Effects of Diabetes Mellitus on Fibrin Clot Structure and Mechanics in a Model of Acute Neutrophil Extracellular Traps (NETs) Formation.

Judith J De Vries,Behiye Özcan,Tamara Hoppenbrouwers,Dingeman C Rijken,Cristina Martinez-Torres,Frank W.G Leebeek,Rezin Majied,Moniek P.M De Maat,Mandy Van Hoek,Gijsje H Koenderink

doi:10.3390/ijms21197107

Abstract

Subjects with diabetes mellitus (DM) have an increased risk of arterial thrombosis, to which changes in clot structure and mechanics may contribute. Another contributing factor might be an increased formation of neutrophil extracellular traps (NETs) in DM. NETs are mainly formed during the acute phase of disease and form a network within the fibrin matrix, thereby influencing clot properties. Previous research has shown separate effects of NETs and DM on clot properties, therefore our aim was to study how DM affects clot properties in a model resembling an acute phase of disease with NETs formation. Clots were prepared from citrated plasma from subjects with and without DM with the addition of NETs, induced in neutrophils by S. aureus bacteria or phorbol myristate acetate (PMA). Structural parameters were measured using scanning electron microscopy, mechanical properties using rheology, and sensitivity to lysis using a fluorescence-based fibrinolysis assay. Plasma clots from subjects with DM had significantly thicker fibers and fewer pores and branch points than clots from subjects without DM. In addition, fibrinolysis was significantly slower, while mechanical properties were similar between both groups. In conclusion, in a model of acute NETs formation, DM plasma shows prothrombotic effects on fibrin clots.

Highlights

Diabetes mellitus (DM) is an increasingly common disease with a prevalence of almost 500 million people worldwide and diabetes mellitus (DM) is directly responsible for 1.6 million deaths per year [1,2]
A wider spread was observed in onset stress of strain-stiffening (σ0), maximum stiffness before rupture (Kmax), stress at Kmax, and stress at point of rupture ((σR) in plasma clots from subjects with DM compared to subjects without DM (Table S3)
We investigated the structural and mechanical parameters of clots prepared from plasma from subjects with and without DM in a model of acute neutrophil extracellular traps (NETs) formation

Summary

Introduction

Diabetes mellitus (DM) is an increasingly common disease with a prevalence of almost 500 million people worldwide and DM is directly responsible for 1.6 million deaths per year [1,2]. Since subjects with DM have more compact clots with smaller pores, which have a decreased susceptibility to fibrinolysis [4,5], it is hypothesized that these different structural and mechanical clot characteristics contribute to the increased thrombotic risk. It has been shown that denser clots with smaller pore sizes are more resistant to fibrinolysis and are associated with arterial thrombosis [6]. NETs are found to influence clot characteristics and fibrinolysis [8,9] and thereby thrombotic risk [10,11]. The formation of NETs was shown to be enhanced in different patient groups with increased pro-coagulant activity [14,15,16] and in patients with DM [17,18]. In a large population study, we did not observe differences in NETs plasma levels between subjects with and without DM in steady state conditions [19]

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