Zavala Jose, O'Brien Bridget, Lichtveld Kim, Sexton Kenneth G, Rusyn Ivan, Jaspers Ilona, Vizuete William
a Department of Environmental Sciences & Engineering , University of North Carolina at Chapel Hill , Chapel Hill , NC, USA .
b Department of Pediatrics , University of North Carolina at Chapel Hill , Chapel Hill , NC, USA , and.
Inhal Toxicol. 2016;28(6):251-9. doi: 10.3109/08958378.2016.1157227.
EpiAirway™ 3-D constructs are human-derived cell cultures of differentiated airway epithelial cells that may represent a more biologically relevant model of the human lung. However, limited information is available on their utility for exposures to air pollutants at the air-liquid interface (ALI).
To assess the biological responses of EpiAirway™ cells in comparison to the responses of A549 human alveolar epithelial cells after exposure to air pollutants at ALI.
Cells were exposed to filtered air, 400 ppb of ozone (O3) or a photochemically aged Synthetic Urban Mixture (SynUrb54) consisting of hydrocarbons, nitrogen oxides, O3 and other secondary oxidation products for 4 h. Basolateral supernatants and apical washes were collected at 9 and 24 h post-exposure. We assessed cytotoxicity by measuring lactate dehydrogenase (LDH) release into the culture medium and apical surface. Interleukin 6 (IL-6) and interleukin 8 (IL-8) proteins were measured in the culture medium and in the apical washes to determine the inflammatory response after exposure.
Both O3 and SynUrb54 significantly increased basolateral levels of LDH and IL-8 in A549 cells. No significant changes in LDH and IL-8 levels were observed in the EpiAirway™ cells, however, IL-6 in the apical surface was significantly elevated at 24 h after O3 exposure.
LDH and IL-8 are robust endpoints for assessing toxicity in A549 cells. The EpiAirway™ cells show minimal adverse effects after exposure suggesting that they are more toxicologically resistant compared to A549 cells. Higher concentrations or longer exposure times are needed to induce effects on EpiAirway™ cells.
EpiAirway™三维构建体是分化气道上皮细胞的人源细胞培养物，可能代表更具生物学相关性的人肺模型。然而，关于它们在气液界面（ALI）暴露于空气污染物时的效用的信息有限。
评估在ALI暴露于空气污染物后，EpiAirway™细胞与A549人肺泡上皮细胞的反应相比的生物学反应。
将细胞暴露于过滤空气、400 ppb的臭氧（O3）或由碳氢化合物、氮氧化物、O3和其他二次氧化产物组成的光化学老化合成城市混合物（SynUrb54）中4小时。在暴露后9小时和24小时收集基底外侧上清液和顶端冲洗液。我们通过测量释放到培养基和顶端表面的乳酸脱氢酶（LDH）来评估细胞毒性。在培养基和顶端冲洗液中测量白细胞介素6（IL-6）和白细胞介素8（IL-8）蛋白，以确定暴露后的炎症反应。
O3和SynUrb54均显著增加了A549细胞中基底外侧的LDH和IL-8水平。然而，在EpiAirway™细胞中未观察到LDH和IL-8水平的显著变化，但是在O3暴露后24小时，顶端表面的IL-6显著升高。
LDH和IL-8是评估A549细胞毒性的可靠指标。EpiAirway™细胞在暴露后显示出最小的不良反应，表明它们比A549细胞具有更高的毒理学抗性。需要更高的浓度或更长的暴露时间才能对EpiAirway™细胞产生影响。
