Effect of metabolic inhibitors on platelet attachment, spreading and aggregation on collagen-coated surfaces .

Misselwitz, Frank; Leytin, Valery; Repin, V. S.

doi:10.1016/0049-3848(87)90285-4

Cited by 14 publications

(8 citation statements)

References 12 publications

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“…thrombin, ADP, thromboxane, serotonin, thrombospondin and platelet activating factors [21,36]. These molecules can induce a progressing activation of further platelets, which could be documented by the measurement of PF4 since the relative concentration of PF4 in platelets exceeds that of plasma by 280,000-fold [6,12,14,23].…”

Section: Discussionmentioning

confidence: 99%

Changes in platelet morphology and function during 24 hours of storage

Braune

Walter

Schulze

et al. 2014

Clinical Hemorheology and Microcirculation

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For in vitro studies assessing the interaction of platelets with implant materials, common and standardized protocols for the preparation of platelet rich plasma (PRP) are lacking, which may lead to non-matching results due to the diversity of applied protocols. Particularly, the aging of platelets during prolonged preparation and storage times is discussed to lead to an underestimation of the material thrombogenicity. Here, we study the influence of whole blood-and PRP-storage times on changes in platelet morphology and function.Blood from apparently healthy subjects was collected according to a standardized protocol and examined immediately after blood collection, four hours and twenty four hours later. The capability of platelets to adhere and form stable aggregates (PFA100, closure time) was examined in sodium citrate anticoagulated whole blood (WB) using the agonists equine type I collagen and epinephrine bitartrate (collagen/epinephrine) as well as equine type I collagen and adenosine-5 -diphosphate (collagen/ADP). Circulating platelets were quantified at each time point. Morphology of platelets and platelet aggregates were visualized microscopically and measured using an electric field multi-channel counting system (CASY). The percentage of activated platelets was assessed by means of P-selectin (CD62P) expression of circulating platelets. Furthermore, platelet factor 4 (PF4) release was measured in platelet poor plasma (PPP) at each time point.Whole blood PFA100 closure times increased after stimulation with collagen/ADP and collagen/epinephrine. Twenty four hours after blood collection, both parameters were prolonged pathologically above the upper limit of the reference range. Numbers of circulating platelets, measured in PRP, decreased after four hours, but no longer after twenty four hours. Mean platelet volumes (MPV) and platelet large cell ratios (P-LCR, 12 fL -40 fL) decreased over time. Immediately after blood collection, no debris or platelet aggregates could be visualized microscopically. After four hours, first debris and very small aggregates occurred. After 24 hours, platelet aggregates and also debris progressively increased. In accordance to this, the CASY system revealed an increase of platelet aggregates (up to 90 m diameter) with increasing storage time. The percentage of CD62P positive platelets and PF4 increased significantly with storage time in resting PRP. When soluble ADP was added to stored PRP samples, the number of activatable platelets decreased significantly over storage time.The present study reveals the importance of a consequent standardization in the preparation of WB and PRP. Platelet morphology and function, particularly platelet reactivity to adherent or soluble agonists in their surrounding milieu, changed rapidly outside the vascular system. This knowledge is of crucial interest, particularly in the field of biomaterial development for cardiovascular applications, and may help to define common standards in the in vitro hemocompatibility testing of biomaterials.

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Section: Discussionmentioning

confidence: 99%

Changes in platelet morphology and function during 24 hours of storage

Braune

Walter

Schulze

et al. 2014

Clinical Hemorheology and Microcirculation

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“…Platelet adhesion to isolated collagens has been studied in many laboratories by a variety of procedures ( [8][9][10][11][12][13][14] and references cited therein). We have chosen in the present investigation to measure adhesion to collagen and collagen-derived fragments immobilized on plastic by adsorption on the plastic surface from solution.…”

Section: Introductionmentioning

confidence: 99%

Platelet-reactive sites in collagens type I and type III. Evidence for separate adhesion and aggregatory sites

Morton¹,

Peachey²,

Barnes³

1989

Biochemical Journal

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The adhesion of human and rabbit platelets to collagens and collagen-derived fragments immobilized on plastic was investigated. Adhesion appeared to be independent of collagen conformation, since similar attachment occurred to collagen (type I) in monomeric form, as fibres or in denatured state. The adhesion of human platelets was stimulated to a variable degree by Mg2+, but rabbit platelet adhesion showed little if any dependence on this cation. Collagens type I, III, V and VI were all able to support adhesion, although that to collagen type V (native) was lower than that to the other collagens. Adhesion to a series of peptides derived from collagens I and III was measured. Attachment did not require the presence of peptides in triple-helical configuration. The extent of adhesion ranged from relatively high, as good as to the intact parent collagen molecule, to little if any adhesive activity beyond the non-specific (background) level. The existence of very different degrees of activity suggests that platelet adhesion is associated with specific structural sites in the collagen molecule. Adhesion in many instances was essentially in accord with the known platelet-aggregatory activity of individual peptides. However, two peptides, alpha 1(I)CB3 and alpha 1(III)CB1,8,10,2, exhibited good adhesive activity although possessing little if any aggregatory activity. Of particular interest, despite its near-total lack of aggregatory activity, adhesion to peptide alpha 1(I)CB3 was as good as that to the structurally homologous peptide alpha 1(III)CB4, in which is located a highly reactive aggregatory site. This implies that platelet adhesion to collagen may involve sites in the collagen molecule distinct from those more directly associated with aggregation.

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“…These could primarily be divided into platelet cytoskeletal and cytoskeleton-associated proteins and metabolic proteins. Both categories are instrumental in platelet activation; activation of platelet surface receptors such as integrin ␣IIb␤3 leads to transduction of signals to the platelet cytoskeleton, its subsequent polymerization, and reorganization, leading to shape change (61), whereas carbohydrate metabolism and glycolysis are thought to be important energy sources for platelet activation (62). Because of their various roles in platelet activation, interference with the function of these proteins through ASinduced modification could have ramifications for the ability of platelets to aggregate.…”

Section: Global Proteome Analysis For Discovery Of Hno-induced Modifimentioning

confidence: 99%

Identification of Nitroxyl-induced Modifications in Human Platelet Proteins Using a Novel Mass Spectrometric Detection Method

Hoffman

Walsh

Rogalski

et al. 2009

Molecular & Cellular Proteomics

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Nitroxyl (HNO) exhibits many important pharmacological effects, including inhibition of platelet aggregation, and the HNO donor Angeli's salt has been proposed as a potential therapeutic agent in the treatment of many diseases including heart failure and alcoholism. Despite this, little is known about the mechanism of action of HNO, and its effects are rarely linked to specific protein targets of HNO or to the actual chemical changes that proteins undergo when in contact with HNO. Here we study the presumed major molecular target of HNO within the body: protein thiols. Cysteine-containing tryptic peptides were reacted with HNO, generating the sulfinamide modification and, to a lesser extent, disulfide linkages with no other long lived intermediates or side products. The sulfinamide modification was subjected to a comprehensive tandem mass spectrometric analysis including MS/MS by CID and electron capture dissociation as well as an MS 3 analysis. These studies revealed a characteristic neutral loss of HS(O)NH 2 (65 Da) that is liberated from the modified cysteine upon CID and can be monitored by mass spectrometry. Upon storage, partial conversion of the sulfinamide to sulfinic acid was observed, leading to coinciding neutral losses of 65 and 66 Da (HS(O)OH). Validation of the method was conducted using a targeted study of nitroxylated glyceraldehyde-3-phosphate dehydrogenase extracted from Angeli's salt-treated human platelets. In these ex vivo experiments, the sample preparation process resulted in complete conversion of sulfinamide to sulfinic acid, making this the sole subject of further ex vivo studies. A global proteomics analysis to discover platelet proteins that carry nitroxyl-induced modifications and a mass spectrometric HNO dose-response analysis of the modified proteins were conducted to gain insight into the specificity and selectivity of this modification. These methods identified 10 proteins that are modified dose dependently in response to HNO, whose functions range from metabolism and cytoskeletal rearrangement to signal transduction, providing for Nitric oxide (NO)1 has emerged as an important physiological signaling molecule, particularly in the vascular, neuronal, and immune systems. NO regulates many processes including platelet function, vascular tone, and leukocyte recruitment mainly through the cGMP second messenger system (1). More recent studies have shown that nitric oxide can react directly with a number of different biological species including metal centers of proteins, nucleophilic amino acid residues (nitrosation/S-nitrosylation), and aromatic amino acid residues (nitration) (2, 3), and the products of these reactions have been analyzed by mass spectrometry (4 -8). The biological relevance of these reactions is slowly coming to light, and NO-mediated S-nitrosylation has now been linked to a number of diseases including diabetes, multiple sclerosis, cystic fibrosis, and asthma (9).Nitroxyl (HNO/NO Ϫ ), an alternative redox form of NO, has only recently begun to draw attention in th...

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Effect of metabolic inhibitors on platelet attachment, spreading and aggregation on collagen-coated surfaces .

Cited by 14 publications

References 12 publications

Changes in platelet morphology and function during 24 hours of storage

Changes in platelet morphology and function during 24 hours of storage

Platelet-reactive sites in collagens type I and type III. Evidence for separate adhesion and aggregatory sites

Identification of Nitroxyl-induced Modifications in Human Platelet Proteins Using a Novel Mass Spectrometric Detection Method

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