Interaction of nanoparticles with cellular compartments and proteins

将合成和表征具有关于尺寸、聚合物类型和含有荧光标记物的表面官能度的限定参数的聚合物纳米颗粒组。通过先进的荧光显微镜技术，我们将详细探索细胞的摄取机制及其对纳米颗粒物理性质（大小，形状）及其表面化学（化学功能，电荷和极性）的依赖性。我们将详细分析纳米颗粒如何在细胞内处理（运输机制，与细胞器的关联，降解），以及细胞对纳米颗粒掺入的反应。这些研究将通过使用时间分辨旋转圆盘共聚焦显微镜对Mailänder/Simmet小组提供的培养细胞进行，并辅以对固定细胞进行超高空间分辨率的4Pi共聚焦实验。通过这些实验，我们的目标是在分子方面实现培养细胞对纳米颗粒的反应的机制描述，这是理解纳米颗粒对整个生物体影响的第一个重要步骤。炎症过程对所有复杂生物体的生存至关重要，并在健康和疾病中起着重要作用。抗原提呈和吞噬单核细胞是炎症和宿主防御的关键。在此背景下，非常重要的是不仅要了解i）独特设计的纳米颗粒和单核细胞之间的相互作用，而且要识别ii）可能的靶分子，iii）基本的生化机制，以及iv）可以被纳米颗粒靶向的细胞功能。

Sets of polymeric nanoparticles of defined parameters concerning size, type of polymer and surface functionality containing a fluorescent marker will be synthesized and characterized. With advanced fluorescence microscopy techniques, we will explore in detail the uptake mechanisms by the cell and their dependencies on the physical properties of the nanoparticle (size, shape) and its surface chemistry (chemical functions, charge, and polarity). We will analyze in detail how nanoparticles are processed within the cell (transport mechanisms, association with organelles, degradation), and responses of the cell to nanoparticle incorporation. These studies will be done by using time-resolved spinning disk confocal microscopy on cultured cells provided by the Mailänder/Simmet groups, complemented by 4Pi-confocal experiments with ultra-high spatial resolution on fixed cells. With these experiments, we aim to achieve a mechanistic description, in molecular terms, of the response of cultured cells to nanoparticles, which is a first important step towards an understanding of the effects of nanoparticles on the entire organism. The inflammatory process is vital to the survival of all complex organisms and plays a fundamental role in health and disease. Antigen-presenting and phagocytosing monocytic cells are pivotal for inflammation and host defence. On this background it is very important to gain insight not only in i) the interaction between distinctly designed nanoparticles and monocytic cells, but also to identify ii) possible target molecules, iii) fundamental biochemical mechanisms, and iv) cell functions that may be targeted by nanoparticles.