Platelet Functions During Extracorporeal Membrane Oxygenation. Platelet–Leukocyte Aggregates Analyzed by Flow Cytometry as a Promising Tool to Monitor Platelet Activation

Alexandre Mansour,Pascale Gaussem,Christilla Bachelot-Loza,Adeline Pontis,Fabienne Nédelec-Gac,Erwan Flécher,Isabelle Gouin-Thibault,Mikael Roussel

doi:10.3390/jcm9082361

Abstract

Extracorporeal membrane oxygenation (ECMO) is an extracorporeal circulation used to manage patients with severe circulatory or respiratory failure. It is associated with both high bleeding and thrombosis risks, mainly as a result of biomaterial/blood interface phenomena, high shear stress, and complex inflammatory response involving the activation of coagulation and complement systems, endothelial cells, leukocytes, and platelets. Besides their critical role in hemostasis, platelets are important players in inflammatory reactions, especially due to their ability to bind and activate leukocytes. Hence, we reviewed studies on platelet function of ECMO patients. Moreover, we addressed the issue of platelet–leukocyte aggregates (PLAs), which is a key step in both platelet and leukocyte activation, and deserves to be investigated in these patients. A reduced expression of GPIb and GPVI was found under ECMO therapy, due to the shedding processes. However, defective platelet aggregation is inconsistently reported and is still not clearly defined. Due to the high susceptibility of PLAs to pre-analytical conditions, defining and strictly adhering to a rigorous laboratory methodology is essential for reliable and reproducible results, especially in the setting of complex inflammatory situations like ECMO. We provide results on sample preparation and flow cytometric whole blood evaluation of circulating PLAs.

Highlights

Extracorporeal circulation is used to manage patients with severe organ dysfunction
Using conditions that minimize in vitro platelet–leukocyte aggregates (PLAs) formation (15 min delay between blood collection and immunolabeling and no delay between immunolabeling and flow cytometric assay), mean PLA levels with sodium citrate and CTAD were, respectively, 17.8 ± 7.4% and 16.9 ± 6.2% (p = 0.7). These results demonstrate that sample preparation before flow cytometry assay is critical, as it induces a significant variability in the PLA levels
Platelets might interact with the inflammation system, resulting in pro-thrombotic effects

Summary

Introduction

Extracorporeal circulation is used to manage patients with severe organ dysfunction. Extracorporeal membrane oxygenation (ECMO) provides gas exchange during severe respiratory failure with veno-venous (VV-ECMO) circuits [1]. Veno-arterial ECMO (VA-ECMO), called extracorporeal life support (ECLS), has been increasingly used in life-threatening circulatory failure refractory in conventional treatments, with the aim of maintaining satisfactory tissue perfusion pending myocardial recovery or heart transplantation. In all these devices, blood comes into sustained contact with a large artificial material surface area and a pump, leading to a complex inflammatory response involving activation of the coagulation system, complement system, endothelial cells, leukocytes, and platelets [1]. Beyond TF-based activation of coagulation, contact activation through artificial surfaces involving notably FXII and FXI, might play an important role in clot formation

Objectives

Results

Conclusion