Piombo Sebastian, Hatch Christopher J, Evangelista Chauncey G, Radom-Aizik Shlomit, Cooper Dan M, Hughes Christopher C W
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.
Pediatric Exercise and Genomics Research Center, Department of Pediatrics, School of Medicine, University of California, Irvine, CA, USA.
J Clin Transl Sci. 2025 Apr 4;9(1):e181. doi: 10.1017/cts.2025.49. eCollection 2025.
Here we present a novel approach to evaluate peripheral blood mononuclear cell vascular adhesion using a microfluidic model designed to approximate the complexity of a human arteriole. While EC monolayer assays are commonly used to investigate leukocyte-EC interactions, we hypothesized that our single channel arteriole (SCA) on a chip would recapitulate the microvasculature more accurately and provide additional insight into the initial stages of atherogenesis.
This model is comprised of stromal cells embedded in a hydrogel surrounding a channel lined by endothelial cells (EC) that has an inner diameter approximating a small arteriole. Under physiologic shear conditions, the EC take on a phenotype distinct from monolayer cultures, including alignment with the direction of flow.
Significant differences were found between the SCA and monolayer cultures in the expression of key EC and stromal cell markers, including ICAM-1, VCAM-1, PDGFB, aSMA, and KLF2. Indeed, flow-induced PDGFB expression likely mediated the recruitment and differentiation of αSMA-positive cells to the vessel wall. Importantly, the vessels were responsive to stimulation by inflammatory mediators, showing both increased leukocyte adhesion and increased permeability. Finally, mechanically mediated protrusion of the vessel wall into the lumen disrupted flow, producing increased shear over the vessel wall.
In summary, our studies demonstrate the utility of the SCA model for studies of small vessel physiology under both normal and disrupted flow and to lay the groundwork for further development into a model for atherosclerosis. Additionally, our data emphasize the advantages of complex 3D assays over more traditional 2D cultures.
在此，我们展示了一种评估外周血单核细胞血管黏附的新方法，该方法使用一种微流控模型，旨在模拟人类小动脉的复杂性。虽然内皮细胞单层分析常用于研究白细胞与内皮细胞的相互作用，但我们推测我们芯片上的单通道小动脉（SCA）将更准确地重现微脉管系统，并为动脉粥样硬化的初始阶段提供更多见解。
该模型由嵌入水凝胶中的基质细胞组成，水凝胶围绕着一个由内皮细胞（EC）内衬的通道，该通道的内径近似于小动脉。在生理剪切条件下，内皮细胞呈现出与单层培养不同的表型，包括与血流方向对齐。
在关键内皮细胞和基质细胞标志物的表达上，发现SCA与单层培养之间存在显著差异，这些标志物包括细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）、血小板衍生生长因子B（PDGFB）、α平滑肌肌动蛋白（αSMA）和 Kruppel样因子2（KLF2）。事实上，流动诱导的PDGFB表达可能介导了αSMA阳性细胞向血管壁的募集和分化。重要的是，这些血管对炎症介质的刺激有反应，表现为白细胞黏附增加和通透性增加。最后，机械介导的血管壁向管腔内的突出扰乱了血流，导致血管壁上的剪切力增加。
总之，我们的研究证明了SCA模型在正常和紊乱血流条件下研究小血管生理学的实用性，并为进一步发展成为动脉粥样硬化模型奠定了基础。此外，我们的数据强调了复杂三维分析相对于更传统二维培养的优势。
