Assessment of biological responses of EpiAirway 3-D cell constructs versus A549 cells for determining toxicity of ambient air pollution.
Assessment of biological responses of EpiAirway 3-D cell constructs versus A549 cells for determining toxicity of ambient air pollution.
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DOI：
10.3109/08958378.2016.1157227
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发表时间：
2016
影响因子：
2.1
通讯作者：
Vizuete W
中科院分区：
文献类型：
--
作者：
Zavala J;O'Brien B;Lichtveld K;Sexton KG;Rusyn I;Jaspers I;Vizuete W
EpiAirway™3-D构建物是分化的呼吸道上皮细胞的人类来源细胞培养，可能代表更具生物学相关性的人类肺模型。然而，关于其在空气-液体界面(ALI)暴露于空气污染物的效用的信息有限。目的：通过比较A549人肺泡上皮细胞和EpiAirway™细胞在ALI暴露于空气污染物后的生物学反应，来评估ALI时EpiAirway细胞的生物学反应。将细胞暴露在过滤后的空气、400 ppb臭氧(O3)或由碳氢化合物、氮氧化物、臭氧和其他二次氧化产物组成的光化学老化的合成城市混合物(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根尖冲洗液。我们通过测量乳酸脱氢酶(LDH)释放到培养液和根尖表面来评估细胞毒性。检测培养上清液和根尖冲洗液中白介素6(IL-6)和白介素8(IL-8)蛋白的含量，以确定暴露后的炎症反应。O_3和SynUrb54均显著增加A549细胞的LDH和IL-8水平。EpiAirway™细胞中乳酸脱氢酶和IL-8水平无明显变化，而根尖表面IL-6水平在臭氧暴露后24 h显著升高。乳酸脱氢酶和IL-8是评估A549细胞毒性的可靠终点。EpiAirway™细胞在暴露后显示出最小的不良反应，这表明它们比A549细胞具有更强的毒理抗性。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。
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 of its 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, 400ppb 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.