Department Biomedical Engineering, Groningen University, University Medical Center Groningen, Antonius Deusinglaan 1, 9713AW Groningen, The Netherlands.
Institute of Metallurgy and Materials Science, Polish Academy of Sciences, Reymonta 25, 30-059 Krakow, Poland.
ACS Sens. 2021 Dec 24;6(12):4349-4359. doi: 10.1021/acssensors.1c01582. Epub 2021 Nov 19.
Free-radical generation is suspected to play a key role in cardiovascular diseases. Another crucial factor is shear stress. Human umbilical vein endothelial cells (HUVECS), which form the lining of blood vessels, require a physiological shear stress to activate many vasoactive factors. These are needed for maintaining vascular cell functions such as nonthrombogenicity, regulation of blood flow, and vascular tone. Additionally, blood clots form at regions of high shear stress within a blood vessel. Here, we use a new method called diamond magnetometry which allows us to measure the dynamics of free-radical generation in real time under shear stress. This quantum sensing technique allows free-radical detection with nanoscale resolution at the single-cell level. We investigate radical formation in HUVECs in a microfluidic environment under different flow conditions typically found in veins and arteries. Here, we looked into free-radical formation before, during, and after flow. We found that the free-radical production varied depending on the flow conditions. To confirm the magnetometry results and to differentiate between radicals, we performed conventional fluorescent reactive oxygen species (ROS) assays specific for superoxide, nitric oxide, and overall ROS.
自由基的产生被怀疑在心血管疾病中起着关键作用。另一个关键因素是切应力。人脐静脉内皮细胞（HUVECS）构成血管的内层，需要生理切应力来激活许多血管活性因子。这些对于维持血管细胞的功能，如非血栓形成、调节血流和血管张力是必要的。此外，血栓在血管内高切应力区域形成。在这里，我们使用一种称为金刚石磁力计的新方法，该方法允许我们在切应力下实时测量自由基生成的动力学。这种量子传感技术允许在单细胞水平上以纳米级分辨率检测自由基。我们在微流环境中研究了不同流动条件下 HUVEC 中的自由基形成，这些条件通常存在于静脉和动脉中。在这里，我们研究了流动前后自由基的形成。我们发现自由基的产生取决于流动条件。为了证实磁力计的结果并区分自由基，我们进行了常规的荧光活性氧（ROS）测定，专门用于超氧化物、一氧化氮和总 ROS。
