Summary
The effect of the heparinoid, pentosan polysulphate (PP) on the proliferative behaviour of cultured bovine endothelial cells (EC) was examined. In addition, the toxicity of the drug towards EC, its influence on prostacyclin production and release, and on cell-associated plasminogen activator activity was determined. At a concentration of 10 μg/ml in the culture medium, PP exerted a growth promoting effect on EC. Increased cell numbers were accompanied by increased 3H-thymidine incorporation into cellular DNA compared with controls, however, final density of the cells was not affected. In contrast, at doses of 1 mg/ml the growth of EC was substantially slowed down. This finding did not reflect cell injury as shown by an unaltered release of 51Cr from the cells. Incubation of PP with EC had no influence on the prostacyclin release from the cells neither on the accumulation of the metabolite in the culture fluid over 24 hours nor on the releasing capacity upon stimulation with arachidonic acid. PP increased the cell-associated plasminogen activator activity in growing cells and counteracted in cultures at final density the inhibitory effect of serum on the intracellular plasminogenactivator activity. Our results suggest that stimulation of the fibrinolytic activity of the endothelium and a growth promoting effect for endothelial cells that may lead to faster coverage of small lesions could contribute to the antithrombotic potency of pentosan polysulphate in vivo.
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Bergqvist D, Efsing HO, Hallböök T, Lindblad B (1980) Prevention of postoperative thromboembolic complications: a prospective comparison between dextran 70, dihydroergotamin heparin and a sulphated polysaccharide. Acta Chir Scand 146: 559–568
Bergqvist D, Ljungnér H (1981) A comparative study of dextran 70 and a sulphated polysaccharide in the prevention of postoperative thromboembolic complications. Br J Surg 68: 449–451
Bergqvist D, Nilsson IM (1981) A sulphated polysaccharide: the effects in vivo and in vitro on the haemostatic system in healthy volunteers. Thromb Res 23: 309–315
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Buonassisi V (1973) Sulfated mucopolysaccharide synthesis and secretion in endothelial cell cultures. Exp Cell Res 76: 363–368
Busch C, Dawes J, Pepper DS, Wasteson A (1980) Binding of platelet factor 4 to cultured human umbilical vein endothelial cells. Thromb Res 19: 129–137
Dawes J, Pumphrey CW, McLaren KM, Prowse CV, Pepper DS (1982) The in vivo release of human platelet factor 4 by heparin. Thromb Res 27: 65–76
Delvos U, Gajdusek C, Sage H, Harker LA, Schwartz SM (1982) Interactions of vascular wall cells with collagen gels. Lab Invest 46: 61–72
Delvos U, Preissner KT, Müller-Berghaus G (1985) Binding of fibrinogen to cultured bovine endothelial cells. Thromb Haemost 53: 26–31
Deutsch DG, Mertz ET (1970) Plasminogen: purification from human plasma by affinity chromatography. Science 170: 1095–1096
Fischer AM, Barrowcliffe TW, Thomas DP (1982) A comparison of pentosan polysulphate (SP 54) and heparin. I: Mechanism of action on blood coagulation. Thromb Haemost 47: 104–108
Fischer AM, Merton RE, Marsh NA, Williams S, Gaffney PJ, Barrowcliffe TW, Thomas DP (1982) A comparison of pentosan polysulphate and heparin. II: Effects on subcutaneous injection. Thromb Haemost 47: 109–113
Fischer AM, Dautzenberg MD, Aurousseau MH, Beguin S, Goudemand J, Hemker HC (1985) Comparison between the effect of pentosan polysulphate, heparin and antithrombin III injections in antithrombin III deficient patients. Thromb Res 37: 295–307
Gajdusek CM (1984) Release of endothelial cell-derived growth factor (ECDGF) by heparin. J Cell Phys 121: 13–21
Gross JL, Moscatelli D, Jaffe EA, Rifkin DB (1982) Plasminogen activator and collagenase production by cultured capillary endothelial cells. J Cell Biol 95: 974–981
Glimelius B, Busch C, Höök M (1978) Binding of heparin on the surface of cultured human endothelial cells. Thromb Res 12: 773–782
Hiebert LM, Jaques LB (1976) Heparin uptake on endothelium. Artery 2: 26–37
Jaffe EA, Hoyer LW, Nachman RL (1973) Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest 52: 2757–2764
Jaffe EA, Weksler BB (1979) Recovery of endothelial cell prostacyclin production after inhibition by low doses of aspirin. J Clin Invest 63: 532–535
Joffe S (1976) Drug prevention of postoperative deep vein thrombosis. Arch Surg 111: 37–40
Kjellen L, Pettersson I, Höök M (1981) Cell-surface heparan sulfate: an intercalated membrane proteoglycan. Proc Natl Acad Sci USA 78: 5371–5375
Kraemer PM (1977) Heparin releases heparan sulfate from the cell surface. Biochem Biophys Res Commun 78: 1334–1340
Levin EG (1983) Latent tissue plasminogen activator produced by human endothelial cells in culture: evidence for an enzyme-inhibitor complex. Proc Natl Acad Sci USA 80: 6804–6808
Levin EG, Loskutoff DJ (1979) Comparative studies on the fibrinolytic activity of cultured vascular cells. Thromb Res 15: 869–878
Levin EG, Loskutoff DJ (1982) Cultured bovine endothelial cells produce both urokinase and tissue-type plasminogen activators. J Cell Biol 94: 631–636
Loskutoff DJ, van Mourik JA, Erickson LA, Lawrence D (1983) Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells. Proc Natl Acad Sci USA 80: 2956–2060
Marsh NA, Gaffney PJ (1983) The effect of pentosan polysulphate (SP 54) on the fibrinolytic system of man. Thromb Haemost 50: 83 (Abstr)
Roth GJ, Stanford N, Majerus PW (1975) Acetylation of prostaglandin synthase by aspirin. Proc Natl Acad Sci USA 72: 3073–3076
Schwartz SM (1978) Selection and characterization of bovine aortic endothelial cells. In Vitro 14: 966–980
Scully MF, Weerasinghe KM, Ellis V, Djazaeri B, Kakkar VV (1983) Anticoagulant and antiheparin activities of a pentosan polysulphate. Thromb Res 31: 87–97
Scully MF, Kakkar VV (1984) The antiheparin effect of a heparinoid, pentosan polysulfate: investigation of a mechanism. Biochem J 218: 657–665
Scully MF, Kakkar VV (1984) Identification of heparin cofactor II as the principle plasma cofactor for the antithrombin activity of pentosan polysulphate (SP 54). Thromb Res 36: 187–194
Thomas DP, Barrowcliffe TW, Merton RE, Stocks J, Dawes J, Pepper DS (1980) In vivo release of anti-Xa clotting activity by a heparin analogue. Thromb Res 17: 831–840
Thornton SC, Mueller SN, Levine EM (1983) Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science 222: 623–625
Vinazzer H, Haas S, Stemberger A (1980) Influence on the clotting mechanism of sodium pentosan polysulphate (SP 54) in comparison to commercial beef lung sodium heparin. Thromb Res 20: 57–68
Weber H, Müller-Berghaus G (1984) Effect of pentosan polysulphate on experimental disseminated intravascular coagulation. Blut 49: 134–135 (Abstr)
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Delvos, U., Paul, J.E. & Müller-Berghaus, G. Effect of the heparinoid pentosan polysulphate (SP 54) on the functional properties of cultured bovine aortic endothelial cells. Blut 51, 127–136 (1985). https://doi.org/10.1007/BF00320026
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DOI: https://doi.org/10.1007/BF00320026