Airborne engineered nanomaterials (ENMs) can readily enter the human body through inhalation potentially leading to adverse health effects such as cardiovascular and pulmonary diseases. Our group has previously utilized and validated an integrated low flow system capable of generating and depositing airborne ENMs directly onto cells at an air-liquid interface (ALI). To further improve this ALI method for an even closer representation of the in vivo system, a co-culture model containing epithelial, endothelial and macrophage cell lines (A549, EA.hy 926, and THP-1 differentiated macrophages) was established and validated for testing ENMs toxicity. In the co-culture model, cells were exposed to citrate-capped gold (Au), 15% silver on silica (Ag-SiO2) and copper oxide (CuO) ENMs under the same protocol (4 h ALI exposure with a target concentration of 3.5 mg/m3) and compared to responses with A549 cells only or THP-1 differentiated cells only. The toxicological profile was assessed by measuring cell viability, reactive oxygen species (ROS) production, lactate dehydrogenase (LDH) release, and interleukin (IL)-8 concentration. Results showed that 15% Ag-SiO2 induced more oxidative stress-related toxicity in the co-culture than in A549 cells alone. Both 15% Ag-SiO2 and CuO exposure produced significantly higher levels of IL-8 in the co-culture compared with A549 cells alone. Citrate-capped Au was largely inert. Further exposures of CuO on macrophages alone provided evidence of cell-cell interaction in the co-culture model. In addition, the co-culture model exhibited a similar response to primary human bronchial epithelial cells in terms of ROS and IL-8 responses after CuO exposure, suggesting a more advanced refinement of the conventional model for in vitro inhalation study.

中文翻译：

---

使用气液界面单一培养和共培养模型比较气雾化工程纳米材料的体外毒性

机载工程纳米材料 (ENM) 很容易通过吸入进入人体，可能导致心血管和肺部疾病等不良健康影响。我们的团队之前已经使用并验证了一种集成的低流量系统，该系统能够在气液界面 (ALI) 处直接生成和沉积机载 ENM 到细胞上。为了进一步改进这种 ALI 方法以更接近地代表体内系统，建立了一个包含上皮细胞、内皮细胞和巨噬细胞系（A549、EA.hy 926 和 THP-1 分化巨噬细胞）的共培养模型，并验证了测试 ENMs 的毒性。在共培养模型中，细胞暴露于柠檬酸盐封端的金 (Au)，15% 银在二氧化硅 (Ag-SiO2) 和氧化铜 (CuO) ENMs 上使用相同的方案（4 小时 ALI 暴露，目标浓度为 3.5 mg/m3）并与仅使用 A549 细胞或 THP-1 分化细胞的反应进行比较只要。通过测量细胞活力、活性氧 (ROS) 产生、乳酸脱氢酶 (LDH) 释放和白细胞介素 (IL)-8 浓度来评估毒理学特征。结果表明，与单独的 A549 细胞相比，15% 的 Ag-SiO2 在共培养物中诱导了更多的氧化应激相关毒性。与单独的 A549 细胞相比，15% Ag-SiO2 和 CuO 暴露在共培养物中产生显着更高水平的 IL-8。柠檬酸盐封端的 Au 在很大程度上是惰性的。单独在巨噬细胞上进一步暴露 CuO 提供了共培养模型中细胞 - 细胞相互作用的证据。此外，