Using Microcontact Printing of Fibrinogen to Control Surface-Induced Platelet Adhesion and Activation

Corum, Lindsey E.; Eichinger, Colin D.; Hsiao, Tony W.; Hlady, Vladimir

doi:10.1021/la201064d

Cited by 54 publications

(50 citation statements)

References 49 publications

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“…[20][21][22][23] In this work, we apply microcontact printing to create micro/nanoscale patterns of matrix proteins that mediate platelet adhesion and activation, to demonstrate that the microenvironmental geometry directly mediates platelet physiology and function. Specifically, we demonstrate that platelet a-granule secretion is spatially regulated at the micro/nanoscale via rearrangement of the actin cytoskeleton.…”

Section: Introductionmentioning

confidence: 99%

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Sakurai

Fitch-Tewfik

Qiu

et al. 2015

Blood

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• The geometric orientation of the underlying matrix regulates platelet a-granule secretion.• On geometrically constrained matrices, platelets selfdeposit additional matrix, providing more cell membrane to extend spreading.Although the biology of platelet adhesion on subendothelial matrix after vascular injury is well characterized, how the matrix biophysical properties affect platelet physiology is unknown. Here we demonstrate that geometric orientation of the matrix itself regulates platelet a-granule secretion, a key component of platelet activation. Using protein microcontact printing, we show that platelets spread beyond the geometric constraints of fibrinogen or collagen micropatterns with <5-mm features. Interestingly, a-granule exocytosis and deposition of the a-granule contents such as fibrinogen and fibronectin were primarily observed in those areas of platelet extension beyond the matrix protein micropatterns. This enables platelets to "self-deposit" additional matrix, provide more cellular membrane to extend spreading, and reinforce platelet-platelet connections. Mechanistically, this phenomenon is mediated by actin polymerization, Rac1 activation, and a IIb b 3 integrin redistribution and activation, and is attenuated in gray platelet syndrome platelets, which lack a-granules, and Wiskott-Aldrich syndrome platelets, which have cytoskeletal defects. Overall, these studies demonstrate how platelets transduce geometric cues of the underlying matrix geometry into intracellular signals to extend spreading, which endows platelets spatial flexibility when spreading onto small sites of exposed subendothelium. (Blood. 2015;126(4):531-538) IntroductionHemostasis begins with platelet adhesion and activation at the site of vascular injury.1 Platelets then secrete their a-and dense granules, leading to further recruitment of other platelets and the formation of a hemostatic plug.2 Extensive research has characterized the underlying biological signaling pathways that govern these processes at the bulk level via in vivo and in vitro techniques.3 However, how platelets interact with their microenvironment at the single-cell level and how these interactions subsequently influence platelet physiology remain poorly understood. For example, although single platelets have long been observed to fill submicron-sized "gaps" in the endothelium, 4 how platelets establish adhesion on such small areas of exposed matrix that is strong enough to withstand the shear forces of the circulation is unknown. Studies of single platelet-matrix interactions may help explain the role of platelets in maintaining vascular integrity and supporting the semipermeable barrier function of the endothelium during homeostatic conditions. 5 Furthermore, because platelets within different regions of the same thrombus exhibit significantly different levels of activation, 6,7 this concept of physical and microenvironmental regulation of platelet physiology at the single-cell level is also important for our overall understanding of clot formation. S...

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Section: Introductionmentioning

confidence: 99%

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Sakurai

Fitch-Tewfik

Qiu

et al. 2015

Blood

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“…A second patch of an immobilized proadhesive protein downstream of the agonist is used to capture primed platelets. In the present study, fibrinogen was used as the protein in the capture region as it has been previously demonstrated to effectively arrest primed platelets; 2,11 however, other capture proteins (e.g., collagen or vWF) could easily be incorporated. This assay would not have the required sensitivity without the phenomenon of platelet margination toward flow chamber walls in laminar flow of whole blood.…”

Section: Discussionmentioning

confidence: 99%

“…We have recently demonstrated that a platelet population allowed to transiently interact with a stimulating surface patch has an increased propensity to activate and adhere downstream. 2,11 This phenomenon is largely a) Author to whom correspondence should be addressed; present address: 20 S 2030 E., Rm. 108A, Salt Lake City, Utah 84112; electronic mail: vladimir.hlady@utah.edu due to the margination of platelets in flowing blood which was utilized here to create a new type of antiplatelet agent assay that takes into account the upstream history of plateletagonist interactions.…”

Section: A Platelet Activation and Adhesionmentioning

confidence: 99%

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Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

2016

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A novel functional assay of antiplatelet drug efficacy was designed by utilizing the phenomena of platelet margination in flowing blood and transient platelet contacts with surface-immobilized platelet agonists. Flow margination enhances transient contacts of platelets with the walls of flow chambers covered with surface-immobilized proteins. Depending on the type and the surface density of the immobilized agonists, such transient interactions could "prime" the marginated platelet subpopulation for enhanced activation and adhesion downstream. By creating an upstream surface patch with an immobilized platelet agonist, platelet flow margination was used to test how effective antiplatelet drugs are in suppressing downstream platelet activation and adhesion. The platelet adhesion downstream was measured by a so-called "capture" patch region close to the distal end of the flow chamber. Platelet adhesion downstream was found to be dose-dependent on the upstream surface coverage of the "priming" patch, with immobilized fibrinogen acting as a platelet agonist. Several antiplatelet agents (acetylsalicylic acid, eptifibatide, and tirofiban) were evaluated for their efficacy in attenuating downstream adhesion after upstream platelet priming. The activation of the platelet population was found to be dependent on both the extent of the upstream agonist stimulus and the antiplatelet drug concentration. Such a relationship provides an opportunity to measure the efficacy of specific antiplatelet agents against the type and concentration of upstream platelet agonists.

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“…However, due to the low blood compatibility of the PDMS surface, a blocking agent (e.g., BSA) has generally been used to coat the PDMS and inhibit cell adsorption. 17,18,30 Accordingly, we next examined the extent of platelet adhesion to native PDMS surfaces, BSA-coated PDMS surfaces, and nanosheet-coated PDMS surfaces in order to validate the applicability of modified PDMS devices for hematological use. Various PDMS plates immersed in PRP were incubated for different hours before the observation.…”

Section: A Characterization Of the Modified Surfacesmentioning

confidence: 99%

Surface modification on polydimethylsiloxane-based microchannels with fragmented poly(l-lactic acid) nanosheets

2015

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Surface modification is a critical issue in various applications of polydimethylsiloxane (PDMS)-based microfluidic devices. Here, we describe a novel method through which PDMS-based microchannels were successfully modified with fragmented poly(L-lactic acid) (PLLA) nanosheets through a simple patchwork technique that exploited the high level of adhesiveness of PLLA nanosheets. Compared with other surface modification methods, our method required neither complicated chemical modifications nor the use of organic solvents that tend to cause PDMS swelling. The experimental results indicated that the modified PDMS exhibited excellent capacity for preventing the adhesion and activation of platelets. This simple yet efficient method can be used to fabricate the special PDMS microfluidic devices for biological, medical, and even hematological purposes. V C 2015 AIP Publishing LLC.

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Using Microcontact Printing of Fibrinogen to Control Surface-Induced Platelet Adhesion and Activation

Cited by 54 publications

References 49 publications

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

Surface modification on polydimethylsiloxane-based microchannels with fragmented poly(l-lactic acid) nanosheets

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