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：组合的培养细胞力学测量系统由用于对生物组织（例如细胞、蛋白质和DNA）成像并检查细胞的力学性质的原子力显微镜和用于对培养细胞中和培养细胞上的受体、细胞骨架和信号分子的分布成像的共聚焦激光显微镜组成。此外，在本研究中使用了提供培养细胞的纳米级微运动测量的电细胞基底阻抗传感（ECIS）系统2。研究成果：单核细胞与内皮细胞之间的相互作用在动脉粥样硬化的早期阶段起着重要的作用，而内皮细胞的微观力学机制可能为我们提供了动脉粥样硬化形成的重要方面。在本研究中，我们评估了（I）ECIS的内皮细胞与细胞和细胞与基底之间的间隙，以及（ii）在施加单细胞后，AFM的内皮细胞微观力学特性。 关于我们 单核细胞THP-1细胞对IL-1b刺激的人脐静脉内皮细胞的应用立即降低了内皮细胞-基质的电阻（细胞-基质间隙增加），而内皮细胞-细胞电阻（细胞-细胞间隙）没有改变。应用单核细胞2小时后，内皮细胞的弹性模量降低，表明内皮细胞的变形能力增加。总之，单核细胞与内皮细胞的相互作用减少了对基质的粘附，增加了内皮细胞的变形能力。此外，还将该系统应用于原发性肺动脉高压（PPH）患者肺动脉平滑肌细胞（PASMC）的纳米级微力学性能的研究。我们阐明PPH患者PASMC对NO和前列环素血管舒张的微弹性改变减少。

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