Li Ya, Xu Min, Zhu Zhu, Xu Feng, Chen Bing
Department of Gastroenterology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of Biological Sample Bank, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Front Bioeng Biotechnol. 2023 Jul 14;11:1236610. doi: 10.3389/fbioe.2023.1236610. eCollection 2023.
Inflammatory bowel disease (IBD) is a chronic relapsing and remitting disease with a rising incidence globally. Circulating exosomes play great roles in IBD pathogenesis through exosomal cargoes, especially impacting the function of endothelial barriers. Transendothelial electrical resistance (TEER) measurement is a widely used non-invasive and label-free strategy to monitor endothelial barrier function . This study established a well-designed microfluidic device to monitor the TEER changes of endothelial cellular barrier on-chip after treated with exosome derived from IBD serum. The chip comprised two layers of microfluidic chambers with top layer for the perfusion of medium to maintain the nutrition and pressure during cell culture, and bottom layer for the extracellular matrix mimic using hydrogel, which are separated by a semipermeable membrane that permitted the formation of endothelial cell barrier. Four electrodes independent from the outlets were integrated to the chip for TEER detection. mouse models mouse models and proteome profiling were performed to finding relevant regulators. With this platform, significant decrease of TEER was detected, indicating that IBD serum exosome impact the endothelial cellular barrier on-chip. mouse models, IBD serum exosome treated group showed great higher DAI scores, shorter colons, more severe histological features, and higher levers of S100A8 expression, promoting the disease progress. Proteome profiling showed that TFRC and ANXA5 have great potentials as novel regulators in IBD. This in-house customized microfluidic chip emulates the endothelial barrier microenvironment and enables the TEER monitoring, and can be used to investigate endothelial barrier function . IBD serum exosome promote the severity of disease.
炎症性肠病（IBD）是一种慢性复发性和缓解性疾病，全球发病率呈上升趋势。循环外泌体通过外泌体货物在IBD发病机制中发挥重要作用，尤其影响内皮屏障的功能。跨内皮电阻（TEER）测量是一种广泛使用的非侵入性且无标记的策略，用于监测内皮屏障功能。本研究建立了一种精心设计的微流控装置，以监测用IBD血清来源的外泌体处理后芯片上内皮细胞屏障的TEER变化。该芯片由两层微流控腔室组成，顶层用于灌注培养基以维持细胞培养期间的营养和压力，底层用于使用水凝胶模拟细胞外基质，两层由半透膜隔开，该半透膜允许形成内皮细胞屏障。四个独立于出口的电极集成到芯片上用于TEER检测。进行小鼠模型和蛋白质组分析以寻找相关调节因子。利用该平台，检测到TEER显著降低，表明IBD血清外泌体影响芯片上的内皮细胞屏障。在小鼠模型中，IBD血清外泌体处理组显示出更高的疾病活动指数（DAI）评分、更短的结肠、更严重的组织学特征以及更高水平的S¹⁰⁰A⁸表达，促进了疾病进展。蛋白质组分析表明，转铁蛋白受体（TFRC）和膜联蛋白A5（ANXA5）作为IBD中的新型调节因子具有很大潜力。这种内部定制的微流控芯片模拟了内皮屏障微环境并能够进行TEER监测，可用于研究内皮屏障功能。IBD血清外泌体促进疾病的严重程度。
