The influence of particulate matter properties on the biophysical entry mechanisms into lung cells

颗粒物特性对肺细胞生物物理进入机制的影响

• 批准号: 448780159
• 负责人: Professor Dr. Alexander Rohrbach
• 金额: --
• 依托单位: Institut für Mikrosystemtechnik (IMTEK)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/448780159?language=en
• 关键词: influence; particulate; matter; properties; biophysical

The pollution of the ambient air with particulate matter (PM), microscopic suspended particles, is an aggravating problem that poses a serious health risk to millions of people. The exposure to elevated levels of PM significantly increases the risk for cardiovascular diseases, lung diseases and cancer. Fine to ultrafine particles with diameters < 1µm can enter lung cells and trigger inflammatory responses. Despite a large number of epidemiological studies, the mechanisms governing the direct interaction between particulate matter and single cells is hardly understood. In particular, the cell mechanical concepts involving binding and uptake are unknown. Hence, investigative optical concepts are needed, which can access cellular responses on scales of nanometers and milliseconds.For this, we will use novel optical technologies and biophysical concepts. In particular, we will use optical tweezers, three-dimensional thermal noise tracking and fast, label-free super resolution microscopy to achieve new insights into the relevant entry mechanisms into cells. Through these tools, we will answer questions about what biophysical principles govern the entry process of particulate matter into cells and how particle properties influence the fate of the particle in contact with the cell, but also how different lung cell types handle particulate matter. With our Photonic Force Microscopy, we will bring particulate matter in the vicinity of lung cells in a reproducible way to measure the change in binding strength and friction of the particle in contact with the cell. These interaction parameters can be extracted from thermal position fluctuations of the particles. The experiments are complemented by live-cell super-resolution microscopy using rotating coherent scattering (ROCS) microscopy, which enables us to observe cytoskeleton reorganization in 100 Hz in response to particulate matter exposure. By using various fluorescence cytotoxicity assays, the cellular response pattern will be correlated with the presence of pro-inflammatory cytokines.The research is devised for two PhD students working closely together on different aspects. The one PhD-student shall investigate the entry of particulate matter into lung cells using photonic force microscopy and characterize the mechanical principles governing the engulfment of different particles by thermal noise tracking. The other PhD student will focus on the investigation of the cytoskeleton response during binding and entry of different particles into lung epithelial cells using ROCS microscopy. With this research project, we expect to provide important additional knowledge to better assess the influence of particulate matter properties on lung diseases, most relevant for the fields of environmental toxicology and pulmonology.

环境空气中的颗粒物（PM），微观悬浮颗粒物的污染是一个严重的问题，对数百万人构成严重的健康风险。暴露于高水平的PM会显著增加患心血管疾病、肺部疾病和癌症的风险。直径< 1µm的细至超细颗粒可进入肺细胞并引发炎症反应。尽管进行了大量的流行病学研究，但颗粒物与单细胞之间直接相互作用的机制仍不清楚。特别是，涉及结合和摄取的细胞力学概念是未知的。因此，需要研究光学概念，它可以访问纳米和毫秒尺度上的细胞响应。为此，我们将使用新的光学技术和生物物理概念。特别是，我们将使用光镊，三维热噪声跟踪和快速，无标记的超分辨率显微镜，以实现对相关进入细胞机制的新见解。通过这些工具，我们将回答有关颗粒物进入细胞过程的生物物理原理以及颗粒特性如何影响与细胞接触的颗粒的命运，以及不同的肺细胞类型如何处理颗粒物的问题。通过我们的光子力显微镜，我们将以可重复的方式将颗粒物质带入肺细胞附近，以测量颗粒与细胞接触时的结合强度和摩擦力的变化。这些相互作用参数可以从粒子的热位置波动中提取。这些实验是由活细胞超分辨率显微镜使用旋转相干散射（ROCS）显微镜，这使我们能够观察到细胞骨架重组在100 Hz的颗粒物暴露。通过使用各种荧光细胞毒性试验，细胞反应模式将与促炎细胞因子的存在相关。该研究是为两名博士生设计的，他们在不同方面密切合作。一名博士生将使用光子力显微镜研究颗粒物质进入肺细胞的情况，并通过热噪声跟踪表征控制不同颗粒吞噬的机械原理。另一名博士生将专注于使用ROCS显微镜研究不同颗粒结合和进入肺上皮细胞期间的细胞骨架反应。通过这项研究项目，我们希望提供重要的额外知识，以更好地评估颗粒物特性对肺部疾病的影响，这与环境毒理学和肺病学领域最相关。