Oxygen-tunable endothelialized microvascular chip to assess hypoxia–reperfusion in sickle cell disease

IF 5.4 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2025-08-20 DOI:10.1039/D5LC00211G
Samantha R. Schad, Joan D. Beckman, Wilbur A. Lam and David K. Wood
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引用次数: 0

Abstract

A better understanding of hypoxia reperfusion (H/R) injury is needed to gain deeper insight into the mechanisms driving sickle cell disease (SCD) pathophysiology. Existing in vivo and in vitro models have yet to fully explain H/R, which is typically associated with harmful inflammatory processes but has also been linked to a protective effect ameliorating subsequent severe vaso-occlusion. To address this need, we developed a novel microfluidic platform that includes three-dimensional endothelial-lined microchannels within an oxygen-tunable environment. These features enable simulation of H/R, red blood cell (RBC) sickling, and vaso-occlusion on-chip. The endothelial network cultured on-chip is physiologically relevant and expresses crucial microvascular features such as 3D lumen structure and expression of functional endothelial markers. We utilized this platform to perform an occlusion assay, evaluating the effects of hypoxic preconditioning on RBC-endothelial interactions contributing to occlusion. Our results demonstrate that both sustained mild hypoxia and cyclic hypoxia endothelial treatment reduce the likelihood of SCD occlusion on-chip. Specifically, average vaso-occlusion rates of 8.89% and 11.78% were observed among endothelialized devices preconditioned to cyclic and sustained hypoxia, respectively, compared to 57.93% and 55.05% for the control groups. Additionally, we leveraged RNA sequencing to identify differential regulation of specific genes contributing to this protective outcome. Of note, hypoxia preconditioning resulted in significant modulation of CYBB, RELN, and SERPINA1. These results offer a better understanding of the mechanistic changes affecting the endothelium during H/R and also offer potential targets for further exploration and therapeutic intervention in SCD.

Abstract Image

氧可调内皮微血管芯片评估镰状细胞病的缺氧再灌注。
为了更深入地了解镰状细胞病(SCD)的病理生理机制,需要更好地了解缺氧再灌注(H/R)损伤。现有的体内和体外模型尚未完全解释H/R,这通常与有害的炎症过程有关,但也与改善随后严重血管闭塞的保护作用有关。为了满足这一需求,我们开发了一种新型的微流控平台,其中包括在氧可调环境中的三维内皮衬里微通道。这些功能使模拟H/R,红细胞(RBC)镰状细胞,血管闭塞芯片。芯片上培养的内皮网络具有生理学相关性,表达了关键的微血管特征,如三维管腔结构和功能性内皮标志物的表达。我们利用这个平台进行闭塞试验,评估缺氧预处理对导致闭塞的红细胞-内皮相互作用的影响。我们的研究结果表明,持续轻度缺氧和循环缺氧内皮治疗可降低芯片上SCD闭塞的可能性。具体而言,在循环缺氧和持续缺氧预处理的内皮化装置中,平均血管闭塞率分别为8.89%和11.78%,而对照组的平均血管闭塞率为57.93%和55.05%。此外,我们利用RNA测序来确定有助于这种保护结果的特定基因的差异调节。值得注意的是,缺氧预处理导致CYBB、RELN和SERPINA1的显著调节。这些结果有助于更好地理解H/R期间影响内皮细胞的机制变化,也为进一步探索和治疗干预SCD提供了潜在的靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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