title: 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.
authors.affiliation: Department of Pharmacy, Drug Design & Discovery Center (D3C), Namur Research Institute for Life Sciences (NARILIS), Namur Thrombosis and Hemostasis Center (NTHC), FUNDP-University of Namur, Namur, Belgium. julie.laloy@fundp.ac.be
authors.forename: Julie
authors.initials: J
authors.lastname: Laloy
authors.forename: Séverine
authors.initials: S
authors.lastname: Robert
authors.forename: Catherine
authors.initials: C
authors.lastname: Marbehant
authors.forename: François
authors.initials: F
authors.lastname: Mullier
authors.forename: Jorge
authors.initials: J
authors.lastname: Mejia
authors.forename: Jean-Pascal
authors.initials: JP
authors.lastname: Piret
authors.forename: Stéphane
authors.initials: S
authors.lastname: Lucas
authors.forename: Bernard
authors.initials: B
authors.lastname: Chatelain
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Dogné
authors.forename: Olivier
authors.initials: O
authors.lastname: Toussaint
authors.forename: Bernard
authors.initials: B
authors.lastname: Masereel
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Rolin
mesh_terms: D000293:Adolescent; D000328:Adult; D001777:Blood Coagulation / Q000187:drug effects*; D001780:Blood Coagulation Tests / Q000379:methods / Q000592:standards*; D002138:Calibration; D003029:Coagulants / Q000633:toxicity; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D049329:Nanostructures / Q000633:toxicity*; D020442:Poloxamer; D015203:Reproducibility of Results; D013917:Thrombin / Q000378:metabolism*; D013925:Thromboplastin / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods / Q000592:standards*
chemical_list: D003029:Coagulants; D020442:Poloxamer; D013925:Thromboplastin; D013917:Thrombin