Generation 2 (G2) - Generation 4 (G4) PAMAM dendrimers disrupt key plasma coagulation parameters

Abstract

The aim of the research was to evaluate the effects of G2 - G4 PAMAM dendrimers on basic plasma haemostasis parameters (Partially Activated Thrombin Time (APTT), Prothrombin Time (PT), Thrombin Time (TT)) as well as the activity of factor X, antithrombin III (AT), protein C and plasmin. Furthermore, tissue factor (TF) synthesis in endothelial cells and viability of smooth muscle cells in the presence of PAMAM dendrimers was investigated. APTT, PT and TT were performed according to the available commercial methods. The activity of factor X was conducted based on deficient plasma factor X. Protein C, AT and plasmin activity were measured spectrophotometrically using chromogenic substrates. Intracellular TF production in human umbilical vein endothelial cells (HUVECs) was measured using immunohistochemical method. Viability of Human Aortal Smooth Muscle cells (hAoSMCs) was established using WST-1 assay. PAMAM dendrimers decreased activity of factor X, and concomitantly prolonged PT and APTT. We also demonstrated shortened TT and increased fibrinogen concentrations in plasma treated with G4 PAMAM dendrimers, suggesting formation of fibrinogen aggregates. G2 - G4 PAMAM dendrimers decreased the activity of both naturally occurring anticoagulants AT and protein C. G2 and G3 PAMAM dendrimers did not affect the proteolytic reaction with plasmin. PAMAM dendrimers were found not to trigger TF production in undisturbed endothelial cells. PAMAM dendrimers, depending on the concentration and generation decreased viability of AoSMCs. The results presented within the current study suggest complex but mostly undesirable effect of G2 - G4 PAMAM dendrimers on plasma haemostasis and underscore the need for further in-depth research.

Keywords: Coagulation; Endothelial cells; Haemostasis; Nanotechnology; PAMAM dendrimers; Smooth muscle cells.