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Microparticles profiling in trauma patients: high level of microparticles induce activation of platelets in vitro

  • Michael CaspersEmail author
  • Nadine Schäfer
  • Matthias Fröhlich
  • Bertil Bouillon
  • Manuel Mutschler
  • Ursula Bauerfeind
  • Marc Maegele
Original Article
  • 34 Downloads

Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusion

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.

Keywords

Primary haemostasis in trauma Microparticles Lethal triad TIC 

Abbreviations

AA

α-Angle

aPTT

Activated partial thromboplastin time

ARDS

Acute respiratory distress syndrome

CFT

Clot formation time

CMMC

Cologne-Merheim Medical Centre

CT

Clotting time

EDMP

Endothelial-derived microparticles

FBC

Full blood count

Fib

Fibrinogen

LT

Lethal triad

MCF

Maximum clot firmness

MOF

Multiple organ failure

MP

Microparticles

PDMP

Platelet-derived microparticles

RDMP

Red blood cells-derived microparticles

SSC

Sideward scatter

TIC

Trauma-induced coagulopathy

TRAP

Thrombin receptor activating peptide

MDMP

Monocytes-derived microparticles

Notes

Acknowledgements

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.

Author contributions

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.

Funding

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.

Ethics approval

The study design and protocol was approved by the Central Ethics Committee of the University Witten/Herdecke (nos. 37/2005 and 168/2014).

Supplementary material

68_2019_1111_MOESM1_ESM.pdf (477 kb)
Suppl. Data Table 1: Results derived from standard coagulation testing and normal blood count. Values are given as mean and standard deviation in parenthesis. If changes are significant compared to baseline they were marked by asterisk. Suppl. Data Table 2: Characteristics and physiological data of our Polytrauma cohort. (PDF 477 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The Institute for Research in Operative Medicine, Faculty of Health, Department of MedicineWitten/Herdecke UniversityCologneGermany
  2. 2.Department of Traumatology, Orthopaedic Surgery and Sports TraumatologyCologne-Merheim Medical Centre (CMMC), Witten/Herdecke UniversityCologneGermany
  3. 3.Department of Haematology and Transfusion Medicine (DTM)Cologne-Merheim Medical Centre (CMMC)CologneGermany

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