title: Covalently immobilized thrombomodulin inhibits coagulation and complement activation of artificial surfaces in vitro.
abstract: Thrombomodulin (TM) serves as the endothelial cell receptor for thrombin and alters its characteristics from pro- to anticoagulant. Additionally, it promotes the formation of activated protein C. We evaluated the conservation of the overall outcome of these functions in recombinant TM linked to artificial surfaces by incubation with human whole blood in vitro. TM was covalently immobilized through poly(ethylene glycol) (PEG) spacers onto thin films of poly(octadecene alt maleic anhydride) covering planar glass substrates. TM binding to the polymer films was achieved after active ester formation at the carboxylic acid terminus of the PEG spacers and thoroughly characterized by HPLC-based amino acid analysis, immunofluorescence and ellipsometry. TM-coated samples were incubated for 3h with freshly drawn whole human blood anticoagulated with heparin (5IU/ml) using in-house developed incubation systems. The substantially reduced activation of blood coagulation (TAT) for TM-coated samples correlates well with the degree of contact activation (bradykinin and FXIIa formation) while no significant effects were observed for the platelet activation (PF4). Further, complement activation (C5a levels), was strongly diminished at the TM-containing surfaces. We conclude that the suggested method for preparation of TM immobilization may serve to prepare model substrates for studies on TM interactions but similarly provides a promising coating strategy for blood contacting medical devices.
authors.affiliation: Department of Biocompatible Materials, Institute of Polymer Research Dresden and The Max Bergmann Center of Biomaterials Dresden, Biocompatible Materials, Hohe Str. 6, 01069 Dresden, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Sperling
authors.forename: K
authors.initials: K
authors.lastname: Salchert
authors.forename: U
authors.initials: U
authors.lastname: Streller
authors.forename: C
authors.initials: C
authors.lastname: Werner
mesh_terms: D000327:Adsorption; D000328:Adult; D000925:Anticoagulants / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects / Q000502:physiology*; D020099:Coated Materials, Biocompatible / Q000008:administration & dosage* / Q000737:chemistry*; D003167:Complement Activation / Q000187:drug effects*; D006801:Humans; D008422:Materials Testing; D011108:Polymers / Q000737:chemistry; D013499:Surface Properties; D018180:Thrombomodulin / Q000008:administration & dosage* / Q000737:chemistry*
chemical_list: D000925:Anticoagulants; D020099:Coated Materials, Biocompatible; D011108:Polymers; D018180:Thrombomodulin