The relationship between the antimicrobial effect of catheter coatings containing silver nanoparticles and the coagulation of contacting blood.

It is well known that surface coatings for medical devices can be made antimicrobial through introduction of silver nanoparticles. By virtue of their extremely large surface-to-volume ratio, the silver particles serve as a depot for sustained release of silver ions, despite the fact that silver is not readily oxidized. Antimicrobial coatings are especially important in connection with indwelling catheters with a high risk of bacterial line infections, such as central venous catheters (CVCs). This study specifically addressed the question what the impact of silver nanoparticles (exposed at the coating's surface) and/or the release of silver ions would be on coagulation of contacting blood. Studies, performed in vitro with fresh platelet-rich blood plasma (PRP) from 5 different healthy volunteer donors, clearly pointed out that: (i) the presence of silver nanoparticles correlates with accelerated thrombin formation upon contact of the coating with PRP; (ii) platelet activation is stronger as a result from the contact with silver nanoparticle-containing coatings as compared to other coatings which are devoid of silver. A series of titration experiments, in which the potential effect of silver ions is mimicked, revealed that the observed activation of blood platelets can be best explained through a collision mechanism. The results suggest that platelets that collide with silver, exposed at the surface, become activated without adhering to the surface. These new results point, rather unexpectedly, at a double effect of the silver nanoparticles in the coating: a strong antimicrobial effect occurs simultaneously with acceleration of the coagulation of contacting blood. This new information is, evidently, most relevant for the development of improved surface coatings for indwelling catheters (such as CVCs) which should combine antimicrobial features and close-to-zero thrombogenicity.

Thrombus formation at the surface of guide-wire models: effects of heparin-releasing or heparin-exposing surface coatings.

PURPOSE: This study was conducted to investigate whether thrombus formation at the surface of guide wires occurs, and--if so--whether this can be suppressed or prevented through incorporation of heparin in the surface coating. MATERIALS AND METHODS: Five guide wire models were examined; three had a polymeric hydrophilic surface coating (90/10 guide wire), which was either heparin-free, impregnated with sodium-heparin (Na-hep), or impregnated with benzalkonium heparin (BAK-hep). The other two guide wires had a coating of polytetrafluoroethylene (PTFE), either without heparin, or impregnated with BAK-hep. Release of heparin, exposure of heparin at the surface of the guide wires, thrombogenicity (under static and flow conditions) and their propensity to attract blood platelets were investigated. RESULTS: The guide wire 90/10 Na-hep releases approximately 150-200 mU active heparin per cm coil within the first few minutes after incubation in buffer. The PTFE BAK-hep shows a relatively slow release of 60-70 mU active heparin per cm coil. The 90/10 BAK-hep showed no released heparin but the most exposed heparin. In a static experiment with human full blood excessive thrombus formation occurred at the heparin-free models, whereas the others remained essentially clean. In a thrombin-generation assay under flow the authors observed strong retardation of thrombin formation in the case of the 90/10 Na-hep guide wire. CONCLUSIONS: The static and dynamic in vitro assays, taken together, show that the 90/10 Na-hep provides a coating with an extremely low level of surface thrombogenicity. Use of a guide wire with a hydrophilic distal coating that releases and exposes sodium heparin may contribute to the safety of diagnostic and therapeutic interventional procedures.

Bioengineering of improved biomaterials coatings for extracorporeal circulation requires extended observation of blood-biomaterial interaction under flow.

Extended use of cardiopulmonary bypass (CPB) systems is often hampered by thrombus formation and infection. Part of these problems relates to imperfect hemocompatibility of the CPB circuitry. The engineering of biomaterial surfaces with genuine long-term hemocompatibility is essentially virgin territory in biomaterials science. For example, most experiments with the well-known Chandler loop model, for evaluation of blood-biomaterial interactions under flow, have been described for a maximum duration of 2 hours only. This study reports a systematic evaluation of two commercial CPB tubings, each with a hemocompatible coating, and one uncoated control. The experiments comprised (i) testing over 5 hours under flow, with human whole blood from 4 different donors; (ii) measurement of essential blood parameters of hemocompatibility; (iii) analysis of the luminal surfaces by scanning electron microscopy and thrombin generation time measurements. The dataset indicated differences in hemocompatibility of the tubings. Furthermore, it appeared that discrimination between biomaterial coatings can be made only after several hours of blood-biomaterial contact. Platelet counting, myeloperoxidase quantification, and scanning electron microscopy proved to be the most useful methods. These findings are believed to be relevant with respect to the bioengineering of extracorporeal devices that should function in contact with blood for extended time.

The effect of high-density-lipoprotein on thrombus formation on and endothelial cell attachement to biomaterial surfaces.

Cardiovascular implants such as vascular grafts fail frequently because they lack genuine blood-compatibility. The blood-contacting surface should simultaneously prevent thrombus formation and promote formation of a confluent endothelial cell layer, to achieve sustained haemostasis. Contact activation and endothelialization are known to be determined by the plasma proteins which adsorb onto virtually all synthetic surfaces almost immediately upon contact with blood. A common approach in blood-compatibility research is, therefore, to use hydrophilic biomaterials, which are sometimes claimed to be "protein-repellent". We report here that, for synthetic polymeric surfaces, hydrophilicity is by no means synonymous to protein-repellency. We discovered that significant amounts of proteins, especially high-density lipoprotein, adsorb to hydrophilic surfaces. Pre-incubation of hydrophilic synthetic surfaces with high-density lipoprotein provides a blood-biomaterial interface, which inhibits thrombin generation and subsequent thrombus formation, and also accommodates overgrowth with a confluent endothelial layer. This approach may open the way to truly functional small-caliber arterial prostheses, and may also be relevant to cardiovascular tissue engineering in which de novo vascular tissues are cultured on or within a biomaterial scaffold.

Human endothelial cell attachment and proliferation on a novel vascular graft prototype.

A new vascular prosthesis prototype was assessed for its ability to support an endothelial cell layer in vitro. A coiled tubular structure, constructed from polymer-coated metallic wires, with an internal diameter of 690 microm, was used. Addition of heparin to the surface coating of the coil strongly enhanced the blood compatibility of the device. A series of coils with five different coatings, increasing in hydrophilicity, was studied. Heparin was added to one series, another series did not contain this anticoagulant drug. Upon contact with blood, a vascular prosthesis will instantaneously adsorb plasma proteins on its surface, and these proteins will influence the behavior of cells binding to the device. When coils were treated with human plasma proteins, mimicking the in vivo situation, human microvascular endothelial cells grew well on all coils studied, irrespective of the hydrophilicity of the underlying coating or the addition of heparin. For control coils, only endothelial cell growth on the most hydrophobic surfaces, and a moderate enhancing effect for heparin, were observed. This novel vascular graft prototype seems well suited for the support of an endothelial cell layer, especially when plasma proteins are adsorbed to its surface, and shows promise for in vivo testing.

Coils and tubes releasing heparin. Studies on a new vascular graft prototype.

Coiled metallic guidewires find widespread use, for instance in interventional cardiology. It is known that release of heparin from the surface of guidewires is advantageous to prevent formation of thrombotic emboli. New coiled tubular structures, having larger inner and outer diameter as compared to guidewires, are presented. In theory these tubes can be used as interposition vascular grafts. Ten coiled tubes with an internal diameter of 690 microm were made. Five different adherent polymeric coatings with increasing hydrophilicity were used. Five tubes contained heparin in the coating and the other five were unheparinised controls. The five tubes containing heparin were studied with respect to heparin release in vitro (amount released, kinetics), and immobilised heparin that is exposed at the surface. All tubes were studied with a direct cell contact assay using 3T3 mouse fibroblast cells, a dynamic thrombin generation test, and endothelial cell growth onto the coils. It was found that the heparinised tubes lead to very little thrombin formation. It is argued that this is due to heparin that is immobilised and exposed at the inner surface of such tubes. Furthermore the coils showed to be cytocompatible and endothelial cells adhere and proliferate well onto the coils. This concept is believed to hold promise for further development of small vascular grafts.

Injectable polymeric microspheres with X-ray visibility. Preparation, properties, and potential utility as new traceable bulking agents.

The copolymer of methyl methacrylate (MMA) and 2-[2',3',5'-triiodobenzoyl]oxoethyl methacrylate (1), ratio 3:1 (mass:mass), was prepared via a free-radical polymerization in bulk. The copolymer (M(w) = 97.8 kD and M(n) = 41.5 kD) was dissolved in chloroform and subsequently transformed into beads with a diameter in the micrometer range, using a solvent evaporation technique. The resulting microbeads were characterized by different techniques, including NMR spectroscopy, differential scanning calorimetry, gel permeation chromatography, and scanning electron microscopy. The latter technique was used as the basis for statistical analysis of the bead size. Typically, an average diameter of 96 microm and a standard deviation of 21 microm were obtained. The beads were also subjected to some preliminary tests regarding cytotoxicity. The copolymer of MMA and 1 contains covalently bound iodine. Therefore, the material is intrinsically radiopaque, i.e., capable of absorbing X-radiation while no contrast additive is needed. Our interest in these microspheres stems primarily from their possible utility as injectable and afterward traceable (radiopaque) bulking agents, e.g., for use in urology for the treatment of female stress incontinence due to sphincter deficiency. As a first test into this direction, a sample of the microbeads was mixed with ethylene glycol, and the resulting suspension was studied with respect to injectability and radiopacity. The results suggest that the radiopaque microbeads may provide access to improved bulking agents. Further modification of the surface may be necessary in order to suppress the migratory aptitude of the radiopaque polymeric microspheres in vivo.