Thrombin inactivation on surfaces with covalently bonded heparin.
About 8000 Daltons porcine mucosa heparin fragments were covalently bonded by end-point attachment to polyethylene. The interaction between the immobilized heparin, added thrombin, and antithrombin III [AT] was investigated. The heparin surface was adsorbed with either albumin, AT dissolved in albumin or Tyrode, or platelet free plasma. Irrespective of the pre-treatment procedure, exposure of the surface to thrombin resulted in the same substantial decrease of thrombin in solution and the same degree of surface-confined thrombin activity. It was concluded that the heparin surface has a large capacity to bind thrombin and that the thrombin inhibitory capacity of high affinity heparin fragments is limited. On exposure of the thrombin-loaded surfaces to defibrinogenated plasma or AT, the surface-confined thrombin was inhibited within 30 seconds. Successive dilutions of plasma or AT decreased the inhibition rate but not the inhibition capacity. It is concluded that inhibition of thrombin adsorbed on the heparin surface occurs as follows: Added AT adheres to high affinity heparin fragments on the surface whereupon adsorbed thrombin migrates in the hydrophilic heparin coating towards the reaction site of AT and becomes inhibited. The inactivated thrombin-AT complex leaves then the surface, thus enabling the process to be repeated.
D000990:Antithrombin III / Q000378:metabolism; D001672:Biocompatible Materials / Q000378:metabolism; D004357:Drug Synergism; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D011095:Polyethylenes / Q000494:pharmacology*; D011485:Protein Binding; D013329:Structure-Activity Relationship; D013499:Surface Properties; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013997:Time Factors
D001672:Biocompatible Materials; D011095:Polyethylenes; D000990:Antithrombin III; D006493:Heparin; D013917:Thrombin