Cloud ALI曝光系统调查了此问题。荧光素加标的盐雾气溶胶的定量荧光光谱法用于确定VITROCELL®Cloud 6和Cloud 12系统中用于剂量控制ALI气溶胶细胞暴露实验的QCM的检出限,精密度和准确性。两个QCM在整个研究剂量范围(200至12,000 ng / cm2)内线性执行,准确度分别为3.4%(云6)和3.8%(云12)。它们的精度(重复性)从大剂量(> 9500 ng / cm2)的2.5%降低到1000 ng / cm2和200 ng / cm2的10%甚至25%的值。对于云6和云12,它们的下限分别为170 ng / cm2和169 ng / cm2。(NM110)ZnO纳米颗粒的剂量反应测量显示起始剂量为3.3μg/ cm2(或0。39平方厘米/平方厘米)的A549肺上皮细胞的细胞活力(WST-1)和细胞毒性(LDH)。如果根据制造商的规范进行操作,Cloud 6和Cloud 12系统的QCM表现出相似的性能,并且是用于ALI细胞暴露实验中的细胞递送颗粒剂量的(准)实时剂量测定的高度灵敏,准确的设备。与本次和先前发表的ALI研究的体外起始剂量进行比较后,发现170 ng / cm2的检出限足以确定所有低(例如聚苯乙烯)或高质量特异性毒性(例如ZnO)颗粒的毒理学起始剂量。和Ag)在这里进行调查。因此,原则上,QCM适用于体外纳米毒理学研究,
更新日期:2020-09-18
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