Effect of Prothrombin Complex Concentrate versus Frozen Plasma on Divalent Cations and Vitamin K Dependent Factor Function: An Ex Vivo Mechanistic Study.

IF 5.2 2区医学 Q1 HEMATOLOGY

Journal of Thrombosis and Haemostasis Pub Date : 2026-10-19 DOI:10.1016/j.jtha.2026.10.006

Min U Jang, Hung A Nguyen, Amir L Butt, Kofi B Vandyck, Kenneth E Stewart, Vibhudutta Awasthi, Kenichi A Tanaka

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引用次数: 0

Abstract

Background: Prothrombin complex concentrate (PCC) has demonstrated hemostatic superiority over frozen plasma (FP) in cardiac surgery, yet the mechanism beyond factor concentration remains unclear.

Objectives: To test ex vivo whether citrate-mediated depletion of ionized calcium (Ca²⁺) and magnesium (Mg²⁺) by FP impairs vitamin K dependent (VKD) factor function, minimized by PCC.

Patients/methods: Heparinized whole blood from healthy volunteers (n=10) was diluted and supplemented with 20% FP or volume-equivalent PCC. Coagulation was assessed by heparinase-modified INTEM (ROTEM) and contact-activated thrombin generation, and validated in cardiopulmonary bypass (CPB) samples (n=6). Factor IX (FIX) and Factor X γ-carboxyglutamate (Gla) domain affinities for Ca²⁺ and Mg²⁺ were measured by MicroScale Thermophoresis (MST).

Results: FP reduced Ca²⁺ from 0.94 ± 0.03 to 0.24 ± 0.01 mmol·L^-1 and Mg²⁺ to undetectable levels. Without exogenous calcium, FP abolished clot formation (clotting time, CT >3600 s) whereas PCC maintained CT (105.4 ± 16.9 s). Low-dose calcium (0.4 mmol·L^-1) partially restored CT (1037 ± 575.5 s); adding MgSO₄ (0.6 mmol·L^-1) improved it to 275.2 ± 56.2 s. Findings held in CPB samples. By MST, the FIX Gla domain bound Mg²⁺ ∼6-fold more tightly than Ca²⁺ (Kd 66.1 ± 8.2 vs 381.2 ± 45.3 μM).

Conclusions: Lowering ionized calcium with FP-derived citrate revealed a functional interaction between magnesium and the Gla domain of VKD factors: at low calcium, restoring magnesium rather than calcium alone recovered thrombin generation and clot formation. PCC preserved the ionic milieu and thrombin generation. Standard coagulation assays potentially underestimate this magnesium dependence because of supraphysiological recalcification.

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凝血酶原复合物浓缩物与冷冻血浆对二价阳离子和维生素K依赖因子功能的影响：体外机制研究。

背景：凝血酶原复合物浓缩物（PCC）在心脏手术中已被证明比冷冻血浆（FP）具有止血优势，但除因子浓度外的机制尚不清楚。目的：在体外测试柠檬酸盐介导的离子钙（Ca2+）和镁（Mg2+）的FP消耗是否会损害维生素K依赖（VKD）因子的功能，而PCC则会使其最小化。患者/方法：将健康志愿者（n=10）的肝素化全血稀释后加入20% FP或等量PCC。采用肝素酶修饰的tem （ROTEM）和接触激活凝血酶生成法评估凝血功能，并在体外循环（CPB）样本中进行验证（n=6）。用微尺度热泳仪（MST）测定了因子IX （FIX）和因子X γ-羧谷氨酸（Gla）结构域对Ca2+和Mg2+的亲和力。结果：FP使Ca2+从0.94±0.03降至0.24±0.01 mmol·L-1， Mg2+降至检测不到的水平。在没有外源性钙的情况下，FP消除了凝块形成（凝块时间，CT值为0 3600 s），而PCC维持了CT值（105.4±16.9 s）。低剂量钙（0.4 mmol·L-1）部分恢复CT(1037±575.5 s)；添加0.6 mmol·L-1的MgSO4使反应时间延长至275.2±56.2 s。CPB样本中的发现。通过MST， FIX Gla结构域结合Mg2+的紧密程度是Ca2+的6倍（Kd为66.1±8.2 vs 381.2±45.3 μM）。结论：用fp衍生的柠檬酸盐降低电离钙揭示了镁与VKD因子的Gla结构域之间的功能相互作用：在低钙条件下，恢复镁而不是单独恢复钙可以恢复凝血酶的生成和凝块的形成。PCC保留了离子环境和凝血酶的生成。由于超生理再钙化，标准凝血试验可能低估了这种镁依赖性。

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

Journal of Thrombosis and Haemostasis 医学-外周血管病

CiteScore

24.30

自引率

3.80%

发文量

321

审稿时长

1 months

期刊介绍： The Journal of Thrombosis and Haemostasis (JTH) serves as the official journal of the International Society on Thrombosis and Haemostasis. It is dedicated to advancing science related to thrombosis, bleeding disorders, and vascular biology through the dissemination and exchange of information and ideas within the global research community. Types of Publications: The journal publishes a variety of content, including: Original research reports State-of-the-art reviews Brief reports Case reports Invited commentaries on publications in the Journal Forum articles Correspondence Announcements Scope of Contributions: Editors invite contributions from both fundamental and clinical domains. These include: Basic manuscripts on blood coagulation and fibrinolysis Studies on proteins and reactions related to thrombosis and haemostasis Research on blood platelets and their interactions with other biological systems, such as the vessel wall, blood cells, and invading organisms Clinical manuscripts covering various topics including venous thrombosis, arterial disease, hemophilia, bleeding disorders, and platelet diseases Clinical manuscripts may encompass etiology, diagnostics, prognosis, prevention, and treatment strategies.