Skip to main content
Top
Published in: BMC Neurology 1/2015

Open Access 01-12-2015 | Research article

The changes in clot microstructure in patients with ischaemic stroke and the effects of therapeutic intervention: a prospective observational study

Authors: Sophia N Stanford, Ahmed Sabra, Lindsay D’Silva, Matthew Lawrence, Roger HK Morris, Sharon Storton, Martyn Rowan Brown, Vanessa Evans, Karl Hawkins, Phylip Rhodri Williams, Simon J Davidson, Mushtaq Wani, John F Potter, Phillip A Evans

Published in: BMC Neurology | Issue 1/2015

Login to get access

Abstract

Background

Stroke is the second largest cause of death worldwide. Hypercoagulability is a key feature in ischaemic stroke due to the development of an abnormally dense clot structure but techniques assessing the mechanics and quality of clot microstructure have limited clinical use. We have previously validated a new haemorheological technique using three parameters to reflect clot microstructure (Fractal Dimension (d f )) ex-vivo, real-time clot formation time (T GP ) and blood clot strength (elasticity at the gel point (G’GP)). We aimed to evaluate these novel clotting biomarkers in ischaemic stroke and changes of clot structure following therapeutic intervention.

Methods

In a prospective cohort study clot microstructure was compared in ischaemic stroke patients and a control group of healthy volunteers. Further assessment took place at 2–4 hours and at 24 hours after therapeutic intervention in the stroke group to assess the effects of thrombolysis and anti-platelet therapy.

Results

75 patients (mean age 72.8 years [SD 13.1]; 47 male, 28 female) with ischaemic stroke were recruited. Of the 75 patients, 32 were thrombolysed with t-PA and 43 were loaded with 300 mg aspirin. The following parameters were significantly different between patients with stroke and the 74 healthy subjects: d f (1.760 ± .053 versus 1.735 ± 0.048, p = 0.003), TGP (208 ± 67 versus 231 ± 75, p = 0.05), G’GP (0.056 ± 0.017 versus 0.045 ± 0.014, p < 0.0001) and fibrinogen (3.7 ± 0.8 versus 3.2 ± 0.5, p < 0.00001). There was a significant decrease in d f (p = 0.02), G’GP (p = 0.01) and fibrinogen (p = 0.01) following the administration of aspirin and for d f (p = 0.003) and fibrinogen (p < 0.001) following thrombolysis as compared to baseline values.

Conclusion

Patients with ischaemic stroke have denser and stronger clot structure as detected by d f and G’GP. The effect of thrombolysis on clot microstructure (d f ) was more prominent than antiplatelet therapy. Further work is needed to assess the clinical and therapeutic implications of these novel biomarkers.
Literature
2.
go back to reference Undas A, Podolec P, Zawilska K, Pieculewicz M, Jedlinski I, Stepien E, et al. Altered fibrin clot structure/function in patients with cryptogenic ischemic stroke. Stroke. 2009;40:1499–501.CrossRefPubMed Undas A, Podolec P, Zawilska K, Pieculewicz M, Jedlinski I, Stepien E, et al. Altered fibrin clot structure/function in patients with cryptogenic ischemic stroke. Stroke. 2009;40:1499–501.CrossRefPubMed
3.
go back to reference Pera J, Undas A, Topor-Madry R, Jagiella J, Klimkowicz-Mrowiec A, Slowik A. Fibrin Clot Properties in Acute Stroke: What Differs Cerebral Hemorrhage From Cerebral Ischemia? Stroke. 2012;43:1412–14.CrossRefPubMed Pera J, Undas A, Topor-Madry R, Jagiella J, Klimkowicz-Mrowiec A, Slowik A. Fibrin Clot Properties in Acute Stroke: What Differs Cerebral Hemorrhage From Cerebral Ischemia? Stroke. 2012;43:1412–14.CrossRefPubMed
4.
go back to reference Scott EM, Ariens RA, Grant PJ. Genetic and environmental determinants of fibrin structure and function: Relevance to clinical disease. Arterioscler Thromb Vasc Biol. 2004;24:1558–66.CrossRefPubMed Scott EM, Ariens RA, Grant PJ. Genetic and environmental determinants of fibrin structure and function: Relevance to clinical disease. Arterioscler Thromb Vasc Biol. 2004;24:1558–66.CrossRefPubMed
5.
go back to reference Collet JP, Allali Y, Lesty C, Tanguy ML, Silvain J, Ankri A, et al. Altered fibrin architecture is associated with hypofibrinolysis and premature coronary atherothrombosis. Arterioscler Thromb Vasc Biol. 2006;26:2567–73.CrossRefPubMed Collet JP, Allali Y, Lesty C, Tanguy ML, Silvain J, Ankri A, et al. Altered fibrin architecture is associated with hypofibrinolysis and premature coronary atherothrombosis. Arterioscler Thromb Vasc Biol. 2006;26:2567–73.CrossRefPubMed
6.
go back to reference Chung I, Lip GY. Virchow’s triad revisited: Blood constituents. Pathophysiol Haemost Thromb. 2003;33:449–54.CrossRefPubMed Chung I, Lip GY. Virchow’s triad revisited: Blood constituents. Pathophysiol Haemost Thromb. 2003;33:449–54.CrossRefPubMed
7.
go back to reference Curry ANG, Pierce JMT. Conventional and near-patient tests of coagulation. Continuing Education in Anaesthesia, Critical Care & Pain. 2007;7:45–50.CrossRef Curry ANG, Pierce JMT. Conventional and near-patient tests of coagulation. Continuing Education in Anaesthesia, Critical Care & Pain. 2007;7:45–50.CrossRef
8.
go back to reference Favaloro EJ, Lippi G. Coagulation update: What’s new in hemostasis testing? Thromb Res. 2011;127 Suppl 2:S13–16.CrossRefPubMed Favaloro EJ, Lippi G. Coagulation update: What’s new in hemostasis testing? Thromb Res. 2011;127 Suppl 2:S13–16.CrossRefPubMed
9.
go back to reference Antovic A. Screening haemostasis-looking for global assays: The overall haemostasis potential (ohp) method-a possible tool for laboratory investigation of global haemostasis in both hypo-and hypercoagulable conditions. Curr Vasc Pharmacol. 2008;6:173–85.CrossRefPubMed Antovic A. Screening haemostasis-looking for global assays: The overall haemostasis potential (ohp) method-a possible tool for laboratory investigation of global haemostasis in both hypo-and hypercoagulable conditions. Curr Vasc Pharmacol. 2008;6:173–85.CrossRefPubMed
10.
go back to reference Rooth E, Wallen NH, Blomback M, He S. Decreased fibrin network permeability and impaired fibrinolysis in the acute and convalescent phase of ischemic stroke. Thromb Res. 2011;127:51–6.CrossRefPubMed Rooth E, Wallen NH, Blomback M, He S. Decreased fibrin network permeability and impaired fibrinolysis in the acute and convalescent phase of ischemic stroke. Thromb Res. 2011;127:51–6.CrossRefPubMed
11.
go back to reference Takahashi A, Kita R, Shinozaki T, Kubota K. Real space observation of 3-d network structure of hydrated fibrin gel. Colloid Polym Sci. 2003;28:832–8.CrossRef Takahashi A, Kita R, Shinozaki T, Kubota K. Real space observation of 3-d network structure of hydrated fibrin gel. Colloid Polym Sci. 2003;28:832–8.CrossRef
12.
go back to reference Weisel JW. The mechanical properties of fibrin for basic scientists and clinicians. Biophys Chem. 2004;112:267–76.CrossRefPubMed Weisel JW. The mechanical properties of fibrin for basic scientists and clinicians. Biophys Chem. 2004;112:267–76.CrossRefPubMed
13.
go back to reference Lawrence MJ, Sabra A, Mills G, Pillai SG, Abdullah W, Hawkins K, Morris RHK, Davidson SJ, D'Silva L, Curtis DJ, Brown MR, Weisel JW, Williams PR, Evans PA. A new biomarker quantifies differences in clot microstructure in patients with venous thromboembolism. Br J Haematol. 2014. doi:10.1111/bjh.13173. Lawrence MJ, Sabra A, Mills G, Pillai SG, Abdullah W, Hawkins K, Morris RHK, Davidson SJ, D'Silva L, Curtis DJ, Brown MR, Weisel JW, Williams PR, Evans PA. A new biomarker quantifies differences in clot microstructure in patients with venous thromboembolism. Br J Haematol. 2014. doi:10.1111/bjh.13173.
14.
go back to reference Lawrence MJ, Kumar S, Hawkins K, Boden S, Rutt H, Mills G, et al. A new structural biomarker that quantifies and predicts changes in clot strength and quality in a model of progressive haemodilution. Thromb Res. 2014;134(2):488–94.CrossRefPubMed Lawrence MJ, Kumar S, Hawkins K, Boden S, Rutt H, Mills G, et al. A new structural biomarker that quantifies and predicts changes in clot strength and quality in a model of progressive haemodilution. Thromb Res. 2014;134(2):488–94.CrossRefPubMed
15.
go back to reference Evans PA, Hawkins K, Morris RHK, Thirumalai N, Munro R, Wakeman L, et al. Gel point and fractal microstructure of incipient blood clots are significant new markers of hemostasis for healthy and anticoagulated blood. Blood. 2010;116:3341–6.CrossRefPubMed Evans PA, Hawkins K, Morris RHK, Thirumalai N, Munro R, Wakeman L, et al. Gel point and fractal microstructure of incipient blood clots are significant new markers of hemostasis for healthy and anticoagulated blood. Blood. 2010;116:3341–6.CrossRefPubMed
16.
go back to reference Evans PA, Lawrence M, Morris RHK, Thirumalai N, Munro R, Wakeman L, et al. Fractal analysis of viscoelastic data with automated gel point location and its potential application in the investigation of therapeutically modified blood coagulation. Rheol Acta. 2010;49:901–8.CrossRef Evans PA, Lawrence M, Morris RHK, Thirumalai N, Munro R, Wakeman L, et al. Fractal analysis of viscoelastic data with automated gel point location and its potential application in the investigation of therapeutically modified blood coagulation. Rheol Acta. 2010;49:901–8.CrossRef
17.
go back to reference Winter HH, Chambon F. Analysis of linear viscoelasticity of a crosslinking polymer at the gel point. J Rheol. 1986;30:367–82.CrossRef Winter HH, Chambon F. Analysis of linear viscoelasticity of a crosslinking polymer at the gel point. J Rheol. 1986;30:367–82.CrossRef
18.
go back to reference Yoon WB, Kim BY, Park JW. Rheological characteristics of fibrinogen-thrombin solution and its effects on surimi gels. J Food Sci. 1999;64:291–4.CrossRef Yoon WB, Kim BY, Park JW. Rheological characteristics of fibrinogen-thrombin solution and its effects on surimi gels. J Food Sci. 1999;64:291–4.CrossRef
19.
go back to reference Hatano S. Experience from a multicentre stroke register: A preliminary report. Bull World Health Organ. 1976;54:541–53.PubMedPubMedCentral Hatano S. Experience from a multicentre stroke register: A preliminary report. Bull World Health Organ. 1976;54:541–53.PubMedPubMedCentral
20.
go back to reference Curtis DJ, Brown MR, Hawkins K, Evans PA, Lawrence MJ, Rees P, et al. Rheometrical and molecular dynamics simulation studies of incipient clot formation in fibrin-thrombin gels: An activation limited aggregation approach. J Non-Newtonian Fluid Mech. 2011;166:932–8.CrossRef Curtis DJ, Brown MR, Hawkins K, Evans PA, Lawrence MJ, Rees P, et al. Rheometrical and molecular dynamics simulation studies of incipient clot formation in fibrin-thrombin gels: An activation limited aggregation approach. J Non-Newtonian Fluid Mech. 2011;166:932–8.CrossRef
21.
go back to reference Undas A, Slowik A, Wolkow P, Szczudlik A, Tracz W. Fibrin clot properties in acute ischemic stroke: Relation to neurological deficit. Thromb Res. 2010;125:357–61.CrossRefPubMed Undas A, Slowik A, Wolkow P, Szczudlik A, Tracz W. Fibrin clot properties in acute ischemic stroke: Relation to neurological deficit. Thromb Res. 2010;125:357–61.CrossRefPubMed
22.
go back to reference Brown MR, Curtis DJ, Rees P, Summers HD, Hawkins K, Evans PA, et al. Fractal discrimination of random fractal aggregates and its application in biomarker analysis for blood coagulation. Chaos, Solitons Fractals. 2012;45:1025–32.CrossRef Brown MR, Curtis DJ, Rees P, Summers HD, Hawkins K, Evans PA, et al. Fractal discrimination of random fractal aggregates and its application in biomarker analysis for blood coagulation. Chaos, Solitons Fractals. 2012;45:1025–32.CrossRef
23.
go back to reference Curtis DJ, Williams PR, Badiei N, Campbell AI, Hawkins K, Evans PA, et al. A study of microstructural templating in fibrin-thrombin gel networks by spectral and viscoelastic analysis. Soft Matter. 2013;9:4883–9.CrossRef Curtis DJ, Williams PR, Badiei N, Campbell AI, Hawkins K, Evans PA, et al. A study of microstructural templating in fibrin-thrombin gel networks by spectral and viscoelastic analysis. Soft Matter. 2013;9:4883–9.CrossRef
24.
go back to reference Simak J, Gelderman MP. Cell membrane microparticles in blood and blood products: potentially pathogenic agents and diagnostic markers. Transfus Med Rev. 2006;20(1):1–26.CrossRefPubMed Simak J, Gelderman MP. Cell membrane microparticles in blood and blood products: potentially pathogenic agents and diagnostic markers. Transfus Med Rev. 2006;20(1):1–26.CrossRefPubMed
25.
go back to reference Wilhelmsen L, Svardsudd K, Korsan-Bengtsen K, Larsson B, Welin L, Tibblin G. Fibrinogen as a risk factor for stroke and myocardial infarction. N Engl J Med. 1984;311:501–5.CrossRefPubMed Wilhelmsen L, Svardsudd K, Korsan-Bengtsen K, Larsson B, Welin L, Tibblin G. Fibrinogen as a risk factor for stroke and myocardial infarction. N Engl J Med. 1984;311:501–5.CrossRefPubMed
26.
go back to reference Di Napoli M, Singh P. Is plasma fibrinogen useful in evaluating ischemic stroke patients?: Why, how, and when. Stroke. 2009;40:1549–52.CrossRefPubMed Di Napoli M, Singh P. Is plasma fibrinogen useful in evaluating ischemic stroke patients?: Why, how, and when. Stroke. 2009;40:1549–52.CrossRefPubMed
27.
go back to reference He S, Blomback M, Yoo G, Sinha R, Henschen-Edman AH. Modified clotting properties of fibrinogen in the presence of acetylsalicylic acid in a purified system. Ann N Y Acad Sci. 2001;936:531–5.CrossRefPubMed He S, Blomback M, Yoo G, Sinha R, Henschen-Edman AH. Modified clotting properties of fibrinogen in the presence of acetylsalicylic acid in a purified system. Ann N Y Acad Sci. 2001;936:531–5.CrossRefPubMed
28.
go back to reference Undas A, Sydor WJ, Brummel K, Musial J, Mann KG, Szczeklik A. Aspirin alters the cardioprotective effects of the factor xiii val34leu polymorphism. Circulation. 2003;107:17–20.CrossRefPubMed Undas A, Sydor WJ, Brummel K, Musial J, Mann KG, Szczeklik A. Aspirin alters the cardioprotective effects of the factor xiii val34leu polymorphism. Circulation. 2003;107:17–20.CrossRefPubMed
29.
go back to reference Bjornsson TD, Schneider DE, Berger H. Aspirin acetylates fibrinogen and enhances fibrinolysis. Fibrinolytic effect is independent of changes in plasminogen activator levels. J Pharmacol Exp Ther. 1989;250:154–61.PubMed Bjornsson TD, Schneider DE, Berger H. Aspirin acetylates fibrinogen and enhances fibrinolysis. Fibrinolytic effect is independent of changes in plasminogen activator levels. J Pharmacol Exp Ther. 1989;250:154–61.PubMed
30.
go back to reference Ceulemans AG, Zgavc T, Kooijman R, Hachimi-Idrissi S, Sarre S, Michotte Y. The dual role of the neuroinflammatory response after ischemic stroke: Modulatory effects of hypothermia. J Neuroinflammation. 2010;7:74.CrossRefPubMedPubMedCentral Ceulemans AG, Zgavc T, Kooijman R, Hachimi-Idrissi S, Sarre S, Michotte Y. The dual role of the neuroinflammatory response after ischemic stroke: Modulatory effects of hypothermia. J Neuroinflammation. 2010;7:74.CrossRefPubMedPubMedCentral
31.
go back to reference Sughrue ME, Mehra A, Connolly Jr ES, D’Ambrosio AL. Anti-adhesion molecule strategies as potential neuroprotective agents in cerebral ischemia: A critical review of the literature. Inflamm Res. 2004;53:497–508.CrossRefPubMed Sughrue ME, Mehra A, Connolly Jr ES, D’Ambrosio AL. Anti-adhesion molecule strategies as potential neuroprotective agents in cerebral ischemia: A critical review of the literature. Inflamm Res. 2004;53:497–508.CrossRefPubMed
32.
go back to reference Bode C, Runge MS, Haber E. Future directions in plasminogen activator therapy. Clin Cardiol. 1990;13:375–81.CrossRefPubMed Bode C, Runge MS, Haber E. Future directions in plasminogen activator therapy. Clin Cardiol. 1990;13:375–81.CrossRefPubMed
33.
go back to reference Tanswell P, Seifried E, Su PC, Feuerer W, Rijken DC. Pharmacokinetics and systemic effects of tissue-type plasminogen activator in normal subjects. Clin Pharmacol Ther. 1989;46:155–62.CrossRefPubMed Tanswell P, Seifried E, Su PC, Feuerer W, Rijken DC. Pharmacokinetics and systemic effects of tissue-type plasminogen activator in normal subjects. Clin Pharmacol Ther. 1989;46:155–62.CrossRefPubMed
34.
go back to reference Noman A, Egred M, Bagnall A, Spyridopoulos I, Jamieson S, Ahmed J. Impact of thrombus aspiration during primary percutaneous coronary intervention on mortality in st-segment elevation myocardial infarction. Eur Heart J. 2012;33:3054–61.CrossRefPubMed Noman A, Egred M, Bagnall A, Spyridopoulos I, Jamieson S, Ahmed J. Impact of thrombus aspiration during primary percutaneous coronary intervention on mortality in st-segment elevation myocardial infarction. Eur Heart J. 2012;33:3054–61.CrossRefPubMed
35.
go back to reference Fokkema ML, Wieringa WG, van der Horst IC, Boersma E, Zijlstra F, de Smet BJ. Quantitative analysis of the impact of total ischemic time on myocardial perfusion and clinical outcome in patients with st-elevation myocardial infarction. Am J Cardiol. 2011;108:1536–41.CrossRefPubMed Fokkema ML, Wieringa WG, van der Horst IC, Boersma E, Zijlstra F, de Smet BJ. Quantitative analysis of the impact of total ischemic time on myocardial perfusion and clinical outcome in patients with st-elevation myocardial infarction. Am J Cardiol. 2011;108:1536–41.CrossRefPubMed
36.
go back to reference De Luca G, Suryapranata H, Ottervanger JP, Antman EM. Time delay to treatment and mortality in primary angioplasty for acute myocardial infarction: Every minute of delay counts. Circulation. 2004;109:1223–5.CrossRefPubMed De Luca G, Suryapranata H, Ottervanger JP, Antman EM. Time delay to treatment and mortality in primary angioplasty for acute myocardial infarction: Every minute of delay counts. Circulation. 2004;109:1223–5.CrossRefPubMed
37.
go back to reference Kramer MC, van der Wal AC, Koch KT, Ploegmakers JP, van der Schaaf RJ, Henriques JP, et al. Presence of older thrombus is an independent predictor of long-term mortality in patients with st-elevation myocardial infarction treated with thrombus aspiration during primary percutaneous coronary intervention. Circulation. 2008;118:1810–16.CrossRefPubMed Kramer MC, van der Wal AC, Koch KT, Ploegmakers JP, van der Schaaf RJ, Henriques JP, et al. Presence of older thrombus is an independent predictor of long-term mortality in patients with st-elevation myocardial infarction treated with thrombus aspiration during primary percutaneous coronary intervention. Circulation. 2008;118:1810–16.CrossRefPubMed
38.
go back to reference Silvain J, Collet JP, Nagaswami C, Beygui F, Edmondson KE, Bellemain-Appaix A, et al. Composition of coronary thrombus in acute myocardial infarction. J Am Coll Cardiol. 2011;57:1359–67.CrossRefPubMedPubMedCentral Silvain J, Collet JP, Nagaswami C, Beygui F, Edmondson KE, Bellemain-Appaix A, et al. Composition of coronary thrombus in acute myocardial infarction. J Am Coll Cardiol. 2011;57:1359–67.CrossRefPubMedPubMedCentral
39.
go back to reference Pretorius E, Oberholzer HM, van der Spuy WJ, Meiring JH. Smoking and coagulation: The sticky fibrin phenomenon. Ultrastruct Pathol. 2010;34:236–9.CrossRefPubMed Pretorius E, Oberholzer HM, van der Spuy WJ, Meiring JH. Smoking and coagulation: The sticky fibrin phenomenon. Ultrastruct Pathol. 2010;34:236–9.CrossRefPubMed
40.
go back to reference Sowers MR, Matthews KA, Jannausch M, Randolph JF, McConnell D, Sutton-Tyrrell K, et al. Hemostatic factors and estrogen during the menopausal transition. J Clin Endocrinol Metab. 2005;90:5942–8.CrossRefPubMed Sowers MR, Matthews KA, Jannausch M, Randolph JF, McConnell D, Sutton-Tyrrell K, et al. Hemostatic factors and estrogen during the menopausal transition. J Clin Endocrinol Metab. 2005;90:5942–8.CrossRefPubMed
41.
go back to reference Lip GYH. Using the CHADS2 and CHA2DS2-VASc Scores for stroke risk prediction as well as the identification of stroke outcomes and cardiac complications in patients with and without atrial fibrillation. Cerebrovasc Dis. 2013;36:281–2.CrossRefPubMed Lip GYH. Using the CHADS2 and CHA2DS2-VASc Scores for stroke risk prediction as well as the identification of stroke outcomes and cardiac complications in patients with and without atrial fibrillation. Cerebrovasc Dis. 2013;36:281–2.CrossRefPubMed
42.
go back to reference Holzer K, Feurer R, Sadikovic S, Esposito L, Bockelbrink A, Sander D, et al. Prognostic value of the ABCD2 score beyond short-term follow-up after transient ischemic attack (TIA)–a cohort study. BMC Neurol. 2010;10:50.CrossRefPubMedPubMedCentral Holzer K, Feurer R, Sadikovic S, Esposito L, Bockelbrink A, Sander D, et al. Prognostic value of the ABCD2 score beyond short-term follow-up after transient ischemic attack (TIA)–a cohort study. BMC Neurol. 2010;10:50.CrossRefPubMedPubMedCentral
43.
go back to reference Prabhakaran S, Jovin TG, Tayal AH, Hussain MS, Nguyen TN, Sheth KN, et al. Posttreatment variables improve outcome prediction after intra-arterial therapy for acute ischemic stroke. Cerebrovasc Dis. 2014;37:356–63.CrossRefPubMedPubMedCentral Prabhakaran S, Jovin TG, Tayal AH, Hussain MS, Nguyen TN, Sheth KN, et al. Posttreatment variables improve outcome prediction after intra-arterial therapy for acute ischemic stroke. Cerebrovasc Dis. 2014;37:356–63.CrossRefPubMedPubMedCentral
Metadata
Title
The changes in clot microstructure in patients with ischaemic stroke and the effects of therapeutic intervention: a prospective observational study
Authors
Sophia N Stanford
Ahmed Sabra
Lindsay D’Silva
Matthew Lawrence
Roger HK Morris
Sharon Storton
Martyn Rowan Brown
Vanessa Evans
Karl Hawkins
Phylip Rhodri Williams
Simon J Davidson
Mushtaq Wani
John F Potter
Phillip A Evans
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Neurology / Issue 1/2015
Electronic ISSN: 1471-2377
DOI
https://doi.org/10.1186/s12883-015-0289-1

Other articles of this Issue 1/2015

BMC Neurology 1/2015 Go to the issue