Mitigation of monocyte driven thrombosis on cobalt chrome surfaces in contact with whole blood by thin film polar/hydrophobic/ionic polyurethane coatings.

Monocytes are active at the crossroads between inflammation and coagulation processes since they can secrete pro-inflammatory cytokines and express tissue factor (TF), a major initiator of coagulation. Cobalt-chrome (CoCr), a metal alloy, used as a biomaterial for vascular stents, has been shown to be potentially pro-thrombotic and pro-inflammatory. Research work with a polymer from a family of degradable-polar hydrophobic ionic polyurethanes (D-PHI), called HHHI, has been shown to exhibit anti-inflammatory responses from human monocytes. We have generated multifunctional polyurethane thin films (MPTF) based on the HHHI chemistry, as a thin coating for CoCr and have evaluated the reactivity of blood with MPTF-coated CoCr. The results showed that the coating of CoCr with MPTF derived from HHHI prevents thrombin generation, reduces coagulation activation, and suppresses fibrin formation in whole blood. Activation of monocytes was also suppressed at the surface of MPTF-coated CoCr and specifically the decrease in thrombin generation was accompanied by a significant decrease in TF and pro-inflammatory cytokine levels. Mass spectroscopy of the adsorbed proteins showed lower levels of fibrinogen, fibronectin and complement C3, C4, and C8 when compared to CoCr. We can conclude that MPTFs reduce the pro-thrombotic and pro-inflammatory phenotype of monocytes and macrophages on CoCr, and prevent clotting in whole blood.

Coating of cobalt chrome substrates with thin films of polar/hydrophobic/ionic polyurethanes: Characterization and interaction with human immunoglobulin G and fibronectin.

Biomaterial implants often lead to specific tissue reactions that could compromise their bio-integration and/or optimal cellular interactions. Polyurethanes (PU) are of particular interest as coatings to mask CoCr's bioactivity, since they are generally more biocompatible than metal substrates, present a broad range of chemistry, and have highly tunable-mechanical properties. In the current work, complex polyvinyl-urethanes (referred to as D-PHI materials) are studied for their surface phase structures: specifically, an original D-PHI polymer (O-D-PHI) and a differential formulation with relatively higher hydrophobic and ionic content (HHHI) are of interest. The PUs are diluted in tetrahydrofuran (THF) to generate thin films which differentially influence the physical and chemical properties of the D-PHI coatings. AFM images over time show the gradual appearance of domains exhibiting crystalline organisation, and whose shape and size were dependent on D-PHI thickness (thin films vs non-solvent cast resin materials). After three weeks, a complete stabilization of the crystal state is observed. The thin coatings are stable in an aqueous and 37 °C environment. The adsorption of two human plasmatic proteins Immunoglobulin G (IgG) and Fibronectin (Fn), involved in inflammation and coagulation, was studied. The exposure of specific protein sequences (IgG-Fab, Fn-Cell Binding Domain and Fn-N-terminal domain) were dramatically reduced on both D-PHI materials when compared to bare metal CoCr. The implications of these findings would be relevant to defining coating strategies used to improve the blood clotting and immune cell reactivity to CoCr implant materials.

Mono vs multilayer fibronectin coatings on polar/hydrophobic/ionic polyurethanes: Altering surface interactions with human monocytes.

Monocyte interactions with materials that are biofunctionalized with fibronectin (Fn) are of interest because of the documented literature which associates this protein with white blood cell function at implant sites. A degradable-polar hydrophobic ionic polyurethane (D-PHI), has been reported to promote an anti-inflammatory response from human monocytes. The aim of the current work was to study the influence of intrinsic D-PHI material chemistry on Fn adsorption (mono and multi-layer structures), and to investigate the influence of such chemistry on the structural state of the Fn, as well as the latter's influence on the activity of human monocytes on the protein coated substrates. Significant differences in Fn adsorption, surface hydrophobicity and the availability of defined peptide sequences (N terminal, C terminal or Cell Binding Domain) for the Fn in mono vs multilayer structures were observed as a function of the changes in intrinsic material chemistry. A D-PHI-formulated polyurethane substrate with subtle changes in anionic and hydrophobic domain content relative to the polar non-ionic urethane/carbonate groups within the polymer matrix promoted the lowest activation of monocytes, in the presence of multi-layer Fn constructs. These results highlight the importance of chemical heterogeneity as a design parameter for biomaterial surfaces, and establishes a desired strategy for controlling human monocyte activity at the surface of devices, when these are coated with multi-layer Fn structures. The latter is an important step towards functionalizing the materials with multi-layer protein drug carriers as interventional therapeutic agents. STATEMENT OF SIGNIFICANCE: The control of the behavior of monocytes, especially migration and activation, is of crucial interest to modulate the inflammatory response at the site of implanted biomaterial. Several studies report the influence of adsorbed serum proteins on the behavior of monocytes on biomaterials. However, few studies show the influence of surface chemical group distribution on the controlled adsorption and the subsequent induced conformation- of mono versus multi-layer assembled structures generated from specific proteins implicated in wound repair. The current research considered the role of Fn adsorption and conformation in thin films while interacting with the intrinsic chemistry of segmented block polyurethanes; and the influence of the former on modulation and activation of human monocytes.