Validation of the calibrated thrombin generation test (cTGT) as the reference assay to evaluate the procoagulant activity of nanomaterials

Abstract

We validated a preclinical toxicological screening assay and provided guidelines to evaluate the potential impact of nanoparticles (NPs) on blood coagulation. Five NPs with various physicochemical properties were studied using several existing methods of clotting times and thrombin generation assays in human normal pool plasma. In both recalcification clotting time (RCT) and calibrated thrombin generation test (cTGT), the NPs exhibited procoagulant activity (SiO₂ ≥ SiC ≥ TiC > CuO > CB) but cTGT was more sensitive and relevant than RCT. Thus, the cTGT appears as a reference assay to investigate the nanoparticle (NP) procoagulant activity in human plasma. It should be used as the reference toxicity test for evaluating the effects of nanomaterials on coagulation cascade. In addition, we also showed that the use of the Pluronic F-108 dispersant and/or the sonication for the NP suspension preparation may mask their procoagulant activity and thus should be avoided.

Keywords::

• Nanoparticles
• procoagulant activity
• thrombin generation
• coagulation cascade
• toxicity

Acknowledgements

O.T. is a Senior Research Associate of the Belgian FRS-FNRS. We thank Dr O. Lozano for its careful reading and correction of the manuscript.

Declaration of interest: This work was financially supported by the Direction Générale des Technologies de la Recherche et de l'Energie (DGTRE) of the Walloon Region of Belgium (Nanotoxico Pole of Excellence, RW/FUNDP research grant No. 516252). The authors report no conflict of interest. The authors alone are responsible for the content and writing of the paper.