A novel in vitro model of trauma-induced endotheliopathy provides a platform for studying mechanisms of coagulopathy

Blood Vessels, Thrombosis & Hemostasis Pub Date : 2025-07-05 DOI:10.1016/j.bvth.2025.100087

Jeries Abu-Hanna , Gang Xu , Gael B. Morrow , Lewis Timms , Naveed Akbar , Mike Laffan , Robin P. Choudhury , Nicola Curry

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Abstract

Trauma-induced coagulopathy (TIC) significantly contributes to trauma-related mortality, driven by dysregulated coagulation and fibrinolysis. Endotheliopathy of trauma (EoT) is central to TIC, yet its underlying mechanisms remain unclear. Current in vitro models fail to replicate the complex trauma environment, including hemorrhagic shock, tissue injury, and inflammation. This study aimed to develop a novel in vitro model of EoT that mimics key TIC features, enabling the investigation of endothelial contributions to TIC. Endothelial colony-forming cells (ECFCs) were exposed to trauma-relevant factors, including epinephrine, tumor necrosis factor α, interleukin 6, high mobility group box 1, hydrogen peroxide, and hypoxia. Endothelial injury markers (syndecan-1 and thrombomodulin), hemostatic protein expression, coagulation, and fibrinolysis were analyzed using enzyme-linked immunosorbent assay, immunofluorescence, global hemostasis assays, and RNA sequencing. Plasma from healthy donors and trauma patients was used to assess clinical relevance. Traumatized ECFCs exhibited progressive dysfunction, with early surface damage and sustained fibrinolytic dysregulation. Transcriptomic analysis showed activation of inflammatory pathways, metabolic shifts, and epigenetic changes. Surface expression of anticoagulant proteins decreased, whereas procoagulant tissue factor increased, heightening thrombogenic potential. Initially, traumatized ECFCs promoted fibrinolysis via thrombomodulin shedding but later secreted antifibrinolytic plasminogen activator inhibitor 1, mimicking the biphasic TIC phenotype. Plasma assays revealed thrombin generation and clot lysis changes similar to trauma patients. This in vitro model successfully replicates EoT and TIC-associated hemostatic imbalances, capturing the time-dependent evolution of endothelial dysfunction. It provides mechanistic insights into TIC and serves as a platform for testing targeted interventions to mitigate endothelial-driven coagulopathy in trauma.

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一种新的体外创伤性内皮病变模型为研究凝血功能障碍的机制提供了一个平台

摘要创伤性凝血功能障碍（TIC）是由凝血和纤溶异常引起的创伤性死亡率的重要因素。创伤性内皮病变（EoT）是TIC的核心，但其潜在机制尚不清楚。目前的体外模型无法复制复杂的创伤环境，包括失血性休克、组织损伤和炎症。本研究旨在建立一种新的体外EoT模型，模拟TIC的关键特征，从而研究内皮细胞对TIC的贡献。内皮集落形成细胞（ECFCs）暴露于创伤相关因素，包括肾上腺素、肿瘤坏死因子α、白细胞介素6、高迁移率组盒1、过氧化氢和缺氧。内皮损伤标志物（syndecan-1和血栓调节蛋白）、止血蛋白表达、凝血和纤维蛋白溶解均采用酶联免疫吸附法、免疫荧光法、整体止血法和RNA测序进行分析。来自健康供体和创伤患者的血浆用于评估临床相关性。创伤后的ecfc表现出进行性功能障碍，早期表面损伤和持续的纤维蛋白溶解失调。转录组学分析显示炎症通路激活、代谢变化和表观遗传变化。抗凝蛋白的表面表达减少，而促凝组织因子增加，提高了血栓形成的潜力。最初，创伤后的ecfc通过血栓调节蛋白脱落促进纤维蛋白溶解，但后来分泌抗纤溶酶原激活物抑制剂1，模拟双期TIC表型。血浆检测显示凝血酶生成和凝块溶解变化与创伤患者相似。该体外模型成功复制了EoT和tic相关的止血失衡，捕捉了内皮功能障碍的时间依赖性演变。它提供了TIC的机制见解，并作为测试靶向干预措施以减轻创伤中内皮驱动的凝血功能障碍的平台。

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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Blood Vessels, Thrombosis & Hemostasis

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