Prothrombotic activity of cytokine-activated endothelial cells and shear-activated platelets in the setting of ventricular assist device support
Section snippets
Methods
To evaluate EC‒platelet prothrombotic interaction mechanisms, the following 3 experimental protocols were performed: Protocol 1: The effect of differing extents of SMPA induced by platelet exposure to 30-, 50-, and 70-dynes/cm2 time-constant shear stress patterns for 10 minutes in a hemodynamic shearing device42 was examined; specifically, we evaluated adhesion of shear-activated platelets to untreated ECs. Platelet prothrombinase activity was also evaluated via the platelet activity state
Shear-mediated platelet activation
The level of SMPA increased progressively with increasing shear stress magnitude. Specifically, PAS values of platelets stimulated in the hemodynamic shearing device (HSD) with different levels of shear stress (30, 50, and 70 dynes/cm2) were significantly higher than basal values measured in control/non-stimulated platelet samples (p < 0.0001; Figure 1A). PAS values of platelets stimulated at 30 and 50 dynes/cm2 were comparable (p > 0.05; Figure 1A), whereas thrombin generation of platelets
Discussion
In this study we have examined the potential synergistic effects of mechanical (shear-mediated) platelet activation (SMPA) and inflammatory (cytokine-mediated) EC activation as to their functional interaction to enhance EC‒platelet surface adhesion and prothrombosis.
We observed that shear-activated platelets bind to ECs regardless of the levels of shear activation and EC activation; that is, they bind to non-activated ECs as well. As such, ECs are a pro-adhesive substrate for shear-activated
Study limitations
In this study human umbilical vein endothelial cells (HUVECs) were utilized as a model for endocardial ECs to examine the effect of cytokine activation. Despite differences between the 2 cell types, HUVECs are a general but reliable model to the study of TNF-α‒mediated activation of endocardial ECs. Indeed, HUVECs have been shown to be responsive to physiologic and/or pathologic stimuli and to express those endothelial markers and signaling molecules we sought to analyze, namely ICAM-1, VCAM-1,
Conclusions
We have provided new mechanistic insights into the prothrombotic interaction of mechanical—shear-mediated—stimulation of platelets with inflammatory activation of ECs. Our findings demonstrate a synergy of this interaction in enhancing overall prothrombosis. Specifically, our results provide evidence that EC activation—cytokine-mediated—combines with shear-mediated platelet activation to drive and sustain platelet adhesion to the inflamed endothelium and enhance prothrombotic activity. We have
Disclosure statement
The authors have no conflicts of interest to disclose. This study was supported by Fondazione CARIPLO (2015-1044 to F.C. and M.J.S.), Fondazione Cariplo and Regione Lombardia (2016-0901 to A.R.), the National Institutes of Health (NIH Cardiovascular Biomedical Engineering Training Grant T32 HL007955 to K.R.A.), the National Institute of Biomedical Imaging and Bioengineering (Quantum Grant Award 5U01EB012487-00 to D.B. and M.J.S.), and the Arizona Center for Accelerated Biomedical
Supplementary data
Supplementary data associated with this article can be found in the online version at www.jhltonline.org/.
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2021, Cardiovascular PathologyCitation Excerpt :The nidus for thrombus formation is suggested to be the smooth-sintered interface of the IC, with the polished smooth surface showing failure of new intima and tissue ingrowth, resulting in largely unorganized thrombus [1]. On top of that, the larger environment of the apex of the left ventricle can be viewed as the milieu where a combination of factors likely contributes to thrombus formation, including (1) blood stasis at the IC tip-myocardium interface, (2) changes in mechanical compliance of the ventricular wall due to the rigid epicardial pump, (3) changes in flow characteristics along the discontinuity gap between the sintered and nonsintered material surfaces of the IC, (4) heat dissipation from the pump through the IC, (5) vibratory mechanical forces transferred from the pump to the IC, (6) malpositioned IC and/or left ventricle remodeling, and (7) endothelial cell inflammation promoting platelet prothrombosis [1,2]. Thrombus growth and accumulation over time can lead to complete IC occlusion.
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- 1
These authors contributed equally to this work.
- 2
These authors contributed equally to this work as senior authors.