In vitro flow based systems to study platelet function and thrombus formation: Recommendations for standardization: Communication from the SSC on Biorheology of the ISTH

Mangin, P; Gardiner, Elizabeth E.; Nesbitt, Warwick Scott; Kerrigan, Steven W.; Korin, Netanel; Lam, Wilbur A.; Panteleev, Mikhail A.

doi:10.1111/jth.14717

Cited by 35 publications

(32 citation statements)

References 28 publications

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“…Collagen coated parallel plate flow chambers are a widely applied modern tool for the hemostasis assessment (20). They are mostly used to mimic micro-vessels, such as arterioles (high blood flow velocity) and post-capillary venules (low blood flow velocity) (21).…”

Section: Granulocytes Crawl Among the Growing Thrombi Under Low Shearmentioning

confidence: 99%

Ex vivoobservation of granulocyte activity during thrombus formation

Martyanov

et al. 2020

Preprint

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Infiltration of growing thrombi by leukocytes, being the key part of the thromboinflammation, is well established in vivo. The study was aimed at the development of an ex vivo simulation of this phenomenon. Thrombus formation in anticoagulated whole blood from healthy volunteers and patients was visualized by fluorescent microscopy in parallel-plate flow chambers with fibrillar collagen type I coverslips.Moving CD66b-positive cells (granulocytes) were observed in hirudinated or recalcified blood under low wall shear rate conditions (<200 s−1). These cells crawled around thrombi in a step-wise manner with an average rate of 70 nm/s. Pre-incubation of blood with leukocyte priming agents lead to a significant increase in average cell velocity. On the contrary, leukocytes from Wiskott-Aldrich syndrome patients demonstrated a 1.5-fold lower average velocity, in line with their impaired actin polymerization.Thereby, the observed features of granulocytes crawling are consistent with the neutrophil chemotaxis phenomenon. We conclude that the proposed ex vivo experimental setting allows us to observe granulocytes activity in near-physiological conditions.

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Section: Granulocytes Crawl Among the Growing Thrombi Under Low Shearmentioning

confidence: 99%

Ex vivoobservation of granulocyte activity during thrombus formation

Martyanov

et al. 2020

Preprint

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“…A number of recommendations for the standardization of flow assays for studies of arterial thrombosis and hemostasis were issued by the Biorheology Scientific and Standardization Committee (SSC) of the International Society on Thrombosis and Haemostasis (ISTH) and other expert groups 3 . These recommendations address the choice of the appropriate flow chamber, standardization of adhesive proteins used to coat the device, and quantification of outcomes.…”

Section: Introductionmentioning

confidence: 99%

Wall shear rates in human and mouse arteries: Standardization of hemodynamics for in vitro blood flow assays: Communication from the ISTH SSC subcommittee on biorheology

Panteleev

Korin

Reesink

et al. 2021

Journal of Thrombosis and Haemostasis

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Hemodynamics play a central role in hemostasis and thrombosis by affecting all aspects linked to platelet functions and coagulation. In vitro flow devices are extensively used in basic research, pharmacological studies, antiplatelet agent screening, and development of diagnostic tools. Because hemodynamic conditions vary tremendously throughout the vascular tree and among different (patho)physiological processes, it is important to use flow conditions based on relevant biorheological reference ranges. Surprisingly, it is particularly difficult to find a concise overview of relevant hemodynamic parameters in various human and mouse vessels. To our knowledge, this is the first time an inventory of flow conditions in healthy, non-diseased, human and mouse vessels has been created. The objective of providing such a repertoire is to aid researchers in the field of hemostasis and thrombosis in choosing rheological conditions relevant in in vitro flow experiments and to promote harmonization of flowbased assays to facilitate comparative evaluations between studies. With reference to the human, we discuss relevant similarities and discrepancies in wall shear rates in the mouse, which are typically one order of magnitude greater in agreement with allometric scaling laws between species. Importantly, we bring the attention of the researchers to the fact that the relevant range of average wall shear rates in human arteries where clinically relevant arterial thrombosis occurs may fall as low as 100 to 200 s −1 , thus significantly overlapping with what are considered "venous" shear rates. The same range for the murine arteries used for arterial thrombosis models may significantly exceed 1000 s −1 reaching values considered to be "pathological."

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“…Given that the work presented here is a proof of concept, optimization of these variables was not a goal at this stage, and a considerable amount of work has already gone into guidelines relating to in vitro clot experiments. 42 Rather, our aim was to determine whether or not we could grow a clot successfully, using a human thrombin–fibrinogen model in a simplified system. We also wanted to see how well we could predict occlusion outcome based on data from the experiment.…”

Section: Discussionmentioning

confidence: 99%

Thrombin–Fibrinogen In Vitro Flow Model of Thrombus Growth in Cerebral Aneurysms

Ngoepe

Pretorius

Shaikh

et al. 2021

TH Open

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Cerebral aneurysms are balloon-like structures that develop on weakened areas of cerebral artery walls, with a significant risk of rupture. Thrombi formation is closely associated with cerebral aneurysms and has been observed both before and after intervention, leading to a wide variability of outcomes in patients with the condition. The attempt to manage the outcomes has led to the development of various computational models of cerebral aneurysm thrombosis. In the current study, we developed a simplified thrombin–fibrinogen flow system, based on commercially available purified human-derived plasma proteins, which enables thrombus growth and tracking in an idealized cerebral aneurysm geometry. A three-dimensional printed geometry of an idealized cerebral aneurysm and parent vessel configuration was developed. An unexpected outcome was that this phantom-based flow model allowed us to track clot growth over a period of time, by using optical imaging to record the progression of the growing clot into the flow field. Image processing techniques were subsequently used to extract important quantitative metrics from the imaging dataset, such as end point intracranial thrombus volume. The model clearly demonstrates that clot formation, in cerebral aneurysms, is a complex interplay between mechanics and biochemistry. This system is beneficial for verifying computational models of cerebral aneurysm thrombosis, particularly those focusing on initial angiographic occlusion outcomes, and will also assist manufacturers in optimizing interventional device designs.

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In vitro flow based systems to study platelet function and thrombus formation: Recommendations for standardization: Communication from the SSC on Biorheology of the ISTH

Cited by 35 publications

References 28 publications

Ex vivoobservation of granulocyte activity during thrombus formation

Ex vivoobservation of granulocyte activity during thrombus formation

Wall shear rates in human and mouse arteries: Standardization of hemodynamics for in vitro blood flow assays: Communication from the ISTH SSC subcommittee on biorheology

Thrombin–Fibrinogen In Vitro Flow Model of Thrombus Growth in Cerebral Aneurysms

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