Intravital Assessment of Blood Platelet Function. A Review of the Methodological Approaches with Examples of Studies of Selected Aspects of Blood Platelet Function

Q: What is the clinical evidence from this study?

Study design: Review. Population: Thrombosis, inflammation, and atherosclerosis in animal models. Intervention: Intravital platelet function assessment protocols (vascular wall injury, systemic agonist injection, and intact vascular wall interaction models).

Q: What does this research mean for the field?

Protocols for intravital assessment of blood platelet function are broadly categorized into three distinct methodological approaches: controlled injury to the vascular wall, systemic agonist injection, and interactions with a mechanically intact vascular wall. Novelty: ClaimNovelty.SYNTHESIS. Consensus alignment: ConsensusAlignment.NEUTRAL.

Dawid Polak; Marcin Talar; Cezary Watała; Tomasz Przygodzki

doi:10.3390/ijms21218334

November 6, 2020International Journal of Molecular SciencesOpen Access

Intravital Assessment of Blood Platelet Function. A Review of the Methodological Approaches with Examples of Studies of Selected Aspects of Blood Platelet Function

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Key result

Intravital platelet function protocols divide into controlled vascular injury, systemic agonists, and intact wall interactions.

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Why the study?

Researchers entering intravital platelet studies may be confused by the heterogeneity of experimental protocols arising from numerous combinations of activation stimuli and measurement approaches.

Key Points

Systematically review and categorize commonly used intravital experimental protocols used to assess platelet activation, adhesion, and thrombus formation in vivo.
Methodological review evaluating in vivo platelet experimental protocols categorized by the trigger used to elicit activation.
Assessed quantification techniques including laser or ultrasound Doppler blood flow measurements and intravital fluorescence microscopy.
Protocols are classified into three core categories: controlled vascular wall injury, systemic agonist injection, and interactions with mechanically intact vascular walls.
Vascular injury models using mechanical disruption or chemical denudation via ferric chloride reliably quantify platelet recruitment, adhesion, and thrombus growth.
Intravital mouse models demonstrated that blockade or genetic deficiency of glycoprotein VI impairs platelet tethering, stable aggregation, and significantly prolongs occlusion time.

Structured PICO

P

Population

Preclinical animal models (including wild-type, transgenic, chimeric, and ApoE-/- mice, rats, and hamsters) used for intravital assessment of blood platelet function in hemostasis, thrombosis, inflammation, and atherosclerosis

I

Intervention

Intravital assessment protocols categorized into three main methodological approaches: (1) controlled vascular wall injury (e.g., chemical denudation with ferric chloride [FeCl3], rose Bengal photostimulation, or focused laser injury), (2) systemic injection of specific agonists (e.g., ADP, collagen, epinephrine, thrombin, U46619), and (3) interaction of platelets with a mechanically intact, activated vascular wall

O

Outcome

Quantification of in vivo platelet activation, tethering, rolling, firm adhesion, aggregation, thrombus formation/architecture, and vessel occlusion (measured via intravital fluorescence/confocal microscopy, laser/ultrasound Doppler flowmetry, or scintillation counts of radiolabeled platelets)

This methodological review systematically categorizes intravital animal models of platelet function into three distinct protocol classes, guiding researchers in selecting appropriate in vivo stimuli and measurement techniques for thrombosis, antiplatelet drug testing, and atherosclerosis studies.

Limitations

High variability in ferric chloride concentrations (ranging from 2.5% to 80%) and application times across experimental protocols.
Controversies and discrepancies regarding the exact underlying mechanisms of ferric chloride-induced vs. laser-induced thrombus generation.
Interspecies differences in receptor structure (such as GPIb and alphaIIb-beta3) limit the direct translation of murine models to human physiology.
Artifactual platelet activation during extracorporeal isolation and radiolabeling procedures in pulmonary thromboembolism models.
High variability in experimental conditions applied to induce vascular injury (e.g., FeCl3 concentrations varying between 2.5% and 80%)
Questions regarding how closely experimental vascular injuries resemble endogenous fissuring or rupture of human atherosclerotic plaques
Divergence between animal models and human platelet physiology (e.g., interspecies differences in vWF A1 domain and alphaIIb-beta3 antagonist sensitivity)
Artifactual platelet activation during collection, isolation, and labeling procedures, as well as high cost and disposal issues with radioisotopes

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Implication

Aids intravital model selection for thrombosis and antiplatelet research; leaves open protocol standardization and clinical translation.

Abstract

Platelet biology owes to intravital studies not only a better understanding of platelets' role in primary hemostasis but also findings that platelets are important factors in inflammation and atherosclerosis. Researchers who enter the field of intravital platelet studies may be confused by the heterogeneity of experimental protocols utilized. On the one hand, there are a variety of stimuli used to activate platelet response, and on the other hand there are several approaches to measure the outcome of the activation. A number of possible combinations of activation factors with measurement approaches result in the aforementioned heterogeneity. The aim of this review is to present the most often used protocols in a systematic way depending on the stimulus used to activate platelets. By providing examples of studies performed with each of the protocols, we attempt to explain why a particular combination of stimuli and measurement method was applied to study a given aspect of platelet biology.

Discussion

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Cite This Study

Polak et al. (2020) conducted a review in Thrombosis, inflammation, and atherosclerosis in animal models. Intravital platelet function assessment protocols (vascular wall injury, systemic agonist injection, and intact vascular wall interaction models) was evaluated. Intravital platelet function protocols are categorized into three approaches: controlled vascular injury, systemic agonist injection, and interactions with the mechanically intact vascular wall.

synapsesocial.com/papers/6a80b68e1c96fcc8ad704455 https://doi.org/10.3390/ijms21218334

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Intravital Assessment of Blood Platelet Function. A Review of the Methodological Approaches with Examples of Studies of Selected Aspects of Blood Platelet Function

Figures

Key Points

Structured PICO

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

Intravital Assessment of Blood Platelet Function. A Review of the Methodological Approaches with Examples of Studies of Selected Aspects of Blood Platelet Function

Figures

Key Points

Structured PICO

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

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