Dynamic measurements of endothelial cell morphology and functions using with a combined system of Atomic Force Microscope and Laser microscope

1.系统设置：联合培养细胞力学测量系统由原子力显微镜和激光共聚焦显微镜组成，原子力显微镜用于对细胞、蛋白质、DNA等生物组织进行成像并检测细胞的力学性质，共聚焦激光显微镜用于成像培养细胞内和细胞上受体、细胞骨架和信号分子的分布。此外，本研究还使用了电细胞-基质阻抗传感系统(ECIS)，该系统可以提供对培养细胞的纳米级微运动测量。研究结果：单核细胞与内皮细胞之间的相互作用被认为在动脉粥样硬化的早期阶段起主要作用，涉及的内皮细胞微观机制可能为动脉粥样硬化的发生提供重要方面。在本研究中，我们使用ecis评估(I)内皮细胞与细胞和细胞与基质之间的间隙，以及(Ii)应用MON…后用原子力显微镜评估内皮细胞的微观力学性质。将单核细胞THP-1细胞应用于IL-1b刺激的人脐静脉内皮细胞，立即降低内皮细胞与底物之间的电阻(细胞与底物之间的间隙增加)，而内皮细胞与细胞间的阻力(细胞与细胞间的间隙)不变。单核细胞作用2小时后内皮细胞弹性模量下降，提示内皮细胞变形性增加。结论：单核细胞与内皮细胞的相互作用降低了与基质的粘附性，增加了内皮细胞的变形性。此外，该测量系统还用于评价原发性肺动脉高压(PPH)患者的肺动脉平滑肌细胞(PASMC)的纳米级细观力学性能。我们阐明了PPH少的患者PASMC对NO和前列环素血管扩张的微弹性改变

1. System setup: The combined cultured cell mechanics measurement system was composed of an atomic force microscope that is for imaging the biological tissues such as cells, proteins and DNA, and examining mechanical properties of cells, and a confocal laser microscope that is for imaging the distributions of receptors, cytoskeleton and signaling molecules in and on the cultured cells. In addition, Electric Cell-substrate Impedance Sensing (ECIS) system that provides nano-order micro-motion measurement of the cultured cell was used in this study2. Research results: The interaction between monocytes and endothelial cells is considered to play a major role at the early stage of atherosclerosis, and the involved endothelial cell micro-mechanics may provide us important aspects of atherogenesis. In the present study, we evaluated (I) the endothelial cell-to-cell and cell-to-substrate gaps with ECIS, and (ii) the endothelial cell micro-mechanical properties with AFM after application of mon … More ocytes to endothelial cellsApplication of monocytic THP-1 cells to IL-1b-stimulated human umbilical vein endothelial cells immediately decreased the electrical resistance of the endothelial cell-to-substrate (increase of the cell-to-substrate gap), while the endothelial cell-to-cell resistance (cell-to-cell gap) did not change. The elastic modulus of the endothelial cells decreased after 2-hour monocytes application, indicating increase of the endothelial cell deformabilityIn conclusion, the interaction of the monocytes to the endothelial cells reduced the adhesiveness to the substrate and increased the deformability of endothelial cells. Those may facilitate migration of monocytes a key process of atherogenesis in the later stageIn addition, the measurement system was applied to the evaluation of the nano-order micromechanical property of pulmonary artery smooth muscles cells (PASMC) from patients with Primary pulmonary hypertension (PPH). We elucidated that Micro-elasticity change in PASMC to vasodilation of NO and prostacyclin is reduced in patients with PPH Less