Role of Shear Stress and tPA Concentration in the Fibrinolytic Potential of Thrombi

Claire S. Whyte; Hadj Ahmed. Mostefai; Kim M. Baeten; Andrew J. Lucking; David E. Newby; Nuala A. Booth; Nicola J. Mutch

doi:10.3390/ijms22042115

Role of Shear Stress and tPA Concentration in the Fibrinolytic Potential of Thrombi

Claire S. Whyte, Hadj Ahmed. Mostefai, Kim M. Baeten, Andrew J. Lucking, David E. Newby, Nuala A. Booth, Nicola J. Mutch^* (Corresponding Author)

^*Corresponding author for this work

University of Edinburgh

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Abstract

The resolution of arterial thrombi is critically dependent on the endogenous fibrinolytic system. Using well-established and complementary whole blood models, we investigated the endogenous fibrinolytic potential of the tissue-type plasminogen activator (tPA) and the intra-thrombus distribution of fibrinolytic proteins, formed ex vivo under shear. tPA was present at physiologically relevant concentrations and fibrinolysis was monitored using an FITC-labelled fibrinogen tracer. Thrombi were formed from anticoagulated blood using a Chandler Loop and from non-anticoagulated blood perfused over specially-prepared porcine aorta strips under low (212 s−1) and high shear (1690 s−1) conditions in a Badimon Chamber. Plasminogen, tPA and plasminogen activator inhibitor-1 (PAI-1) concentrations were measured by ELISA. The tPA–PAI-1 complex was abundant in Chandler model thrombi serum. In contrast, free tPA was evident in the head of thrombi and correlated with fibrinolytic activity. Badimon thrombi formed under high shear conditions were more resistant to fibrinolysis than those formed at low shear. Plasminogen and tPA concentrations were elevated in thrombi formed at low shear, while PAI-1 concentrations were augmented at high shear rates. In conclusion, tPA primarily localises to the thrombus head in a free and active form. Thrombi formed at high shear incorporate less tPA and plasminogen and increased PAI-1, thereby enhancing resistance to degradation.

Original language	English
Article number	2115
Number of pages	13
Journal	International Journal of Molecular Sciences
Volume	22
Issue number	4
DOIs	https://doi.org/10.3390/ijms22042115
Publication status	Published - 20 Feb 2021

Bibliographical note

Funding: This research was funded by British Heart Foundation PG/08/127/26517 awarded to N.A.B. and D.E.N. C.S.W. and N.J.M. were supported by the British Heart Foundation project grants (PG/15/82/31721 and PG/20/17/35050). D.E.N. and A.J.L. were supported by the British Heart
Foundation project grant PG/04/131/18118. D.E.N. is supported by the British Heart Foundation (CH/09/002, RE/18/5/34216, RG/16/10/32375) and is the recipient of a Wellcome Trust Senior Investigator Award (WT103782AIA).

Keywords

plasminogen activator inhibitor-1
fibrinolysis
tissue plasminogen activator
thrombi
shear
Shear
Tissue plasminogen activator
Plasminogen activator inhibitor-1
Fibrinolysis
Thrombi

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title = "Role of Shear Stress and tPA Concentration in the Fibrinolytic Potential of Thrombi",

abstract = "The resolution of arterial thrombi is critically dependent on the endogenous fibrinolytic system. Using well-established and complementary whole blood models, we investigated the endogenous fibrinolytic potential of the tissue-type plasminogen activator (tPA) and the intra-thrombus distribution of fibrinolytic proteins, formed ex vivo under shear. tPA was present at physiologically relevant concentrations and fibrinolysis was monitored using an FITC-labelled fibrinogen tracer. Thrombi were formed from anticoagulated blood using a Chandler Loop and from non-anticoagulated blood perfused over specially-prepared porcine aorta strips under low (212 s−1) and high shear (1690 s−1) conditions in a Badimon Chamber. Plasminogen, tPA and plasminogen activator inhibitor-1 (PAI-1) concentrations were measured by ELISA. The tPA–PAI-1 complex was abundant in Chandler model thrombi serum. In contrast, free tPA was evident in the head of thrombi and correlated with fibrinolytic activity. Badimon thrombi formed under high shear conditions were more resistant to fibrinolysis than those formed at low shear. Plasminogen and tPA concentrations were elevated in thrombi formed at low shear, while PAI-1 concentrations were augmented at high shear rates. In conclusion, tPA primarily localises to the thrombus head in a free and active form. Thrombi formed at high shear incorporate less tPA and plasminogen and increased PAI-1, thereby enhancing resistance to degradation.",

keywords = "plasminogen activator inhibitor-1, fibrinolysis, tissue plasminogen activator, thrombi, shear, Shear, Tissue plasminogen activator, Plasminogen activator inhibitor-1, Fibrinolysis, Thrombi",

author = "Whyte, {Claire S.} and Mostefai, {Hadj Ahmed.} and Baeten, {Kim M.} and Lucking, {Andrew J.} and Newby, {David E.} and Booth, {Nuala A.} and Mutch, {Nicola J.}",

note = "Funding: This research was funded by British Heart Foundation PG/08/127/26517 awarded to N.A.B. and D.E.N. C.S.W. and N.J.M. were supported by the British Heart Foundation project grants (PG/15/82/31721 and PG/20/17/35050). D.E.N. and A.J.L. were supported by the British Heart Foundation project grant PG/04/131/18118. D.E.N. is supported by the British Heart Foundation (CH/09/002, RE/18/5/34216, RG/16/10/32375) and is the recipient of a Wellcome Trust Senior Investigator Award (WT103782AIA).",

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AU - Whyte, Claire S.

AU - Mostefai, Hadj Ahmed.

AU - Baeten, Kim M.

AU - Lucking, Andrew J.

AU - Newby, David E.

AU - Booth, Nuala A.

AU - Mutch, Nicola J.

N1 - Funding: This research was funded by British Heart Foundation PG/08/127/26517 awarded to N.A.B. and D.E.N. C.S.W. and N.J.M. were supported by the British Heart Foundation project grants (PG/15/82/31721 and PG/20/17/35050). D.E.N. and A.J.L. were supported by the British Heart Foundation project grant PG/04/131/18118. D.E.N. is supported by the British Heart Foundation (CH/09/002, RE/18/5/34216, RG/16/10/32375) and is the recipient of a Wellcome Trust Senior Investigator Award (WT103782AIA).

PY - 2021/2/20

Y1 - 2021/2/20

N2 - The resolution of arterial thrombi is critically dependent on the endogenous fibrinolytic system. Using well-established and complementary whole blood models, we investigated the endogenous fibrinolytic potential of the tissue-type plasminogen activator (tPA) and the intra-thrombus distribution of fibrinolytic proteins, formed ex vivo under shear. tPA was present at physiologically relevant concentrations and fibrinolysis was monitored using an FITC-labelled fibrinogen tracer. Thrombi were formed from anticoagulated blood using a Chandler Loop and from non-anticoagulated blood perfused over specially-prepared porcine aorta strips under low (212 s−1) and high shear (1690 s−1) conditions in a Badimon Chamber. Plasminogen, tPA and plasminogen activator inhibitor-1 (PAI-1) concentrations were measured by ELISA. The tPA–PAI-1 complex was abundant in Chandler model thrombi serum. In contrast, free tPA was evident in the head of thrombi and correlated with fibrinolytic activity. Badimon thrombi formed under high shear conditions were more resistant to fibrinolysis than those formed at low shear. Plasminogen and tPA concentrations were elevated in thrombi formed at low shear, while PAI-1 concentrations were augmented at high shear rates. In conclusion, tPA primarily localises to the thrombus head in a free and active form. Thrombi formed at high shear incorporate less tPA and plasminogen and increased PAI-1, thereby enhancing resistance to degradation.

AB - The resolution of arterial thrombi is critically dependent on the endogenous fibrinolytic system. Using well-established and complementary whole blood models, we investigated the endogenous fibrinolytic potential of the tissue-type plasminogen activator (tPA) and the intra-thrombus distribution of fibrinolytic proteins, formed ex vivo under shear. tPA was present at physiologically relevant concentrations and fibrinolysis was monitored using an FITC-labelled fibrinogen tracer. Thrombi were formed from anticoagulated blood using a Chandler Loop and from non-anticoagulated blood perfused over specially-prepared porcine aorta strips under low (212 s−1) and high shear (1690 s−1) conditions in a Badimon Chamber. Plasminogen, tPA and plasminogen activator inhibitor-1 (PAI-1) concentrations were measured by ELISA. The tPA–PAI-1 complex was abundant in Chandler model thrombi serum. In contrast, free tPA was evident in the head of thrombi and correlated with fibrinolytic activity. Badimon thrombi formed under high shear conditions were more resistant to fibrinolysis than those formed at low shear. Plasminogen and tPA concentrations were elevated in thrombi formed at low shear, while PAI-1 concentrations were augmented at high shear rates. In conclusion, tPA primarily localises to the thrombus head in a free and active form. Thrombi formed at high shear incorporate less tPA and plasminogen and increased PAI-1, thereby enhancing resistance to degradation.

KW - plasminogen activator inhibitor-1

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KW - tissue plasminogen activator

KW - thrombi

KW - shear

KW - Shear

KW - Tissue plasminogen activator

KW - Plasminogen activator inhibitor-1

KW - Fibrinolysis

KW - Thrombi

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U2 - 10.3390/ijms22042115

DO - 10.3390/ijms22042115

M3 - Article

C2 - 33672724

SN - 1422-0067

VL - 22

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

IS - 4

M1 - 2115

ER -

Role of Shear Stress and tPA Concentration in the Fibrinolytic Potential of Thrombi

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Keywords

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