96-well plate-based aggregometry

Chan, Melissa; Armstrong, Paul C; Warner, Timothy D.

doi:10.1080/09537104.2018.1445838

Cited by 32 publications

(32 citation statements)

References 36 publications

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“…While 96-well LTA is not validated for this purpose, the test is based on the exact same principle as classical LTA. Of relevance, 96-well LTA has been shown to be sensitive to common platelet inhibitors as aspirin [24], ADP-receptor antagonists and glycoprotein IIb/IIIa inhibitors [47]. The cross-sectional design of the study limits the present study to examining associations and not causalities.…”

Section: Discussionmentioning

confidence: 95%

Platelet aggregation is not altered among men with diabetes mellitus

et al. 2019

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Aims: Platelets are pivotal in arterial thrombosis, and platelet hyperresponsiveness may contribute to the increased incidence of cardiovascular events in diabetes mellitus. Consequently, we hypothesized that increased in vitro platelet aggregation responses exist in men with diabetes mellitus. Methods: The Danish Cardiovascular Screening Trial (DANCAVAS) is a communitybased cardiovascular screening trial including men aged 65-74 years. Platelet aggregation was tested using 96-well light transmission aggregometry with thrombin receptor-activating peptide (TRAP), adenosine diphosphate, collagen type-1, arachidonic acid, and proteaseactivated receptor-4 in three concentrations. Further, cardiovascular risk factors and coronary artery calcification (CAC), estimated by CT-scans and ankle-brachial index were obtained. Results: Included were 720 men aged 65-74 years, 110 with diabetes mellitus. Overall, there was no difference in platelet aggregation among men with versus without diabetes mellitus when adjusting for, or excluding platelet inhibitor treatment and men with established cardiovascular disease (CVD). This was true for all agonists, e.g. 10 µM TRAP-induced platelet aggregation of median 69 % (IQR 53-75) versus 70% (IQR 60-76) in men with versus without diabetes mellitus. Platelet aggregation did not correlate with HbA1c, or CAC. Men with diabetes mellitus displayed higher CAC, median 257 Agatston Units (IQR 74-1141) versus median 111 Agatston Units (IQR 6-420) in the remaining individuals, p<0.0001.

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Section: Discussionmentioning

confidence: 95%

Platelet aggregation is not altered among men with diabetes mellitus

et al. 2019

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“…Furthermore, promising results are accomplished with platelet aggregation studies under flow conditions [28]. Advantages of these new methods are the use of whole blood and low sample preparation, but they also require trained technicians and specialized equipment [29][30][31][32][33][34]. Likewise, standardization of the traditional LTA method is still essential here.…”

Section: Discussionmentioning

confidence: 99%

Harmonizing light transmission aggregometry in the Netherlands by implementation of the SSC-ISTH guideline

et al. 2020

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Light transmission aggregometry (LTA) is considered the gold standard method for evaluation of platelet function. However, there are a lot of variation in protocols (pre-analytical procedures and agonist concentrations) and results. The aim of our study was to establish a national LTA protocol, to investigate the effect of standardization and to define national reference values for LTA. The SSC guideline was used as base for a national procedure. Almost all recommendations of the SSC were followed e.g. no adjustment of PRP, citrate concentration of 109 mM, 21 needle gauge, fasting, resting time for whole blood and PRP, centrifugation time, speed and agonists concentrations. LTA of healthy volunteers was measured in a total of 16 hospitals with 5 hospitals before and after standardization. Results of more than 120 healthy volunteers (maximum aggregation %) were collected, with participating laboratories using 4 different analyzers with different reagents. Use of low agonist concentrations showed high variation before and after standardization, with the exception of collagen. For most high agonist concentrations (ADP, collagen, ristocetin, epinephrine and arachidonic acid) variability in healthy subjects decreased after standardization. We can conclude that a standardized Dutch protocol for LTA, based on the SSC guideline, does not result in smaller variability in healthy volunteers for all agonist concentrations.

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“…Platelet aggregometry was performed using a modification of the previously published technique [ 28 ]. Following platelet incubation with TE constructs, 200 µL of the platelet suspension was transferred into a 96-well plate and then placed into a plate reader prewarmed to 37 °C (BioTek Synergy 2 microplate, Winooski, VT, USA).…”

Section: Methodsmentioning

confidence: 99%

The Combination of Tissue-Engineered Blood Vessel Constructs and Parallel Flow Chamber Provides a Potential Alternative to In Vivo Drug Testing Models

et al. 2021

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Cardiovascular disease is a major cause of death globally. This has led to significant efforts to develop new anti-thrombotic therapies or re-purpose existing drugs to treat cardiovascular diseases. Due to difficulties of obtaining healthy human blood vessel tissues to recreate in vivo conditions, pre-clinical testing of these drugs currently requires significant use of animal experimentation, however, the successful translation of drugs from animal tests to use in humans is poor. Developing humanised drug test models that better replicate the human vasculature will help to develop anti-thrombotic therapies more rapidly. Tissue-engineered human blood vessel (TEBV) models were fabricated with biomimetic matrix and cellular components. The pro- and anti-aggregatory properties of both intact and FeCl3-injured TEBVs were assessed under physiological flow conditions using a modified parallel-plate flow chamber. These were perfused with fluorescently labelled human platelets and endothelial progenitor cells (EPCs), and their responses were monitored in real-time using fluorescent imaging. An endothelium-free TEBV exhibited the capacity to trigger platelet activation and aggregation in a shear stress-dependent manner, similar to the responses observed in vivo. Ketamine is commonly used as an anaesthetic in current in vivo models, but this drug significantly inhibited platelet aggregation on the injured TEBV. Atorvastatin was also shown to enhance EPC attachment on the injured TEBV. The TEBV, when perfused with human blood or blood components under physiological conditions, provides a powerful alternative to current in vivo drug testing models to assess their effects on thrombus formation and EPC recruitment.

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96-well plate-based aggregometry

Cited by 32 publications

References 36 publications

Platelet aggregation is not altered among men with diabetes mellitus

Platelet aggregation is not altered among men with diabetes mellitus

Harmonizing light transmission aggregometry in the Netherlands by implementation of the SSC-ISTH guideline

The Combination of Tissue-Engineered Blood Vessel Constructs and Parallel Flow Chamber Provides a Potential Alternative to In Vivo Drug Testing Models

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