title: Thrombin generation and clot formation in methylene blue-treated plasma and cryoprecipitate.
abstract: BACKGROUND
  Methylene blue (MB) treatment of plasma is known to reduce the activity of clotting factors, but its effect on thrombin generation and clot formation is not well documented.
  
  
  METHODS
  Individual clotting factors and inhibitors and global tests of thrombin generation and clot formation using rotational thrombelastometry (ROTEM) were assessed in a paired study of standard or MB plasma and cryoprecipitate (n = 20 each).
  
  
  RESULTS
  MB treatment resulted in a 10 percent reduction in endogenous thrombin potential and 30 percent decrease in peak thrombin as well as the expected 20 to 35 percent loss of Factor (F)VIII, fibrinogen, and FXI activity. MB treatment had no effect on the rate of clot formation and increased the clot firmness by 20 percent as assessed by ROTEM. There were minimal further changes in either coagulation factor levels or thrombin generation when thawed plasma was stored for an additional 24 hours. FVIII and fibrinogen content of MB cryoprecipitate was reduced by 30 and 40 percent, respectively, but this was not associated with altered clot time or rate of clot formation by ROTEM and only an 8 percent decrease in clot firmness.
  
  
  CONCLUSIONS
  It is concluded that MB treatment is associated with a reduction in the thrombin-generating capacity of plasma, but has very little effect on the strength of clot formation as assessed by thrombelastometry. The thrombin-generating capacity of standard and MB plasma is relatively unaltered by subsequent storage of thawed plasma at 4 degrees C for 24 hours.
authors.affiliation: NHS Blood & Transplant, Long Road, Brentwood, Cambridge, UK. cardigan@nbs.nhs.uk
authors.forename: Rebecca
authors.initials: R
authors.lastname: Cardigan
authors.forename: Katherine
authors.initials: K
authors.lastname: Philpot
authors.forename: Philip
authors.initials: P
authors.lastname: Cookson
authors.forename: Roger
authors.initials: R
authors.lastname: Luddington
mesh_terms: D000465:Algorithms; D001769:Blood / Q000187:drug effects*; D001777:Blood Coagulation / Q000187:drug effects* / Q000502:physiology; D001780:Blood Coagulation Tests; D001793:Blood Preservation / Q000009:adverse effects / Q000379:methods; D003080:Cold Temperature; D005169:Factor VIII / Q000187:drug effects* / Q000378:metabolism; D005340:Fibrinogen / Q000187:drug effects* / Q000378:metabolism; D006401:Hematologic Agents / Q000494:pharmacology; D006801:Humans; D008751:Methylene Blue / Q000494:pharmacology*; D055550:Protein Stability / Q000187:drug effects; D013917:Thrombin / Q000378:metabolism*
chemical_list: D006401:Hematologic Agents; C026912:cryoprecipitate coagulum; D005169:Factor VIII; D005340:Fibrinogen; D013917:Thrombin; D008751:Methylene Blue