Quartz crystal microbalances (QCM) are suitable for real-time dosimetry in nanotoxicological studies using VITROCELL®Cloud cell exposure systems
Quartz crystal microbalances (QCM) are suitable for real-time dosimetry in nanotoxicological studies using VITROCELL®Cloud cell exposure systems
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DOI：
10.1186/s12989-020-00376-w
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发表时间：
2020-09-16
影响因子：
10
通讯作者：
Schmid, Otmar
中科院分区：
文献类型：
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作者：
Ding, Yaobo;Weindl, Patrick;Schmid, Otmar
背景 准确了解细胞/组织传递的剂量在吸入毒理学研究中起着关键作用，因为它是危险评估和体外剂量反应转化为体内剂量反应的关键参数。传统上，体内和体外肺模型的（纳米）颗粒毒理学研究依赖于硅计算或离线剂量测定分析方法。与水下细胞培养条件下的传统体外测试相比，更生理的气液界面 (ALI) 条件为使用石英晶体微天平 (QCM) 进行实时剂量测定提供了可能性。然而，尚不清楚 QCM 是否足够灵敏以进行纳米毒理学研究。我们针对两种商用 VITROCELL (R) Cloud ALI 曝光系统调查了这个问题。结果 添加荧光素的盐水气溶胶的定量荧光光谱用于确定 VITROCELL (R) 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 的检测限、精密度和准确度。两种 QCM 在整个研究剂量范围（200 至 12,000 ng/cm(2)）内均呈线性表现，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/cm(2)）的2.5%分别下降到1000 ng/cm（2）和200 ng/cm（2）剂量的10%甚至25%。 Cloud 6 和 Cloud 12 的检测下限分别为 170 ng/cm(2) 和 169 ng/cm(2)。 (NM110) ZnO 纳米颗粒的剂量反应测量显示，A549 肺上皮细胞的细胞活力 (WST-1) 和细胞毒性 (LDH) 的起效剂量为 3.3 μg/cm(2)（或 0.39 cm(2)/cm(2)）。结论 Cloud 6 和 Cloud 12 系统的 QCM 显示出相似的性能，并且如果按照制造商规范操作，则它们是用于在 ALI 细胞暴露实验中对细胞传递的粒子剂量进行（准）实时剂量测定的高度灵敏、准确的设备。与本次和之前发表的 ALI 研究的体外起效剂量进行比较表明，170 ng/cm(2) 的检测限足以确定此处研究的所有具有低质量特异性毒性（例如聚苯乙烯）或高质量特异性毒性（例如 ZnO 和 Ag）的颗粒类型的毒理学起效剂量。因此，原则上 QCM 适合体外纳米毒理学研究，但应在本研究中描述的特定暴露条件下对每个 QCM 和 ALI 暴露系统进行研究。
Background 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 onin siliocomputational 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 (R) Cloud ALI exposure systems. Results Quantitative fluorescence spectroscopy of fluorescein-spiked saline aerosol was used to determine detection limit, precision and accuracy of the QCMs implemented in a VITROCELL (R) 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/cm(2)) 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/cm(2)) to values of 10% and even 25% for doses of 1000 ng/cm(2)and 200 ng/cm(2), respectively. Their lower detection limit was 170 ng/cm(2)and 169 ng/cm(2)for the Cloud 6 and Cloud 12, respectively. Dose-response measurements with (NM110) ZnO nanoparticles revealed an onset dose of 3.3 mu g/cm(2)(or 0.39 cm(2)/cm(2)) for both cell viability (WST-1) and cytotoxicity (LDH) of A549 lung epithelial cells. Conclusions 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/cm(2)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.