Comparison of human primary lung and 3D co-culture with respect to adaptive response towards Bariumsulphate nanoparticle aerosol

人原肺和 3D 共培养对硫酸钡纳米颗粒气溶胶适应性反应的比较

• 批准号: 397981139
• 负责人: Professorin Dr. Heidi Foth
• 金额: --
• 依托单位: Institut für Umwelttoxikologie (aufgelöst)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/397981139?language=en
• 关键词: Comparison; human; primary; lung; 3D

Despite all progress in toxicological risk assessment there are still substantial information deficits, in particular concerning assessment of subtoxic and repeated exposure scenarios as well as in the understanding in the stress response due to particle overload and the potential of adaptive response towards low dose exposure. It is still not understood whether the differences concerning sensitivity and variability between human tissue and other species or between tumor cell lines and primary cells are substantial. The designed project aims to determine the mobility and cellular effects of Bariumsulphate and Titandioxide nanoparticle (positive control) on respiratory tissue of native human lung (L-MOC) as well as on 3D co-cultures of human lung cells and endothelial cells. The biological material will be exposed towards subtoxic amount of nanoparticle aerosols (BaSO4, TiO2) in order to avoid particle overload. The exposure scenario will be applied repeatedly and effect will be monitored over several weeks. The project will focus on following tasks: 1) Is there any difference between 3D Configuration of cell culture inserts (MatriGrids®) and 2D culture systems with respect of deposition of nanoparticles? Furthermore does the cellular response differ accordingly?2) Is the sensitivity concerning stress reaction the same in primary lung cells compared to permanent cell line A549 when both cell types are maintained in 3D co-culture conditions. 3) Does the sensitivity of peripheral (respiratory) lung cells change between physiological morphology in L-MOC compared to 3D co-culture configuration?The nanoparticle aerosols will be applied as single or repeated dose in an exposure chamber with MatriGrids® culture inserts (expertise TILUM) on primary lung tissue (expertise MLU). Monitoring time of cellular responses will be extended up to four weeks. The parameters of interest are cytotoxic effects (viability, marker for reactive oxygen species, level of glutathione, expression and release of cytokines), markers for epithelial morphology and cell to cell contacts. The special focus will be laid on low subtoxic exposure levels and on monitoring of cellular reactions over several weeks. The aim is to determine which effects are direct particle associated and which are induced by mediators of inflammation. The project will combine the expertise from engineering science and toxicology in order to establish and control conditions of aerosol generation and application on cell culture. The exposure chambers and MatriGrids® 3D inserts are established at TIULM and will be optimized according to the needs of the projects. The results will provide the bases to derive NOAEL for BaSO4 to enable risk assessment for nanoparticles for in vivo situation neither to overestimate nor underestimate.

尽管在毒理学风险评估方面取得了所有进展，但仍然存在大量信息不足，特别是在评估亚毒性和重复接触情景方面，以及在理解颗粒过载引起的应激反应和对低剂量接触的适应性反应潜力方面。目前还不清楚人体组织和其他物种之间或肿瘤细胞系和原代细胞之间的敏感性和变异性差异是否很大。该设计项目旨在确定硫酸钡和二氧化钛纳米颗粒（阳性对照）对天然人肺（L-MOC）呼吸组织以及人肺细胞和内皮细胞的3D共培养物的迁移率和细胞效应。生物材料将暴露于毒性较低的纳米颗粒气溶胶（BaSO 4、TiO 2）中，以避免颗粒过载。将重复应用暴露场景，并在数周内监测效果。本项目将重点开展以下工作：1） 细胞培养插入物（MatriGrids®）的3D配置和2D培养系统在纳米颗粒沉积方面是否有任何差异？此外，细胞反应是否相应地不同？（二） 当两种细胞类型维持在3D共培养条件下时，与永久细胞系A549相比，原代肺细胞中关于应激反应的灵敏度是否相同。第三章 与3D共培养配置相比，外周（呼吸）肺细胞的灵敏度在L-MOC中的生理形态之间是否发生变化？纳米颗粒气雾剂将在具有MatriGrids®培养插入物（expertise TILUM）的暴露室中以单次或重复剂量应用于原发性肺组织（expertise MLU）。细胞反应的监测时间将延长至四周。感兴趣的参数是细胞毒性效应（活力、活性氧物质标记物、谷胱甘肽水平、细胞因子的表达和释放）、上皮形态标记物和细胞与细胞接触。特别重点将放在低亚毒性接触水平和监测几周内的细胞反应。目的是确定哪些效应是直接与颗粒相关的，哪些是由炎症介质诱导的。该项目将联合收割机结合工程科学和毒理学的专业知识，以建立和控制气溶胶产生和应用于细胞培养的条件。暴露室和MatriGrids® 3D插件在TIULM建立，并将根据项目需求进行优化。这些结果将为推导BaSO 4的NOAEL提供依据，以使纳米颗粒在体内情况下的风险评估既不会高估也不会低估。