A Rapid-Patterning 3D Vessel-on-Chip for Imaging and Quantitatively Analyzing Cell-Cell Junction Phenotypes.
A Rapid-Patterning 3D Vessel-on-Chip for Imaging and Quantitatively Analyzing Cell-Cell Junction Phenotypes.
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DOI：
10.3390/bioengineering10091080
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发表时间：
2023-09-13
影响因子：
4.6
通讯作者：
Stroka, Kimberly M.
中科院分区：
文献类型：
--
作者：
Yan, Li;Dwiggins, Cole W.;Gupta, Udit;Stroka, Kimberly M.
血脑屏障（blood-brain barrier，BBB）是调节脑与外周血之间分子交换的动态界面。血脑屏障的通透性主要由脑内皮细胞上的连接蛋白调节。体外血脑屏障模型在研究脑内生理功能、病理和药物传递机制方面显示出巨大的潜力。然而，很少有研究已经证明了通过定量分析3D微血管中的连接呈现来监测和评价屏障完整性的能力。本研究的目的是制作一个简单的芯片上的血管，它允许在3D微血管中的连接呈现严格的定量研究。为此，我们开发了一种快速协议，用聚二甲基硅氧烷和微针创建3D微血管。我们建立了一个简单的血管芯片模型，内衬人iPSC衍生的脑微血管内皮样细胞（iBMEC样细胞）。然后，血管结构的3D图像可以被“展开”并转换为2D图像，用于细胞-细胞连接表型的定量分析。我们的研究结果表明，三维圆柱形结构改变了紧密连接蛋白的表型，沿着细胞的形态。此外，我们的3D模型中的细胞-细胞连接完整性被肿瘤坏死因子α破坏。这项工作提出了一个“快速简便”的3D血管芯片模型和分析管道，同时允许在各种微环境条件和治疗下筛选和评估内皮细胞的细胞-细胞连接完整性的能力。
The blood-brain barrier (BBB) is a dynamic interface that regulates the molecular exchanges between the brain and peripheral blood. The permeability of the BBB is primarily regulated by the junction proteins on the brain endothelial cells. In vitro BBB models have shown great potential for the investigation of the mechanisms of physiological function, pathologies, and drug delivery in the brain. However, few studies have demonstrated the ability to monitor and evaluate the barrier integrity by quantitatively analyzing the junction presentation in 3D microvessels. This study aimed to fabricate a simple vessel-on-chip, which allows for a rigorous quantitative investigation of junction presentation in 3D microvessels. To this end, we developed a rapid protocol that creates 3D microvessels with polydimethylsiloxane and microneedles. We established a simple vessel-on-chip model lined with human iPSC-derived brain microvascular endothelial-like cells (iBMEC-like cells). The 3D image of the vessel structure can then be “unwrapped” and converted to 2D images for quantitative analysis of cell–cell junction phenotypes. Our findings revealed that 3D cylindrical structures altered the phenotype of tight junction proteins, along with the morphology of cells. Additionally, the cell–cell junction integrity in our 3D models was disrupted by the tumor necrosis factor α. This work presents a “quick and easy” 3D vessel-on-chip model and analysis pipeline, together allowing for the capability of screening and evaluating the cell–cell junction integrity of endothelial cells under various microenvironment conditions and treatments.
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