Measurement of adhesion of human platelets in plasma to protein surfaces in microplates

Eriksson, Anna; Whiss, Per A

doi:10.1016/j.vascn.2005.06.002

Cited by 53 publications

(55 citation statements)

References 35 publications

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“…It has previously been shown that for absorbance values between 0 and 1, the platelet count correlates well with the amount of product developed [18]. When performing the same type of analysis on one single patient with high platelet count we found that linear relationships between absorbance and platelet amount can be expected for absorbance values as high as 1.9 (r 2 = 0.98).…”

Section: Platelet Adhesion Assaysupporting

confidence: 55%

“…In the cases where we observed a significant difference in the increase in platelet adhesion (1 μmol/L ADP to collagen; 10 μmol/L ADP to albumin, collagen and fibrinogen; 1 mg/ml ristocetin to collagen and fibrinogen; 0.1 μmol/L adrenaline + 10 μmol/L LPA to albumin), we performed linear regression analyses for the amount of ET-platelets added to wells versus platelet adhesion. Platelet amount was estimated by measurements of acid phosphatase activity, which correlates well with the actual platelet count [18]. However, there were no significant correlations between platelet amount and adhesion since r 2 -values ranged from 0.0099 to 0.094 for the seven different stimuli.…”

Section: Influence Of Platelet Amount History Of Thrombohemorrhagic mentioning

confidence: 99%

“…Platelet adhesion was measured as previously described [18,21]. Briefly, protein-coated microplates were washed twice in 0.9% NaCl by plate inversion followed by addition of 50 μL diluted PRP and 50 μL platelet activator or solvent.…”

Section: Platelet Adhesion Assaymentioning

confidence: 99%

“…Median age at blood sampling -years (range) 67 Median age at diagnosis a -years (range) 61 Median time from diagnosis to blood sampling a -years (range) 7 Treatment at blood sampling -n (%) 27: 82-90 on the assay used, both increased and decreased activity of ET-platelets have been reported [17]. In the present study, we used a novel assay of static platelet adhesion [18] to investigate platelet function in ET patients. The study used a screening approach and investigated adhesion to albumin, collagen, fibrinogen, and fibronectin in the presence of soluble activators such as adenosine 5'-diphosphate (ADP), adrenaline, ristocetin and L-α-lysophosphatidic acid (oleoyl-sn-glycero-3-phosphate, LPA).…”

Section: Introductionmentioning

confidence: 99%

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Enhanced platelet adhesion in essential thrombocythemia after in vitro activation

Eriksson¹,

Lotfi²,

Whiss³

2010

Turk J Hematol

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Objective: Essential thrombocythemia (ET) is a chronic myeloproliferative disorder characterized by elevated platelet counts and increased risk of thrombosis. Ex vivo data suggest increased platelet reactivity in agreement with the increased thrombosis risk, while in vitro tests often detect decreased platelet activity. The present study aimed to investigate adhesion of ET-platelets in vitro, which is an aspect of platelet function that has been addressed in only a few studies on ET patients. Material and Methods: The study included 30 ET patients and 14 healthy controls. Platelet adhesion was measured with a static platelet adhesion assay. Results: The main finding was that ET-platelets were more readily activated by adhesion-inducing stimuli in vitro than control platelets. This was particularly evident in elderly patients and when using multiple stimuli, such as surfaces of collagen or fibrinogen combined with addition of adenosine 5'-diphosphate or ristocetin. Such multiple stimuli resulted in adhesion above the control mean +2 standard deviations for approximately 50% of the patients. Conclusion:The results are in accordance with the concept of increased platelet activity in ET, but opposite to most other in vitro studies. We suggest that the conditions in the adhesion assay might mimic the in vivo situation regarding the presence of chronic platelet activation. ÖzetAmaç: Esansiyel trombositemi (ET) platelet sayısının artması ve yüksek tromboz riski ile karakterize kronik bir myeloproliferatif bozukluktur. Ex vivo veriler tromboz riskine uygun olarak artan platelet reaktivitesini öne sürerken in vitro testler sıklıkla platelet aktivitesinde azalma tespit etmektedir. Bu çalışmanın amacı ET-hastalarında az sayıda çalışmaya dahil edilmiş bir platelet fonksiyonu konusu olan ET-plateletleri adezyonunun in vitro incelenmesidir. Yöntem ve Gereçler: Çalışmaya 30 ET hastası ile 14 sağlıklı kontrol dahil edilmiştir. Statik platelet adezyonu tayini ile platelet adezyonu ölçülmüştür. Bulgular: Temel bulgu ET plateletlerinin, in vitro adezyon indüklenmiş uyaranlar ile kontrol plateletlerinden daha kolay aktive olduğu olmuştur. Bu durum özellikle yaşlı hastalarda ve adenosin 5-difosfat ya da ristosetin eklenerek kombine edilmiş kolajen ya da fibrinojen yüzeyler gibi çoklu uyaran kullanıldığında barizdir. Bu gibi çoklu uyaran hastaların yaklaşık %50'sinde kontrol değeri + 2 standart sapmanın üzerinde adezyon sonucunu vermiştir.

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Section: Platelet Adhesion Assaysupporting

confidence: 55%

Section: Influence Of Platelet Amount History Of Thrombohemorrhagic mentioning

confidence: 99%

Section: Platelet Adhesion Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Enhanced platelet adhesion in essential thrombocythemia after in vitro activation

Eriksson¹,

Lotfi²,

Whiss³

2010

Turk J Hematol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Platelet adhesion to collagen coated surfaces was performed as previously described (Eriksson and Whiss, 2005). In short, microplates were coated with 0.1 mg/mL bovine collagen (RnDSystems, Abingdon, UK) or 2 mg/mL fibrinogen (American Diagnostica Inc., Greenwich, CN).…”

Section: Platelet Adhesion To Collagen Coated Surfacesmentioning

confidence: 99%

C1q induces a rapid up-regulation of P-selectin and modulates collagen- and collagen-related peptide-triggered activation in human platelets

et al. 2010

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, C1q induces a rapid up-regulation of P-selectin and modulates collagen-and collagen-related peptidetriggered activation in human platelets, 2010, Immunobiology, (215), 12, 987-995 The aim of this study was to further characterize the effects of C1q on platelets, by quantifying the platelet surface expression of P-selectin (CD62P) and monitoring the formation of platelet-neutrophil aggregates. Using flow cytometry, we found that C1q dosedependently triggered a rapid but moderate and transient up-regulation of P-selectin already within 5 seconds of C1q exposure. Pre-incubation with an antibody directed against gC1qR significantly inhibited (with 57 % compared to control) the up-regulation, whereas an antibody towards the αIIβI-integrin showed no effect. Stimulation with C1q did not change the cytosolic calcium-levels, as measured with the fluorescent ratiometric probe Fura-2, however, a protein kinase C inhibitor (GF109203x) blocked the C1q-induced P-selectin expression. Furthermore, pre-incubation of platelets with C1q diminished both the collagen as well as the collagen-related peptide induced up-regulation of P-selectin, most evident after 90 seconds of stimulation. This indicates that C1q may regulate platelet activation via the GPVI receptor, which is a novel finding. Moreover, C1q antagonized the collagen-induced formation of platelet-neutrophil aggregates, indicating a reduced interaction between platelet P-selectin and neutrophil P-selectin glycoprotein ligand-1(PSGL-1/CD162). In summary, C1q induces a moderate rapid platelet P-selectin expression, modulates subsequent collagen and collagen-related peptide stimulation of platelets, and inhibits the formation of platelet-4 neutrophil aggregates. These immuno-regulatory effects of C1q may have a crucial role in innate immunity and inflammation.5

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In Vitro Thrombogenicity Testing of Biomaterials

Braune

Latour

Reinthaler

et al. 2019

Adv Healthcare Materials

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reliable, and reproducible. [20][21][22] This is, not least, important for realizing interstudy comparisons of the thrombogenicity of different materials. According, e.g., to the regulation (EU) 2017/745 of the European parliament and of the council on medical devices, in vitro and in vivo tests are a mandatory parts within the preclinical product verification and validation (see Annex II: Technical documentation, 6.1 Preclinical and clinical data). [23] Beyond the results of these tests, also detailed information concerning the test design, respective protocols and data analysis are obligatory to meet the requirements of the regulation, particularly those formulated in the ISO standard 10993 Part 4. [24,25] Since the regulatory agencies provide recommendations rather than protocols or standard operation procedures, accepted standardizations regarding the preanalytical handling of the blood, test conditions, test parameters, reference materials, static or dynamic conditions, flow conditions, and finally, which type of cells and donors should be included is lacking. Also, standard operating procedures how the testing should be performed are lacking. [9] This has led to a situation that laboratories perform in vitro assays under substantially different test conditions, including the preanalytical characterization of blood samples, the application of blood from different species, varying anticoagulation and stabilization or storage times as well as test setups. Taking these variations into account, interlaboratory or interstudy comparisons are impossible. [8,[26][27][28] Here, we review described test methods and procedures for the assessment of the thrombogenicity of materials, prerequisites for a reproducible in vitro testing, applied test system and test parameters for the characterization of the interaction of blood components/cells and the implant. We further review and outline recent approaches that may support realizing reliable, reproducible, and laboratory independent tests.

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Measurement of adhesion of human platelets in plasma to protein surfaces in microplates

Cited by 53 publications

References 35 publications

Enhanced platelet adhesion in essential thrombocythemia after in vitro activation

Enhanced platelet adhesion in essential thrombocythemia after in vitro activation

C1q induces a rapid up-regulation of P-selectin and modulates collagen- and collagen-related peptide-triggered activation in human platelets

In Vitro Thrombogenicity Testing of Biomaterials

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