Platelet Compatibility of an Artificial Surface Modified with Functionally Active Heparin

Summary

Platelet compatibility after coating an artificial material with functionally active heparin was investigated. Blood was circulated in uncoated or heparin coated PVC tubing. In one hour platelet counts decreased from 155 (113-184) ×10⁹/l to 124 (100-148) ×10⁹/l with uncoated compared to 164 (132-192) ×10⁹/l with heparin coated tubing (intergroup p = 0.02). β-thromboglobulin increased from 116 (80-148) μg/l to 1039 (757-1298) μg/l with uncoated and to 352 (229-638) μg/l with heparin coated tubing (intergroup p = 0.005). Platelet counts and β-thromboglobulin correlated closely with complement activation. Solid-phase enzyme immunoassay demonstrated substantial deposition of CD42a/GPIbIX and CD61/GPIIIa on uncoated, but not on heparin coated tubing (intergroup p <0.0005). Scanning electron microscopy demonstrated activated platelets and aggregates on uncoated in contrast to heparin coated tubing, where scattered, unactivated platelets were found. Changes in P-selectin and microparticles were minor. In conclusion, this heparin surface substantially improved platelet compatibility. Markers of choice for in vitro evaluation were platelet counts, β-thromboglobulin and platelet deposition.

Key words

Platelets - heparin - complement - artificial surface - β-thromboglobulin - P-selectin - microparticles - flow cytometry - biocompatibility

• References

• 1 Williams DF. (ed). Definitions in biomaterials. Amsterdam – Oxford: Elsevier; 1987
• 2 Wan S, Leclerc JL, Vincent JL. Inflammatory response to cardiopulmonary bypass: Mechanisms involved and possible therapeutic strategies. Chest 1997; 112: 676-92.
• 3 Hall RI, Smith MS, Rocker G. The systemic inflammatory response to cardiopulmonary bypass: Pathophysiological, therapeutic, and pharmacological considerations. Anesth Analg 1997; 85: 766-82.
• 4 Larm O, Larsson R, Olsson P. A new non-thrombogenic surface prepared by selective covalent binding of heparin via a modified reducing terminal residue. Biomat Med Dev Art Org 1983; 11: 161-73.
• 5 Tong SD, Rolfs MR, Hsu LC. Evaluation of Duraflo II heparin immobilized cardiopulmonary bypass circuits. Asaio Trans 1990; 36: M654-M656.
• 6 Cheung AK, Parker CJ, Janatova J, Brynda E. Modulation of complement activation on hemodialysis membranes by immobilized heparin. J Am Soc Nephrol 1992; 2: 1328-37.
• 7 Takano H, Nakano S, Kadoba K, Kaneko M, Miyamoto Y, Ohtake S, Matsuwaka R, Chang JC, Amemiya A, Matsuda H. Evaluation of the bio-compatibility of a new method for heparin coating of a cardiopulmonary bypass circuit. ASAIO J 1992; 38: M390-M394.
• 8 Mollnes TE, Riesenfeld J, Garred P, Nordström E, Høgåsen K, Fosse E, Götze O, Harboe M. A new model for evaluation of biocompatibility: combined determination of neoepitopes in blood and on artificial surfaces demonstrates reduced complement activation by immobilization of heparin. Artif Organs 1995; 19: 909-17.
• 9 Garred P, Mollnes TE. Immobilized heparin inhibits the increase in leukocyte surface expression of adhesion molecules. Artif Organs 1997; 21: 293-9.
• 10 Videm V, Mollnes TE, Fosse E, Mohr B, Bergh K, Hagve TA, Aasen AO, Svennevig JL. Heparin-coated cardiopulmonary bypass equipment. I. Bio-compatibility markers and development of complications in a high-risk population. J Thorac Cardiovasc Surg 1999; 117: 794-802.
• 11 Garred P, Mollnes TE, Lea T. Quantification in enzyme-linked immunosorbent assay of a C3 neoepitope expressed on activated human complement factor C3. Scand J Immunol 1988; 27: 329-35.
• 12 Braet F, de Zanger R, Wisse E. Drying cells for SEM, AFM and TEM by hexamethyldisilazane: a study on hepatic endothelial cells. J Microscopy 1997; 186: 84-7.
• 13 Sanchez J, Elgue G, Riesenfeld J, Olsson P. Studies of adsorption, activation, and inhibition of factor XII on immobilized heparin. Thromb Res 1998; 89: 41-50.
• 14 Elgue G, Blombäck M, Olsson P, Riesenfeld J. On the mechanism of coagulation inhibition on surfaces with end point immobilized heparin. Thromb Haemost 1993; 70: 289-93.
• 15 Maillet F, Petitou M, Choay J, Kazatchkine MD. Structure-function relationships in the inhibitory effect of heparin on the complement activation: Independency of the anti-coagulant and anti-complementary sites on the heparin molecule. Mol Immunol 1988; 25: 917-23.
• 16 Rinder CS, Rinder HM, Smith BR, Fitch JCK, Smith MJ, Tracey JB, Matis LA, Squinto SP, Rollins SA. Blockade of C5a and C5b-9 generation inhibits leukocyte and platelet activation during extracorporeal circulation. J Clin Invest 1995; 96: 1564-72.
• 17 Moen O, Fosse E, Brockmeier V, Andersson C, Mollnes TE, Høgåsen K, Venge P. Disparity in blood activation by two different heparin-coated cardiopulmonary bypass systems. Ann Thorac Surg 1995; 60: 1317-23.
• 18 Øvrum E, Mollnes TE, Fosse E, Holen EA, Tangen G, Abdelnoor M, Ringdal ML, Øystese R, Venge P. Complement and granulocyte activation in two different types of heparinized extracorporeal circuits. J Thorac Cardiovasc Surg 1995; 110: 1623-32.
• 19 Moen O, Fosse E, Bråten J, Andersson C, Høgåsen K, Mollnes TE, Venge P, Kierulf P. Differences in blood activation related to roller/centrifugal pumps and heparin-coated/uncoated surfaces in a cardiopulmonary bypass model circuit. Perfusion 1996; 11: 113-23.
• 20 Wendel HP, Heller W, Gallimore MJ, Hoffmeister HE. Heparin-coated oxygenators significantly reduce contact system activation in an in vitro cardiopulmonary bypass model. Blood Coagulat Fibrinol 1994; 5: 673-8.
• 21 Videm V, Mollnes TE, Garred P, Svennevig JL. Biocompatibility of extracorporeal circulation. In vitro comparison of heparin-coated and uncoated oxygenator circuits. J Thorac Cardiovasc Surg 1991; 101: 654-60.
• 22 Fukutomi M, Kobayashi S, Niwaya K, Hamada Y, Kitamura S. Changes in platelet, granulocyte, and complement activation during cardiopulmonary bypass using heparin-coated equipment. Artif Organs 1996; 20: 767-76.
• 23 Wagner WR, Johnson PC, Thompson KA, Marrone GC. Heparin-coated cardiopulmonary bypass circuits: Hemostatic alterations and postoperative blood loss. Ann Thorac Surg 1994; 58: 734-40.
• 24 Videm V, Svennevig JL, Fosse E, Semb G, Østerud B, Mollnes TE, Nielsen H. Reduced complement activation with heparin-coated oxygenator and tubings in coronary bypass operations. J Thorac Cardiovasc Surg 1992; 103: 806-13.
• 25 Svennevig JL, Geiran OR, Karlsen H, Pedersen T, Mollnes TE, Kongsgård U, Frøysaker T. Complement activation during extracorporeal circulation – In vitro comparison of Duraflo-II heparin-coated and uncoated oxygenator circuits. J Thorac Cardiovasc Surg 1993; 106: 466-72.
• 26 Boonstra PW, Gu YJ, Akkerman C, Haan J, Huyzen R, van Oeveren W. Heparin coating of an extracorporeal circuit partly improves hemostasis after cardiopulmonary bypass. J Thorac Cardiovasc Surg 1994; 107: 289-92.
• 27 Palatianos GM, Dewanjee MK, Smith W, Novak S, Hsu LC, Kapadvanjwala M, Sfakianakis GN, Kaiser GA. Platelet preservation during cardio-pulmonary bypass with iloprost and Duraflo-II heparin-coated surfaces. Asaio Trans 1991; 37: 620-2.
• 28 Plotz FB, van Oeveren W, Aloe LS, Riley MK, Hultquist KA, Bartlett RH, Wildevuur CR. Prophylactic administration of tranexamic acid preserves platelet numbers during extracorporeal circulation in rabbits. Asaio Trans 1991; 37: M416-7.
• 29 Chronos N, Robinson K, King S, Lunn A, White D, Kelly A, Harker L. Heparin-coated Palmaz-Schatz stents are highly thromboresistant: A baboon A-V shunt study. J Am Coll Cardiol 1996; 27 Suppl A 84A-5A.
• 30 Sims PJ, Faioni EM, Wiedmer T, Shattil SJ. Complement proteins C5b-9 cause release of membrane vesicles from the platelet surface that are enriched in the membrane receptor for coagulation factor Va and express prothrombinase activity. J Biol Chem 1988; 263: 18205-12.