ImmuONE Ltd, Science Building, College Lane, Hatfield, Herts AL10 9AB, UK; Centre for Topical Drug Delivery and Toxicology, University of Hertfordshire, College Lane Campus, Hatfield, Herts AL10 9AB, UK.
Thornton & Ross Ltd, Linthwaite, Huddersfield HD7 5QH, UK; School of Clinical and Applied Sciences, Leeds Becket University, City Campus, Woodhouse Lane, Leeds LS1 3HE, UK.
Toxicol In Vitro. 2023 Feb;86:105506. doi: 10.1016/j.tiv.2022.105506. Epub 2022 Oct 27.
Assessing the safety of inhaled substances in the alveolar region of the lung requires an understanding of how the respired material interacts with both physical and immunological barriers. Human alveolar-like macrophages in vitro provide a platform to assess the immunological response in the airways and may better inform the understanding of a response to an inhaled challenge being adaptive or adverse. The aim of this study was to determine if a morphometric phenotyping approach could discriminate between different inhaled nicotine products and indicate the potential mechanism of toxicity of a substance. Cigarette smoke (CS) and e-liquids extracted into cell culture medium were applied to human alveolar-like macrophages in mono-culture (ImmuONE™) and co-culture (ImmuLUNG™) to test the hypothesis. Phenotype profiling of cell responses was highly reproducible and clearly distinguished the different responses to CS and e-liquids. Whilst the phenotypes of untreated macrophages were similar regardless of culture condition, macrophages cultured in the presence of epithelial cells were more sensitive to CS-induced changes related to cell size and vacuolation processes. This technique demonstrated phenotypical observations typical for CS exposure and indicative of the established mechanisms of toxicity. The technique provides a rapid screening approach to determine detailed immunological responses in the airways which can be linked to potentially adverse pathways and support inhalation safety assessment.
评估肺部肺泡区域吸入物质的安全性需要了解呼吸物质与物理和免疫屏障的相互作用方式。体外人肺泡样巨噬细胞为评估气道免疫反应提供了一个平台，并且可以更好地了解对吸入性挑战的反应是适应性的还是有害的。本研究旨在确定形态计量表型分析方法是否可以区分不同的吸入尼古丁产品，并指出物质毒性的潜在机制。将香烟烟雾（CS）和细胞培养液中提取的电子烟液应用于单核培养（ImmuONE™）和共培养（ImmuLUNG™）中的人肺泡样巨噬细胞，以验证假设。细胞反应的表型分析具有高度的可重复性，可清楚地区分 CS 和电子烟液的不同反应。虽然未处理的巨噬细胞的表型在培养条件下相似，但在存在上皮细胞的情况下培养的巨噬细胞对 CS 诱导的与细胞大小和空泡形成过程相关的变化更敏感。该技术显示出与 CS 暴露相关的典型表型观察结果，表明了已建立的毒性机制。该技术提供了一种快速筛选方法，可确定气道中详细的免疫反应，这些反应可与潜在的有害途径相关联，并支持吸入安全性评估。
