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，前者用于对细胞、蛋白质、DNA等生物组织进行成像，并检测细胞的力学特性；后者用于对培养细胞内外的受体、细胞骨架和信号分子的分布进行成像。此外，本研究还使用了能对培养细胞进行纳米级微运动测量的ECIS （electro cell -substrate Impedance Sensing）系统2。研究结果：单核细胞与内皮细胞的相互作用被认为在动脉粥样硬化早期起主要作用，内皮细胞参与的微观力学可能为我们提供动脉粥样硬化发生的重要方面。在本研究中，我们用ECIS评估了(I)内皮细胞与细胞之间和细胞与底物之间的间隙，以及（ii）应用单核细胞到内皮细胞后，用AFM评估了内皮细胞的微观力学特性。更多的细胞到内皮细胞。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，内皮细胞到底物的电阻立即降低（细胞到底物的间隙增加）。而内皮细胞间阻力（细胞间间隙）没有变化。单核细胞作用2小时后，内皮细胞的弹性模量下降，表明内皮细胞的变形能力增强。由此可见，单核细胞与内皮细胞的相互作用降低了内皮细胞与底物的黏附性，增强了内皮细胞的变形能力。这些可能会促进单核细胞的迁移，这是后期动脉粥样硬化的关键过程。此外，该测量系统被用于评价原发性肺动脉高压（PPH）患者肺动脉平滑肌细胞（PASMC）的纳米级微力学特性。我们阐明了PPH Less患者PASMC对NO和前列环素血管舒张的微弹性变化降低

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