Measurement of whole blood thrombus formation using parallel-plate flow chambers – a practical guide

Kruchten, Roger van; Cosemans, Judith M.E.M.; Heemskerk, Johan W. M.

doi:10.3109/09537104.2011.630848

Cited by 126 publications

(127 citation statements)

References 88 publications

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“…Under flow conditions, platelets adhesion is affected by rheological conditions such as shear rate, presence of red blood cells, red blood cell deformability, and viscosity of the medium. In this multitude of conditions, platelet adhesion can be evaluated by using microfluidic devices for example biochip containing several different adhesion molecules [152,153] . Platelet spreading tests, using fluorescence microscopy or scanning electron microscopy are frequently employed [154] .…”

Section: Future Perspectivesmentioning

confidence: 99%

Assessment of platelet function: Laboratory and point-of-care methods

Paniccia¹

2014

WJTM

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In the event of blood vessel damage, human platelets are promptly recruited on the site of injury and, after their adhesion, activation and aggregation, prevent blood loss with the formation of a clot. The consequence of abnormal regulation can be either hemorrhage or the development of thrombosis. Qualitative and/or quantitative defects in platelets promote bleeding, whereas the residual reactivity of platelets, despite antiplatelet therapies, play an important role in promoting arterial thrombotic complications. Platelet function is traditionally assessed to investigate the origin of a bleeding syndrome, to predict the risk of bleeding prior surgery or during pregnancy or to monitor the efficacy of antiplatelet therapy in thrombotic syndromes that, now, can be considered a new discipline. "Old" platelet function laboratory tests such as the evaluation of bleeding time and the platelet aggregation analysis in platelet-rich plasma are traditionally utilized to aid in the diagnosis and management of patients with platelet and hemostatic disorders and used as diagnostic tools both in bleeding and thrombotic diathesis in specialized laboratories. Now, new and renewed automated systems have been introduced to provide a simple, rapid assessment of platelet function including point of care methods. These new methodologies are also suitable for being used in non-specialized laboratories and in critical area for assessing platelet function in whole blood without the requirement of sample processing. Some of these methods are also beginning to be incorporated into routine clinical use and can be utilized as not only as first panel for the diagnosis of platelet dysfunction, but also for monitoring anti-platelet therapy and to potentially assess risk of both bleeding and/or thrombosis.© 2014 Baishideng Publishing Group Inc. All rights reserved.Key words: Platelets; Method; Test; Point of care testing; Laboratory assessment; Bleeding; Thrombosis; Platelet function Core tip: This review discussed the scenario of available platelet function laboratory and point-of-care methods suitable in different clinical setting. As this matter has become of crucial importance in the bleeding management and for monitoring antiplatelet therapies, improved ability to assess platelet function in a timely and efficient manner is essential. Traditional platelet function methods, requiring a fair degree of expertise, have been limited to specialized laboratory. Many efforts have been carried out for improving platelet function assays for centralized laboratory, such as different point-of-care testing methodologies have been developed. Moreover, different guidelines and recommendations for their method standardization are growing.

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Section: Future Perspectivesmentioning

confidence: 99%

Assessment of platelet function: Laboratory and point-of-care methods

Paniccia¹

2014

WJTM

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“…The vessel wall is modeled by either coating of biomolecules or growth of cells on one surface of the flow chamber 3 . Particles [4][5][6][7] or cells [8][9][10][11][12][13][14][15][16] are then flowed in at desired range of flow rates to quantify the number of adhering particles under various shear rates.…”

Section: Introductionmentioning

confidence: 99%

Generation of Shear Adhesion Map Using SynVivo Synthetic Microvascular Networks

Prabhakarpandian

Pant

2014

JoVE

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Cell/particle adhesion assays are critical to understanding the biochemical interactions involved in disease pathophysiology and have important applications in the quest for the development of novel therapeutics. Assays using static conditions fail to capture the dependence of adhesion on shear, limiting their correlation with in vivo environment. Parallel plate flow chambers that quantify adhesion under physiological fluid flow need multiple experiments for the generation of a shear adhesion map. In addition, they do not represent the in vivo scale and morphology and require large volumes (~ml) of reagents for experiments. In this study, we demonstrate the generation of shear adhesion map from a single experiment using a microvascular network based microfluidic device, SynVivo-SMN. This device recreates the complex in vivo vasculature including geometric scale, morphological elements, flow features and cellular interactions in an in vitro format, thereby providing a biologically realistic environment for basic and applied research in cellular behavior, drug delivery, and drug discovery. The assay was demonstrated by studying the interaction of the 2 µm biotin-coated particles with avidin-coated surfaces of the microchip. The entire range of shear observed in the microvasculature is obtained in a single assay enabling adhesion vs. shear map for the particles under physiological conditions. Video LinkThe video component of this article can be found at

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“…The extent of variation in construction, usage and analysis results in a high degree of variation and complicates comparison and verification of the results (81). However, there are recommendations for design and use aimed at standardizing the method (81,92,93).…”

Section: In Vitro Flow Chambersmentioning

confidence: 99%

“…= 100 ℎ ( In the equation g is the shear rate (s -1 ), Q is the volumetric flow rate (mL/min), h is the chamber height (mm) and w is the chamber width (mm). The equation is based on a number of assumptions; the flow is parabolic, the fluid is Newtonian and in addition that the width of the channel is greater than the height (92). When doing these calculations blood is assumed to be Newtonian, i.e.…”

Section: Shear Rate Settingsmentioning

confidence: 99%

“…In literature hirudin is stated as a preferable option as the effect on the calcium balance is low and therefore the anticoagulation itself has a low impact on platelet adhesion and aggregation under flow (92). Another advantage is that the anticoagulant is in powder form and will not dilute or change the haematocrit of the sample (red blood cell -plasma ratio).…”

Section: Blood Collection Anticoagulation and Labellingmentioning

confidence: 99%

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Counting and Tracking: Development and Use of New Methods for Detailed Analysis of Thrombus Formation

Tunströmer¹

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Blood platelets are a part of the complex system called haemostasis aimed at ensuring our blood's continuous transport of oxygen and nutrients throughout the body. The transport is ensured by limiting blood loss due to vessel injury and in this process, the platelets form a plug in the damaged area, reinforced by the formation of fibrin. Similar mechanisms may cause thrombus formation, often triggered by atherosclerotic plaque rupture, causing vessel occlusion, embolism or ischemia, which may cause irreversible damage to the heart or the brain.Platelet research is crucial for improved prevention and treatment of thrombotic disorders. For such research, flow chambers are an interesting tool for studies of platelet adhesion, aggregation and thrombus formation under similar flow conditions as in the blood vessels, which is important, as the flow affects the mechanisms involved in both haemostasis and thrombosis. Flow chambers can be designed for specific purposes, such as for the study of haemostasis at specific flow conditions or to evaluate drugs or biomaterials. In this thesis, our aim has been to improve the usefulness of in-vitro flow chambers and develop a more robust and informative image analysis of such experiments.Initially, we introduced an internal control within each flow chamber experiment, thereby reducing the experimental variance caused by unknown factors. Furthermore, control and sample were thus exposed to identical experimental settings. By using platelet count as quantification of thrombus formation we introduce a method of analysis with increased or similar sensitivity to today's standards. The platelet count method facilitated comparison of results obtained in different types of flow chambers by an absolute scale of measurement, independent of user settings. The platelet count method was further developed so that additional parameters could be analysed, providing more information about each individual platelet and the overall thrombus. The parameters analysed included platelet stability, height, movement and contraction. The method was used to evaluate how the pharmacokinetics of a reversible (ticagrelor) and irreversible (prasugrel) platelet ADP-receptor inhibitor affected the overall thrombus formation. Especially, how a non-inhibited platelet fraction, formed between drug administrations of irreversible inhibitors, affected thrombus formation. In addition, we sought to understand the regulation of the thrombin receptor, PAR1, expression in cancer cells. We found the microRNA miR20b to be antioncogenic through its downregulation of PAR1 expression.This thesis contains numerous flow chamber experiments. However, for further use and full potential of the method increased standardisation is important. Our work regarding the quantification and analysis of flow chamber experiments will contribute to a more robust analysis and maybe even more important, provide new and detailed information on thrombus formation. Populärvetenskaplig sammanfattningVåra celler är i konstant behov av syre och...

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Measurement of whole blood thrombus formation using parallel-plate flow chambers – a practical guide

Cited by 126 publications

References 88 publications

Assessment of platelet function: Laboratory and point-of-care methods

Assessment of platelet function: Laboratory and point-of-care methods

Generation of Shear Adhesion Map Using SynVivo Synthetic Microvascular Networks

Counting and Tracking: Development and Use of New Methods for Detailed Analysis of Thrombus Formation

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