Thrombogenesis with continuous blood flow in the inferior vena cava. A novel mouse model.
Several rodent models have been used to study deep venous thrombosis (DVT). However, a model that generates consistent venous thrombi in the presence of continuous blood flow, to evaluate therapeutic agents for DVT, is not available. Mice used in the present study were wild-type C57BL/6 (WT), plasminogen activator inhibitor-1 (PAI-1) knock out (KO) and Delta Cytoplasmic Tail (DCT). An electrolytic inferior vena cava (IVC) model (EIM) was used. A 25G stainless-steel needle, attached to a silver coated copper wire electrode (anode), was inserted into the exposed caudal IVC. Another electrode (cathode) was placed subcutaneously. A current of 250 muAmps over 15 minutes was applied. Ultrasound imaging was used to demonstrate the presence of IVC blood flow. Analyses included measurement of plasma soluble P-selectin (sP-Sel), thrombus weight (TW), vein wall morphometrics, P-selectin and Von Willebrand factor (vWF) staining, transmission electron microscopy (TEM), scanning electron microscopy (SEM); and the effect of enoxaparin on TW was evaluated. A current of 250 muAmps over 15 minutes consistently promoted thrombus formation in the IVC. Plasma sP-Sel was decreased in PAI-1 KO and increased in DCT vs. WT (WT/PAI-1: p=0.003, WT/DCT: p=0.0002). Endothelial activation was demonstrated by SEM, TEM, P-selectin and vWF immunohistochemistry and confirmed by inflammatory cell counts. Ultrasound imaging demonstrated thrombus formation in the presence of blood flow. Enoxaparin significantly reduced the thrombus size by 61% in this model. This EIM closely mimics clinical venous disease and can be used to study endothelial cell activation, leukocyte migration, thrombogenesis and therapeutic applications in the presence of blood flow.
Diaz
José A
JA
Department of Surgery, Section of Vascular Surgery, University of Michigan, Ann Arbor, Michigan 48109-0654, USA. josediaz@med.umich.edu
Hawley
Angela E
AE
Alvarado
Christine M
CM
Berguer
Alexandra M
AM
Baker
Nichole K
NK
Wrobleski
Shirley K
SK
Wakefield
Thomas W
TW
Lucchesi
Benedict R
BR
Myers
Daniel D
DD
D000818:Animals; D001777:Blood Coagulation / Q000235:genetics; D004195:Disease Models, Animal; D004572:Electrolysis; D042783:Endothelial Cells / Q000378:metabolism; D017984:Enoxaparin / Q000494:pharmacology; D005343:Fibrinolytic Agents / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D020556:Neutrophil Infiltration; D019007:P-Selectin / Q000097:blood / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000172:deficiency / Q000235:genetics; D012039:Regional Blood Flow; D019851:Thrombophilia / Q000097:blood / Q000235:genetics / Q000503:physiopathology; D013997:Time Factors; D018615:Ultrasonography, Doppler, Color; D014682:Vena Cava, Inferior / Q000000981:diagnostic imaging / Q000293:injuries / Q000378:metabolism / Q000503:physiopathology* / Q000648:ultrastructure; D020246:Venous Thrombosis / Q000097:blood / Q000175:diagnosis / Q000235:genetics / Q000503:physiopathology* / Q000517:prevention & control; D014841:von Willebrand Factor / Q000378:metabolism
D017984:Enoxaparin; D005343:Fibrinolytic Agents; D019007:P-Selectin; D017395:Plasminogen Activator Inhibitor 1; D014841:von Willebrand Factor