Microparticles profiling in trauma patients: high level of microparticles induce activation of platelets in vitro
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Trauma-induced coagulopathy (TIC) is recognised as an own clinical entity which includes all components of haemostasis following rapidly tissue injury, hypoperfusion and shock. Microparticles (MP) are known to be released in large quantities from different cell types after trauma. The present study aimed to perform a phenotypic MP profiling after major trauma and to elucidate potential procoagulative function of MP under simulated conditions of lethal triad.
For MP isolation, 20 trauma patients (median ISS 24) were included. To produce a Standard MP Phenotype Profile after trauma, samples were pooled, extracted and concentrated by using an ultracentrifuge protocol. Specific cell surface markers were measured by flow cytometry. Our Standard MP Phenotype Profile was subsequently added in high and low concentration to an in vitro lethal triad assay, simulating coagulopathy via induced hypothermia, dilution and acidosis. A comprehensive analysis of coagulation function was performed.
Within our Standard MP Phenotype Profile, PDMP (56%) were found as predominant phenotype followed by EDMP (33%) and MDMP (11%). EDMP characterized by CD144, CD62E and Annexin were determined most frequently but also EDMP expressing CD62P. In addition, tissue factor (TF) was expressed on all MP entities (EDMP 63%, PDMP 30%, MDMP 7%). Within our lethal triad simulation assay, the addition of low and high concentrated MP did not cause any significant alteration in standard coagulation assays, coagulation initiation, clot kinetics or stability. Addition of high concentrated MP increased platelet function and P-selectin expression significantly.
Our data confirm the assumption that there is a characteristic MP phenotype pattern in trauma, which may alter haemostatic capacity at least in part mediated via augmenting in primary haemostasis resulting in an improved contribution of platelets to clot formation. There are indications that expression of selectins on MP surface is involved in this activation process, but this pathway needs to be investigated in more detail.
KeywordsPrimary haemostasis in trauma Microparticles Lethal triad TIC
Activated partial thromboplastin time
Acute respiratory distress syndrome
Clot formation time
Cologne-Merheim Medical Centre
Full blood count
Maximum clot firmness
Multiple organ failure
Red blood cells-derived microparticles
Thrombin receptor activating peptide
We thank Verena Köster, Wilma Groß-Holz and Arthur Camphausen for technical support. Furthermore, we would like to thank Ewa Stürmer for assistance in organizational issues.
MC, NS and MF carried out the laboratory procedure. BB and MM participated in the study design and coordination of the study and the inclusion of trauma patients. MC wrote the initial manuscript and performed the statistical analysis. All authors read and approved the final manuscript. MM conceived the study and supported the whole process as a senior author.
This study was financially supported by the research funding of the University of Witten/Herdecke. An additional ROTEM delta® had been provided by TEM International GmbH for this study.
Compliance with ethical standards
Conflict of interest
Marc Maegele has received honoraria for lectures and travel fees from AstraZeneca, CSL Behring LFB Biomedicaments France, and TEM International/ IL-Werfen. For the remaining authors, no conflicts were declared.
Consent for publication
For MP isolation, 20 trauma patients admitted to our trauma centre (Cologne-Merheim Medical Centre, CMMC) (median ISS 24, range ISS 18–36) were included meeting the following inclusion criteria: (1) adult patient (age ≥ 18 years), (2) injury severity score (ISS) ≥ 16, (3) less than 2 h between injury and hospital admission, (4) less than 2000 ml of pre-hospital fluid administration, (5) no hospital transfer, (6) exclusion of patients with severe liver disease, bleeding abnormalities and/or anticoagulant medication (excluding aspirin), (7) patient’s/consultee’s informed written consent.
For simulating in vitro the lethal triad conditions, the participation of five healthy blood donors from the Institute for Transfusion Medicine (CMMC) was requested. All donors and patients gave their informed and written consent to participating in this study.
The study design and protocol was approved by the Central Ethics Committee of the University Witten/Herdecke (nos. 37/2005 and 168/2014).
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