Document Detail


Localization of topically applied TFPI binding sites in an intimectomized microvessel.
MedLine Citation:
PMID:  7740496     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Tissue factor pathway inhibitor, TFPI, has been shown to be highly effective as a topically applied antithrombotic in an arterial model of vascular thrombosis. To elucidate the mechanism and site of TFPI action, recombinant TFPI was conjugated to 30 nm diameter gold particles and used to localize the sites of TFPI binding in a traumatized microvessel by transmission electron microscopy. The model, the central artery of the rabbit ear, was transected, denuded of endothelial lining (intimectomized), and re-anastomosed. Prior to the restoration of blood flow, TFPI-gold or unconjugated gold particles in solution were applied by irrigation to the intimectomized vessel lumen. After 10 min of blood flow, the artery was harvested for electron microscopy. TFPI-gold binding was localized to the fine strands of fibrin that lined the lumen of the intimectomized section of the artery. Little or no binding was found on platelets, exposed smooth muscle, cell membrane fragments, or uninjured vessel segments. The TFPI-gold binding could be competed with native TFPI. TFPI-gold was inhibitory, although less potent than native TFPI, in a prothrombin time assay. Unconjugated gold exhibited very little binding in the vascular model. Hence, the TFPI-gold conjugate behaved like native TFPI. Our observations have identified the fibrin complex as an in vivo binding site for TFPI and suggest that this is an in vivo site of action for TFPI as a topical antithrombotic agent.
Authors:
R L Ornberg; E G Deune; M R Ozbek; T C Wun; R K Khouri
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Thrombosis and haemostasis     Volume:  73     ISSN:  0340-6245     ISO Abbreviation:  Thromb. Haemost.     Publication Date:  1995 Jan 
Date Detail:
Created Date:  1995-06-05     Completed Date:  1995-06-05     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  7608063     Medline TA:  Thromb Haemost     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  55-8     Citation Subset:  IM    
Affiliation:
Monsanto Company, St. Louis, MO 63167, USA.
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MeSH Terms
Descriptor/Qualifier:
Administration, Topical
Anastomosis, Surgical
Animals
Arteries / injuries,  metabolism*,  surgery,  ultrastructure
Binding Sites
Binding, Competitive
Ear, External / blood supply
Endothelium, Vascular / injuries
Fibrin / metabolism*
Gold
Humans
Lipoproteins / administration & dosage,  metabolism*
Microscopy, Electron
Microsurgery
Muscle, Smooth, Vascular / metabolism,  ultrastructure
Rabbits
Recombinant Fusion Proteins / administration & dosage,  metabolism
Thrombosis / etiology
Vascular Surgical Procedures / adverse effects
Chemical
Reg. No./Substance:
0/Lipoproteins; 0/Recombinant Fusion Proteins; 0/lipoprotein-associated coagulation inhibitor; 7440-57-5/Gold; 9001-31-4/Fibrin

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