Kinetics of Platelet Adhesion to Protein-Coated Surface in Whole Blood Samples at High Flow Rates

Avtaeva, Yuliya; Melnikov, Ivan; Okhota, Sergey; Zozulya, Nadezhda; Gabbasov, Zufar

doi:10.1007/s10517-020-04856-z

Cited by 5 publications

(4 citation statements)

References 14 publications

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“…The microfluidic optical system consisted of a disposable flow chamber with optical surface, specialised optical prism system with semiconductor laser (λ=650 nm) with a photodetector (Hamamatsu), a computer-controlled peristaltic pump (Instech Laboratories, Plymouth Meeting, PA), and an analogue-to-digital con-verter connecting the system to a computer. We have previously validated the method by measuring platelet adhesion to fibrinogen-coated surfaces in platelet rich plasma [1] and whole blood [2]. Shear stress-induced adhesion of blood cells to type I collagen-coated surface of a perfusion chamber was recorded during 15-min circulation of whole blood samples from healthy volunteers at shear rate of 2300 sec -1 .…”

Section: Methodsmentioning

confidence: 99%

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

et al. 2021

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We studied the contribution of von Willebrand factor (vWF) into blood cell adhesion to collagen-coated surfaces in whole blood of healthy volunteers. Adhesion of blood cells to collagen I was measured at shear rate of 2300 sec -1 . The interaction of platelet GPIIb/IIIa receptor with vWF was blocked with monoclonal anti-GPIIb/IIIa antibodies. The degree of cell adhesion was quantified by measuring the intensity of scattered light after 15-min perfusion: in samples with blocked GPIIb/IIIa it decreased to 0.39±0.13 V vs 0.06±0.03 V in control samples (p=0.002). Under a fluorescence microscope, intensively stained structures consisting of vWF, platelets, and leukocytes attached to the collagen surface were observed. After blockade of GPIIb/IIIa, these structures were absent. Leukocyte recruitment at high shear rates is a time-dependent process sensitive to complex interaction of vWF, leukocytes, and platelets, in which the platelet GPIIb/IIIa receptor is essential.

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Section: Methodsmentioning

confidence: 99%

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

et al. 2021

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“…In this case, it is also necessary to determine the involvement of other proteins in clot formation, such as vWF, which has additional receptors capable to bind inactive platelets from the bloodstream. [50,51]…”

Section: Proposed Crystallography-induced Hpf Conformation Mechanismmentioning

confidence: 99%

How Crystallographic Orientation‐Induced Fibrinogen Conformation Affects Platelet Adhesion and Activation on TiO₂

Struczyńska

Firkowska‐Boden

Levandovsky

et al. 2023

Adv Healthcare Materials

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Control of protein adsorption is essential for successful integration of healthcare materials into the body. Human plasma fibrinogen (HPF), especially its conformation is a key upstream regulator for platelet behavior and thus pathological clot formation at the blood‐biomaterial interface. A previous study by the authors revealed that the conformation of adsorbed HPF can be controlled by rutile surface crystallographic orientation. Therefore, it is hypothesized that pre‐adsorbed HPF on specific rutile orientation can regulate platelets adhesion and activation. Here, it is shown that platelets exposed to the four low index (110), (100), (101), (001) facets of TiO2 (rutile) exhibit surface‐specific behavior. Scanning electron microscopy (SEM) observations of platelets morphology and P‐selectin expression measurement revealed that on (110) facets, platelets adhesion and activation are suppressed. In contrast, extensive surface coverage by fully activated platelets is observed on (001) facets. Platelets' behavior has been linked to the HPF conformation and thereby availability of platelet‐binding sequences. Atomic force microscopy (AFM) imaging supported by immunochemical analysis shows that on (110) facets, HPF is adsorbed in trinodular conformation rendering the γ400‐411 platelet‐binding sequence inaccessible. This research has potential implications on the bioactivity of different materials crystal facets, reducing the risk of pathological clot formation and thromboembolic complications.

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Section: Mainmentioning

confidence: 99%

“…But fluorophore labeling in whole blood can be challenging. Finally, laser optical imaging has been used for obtaining the number of platelet interactions that occur with a protein substrate in a microfluidic channel, 18 but this method is limited to a single numeric output (intensity of light scattered over time), and has additional experimental setup complexity. Here, we describe a practical, accessible, and easily adaptable microscopy method that enables real-time imaging of dynamics of cellular interactions under whole blood flow in vitro by completely eliminating the need for blood sample dilution.…”

Section: Mainmentioning

confidence: 99%

Motion Blur Microscopy

Goreke,

Gonzales,

Shipley

et al. 2023

Preprint

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Imaging and characterizing the dynamics of cellular adhesion in blood samples is of fundamental importance in understanding biological function.In vitromicroscopy methods are widely used for this task, but typically require diluting the blood with a buffer to allow for transmission of light. However whole blood provides crucial mechanical and chemical signaling cues that influence adhesion dynamics, which means that conventional approaches lack the full physiological complexity of living microvasculature. We propose to overcome this challenge by a newin vitroimaging method which we call motion blur microscopy (MBM). By decreasing the source light intensity and increasing the integration time during imaging, flowing cells are blurred, allowing us to identify adhered cells. Combined with an automated analysis using machine learning, we can for the first time reliably image cell interactions in microfluidic channels during whole blood flow. MBM provides a low cost, easy to implement alternative to intravital microscopy, thein vivoapproach for studying how the whole blood environment shapes adhesion dynamics. We demonstrate the method’s reproducibility and accuracy in two example systems where understanding cell interactions, adhesion, and motility is crucial—sickle red blood cells adhering to laminin, and CAR-T cells adhering to E-selectin. We illustrate the wide range of data types that can be extracted from this approach, including distributions of cell size and eccentricity, adhesion times, trajectories and velocities of adhered cells moving on a functionalized surface, as well as correlations among these different features at the single cell level. In all cases MBM allows for rapid collection and processing of large data sets, ranging from thousands to hundreds of thousands of individual adhesion events. The method is generalizable to study adhesion mechanisms in a variety of diseases, including cancer, blood disorders, thrombosis, inflammatory and autoimmune diseases, as well as providing rich datasets for theoretical modeling of adhesion dynamics.

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Kinetics of Platelet Adhesion to Protein-Coated Surface in Whole Blood Samples at High Flow Rates

Cited by 5 publications

References 14 publications

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

How Crystallographic Orientation‐Induced Fibrinogen Conformation Affects Platelet Adhesion and Activation on TiO₂

Motion Blur Microscopy

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Kinetics of Platelet Adhesion to Protein-Coated Surface in Whole Blood Samples at High Flow Rates

Cited by 5 publications

References 14 publications

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

Complex Interaction of Platelets, von Willebrand Factor and Leukocytes, in Whole Blood at High Shear Rates Is Mediated by Platelet GPIIb/IIIa Receptor

How Crystallographic Orientation‐Induced Fibrinogen Conformation Affects Platelet Adhesion and Activation on TiO2

Motion Blur Microscopy

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Product

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How Crystallographic Orientation‐Induced Fibrinogen Conformation Affects Platelet Adhesion and Activation on TiO₂