Facile conjugation of heparin onto titanium surfaces via dopamine inspired coatings for improving blood compatibility

Abstract

Immobilization of heparin on biomaterials surface has been proven to be an effective strategy for preventing thrombus formation. However, the procedures of most immobilization methods (physical adsorption, covalent linkage, electrostatic interaction) are complicated and time-consuming. In the present study, heparin with various concentrations immobilized on a titanium (Ti) substrate via polydopamine layer for improving blood compatibility was investigated. Water contact angle measurement showed that the immobilization of heparin resulted in an increase of the hydrophilicity. X-ray photoelectron spectroscopy (XPS) and Toluidine Blue O (TBO) tests displayed that the heparin molecules were successfully immobilized on Ti surface. The evaluations of blood compatibility (hemolysis rate, APTT, platelet adhesion and activation, fibrinogen conformational change) showed that the immobilization of heparin decreased hemolysis rate, prolonged blood coagulation time, reduced platelets adhesion and activation, and induced less fibrinogen conformational change. Moreover, a significant inhibition of blood coagulation and platelet adhesion was obtained when the heparin concentration was higher than 5 mg/mL, indicating that only with a certain surface densities could heparin perform its anticoagulant properties well. The results suggest that the immobilization of heparin via polydopamine layer can confer excellent antithrombotic properties, and the heparin immobilization method via polydopamine layer provides an alternative approach for other biomolecules immobilization on biomaterials surface. Thus it is envisaged that this method will be potentially useful for the surface modification of blood-contacting biomaterials.

Keywords

heparin / dopamine / titanium / cardiovascular biomaterials / blood compatibility

References

[1] Weber N, Wendel H P, Ziemer G Hemocompatibility of Heparin-Coated Surfaces and the Role of Selective Plasma Protein Adsorption[J]. Biomaterials, 2002, 23: 429-439.

[2] Huang N, Yang P, Leng Y X, . Hemocompatibility of Titanium Oxide Films[J]. Biomaterials, 2003, 24: 2 177-2 187.

[3] Guarnieri D, De Capu A, Ventre M, . Covalently Immobilized RGD Gradient on PEG Hydrogel Scaffold Influences Cell Migration Parameters[J]. Acta Biomaterialia, 2010, 6: 2 532-2 539.

[4] Li GC, Yang P, Qin W, . The effect of Coimmobilizing Heparin and Fibronectin on Titanium on Hemocompatibility and Endothelialization[J]. Biomaterials, 2011, 32: 4691-4703.

[5] Hersel U, Dahmen C, Kessler H RGD Modified Polymers: Biomaterials for Stimulated Cell Adhesion and Beyond[J]. Biomaterials, 2003, 24: 4 385-4 415.

[6] Zhu AP, Zhang M, Wu J, . Covalent Immobilization of Chitosan/ Heparin Mixture with a Photosensitive Hetero-bifunctional Crosslinking Reagent on PLA Surface[J]. Biomaterials, 2002, 23: 4 657-4 665.

[7] Yin YB, Wise SG, Nosworthy NJ, . Covalent Immobilisation of Tropoelastin on a Plasma Deposited Interface for Enhancement of Endothelialisation on Metal Surfaces[J]. Biomaterials, 2009, 30: 1 675-1 681.

[8] Bakker WW, van der Lei B, Nieuwenhuis P, . Reduced Thrombogenicity of Artificial Materials by Coating with ADPase[J]. Biomaterials, 1991, 12: 603-606.

[9] Liu Q, Ding J, Mante FK, . The Role of Surface Functional Groups in Calcium Phosphate Nucleation on Titanium Foil: A Self-assembled Monolayer Technique[J]. Biomaterials, 2002, 23: 3 103-3 111.

[10] Fu J, Ji J, Yuan W, . Construction of Anti-adhesive and Antibacterial Multilayer Films via Layer-by-layer Assembly of Heparin and Chitosan[J]. Biomaterials, 2005, 26: 6 684-6 692.

[11] Meng S, Liu Z, Shen L, . The Effect of a Layer-by-layer Chitosan-heparin Coating on the Endothelialization and Coagulation Properties of a Coronary Stent System[J]. Biomaterials, 2009, 30: 2 276-2 283.

[12] Muller R, Abke J, Schnell E, . Influence of Surface Pretreatment of Titanium- and Cobalt-based Biomaterials on Covalent Immobilization of Fibrillar Collagen[J]. Biomaterials, 2006, 27: 4 059-4 068.

[13] Leon MB Late Thrombosis a Concern with Drug-eluting Stents[J]. J. Interv. Cardiol, 2007, 20: 26-29.

[14] Lee H, Dellatore SM, Miller WM, . Mussel-inspired Surface Chemistry for Multifunctional Coatings[J]. Science, 2009, 318: 426-430.

[15] Yu M, Hwang J, Deming TJ Role of l-3, 4-dihydroxyphenylalanine in Mussel Adhesive Proteins[J]. J. Am. Chem. Soc, 1999, 121: 5 825-5 826.

[16] Burzio LA, Waite JH Cross-linking in Adhesive Quinoproteins: Studies with Model Decapeptides[J]. Biochemistry, 2000, 39: 1 147-1 153.

[17] Lee H, Rho J, Messersmith PB Facile Conjugation of Biomolecules onto Surfaces via Mussel Adhesive Protein Inspired Coatings[J]. Adv. Mater., 2009, 21: 431-434.

[18] Zhu L P, Yu J, Xu YY, . Surface Modification of PVDF Porous Membranes via Poly(DOPA) Coating and Heparin Immobilization[J]. Colloids Surf. B: Biointerfaces, 2009 152-155.

[19] Jiang JH, Zhu LP, Li XL, . Surface Modification of PE Porous Membranes Based on the Strong Adhesion of Polydopamine and Covalent Immobilization of Heparin[J]. J. Membr. Sci., 2010, 364: 194-202.

[20] Bae IH, Park IK, Park DS, . Thromboresistant and Endothelialization Effects of Dopamine-mediated Heparin Coating on a Stent Material Surface[J]. J. Mater. Sci: Mater. Med., 2012, 23: 1 259-1 269.

[21] You I, Kang SM, Byun Y, . Enhancement of Blood Compatibility of Poly(urethane) Substrates by Mussel-Inspired Adhesive Heparin Coating[J]. Bioconjugate Chem., 2011, 22: 1 264-1 269.

[22] Poh CK, Shi ZL, Lim TY, . The Effect of VEGF Functionalization of Titanium on Endothelial Cells in Vitro[J]. Biomaterials, 2010 1 578-1 585.

[23] Zhu LP, Jiang JH, Zhu BK, . Immobilization of Bbovine Serum Albumin onto Porous Polyethylene Membranes Using Stronglyattached Polydopamine as a Spacer[J]. Colloids Surf. B: Biointerfaces, 2011, 86: 111-118.

[24] Yan P, Wang J, Wang L, . The in-vitro Biomineralization and Cytocompatibility of Polydopamine Coated Carbon Nanotubes[J]. Appl. Surf. Sci., 2011, 257: 4 849-4 856.

[25] Kang K, Choi I S, Namb Y A Biofunctionalization Scheme for Neural Interfaces Using Polydopamine Polymer[J]. Biomaterials, 2011, 32: 6 374-6 380.

[26] Ku SH, Park ChB Human Endothelial Cell Growth on Mussel-Inspired Nanofiber Scaffold for Vascular Tissue Engineering[J]. Biomaterials, 2010, 31: 9 431-9 437.

[27] Kang J, Sakuragi M, Shibata A, . Immobilization of Epidermal Growth Factor on Titanium and Stainless Steel Surfaces via Dopamine Treatment[J]. Mat. Sci. Eng. C, 2012, 32: 2 552-2 561.

[28] Sever M, Wilker J Synthesis of Peptides Containing DOPA (3,4-dihydrocyphenylalanine) [J]. Tetrahedron, 2001, 57: 6 139-6 146.

[29] Wang J, Liu C, Lu X, . Co-polypeptides of 3,4-dihydroxy-phenylalanine and L-lysine to Mimic Marine Adhesive Protein[J]. Biomaterials, 2007, 28: 3 456-3 468.

[30] Gong YK, Liu LP, Messersmith PB Doubly Biomimetic Catecholic Phosphorylcholine Copolymer: A Platform Strategy for Fabricating Antifouling Surfaces[J]. Macromol. Biosci., 2012, 12: 979-985.

[31] Rigberg D, Tulloch A, Chun Y, . Thin-film Nitinol (NiTi): a Feasibility Studyfor a Novel Aortic Stent Graft Material[J]. J. Vasc. Surg., 2009, 50: 375-380.

[32] Chuang TW, Masters KS Regulation of Polyurethane Hemocompatibility and Endothelialization by Tethered Hyaluronic Acid Oligosaccharides[J]. Biomaterials, 2009, 30: 5 341-5 351.

[33] Li LH, Tu M, Mou SS, . Preparation and Blood Compatibility of Polysiloxane/Liquid-crystal Composite Membranes[J]. Biomaterials, 2001, 22: 2 595-2 599.

[34] Lagergren M, Olsson P, Swedenborg J Inhibited Platelet Adhesion: a Non-thrombogenic Characteristic of a Heparin-coated Surface[J]. J. Surgery., 1974, 75: 643-650.

[35] Yang MC, Lin WC Surface Modification and Blood Compatibility of Polyacrylonitrile Membrane with Immobilized Chitosan-heparin Conjugate[J]. J. Polym. Res, 2002, 9: 201-206.

[36] Grasel TG, Goodman SL, Cooper SL, . Platelet Shape Change and Cytoskeletal Reorganization on Polyurethaneureas[J]. J. Biomed. Mater. Res., 1989, 23: 105-123.

[37] Kloczewiak M, Timmons S, Lukas TJ, . Platelet Receptor Recognition Site on Human-fibrinogen e Synthesis and Structurefunction Relationship of Peptides Corresponding to the Carboxyterminal Segment of the Gamma-chain[J]. Biochem. J., 1984, 23: 1 767-1 774.