In vivo dynamic real-time monitoring and quantification of platelet-thrombus formation: use of a local isotope detector

Arterioscler Thromb Vasc Biol. 2000 Mar;20(3):860-5. doi: 10.1161/01.atv.20.3.860.

Abstract

Current methods for monitoring thrombosis and thrombus growth are invasive and provide only single-time-point data. Animal models rely mainly on flow changes as a surrogate of thrombus formation. Our aim was to validate a unique potentially noninvasive system to detect and quantify dynamic thrombus formation in vivo by using a porcine model of carotid artery injury. Thrombus growth was monitored by deposition of autologous (111)In-labeled platelet activity over the injured artery by use of miniaturized gamma detectors and Doppler blood flow. Counts were recorded at 2-minute intervals for 2 hours. The technique was validated by comparing standard antithrombotic agents against controls. Platelet recruitment was detected before significant change in flow. Thrombus formation, calculated as the area under the curve (platelets x minutes x 10(6)), was greatest for control animals (11.7+/-1.28), followed by animals treated with aspirin (6.13+/-0.91, P<0.05), heparin (2.45+/-0.34, P<0.05), and hirudin (0.2+/-0.01, P<0.01 compared with heparin). The rate of platelet deposition was assessed as the slope of the curve in the first 30 minutes (platelets x 10(6) per minute) for the following treatment groups of animals: control, 3.53+/-0.34; aspirin, 1.67+/-0. 34 (P<0.01); heparin, 1.55+/-0.3 (P<0.01); and hirudin, 0.25+/-0.03 (P<0.001). There was no statistical difference between heparin and aspirin treatments. Change in flow was assessed as reduction from baseline: control, >99+/-0.34%; aspirin, 39+/-9.1%; heparin, 36+/-12. 5%; and hirudin, 17+/-5.4%. There was no statistical difference between the aspirin- and heparin-treated groups. Morphometric analysis revealed >99+/-0.63% occlusion of the luminal area with thrombus for the control group, 43+/-14.3% for the aspirin-treated group, 30+/-5.6% for the heparin-treated group, and <10+/-1.8% for the hirudin-treated group. Assessment of platelet-thrombus formation with this technique was more sensitive than change in flow in determining antithrombotic efficacy, and thrombus formation was detected earlier. This study validates a new quantitative, sensitive, potentially noninvasive, portable, in vivo monitoring of dynamic thrombus growth, which appears applicable to phase II studies in humans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / pharmacology
  • Animals
  • Anticoagulants / pharmacology
  • Aspirin / pharmacology
  • Blood Platelets / drug effects
  • Blood Platelets / physiology*
  • Carotid Arteries / diagnostic imaging
  • Carotid Arteries / physiology*
  • Carotid Artery Injuries / diagnostic imaging
  • Carotid Artery Injuries / physiopathology
  • Computer Systems*
  • Fibrinolytic Agents / pharmacology
  • Gene Expression / drug effects
  • Heparin / pharmacology
  • Indium Radioisotopes
  • Partial Thromboplastin Time
  • Plasminogen Activator Inhibitor 1 / genetics
  • RNA, Messenger / analysis
  • Radionuclide Imaging / instrumentation
  • Radionuclide Imaging / methods
  • Swine
  • Thrombosis / diagnostic imaging*
  • Thrombosis / physiopathology*
  • Ultrasonography, Doppler
  • Uridine Diphosphate / pharmacology
  • Uridine Triphosphate / pharmacology

Substances

  • Anticoagulants
  • Fibrinolytic Agents
  • Indium Radioisotopes
  • Plasminogen Activator Inhibitor 1
  • RNA, Messenger
  • Uridine Diphosphate
  • Adenosine Diphosphate
  • Adenosine Triphosphate
  • Heparin
  • Aspirin
  • Uridine Triphosphate