title: Model thrombi formed under flow reveal the role of factor XIII-mediated cross-linking in resistance to fibrinolysis.
abstract: BACKGROUND
  Activated factor XIII (FXIIIa), a transglutaminase, introduces fibrin-fibrin and fibrin-inhibitor cross-links, resulting in more mechanically stable clots. The impact of cross-linking on resistance to fibrinolysis has proved challenging to evaluate quantitatively.
  
  
  METHODS
  We used a whole blood model thrombus system to characterize the role of cross-linking in resistance to fibrinolytic degradation. Model thrombi, which mimic arterial thrombi formed in vivo, were prepared with incorporated fluorescently labeled fibrinogen, in order to allow quantification of fibrinolysis as released fluorescence units per minute.
  
  
  RESULTS
  A site-specific inhibitor of transglutaminases, added to blood from normal donors, yielded model thrombi that lysed more easily, either spontaneously or by plasminogen activators. This was observed both in the cell/platelet-rich head and fibrin-rich tail. Model thrombi from an FXIII-deficient patient lysed more quickly than normal thrombi; replacement therapy with FXIII concentrate normalized lysis. In vitro addition of purified FXIII to the patient's preprophylaxis blood, but not to normal control blood, resulted in more stable thrombi, indicating no further efficacy of supraphysiologic FXIII. However, addition of tissue transglutaminase, which is synthesized by endothelial cells, generated thrombi that were more resistant to fibrinolysis; this may stabilize mural thrombi in vivo.
  
  
  CONCLUSIONS
  Model thrombi formed under flow, even those prepared as plasma 'thrombi', reveal the effect of FXIII on fibrinolysis. Although very low levels of FXIII are known to produce mechanical clot stability, and to achieve γ-dimerization, they appear to be suboptimal in conferring full resistance to fibrinolysis.
authors.affiliation: Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK.
authors.forename: N J
authors.initials: NJ
authors.lastname: Mutch
authors.forename: J S
authors.initials: JS
authors.lastname: Koikkalainen
authors.forename: S R
authors.initials: SR
authors.lastname: Fraser
authors.forename: K M
authors.initials: KM
authors.lastname: Duthie
authors.forename: M
authors.initials: M
authors.lastname: Griffin
authors.forename: J
authors.initials: J
authors.lastname: Mitchell
authors.forename: H G
authors.initials: HG
authors.lastname: Watson
authors.forename: N A
authors.initials: NA
authors.lastname: Booth
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry / Q000494:pharmacology*; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005176:Factor XIII / Q000737:chemistry / Q000502:physiology*; D005342:Fibrinolysis; D005353:Fibronectins / Q000737:chemistry; D019204:GTP-Binding Proteins / Q000737:chemistry; D006801:Humans; D020128:Inhibitory Concentration 50; D010960:Plasminogen Activators / Q000737:chemistry; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D013927:Thrombosis / Q000378:metabolism*; D013997:Time Factors; D011503:Transglutaminases / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D005353:Fibronectins; D005176:Factor XIII; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D010960:Plasminogen Activators; D019204:GTP-Binding Proteins