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.

微粒物质（PM）的环境空气污染是微观悬浮颗粒，是一个汇总的问题，对数百万人构成了严重的健康风险。 PM升高的暴露显着增加了心血管疾病，肺部疾病和癌症的风险。直径<1µm的细细到超细颗粒可以进入肺部细胞并触发炎症反应。尽管有大量的流行病学研究，但几乎不了解管理特定物质与单个细胞之间直接相互作用的机制。特别是，细胞机械概念涉及结合和摄取是未知的。因此，需要调查性光学概念，它可以访问纳米和毫秒的尺度上的细胞反应。为此，我们将使用新颖的光学技术和生物物理概念。特别是，我们将使用光学镊子，三维热噪声跟踪和快速，无标签的超级分辨率显微镜来实现对相关进入机制的新见解。通过这些工具，我们将回答有关哪些生物物理原理控制特定物质进入细胞的问题以及粒子特性如何影响与细胞接触的粒子的命运，以及不同的肺细胞类型如何处理特定物质的问题。使用我们的光子力显微镜，我们将以可重复的方式在肺部细胞附近带来特殊物质，以测量与细胞接触的粒子的结合强度和摩擦的变化。这些相互作用参数可以从颗粒的热位置波动中提取。实验是通过使用旋转相干散射（ROC）显微镜实时超分辨率显微镜完成的，这使我们能够观察100 Hz的细胞骨架重组，以响应颗粒物暴露。通过使用各种荧光细胞毒性刺客，细胞反应模式将与促炎性细胞因子的存在相关。该研究是针对两个在不同方面紧密合作的博士学位学生的研究。一位博士学位学生应使用光子力显微镜研究颗粒物的进入肺细胞，并通过热噪声跟踪来表征控制不同颗粒的机械原理。另一位博士生将使用ROCS显微镜在不同颗粒进入和进入肺上皮细胞期间对细胞骨架反应的研究。通过该研究项目，我们希望提供重要的额外知识，以更好地评估颗粒物质对肺部疾病的影响，这与环境毒理学和肺病学领域最相关。