Abstract:
Experimental systems allowing aerosol exposure (AE) of cell cultures at the air-liquid-interface (ALI) are increasingly being used to assess the toxicity of inhaled contaminants as they are more biomimetic than standard methods using submerged cultures, however, they require detailed characterisation before use. An AE-ALI system combining aerosol generation with a CULTEX® exposure chamber was characterised with respect to particle deposition and the cellular effects of filtered air (typical control) exposures. The effect of system parameters (electrostatic precipitator voltage, air flowrate to cells and insert size) on deposition efficiency and spatial distribution were investigated using ICP-MS and laser ablation ICP-MS, for an aerosol of CeO2 nanoparticles. Deposition varied with conditions, but appropriate choice of operating parameters produced broadly uniform deposition at suitable levels. The impact of air exposure duration on alveolar cells (A549) and primary small airway epithelial cells (SAECs) was explored with respect to LDH release and expression of selected genes. Results indicated that air exposures could have a significant impact on cells (e.g., cytotoxicity and expression of genes, including CXCL1, HMOX1, and SPP1) at relatively short durations (from 10 mins) and that SAECs were more sensitive. These findings indicate that detailed system characterisation is essential to ensure meaningful results.
摘要:
允许在气-液界面(ALI)进行细胞培养物的气溶胶暴露(AE)的实验系统越来越多地用于评估吸入污染物的毒性,因为它们比使用浸没培养物的标准方法更具仿生作用。然而,它们在使用前需要详细的表征。结合气溶胶产生与CULTEX®暴露室的AE-ALI系统的特征在于颗粒沉积和过滤空气(典型对照)暴露的细胞效应。系统参数的影响(静电除尘器电压,使用ICP-MS和激光烧蚀ICP-MS研究了到细胞的空气流量和插入物尺寸)对沉积效率和空间分布的影响,用于CeO2纳米颗粒的气溶胶。沉积随条件而变化,但适当选择的操作参数产生广泛均匀的沉积在适当的水平。在LDH释放和所选基因表达方面,探索了空气暴露持续时间对肺泡细胞(A549)和原发性小气道上皮细胞(SAEC)的影响。结果表明,空气暴露可能对细胞产生重大影响(例如,细胞毒性和基因表达,包括CXCL1,HMOX1和SPP1),持续时间相对较短(从10分钟起),并且SAEC更敏感。这些发现表明,详细的系统表征对于确保有意义的结果至关重要。