Thrombosis and Haemostasis, Table of Contents Thromb Haemost 2004; 92(05): 980-985DOI: 10.1160/TH04-02-0119 Blood Coagulation, Fibrinolysis and Cellular Haemostasis Schattauer GmbH Ultrasound affects distribution of plasminogen and tissuetype plasminogen activator in whole blood clots in vitro Authors Author Affiliations Branka Devcic-Kuhar * 1 Institute of General Physics, Vienna University of Technology, Austria Stefan Pfaffenberger * 2 Department of Internal Medicine II, University of Vienna, Austria Lisa Gherardini 3 Department of Pharmacology, Conway Institute, University College Dublin, Ireland Christoph Mayer 4 Department of Dermatology, University of Vienna, Austria Martin Gröschl 1 Institute of General Physics, Vienna University of Technology, Austria Christoph Kaun 2 Department of Internal Medicine II, University of Vienna, Austria Ewald Benes 1 Institute of General Physics, Vienna University of Technology, Austria Erwin Tschachler 4 Department of Dermatology, University of Vienna, Austria Kurt Huber 2 Department of Internal Medicine II, University of Vienna, Austria Gerald Maurer 2 Department of Internal Medicine II, University of Vienna, Austria Johann Wojta ‡ 2 Department of Internal Medicine II, University of Vienna, Austria Michael Gottsauner-Wolf ‡ 2 Department of Internal Medicine II, University of Vienna, Austria Recommend Article Abstract Buy Article(opens in new window) Summary Ultrasound of 2 MHz frequency and 1.2 W/cm2 acoustic intensity was applied to examine the effect of sonication on recombinant tissue-type plasminogen activator (rt-PA)-induced thrombolysis as well as on the distribution of plasminogen and t-PA within whole blood clots in vitro. Thrombolysis was evaluated quantitatively by measuring clot weight reduction and the level of fibrin degradation product D-dimer (FDP-DD) in the supernatant. Weight reduction in the group of clots treated both with ultrasound and rt-PA was 35.2% ± 6.9% which is significantly higher (p<0.0001) than in the group of clots treated with rt-PA only (19.9% ± 4.3%). FDP-DD level in the supernatants of the group treated with ultrasound and rt-PA increased sevenfold compared to the group treated with rt-PA alone, (14895 ± 2513 ng/ml vs. 2364 ± 725 ng/ml). Localization of fibrinolytic components within the clots was accomplished by using gel-entrapping technique and immunohistochemistry. Spatial distributions of t-PA and plasminogen showed clearly that ultrasound promoted the penetration of rt-PA into thrombi significantly (p<0.0001), and broadened the zone of lysis from 8.9 ± 2.6 µm to 21.2 ± 7.2 µm. We speculate that ultrasound enhances thrombolysis by affecting the distribution of rt-PA within the clot. Keywords KeywordsThrombolysis - ultrasound - sonothrombolysis - immunohistochemistry - fibrinolytic enzymes Full Text References References 1 Collet J-P, Park D, Lesty C. et al. Influence of fibrin network conformation and fibrin fiber diameter on fibrinolysis speed. Arterioscler Thromb Vasc Biol 2000; 20: 1354-61. 2 Diamond SL, Anand S. Inner clot diffusion and permeation during fibrinolysis. Biophysical J 1993; 65: 2622-43. 3 Zidansek A, Blinc A. the influence of transport parameters and enzyme-kinetics of the fibrinolytic system on thrombolysis – mathematical-modeling of 2 idealized cases. Thromb Haemost 1991; 65: 553-9. 4 Blinc A, Keber D, Lahajnar G. et al. Lysing patterns of retracted blood-clots with diffusion or bulk flow transport of plasma with urokinase into clots – a magnetic-resonance-imaging study in vitro. Thromb Haemost 1992; 68: 667-671. 5 Sakharov DV, Nagelkerke JF, Rijken DC. Rearrangements of the fibrin network and spatial distribution of fibrinolytic components during plasma clot lysis. Study with confocal microscopy. J Biol Chem 1996; 271: 2133-8. 6 Devcic-Kuhar B, Pfaffenberger S, Gröschl M. et al. M. In vitro thrombolysis enhanced by standing and traveling ultrasound wave fields. Ultrasound Med Biol 2002; 28: 1181-7. 7 Pfaffenberger S, Devcic-Kuhar B, El-Rabadi K. et al. 2 MHz ultrasound enhances t-PAmediated thrombolysis: Comparison of continuous versus pulsed ultrasound and standing versus traveling acoustic waves. Thromb Haemost 2003; 89: 583-9. 8 Blinc A, Francis CW, Trudnowski JL. et al. Characterization of ultrasound-potentiated fibrinolysis in vitro. Blood 1993; 81: 2636-43. 9 Francis CW. Ultrasound-enhanced thrombolysis. Echocardiography 2001; 18: 239-46. 10 Harpaz D, Chen X, Francis CW. et al. Ultrasound accelerates urokinase-induced thrombolysis and reperfusion. Am Heart J 1994; 127: 1211-9. 11 Luo H, Nishioka T, Fishbein MC. et al. Transcutaneous ultrasound augments lysis of arterial thrombi in vivo. Circulation 1996; 94: 775-8. 12 Lauer CG, Burge R, Tang DB. et al. Effect of ultrasound on tissue-type plasminogen activator-induced thrombolysis. Circulation 1992; 86: 1257-64. 13 Nilsson A-M, Odselius R, Roijer A. et al. Proand antifibrinolytic effects of ultrasound on streptokinase-induced thrombolysis. Ultrasound Med Biol 1995; 21: 833-40. 14 Ollson SB, Johansson B, Nilsson A-M. et al. Enhancement of thrombolysis by ultrasound. Ultrasound Med Biol 1994; 20: 375-82. 15 Rosenschein U, Furman V, Kerner E. et al. Ultrasound imaging-guided noninvasive ultrasound thrombolysis. Circulation 2000; 102: 238-45. 16 Braaten J, Goss RA, Francis CW. Ultrasound reversibly dissagregates fibrin fibers. Thromb Haemost 1997; 78: 1063-8. 17 Francis CW, Blinc A, Lee S. et al. Ultrasound accelerates transport of recombinant tissue plasminogen activator into clots. Ultrasound Med Biol 1995; 21: 419-24. 18 Kollmann C, Bezemer RA, Fredfeldt KE. et al. Ein Testobjekt für die apparative Qualitätssicherung bei Ultraschall-Doppler (Duplex) Geräten, ausgehend vom Normenentwurf IEC 61685. Ultraschall Med 1999; 20: 248-57. 19 Kimura M, Iijima S, Kobayashi K. et al. Evaluation of the thrombolytic effect of tissue-type plasminogen activator with ultrasonic irradiation: in vitro experiment involving assay of the fibrin degradation products from the clot. Biol Pharm Bull 1994; 17: 126-30. 20 Gherardini L, Radel S, Sielemann S. et al. A study of the spatial organisation of microbial cells in a gel matrix subjected to treatment with ultrasound standing waves. Bioseparation 2002; 10: 153-62. 21 Cintas P, Le Traon AP, Larrue V. High rate of recanalization of middle cerebral artery occlusion during 2-MHz Transcranial Color-Coded Doppler continuous monitoring without thrombolytic drug. Stroke 2002; 33: 626-8. 22 Siddiqi F, Odrljin TM, Fay PJ. et al. Binding of tissue-plasminogen activator to fibrin: effect of ultrasound. Blood 1998; 91: 2019-25. 23 Sakharov DV, Rijken DC. Superficial accumulation of plasminogen during plasma clot lysis. Circulation 1995; 92: 1883-90.
