Topically Administered Macromolecular Heparin Proteoglycans Inhibit Thrombus Growth in Microvascular Anastomoses

Summary

Previously, during blood perfusion over collagen-coated surfaces; soluble or immobilized heparin proteoglycans (HEP-PG) have been shown to block thrombus growth. Our aim was to study the antithrombotic effect of locally applied unfractionated heparin (UFH, 1 mg/ml), or rat mast cell-derived HEP-PG (MW 750 kD, 10 µg/ml) compared with saline in early (10 min) and late (3 days) thrombus formation upon anastomosis of rat common femoral arteries. In both semiquantitative scanning electron microscopy (SEM) and quantitative platelet Indium 111 -labeling HEP-PG inhibited thrombus growth in comparison with saline. At 10 min, the extent of thrombosis (scale 1-4) in SEM followed the order: saline (3.2+/-0.8) > UFH (2.8+/-1.0) > HEP-PG (1.8+/-0.8), and also Indium111-positive platelets (106) accumulated on the anastomosed vessel in the same order 14.2 +/-7.2, 10.3 +/-5.0, and 7.7 +/-3.1 (saline vs. HEP-PG, p = 0.03 and 0.05, respectively). At 3 days all HEP-PGtreated vessels remained patent with only small mural thrombi, whereas 2/7 saline and 1/7 UFH-treated anastomoses occluded and showed more thrombosis overall. We conclude that locally administered HEPPG inhibit arterial thrombus growth in anastomosed small-sized arteries and could prevent thrombotic complications in (micro)vascular surgery and arteriovenous shunts.

Keywords

Thrombosis - platelets - heparin proteoglycans - microvascular surgery

• References

• 1 Percival NJ, Sykes PJ, Earley MJ. Free flap surgery: the Welsh regional unit experience. Br J Plast Surg 1989; 42: 435-40.
• 2 Khouri RK. Avoiding free flap failure. Clin Plast Surg 1992; 09: 773-81.
• 3 Davies MG, Hagen P-O. Pathophysiology of vein graft failure: a review. Eur J Vasc Endovasc Surg 1995; 09: 7-18.
• 4 Maraganore JM. Thrombin, thrombin inhibitors, and the arterial thrombotic process. Thromb Haemost 1993; 70: 208-11.
• 5 Khouri RK, Cooley BC, Kenna DM, Edstrom LE. Thrombosis of microvascular anastomoses in traumatized vessels: fibrin versus platelets. Plast Reconstr Surg 1990; 86: 110-7.
• 6 Ruggeri ZM. Mechanisms initiating platelet thrombus formation. Thromb Haemost 1997; 78: 611-6.
• 7 Bertina RM, van Tilburg NH, de Fouw NJ, Haverkate F. Thrombin, a link between coagulation activation and fibrinolysis. Ann NY Acad Sci 1992; 667: 239-48.
• 8 Acland R. Thrombus formation in microvascular surgery: an experimental study of the effects of surgical trauma. Surgery 1973; 73: 66-771.
• 9 Davies MJ, Thomas AC. Plaque fissuring: the cause of acute myocardial infarction, sudden ischemic death and crescendo angina. Br Heart J 1985; 53: 363-70.
• 10 Johnson PC, Dickson CS, Garrett KO, Sheppeck RA, Bentz ML. The effect of microvascular anastomosis configuration on initial platelet deposition. Plast Reconstr Surg 1993; 91: 522-7.
• 11 Wieslander JB, Salemark L, Dougan P. Hydroxyethyl starch increases patency and reduces thrombus formation following arteriotomy/intimectomy in small arteries: an experimental study in the rabbit. J Reconstr Microsurg 1990; 06: 357-61.
• 12 Rooks MD, Rodriquez J, McNaughton M, Turnidge K, Zusmanis K, Hutton W. Refinement of the rat crush-avulsion femoral artery injury model. Microsurgery 1993; 14: 130-4.
• 13 Chen L-E, Seaber AV, Urbaniak JR. Thrombosis and thrombolysis in crushed arteries with or without anastomosis: a new microvascular thrombosis model. J Reconstr Microsurg 1996; 12: 31-8.
• 14 Sigurbjörnsson B, Hallberg E, Andrén-Sandberg Å, Ribbe E. Endothelialization of anastomoses in the aorta and inferior vena cava of the rat. Microsurgery 1994; 15: 787-94.
• 15 Johnson PC, Barker JH. Thrombosis and antithrombotic therapy in microvascular surgery. Clin Plast Surg 1992; 19: 799-807.
• 16 Cooley BC, Gould JS. Topically applied antithrombotic agents offer a new therapeutic approach to the prevention of microvascular thrombosis. Microsurgery 1991; 12: 281-7.
• 17 Salemark L. International survey of current microvascular practices in free tissue transfer and replantation surgery. Microsurgery 1991; 12: 308-11.
• 18 Kroll SS, Miller MJ, Reece GP, Baldwin BJ, Robb GL, Bengtson BP, Phillips MD, Kim D, Schusterman MA. Anticoagulants and hematomas in free flap surgery. Plast Reconstr Surg 1995; 96: 643-7.
• 19 Arnljots B, Bergqvist D. Inhibition of heparin-resistant microarterial thrombosis by recombinant hirudin: a specific thrombin inhibitor. Plast Reconstr Surg 1995; 95: 894-900.
• 20 Ozbek MR, Brown DM, Deune EG, Lantieri LA, Kania NM, Pasia EN, Cooley BC, Wun T-C, Khouri RK. Topical tissue factor pathway inhibitor improves free-flap survival in a model simulating free-flap errors. J Reconstr Microsurg 1995; 11: 185-8.
• 21 Arnljots B, Ezban M, Hedner U. Prevention of experimental arterial thrombosis by topical administration of active site-inactivated factor VIIa. J Vasc Surg 1997; 25: 341-6.
• 22 Sørensen HB, Kristensen AT, Ravn HB, Fuglsang J, Hjortdal VE. Local application of FFR-rFVIIa reduces thrombus formation at arterial anastomosis in rats. Microsurgery 1999; 19: 369-73.
• 23 Lassila R, Lindstedt K, Kovanen PT. Native macromolecular heparin proteoglycans exocytosed from simulated rat serosal mast cells strongly inhibit platelet-collagen interactions. Arterioscler Thromb Vasc Biol 1997; 17: 3578-87.
• 24 Wang Y, Kovanen PT. Heparin proteoglycans released from rat serosal mast cells inhibit proliferation of rat aortic smooth muscle cells in culture. Circ Res 1999; 84: 74-83.
• 25 Cox GW, Runnels S, Hsu HS-H, Das SK. A comparison of heparinised saline irrigation solutions in a model of microvascular thrombosis. Br J Plast Surg 1992; 45: 345-8.
• 26 Catalfamo JL, Dodds WJ. Isolation of platelets from laboratory animals. In: Hawiger J. ed. Methods in Enzymology. San Diego, CA: Academic Press, Inc; 1989: 27-34.
• 27 Arnljots B, Dougan P, Bergqvist D. Antithrombotic and platelet activating effects of heparin in prevention of microarterial thrombosis. Plast Reconstr Surg 1997; 99: 1122-8.
• 28 Kauhanen P, Kovanen PT, Lassila R. Co-immobilized native macromolecular heparin proteoglycans strongly inhibit platelet-collagen interactions in flowing blood. Arterioscler Thromb Vasc Biol 2000; 20: e113-e119.
• 29 Kauhanen P, Kovanen PT, Reunala T, Lassila R. Effects of skin mast cells on bleeding time and coagulation activation at the site of platelet plug formation. Thromb Haemost 1998; 79: 843-7.
• 30 Stockmans F, Deckmyn H, Gruwez J, Vermylen J, Ackland R. Continuous quantitative monitoring of mural, platelet-dependent, thrombus kinetics in the crushed rat femoral vein. Thromb Haemost 1991; 65: 425-31.
• 31 Nightingale G, Fogdestam I, O´Brien BMC. Scanning electron microscope study of experimental microvascular anastomoses in the rabbit. Br J Plast Surg 1980; 33: 283-98.
• 32 Yu-dong G, Ji-feng L, Ji-fu J, Ci-sheng Z, Pei-zhu T. Scanning electron microscopic observations on the endothelial healing mechanism of vessels. J Reconstr Microsurg 1989; 05: 327-30.
• 33 Lassila R, Badimon JJ, Vallabhajosula S, Badimon L. Dynamic monitoring of platelet deposition on severely damaged vessel wall in flowing blood. Arteriosclerosis 1990; 10: 306-15.
• 34 Johnson PC. Platelet-mediated thrombosis in microvascular surgery: New knowledge and strategies. Plast Reconstr Surg 1990; 86: 359-67.