Abu-Hanna Jeries, Xu Gang, Morrow Gael B, Timms Lewis, Akbar Naveed, Laffan Mike, Choudhury Robin P, Curry Nicola
Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
Oxford Haemophilia and Thrombosis Centre, Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Blood Vessel Thromb Hemost. 2025 Jul 5;2(4):100087. doi: 10.1016/j.bvth.2025.100087. eCollection 2025 Nov.
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.
创伤诱导的凝血病（TIC）显著导致创伤相关的死亡率，其由凝血和纤维蛋白溶解失调驱动。创伤性内皮病变（EoT）是TIC的核心，但其潜在机制仍不清楚。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。本研究旨在开发一种新型的EoT体外模型，该模型模拟关键的TIC特征，从而能够研究内皮细胞对TIC的作用。将内皮祖细胞（ECFCs）暴露于与创伤相关的因素中，包括肾上腺素、肿瘤坏死因子α、白细胞介素6、高迁移率族蛋白B1、过氧化氢和缺氧。使用酶联免疫吸附测定、免疫荧光、整体止血测定和RNA测序分析内皮损伤标志物（多配体蛋白聚糖-1和血栓调节蛋白）、止血蛋白表达、凝血和纤维蛋白溶解。使用健康供体和创伤患者的血浆来评估临床相关性。受创伤的ECFCs表现出进行性功能障碍，早期有表面损伤和持续的纤维蛋白溶解失调。转录组分析显示炎症途径激活、代谢转变和表观遗传变化。抗凝蛋白的表面表达降低，而促凝组织因子增加，从而增加血栓形成潜力。最初，受创伤的ECFCs通过血栓调节蛋白脱落促进纤维蛋白溶解，但后来分泌抗纤维蛋白溶解的纤溶酶原激活物抑制剂1，模拟双相TIC表型。血浆测定显示凝血酶生成和凝块溶解变化与创伤患者相似。这种体外模型成功地复制了EoT和与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。它为TIC提供了机制性见解，并作为一个平台来测试有针对性的干预措施，以减轻创伤中内皮驱动的凝血病。
