Mechanical Response of Individual Living Epithelial and Endothelial Cells to in situ Biochemical Signal Exposure and Direct Drug Delivery

活体上皮细胞和内皮细胞对原位生化信号暴露和直接给药的机械反应活细胞的机械特性对细胞的功能和健康起着重要的作用。通过应用光学和原子力显微镜(AFM)的最新进展，将独特地研究单个细胞和细胞连接的纳米力学性质。具体地说，内皮细胞(例如，那些衬里血管的细胞)的机械特性将在原位暴露于血管内皮生长因子的过程中表征，血管内皮生长因子是一种已知的破坏细胞间黏附的细胞信号。另外，HaCaT角质形成细胞(皮肤细胞)将在暴露于EGF的过程中进行测量，EGF可促进细胞生长和增殖。最后，通过将生物素化的血管内皮生长因子连接到原子力显微镜尖端，直接将这种细胞信号分子传递到上皮细胞内的特定位置，将展示有针对性的“药物输送”。这种新的局部生化传递最终将扩展到其他生长因子和配体，包括信号素和过氧化体增殖物的信号通路。活细胞的纳米机械测量将提供对人类疾病的过程和潜在的中介，如急性和慢性炎症，糖尿病视网膜病变，中风和心肌梗死后的组织损伤，以及癌细胞的增殖和转移。他们将与康涅狄格州大学、美国国立卫生研究院和其他地方的学术和临床合作者一起表演。更直接的是，“NanoVan”的推广工作包括在当地学校、博物馆、图书馆等地准备和展示纳米概念的移动手势演示。还将提供交互式虚拟现实课程(“NAnimations”)，供互联网分发、教授原子力显微镜及其在生物机械测量和药物输送中的应用。

Mechanical Response of Individual Living Epithelial and Endothelial Cells to in situ Biochemical Signal Exposure and direct Drug DeliveryThe mechanical properties of living cells play an important role in cell function and health. By applying recent advances in coupled optical and Atomic Force Microscopy (AFM), the nanomechanical properties of individual cells and cell junctions will be uniquely investigated. Specifically, the mechanical properties of endothelial cells (e.g. those lining blood vessels) will be characterized during in situ exposure to VEGF, a cell signal known to break down intercellular adhesion. Separately, HaCaT keratinocytes (skin cells) will be measured during exposure to EGF, known to promote cell growth and proliferation. Finally, targeted 'drug delivery' will be demonstrated by attaching biotinylated VEGF to the AFM tip for direct delivery of this cell-signaling molecule to specific positions within epithelial cells. This novel local biochemical delivery will eventually be extended to other growth factors and ligands, including signaling pathways for semaphorins and peroxisome proliferators.The nanomechanical measurements of living cells will provide insight into the processes, and potentially the mediation, of human diseases such as acute and chronic inflammation, diabetic retinopathy, tissue damage following stroke and myocardial infarction, and the proliferation and metastasization of cancerous cells. They will be performed with academic and clinical collaborators at UConn, NIH, and elsewhere. More immediately, the 'NanoVan' outreach effort involves preparing and showing mobile, hand's on demonstrations of nanoscale concepts at local schools, museums, libraries, etc. Interactive virtual reality lessons ('Nanimations') will also be presented for internet distribution, teaching atomic force microscopy and its use in bio-mechanical measurements and drug delivery.