Enhanced Blood Compatibility Research Articles

Antheraea assama Silk Fibroin‐Based Functional Scaffold with Enhanced Blood Compatibility for Tissue Engineering Applications

AbstractThe architecture and surface chemistry of a scaffold determine its utility in tissue engineering (TE). Conventional techniques have limitations in fabricating a scaffold with control over both architecture and surface chemistry. To ameliorate this, in this report, we demonstrate the fabrication of an Antheraea assama silk fibroin (AASF)‐based functional scaffold. AASF is a non‐mulberry variety having superior qualities to mulberry SF and is largely unexplored in the context of TE. First, a 3D scaffold with biomimetic architecture is fabricated. The scaffold is subsequently made blood compatible by modifying the surface chemistry through a simple sulfation reaction. EDX and FTIR analysis demonstrate the successful sulfation of the scaffold. SEM observations reveal that sulfation has no any effect on the scaffold architecture. TGA reveals that it has increased thermal stability. The sulfation reaction significantly improves the overall hydrophilicity of the scaffold, as is evident from the increase in water holding capacity; this possibly enhances the blood compatibility. The enhancement in blood compatibility of the sulfated scaffold is determined from in vitro haemolysis, protein adsorption and platelet adhesion studies. The sulfated scaffold is non‐toxic and supports cell adhesion and growth, as revealed by indirect and direct contact‐based in vitro cytotoxicity assays. This study reveals that the AASF‐based functional scaffold, which has biomimetic architecture and blood‐compatible surface chemistry, could be suitable for TE applications.

Enhanced blood compatibility of polyurethane functionalized with sulfobetaine

The purpose of this work is to develop a biocompatible polyurethane surface by the tailoring of sulfobetaine. The polyurethane film was first grafted polymerization with acrylic acid by ozonization, followed by immobilizing sulfobetaine via two routes: (i) synthesize primary amine group terminated sulfobetaine and then couple onto the surface of polyurethane using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC); (ii) couple the primary amine group terminated tertiary amine onto the surface of polyurethane primarily using EDC and then form sulfobetaine through a ring-opening reaction between tertiary amine and 1,3-propanesultone (PS). The reaction process was monitored with attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). The blood compatibility was evaluated by a hemolytic test and platelet-rich plasma (PRP) adhesion experiment. Little hemolysis took place on the surface of polyurethane grafted with sulfobetaine. Platelets adhering on the surface of grafted polyurethane decreased greatly as compared to the original after 1 and 3 h of incubation with PRP.

Surface Modification of Polyethylene by Heparin for Improvement of Antithrombogenicity

The purpose of this paper was to enhance blood compatibility ofpolyethylene (PE) film. PE film pretreated by argon plasma wassubjected to ultraviolet (UV) -induced graft polymerization withAcrylic acid(AAc) (AAc-grafted PE films, PE-g-PAAc) withoutphoto-initiator, then heparin was covalently immobilized on the PEsurface (PE-g-HPAAc). The surface properties and microstructure ofPE-g-PAAc and PE-g-HPAAc were studied by static contact anglemeasurement, atomic force microscope (AFM), X-ray photoelectronspectroscopy (XPS) and Attenuated total reflectance Fourier transferinfrared spectroscopy (ATR-FTIR). It was confirmed that AAc andheparin were successfully immobilized onto the surface of PE film.Results of platelet adhesion experiments indicated that theantithrombogenicity of the modified PE film was remarkably improved.

Behaviors of Platelets Adherent on Si-N(H) Surface Prepared from Ammonia Plasma-Implanted Silicon

Ammonia implanted silicon was performed by using plasma immersion ion implantation (PIII) to form a silicon nitride films. Blood compatibility of the prepared samples was investigated by platelets adhesion testing. It showed less activation i.e. lower thrombosis risks occurs on the prepared silicon nitride films than control silicon sample. The enhanced blood compatibility of the material is attributed to the modified surface properties such as hydrophilicity from thermodynamic adsorption perspective, which is related to surface chemical bonding states achieved by PIII process.

Surface modification of polymeric biomaterials: Utilization of cyclodextrins for blood compatibility improvement

A novel modified polymeric biomaterial surface using cyclodextrins (CDs) for improved blood compatibility was studied. Plasticized poly(vinyl chloride) (PVC-P) was selected for modification and polyethylene was used as a reference material. The modification was achieved by polymer blending. Fibrinogen and albumin adsorption were utilized as indices for the assessment of the blood compatibility. Surface characterization confirmed that CDs were able to accumulate at the PVC surface and alter the surface properties. The combination of other hydrophilic polymers such as poly(ethylene oxide) (PEO) and PEO/poly(propylene oxide) (PPO) copolymers, such as Pluronic F68 (F68), with CDs were also investigated. These modified materials have a remarkable protein-resistant surface. The combination of B-cyclodextrin (B-CD)/PEO and B-CD/F68 in certain feeding ratio are synergistic in producing enhanced blood compatibility.

Synthesis of biodegradable PU/PEGDA IPNs having micro-separated morphology for enhanced blood compatibility

New biodegradable hydrophobic polyurethane (PU)/hydrophilic poly (ethylene glycol) diacrylate (PEGDA) IPN was simultaneously synthesized with changing the molecular weight of PEGDA to investigate the effect of crosslinking density on the degree of phase separation. PU was modified using biodegradable poly(e-caprolactone)diol and the hydroxy group of PEG was substituted to crosslinkable acrylate group having double bond, which induce photo-polymerization. The sturucture of PEGDA was confirmed by NMR. Because the reaction rate of PEGDA was faster than that of PU, the continuous matrix of the micro-separated PU/PEGDA IPNs having amphiphilic character was made of hydrophilic PEGDA-rich phase. All IPNs have sea-island morphology resulting from the suppressed phase separation. The effect of the degree of phase separation on blood compatibility was investigated.

Surface modification of polyethylene by plasma pretreatment and UV‐induced graft polymerization for improvement of antithrombogenicity

AbstractThe purpose of this study was to enhance blood compatibility of polyethylene (PE) films. Glycidyl methacrylate (GMA) was grafted onto the surface of PE by Ar plasma pretreatment and UV‐induced graft polymerization without photo‐initiator, then heparin was immobilized onto the poly (glycidyl methacrylate) segments. The surface compositions and microstructure of GMA graft polymerized PE films were studied by X‐ray photoelectron spectroscopy (XPS) and Attenuated Total Reflectance Fourier Transfer Infrared (ATR‐FTIR) spectroscopy. It was confirmed that heparin was successfully immobilized onto the surface of PE films by XPS analysis. The antithrombogenicity of the samples was determined by the activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and plasma recalcification time (PRT) tests and platelet adhesion experiment. Results indicated that the antithrombogenicity of modified PE was improved remarkably. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 93: 2014–2018, 2004

Surface-localized release of nitric oxide via sol-gel chemistry.

The release of nitric oxide (NO) from polymers has proven to be highly effective at inhibiting platelet adhesion and thus enhancing the blood compatibility of medical implants. Micropatterning techniques were used to design surfaces that release NO while preserving the underlying substrate for other applications (e.g., sensors). Micropatterned NO-releasing substrates based on aminosilane-containing methyltrimethoxysilane sol-gels were prepared and characterized in terms of stability, NO surface flux, and resistance to in vitro platelet adhesion. We have found that surface-localized NO release from substrates modified with sol-gel micropatterns exhibit enhanced blood compatibility relative to controls.

Enhanced blood compatibility of polymers grafted by sulfonated PEO via a negative cilia concept

In our laboratory sulfonated PEO (PEO-SO 3) was designed as a “negative cilia model” to investigate a synergistic effect of PEO and negatively charged SO 3 groups. PEO-SO 3 itself exhibited a heparin-like anticoagulant activity of 14% of free heparin. Polyurethane grafted with PEO-SO 3 (PU-PEO-SO 3) increased the albumin adsorption to a great extent but suppressed other proteins, while PU-PEO decreased the adsorption of all the proteins. The platelet adhesion was decreased on PU-PEO but least on PU-PEO-SO 3 to demonstrate an additional effect of SO 3 groups. The enhanced blood compatibility of PU-PEO-SO 3 in the ex vivo rabbit and in vivo canine implanting tests was confirmed. Furthermore, PU-PEO-SO 3 exhibited an improved biostability and suppressed calcification in addition to the enhanced antithrombogenicity. The in vivo antithrombogenicity and biostability were improved in the order of PU<PU-PEO<PU-PEO-SO 3. The calcium amounts deposited was decreased in the order of PU>PU-PEO>PU-PEO-SO 3 in spite of the possible attraction between negative SO 3 groups and positive calcium ions. The bioprosthetic tissue (BT) was grafted with H 2N-PEO-SO 3 via glutaraldehyde (GA) residues after conventional GA fixation. BT-PEO-SO 3 also displayed the decreased calcification by in vivo animal models. The application of PEO-SO 3 was extended by designing amphiphilic copolymers containing PEO-SO 3 moiety and hydrophobic long alkyl groups as anchors. The superior effect of PEO-SO 3 groups on thromboresistance compared to PEO was confirmed also in the case of copolymers coated or blended with other polymers and the systems coupled by UV irradiation, photoreaction or gold/sulfur or silane coupling technology, and therefore it might be very useful for the medical devices.

Improving the Blood Compatibility of Ion-Selective Electrodes by Employing Poly(MPC-co-BMA), a Copolymer Containing Phosphorylcholine, as a Membrane Coating

The hydrogelpoly(2-methacryloyloxyethylphosphorylcholine-co-butyl methacrylate), or poly(MPC-co-BMA), was used as a coating for polyurethane- and poly(vinyl chloride)-based membranes to develop ion-selective electrodes (ISEs) with enhanced blood compatibility. Adverse interactions of poly(MPC-co-BMA) with blood were diminished due to the phosphorylcholine functionalities of the hydrogel, which mimic the phospholipid polar groups present on the surface of many cell membranes. As demonstrated by immunostaining, hydrogel-coated PVC membranes soaked in platelet-rich plasma showed less adhesion and activation of platelets than uncoated PVC membranes, indicating an improvement in biocompatibility owing to the hydrogel. Furthermore, little differences in the potentiometric response characteristics, e.g., slope, detection limit, and selectivity, of ISEs employing uncoated and coated membranes were observed.

Controlling the morphology of polyurethane/polystyrene interpenetrating polymer networks for enhanced blood compatibility

Polyurethane (PU)/polystyrene (PS) IPNs were simultaneously synthesized at 80°C, controlling the reaction kinetics to change the morphology. Polymerization kinetics of styrene was controlled by the content of initiator, and that of polyurethane by the catalyst concentration. The effect of the initiator and the catalyst on the polymerization rate was analyzed by NMR spectroscopy and FTIR. Gelation time was also measured by using the advanced rheometric expansion system (ARES). Samples with sea-and-island morphology were obtained, when the polymerization rate of PS was relatively slow, and the phase separation time was long. When the polymerization rate of PS was relatively fast, and the phase separation time was short, cocontinuous morphology was obtained. The degree of phase separation and surface roughness decreased, as the rate of PU network formation was increased, and the phase-continuity was increased. The in vitro blood-compatibility tests showed that the surface roughness was an important factor on the adsorption of fibrinogens and platelets. A large amount of fibrinogens and platelets were adsorbed on the relatively rough surface of samples showing sea-island morphology. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 379–387, 2002; DOI 10.1002/app.10358

Controlling the morphology of polyurethane/polystyrene interpenetrating polymer networks for enhanced blood compatibility

AbstractPolyurethane (PU)/polystyrene (PS) IPNs were simultaneously synthesized at 80°C, controlling the reaction kinetics to change the morphology. Polymerization kinetics of styrene was controlled by the content of initiator, and that of polyurethane by the catalyst concentration. The effect of the initiator and the catalyst on the polymerization rate was analyzed by NMR spectroscopy and FTIR. Gelation time was also measured by using the advanced rheometric expansion system (ARES). Samples with sea‐and‐island morphology were obtained, when the polymerization rate of PS was relatively slow, and the phase separation time was long. When the polymerization rate of PS was relatively fast, and the phase separation time was short, cocontinuous morphology was obtained. The degree of phase separation and surface roughness decreased, as the rate of PU network formation was increased, and the phase‐continuity was increased. The in vitro blood‐compatibility tests showed that the surface roughness was an important factor on the adsorption of fibrinogens and platelets. A large amount of fibrinogens and platelets were adsorbed on the relatively rough surface of samples showing sea‐island morphology. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 379–387, 2002; DOI 10.1002/app.10358

Improved blood compatibility and decreased VSMC proliferation of surface-modified metal grafted with sulfonated PEG or heparin

Although the technique of coronary stenting has remarkably improved long-term results in recent years, (sub)acute thrombosis and late restenosis still remain problems to be solved. Metallic surfaces were regarded as thrombogenic, due to their positive surface charges, and stenosis resulted from the activation and proliferation of vascular smooth muscle cells (VSMCs). In this study, a unique surface modification method for metallic surfaces was studied using a self-assembled monolayer (SAM) technique. The method included the deposition of thin gold layers, the chemisorption of disulfides containing functional groups, and the subsequent coupling of PEG derivatives or heparin utilizing the functional groups of the disulfides. All the reactions were confirmed by ATR-FTIR and XPS. The surface modified with sulfonated PEG (Au-S-PEG-SO3) or heparinized PEG (Au-S-PEG-Hep) exhibited decreased static contact angles and therefore increased hydrophilicity to a great extent, which resulted from the coupling of PEG and the ionic groups attached. In vitro fibrinogen adsorption and platelet adhesion onto the Au-S-PEG-SO3 or Au-S-PEG-Hep surfaces decreased to a great extent, indicating enhanced blood compatibility. This decreased interaction of the modified surfaces should be attributed to the non-adhesive property of PEG and the synergistic effect of sulfonated PEG. The effect of the surface modification on the adhesion and proliferation of VSMCs was also investigated. The modified Au-S-PEG-SO3 or Au-S-PEG-Hep surfaces also exhibited decreased adhesion of VSMCs, while the deposited gold layer itself was effective. The enhanced blood compatibility and the decreased adhesion of VSMCs on the modified metallic surfaces may help to decrease thrombus formation and suppress restenosis. It would therefore be very useful to apply these modified surfaces to stents for improved functions. A long-term in vivo study using animal models is currently under way.

PEO-grafting on PU/PS IPNs for enhanced blood compatibility—effect of pendant length and grafting density

Polyurethane (PU) homopolymers and PU/polystyrene (PS) interpenetrating polymer networks (IPNs) were successfully synthesized changing the length of the pendant poly(ethylene oxide) (PEO) chains and the grafting density of PEO chains. All the PU/PS IPNs had the microphase-separated structures in which the PS-rich phase domains were dispersed in the matrix of the PU-rich phase. The domain size decreased a little, as the degree of grafting with PEO chains was increased. The water swelling ratio increased, and the interfacial energy decreased, as the length of the pendant PEO chains, and the grafting density of PEO chains of the PEO-grafted PU/PS IPNs were increased, since the mobile hydrophilic pendant PEO chains effectively induced and absorbed the water, when they were contacted with water. The hydrophilic and highly concentrated pendant PEO chains could easily prohibit the adhesion of the fibrinogens and the platelets on the surface, and the blood compatibility of IPNs was enhanced by increasing of grafting with PEO chains. The adsorption of the fibrinogens and the platelets was suppressed, as the length of pendant PEO chains, and the grafting density were increased.

In vivo biostability and calcification-resistance of surface-modified PU-PEO-SO3.

To examine the biostability and calcification-resistance of polyurethanes (PUs), the surface of PU was grafted with hydrophobic perfluorodecanoic acid (PFDA) (PU-PFDA), hydrophilic polyethyleneoxide (PEO) (PU-PEO1000), and further negatively charged sulfonate groups (PU-PEO1000-SO3). An in vivo animal test was conducted by subcutaneous implantation in rats during 2, 4 and 6 months. A scanning electron microscope study demonstrated that the degree of surface cracking on explanted PUs was increased in the following order: PU-PFDA > PU > PU-PEO1000 > PU-PEO1000-SO3. In the results of energy dispersive x-ray analysis and inductively coupled plasma atomic emission spectrometry, the deposition of calcium was found abundantly, but that of phosphorus was hardly in existence in all implanted PUs, suggesting that this calcium compound is not a hydroxyapatite. The calcium contents, regardless of implantation time, were also increased in the same order (PU-PFDA > PU > PU-PEO1000 > PU-PEO1000-SO3). After 6 months implantation, no severe tissue reactions were observed and calcification almost occurred on polymer surfaces in all implants. Such superior biostability and anticalcification of PU-PEO1000-SO3 might be attributed to synergistic effects of its excellent surface smoothness, sulfonate acid (SO3-) groups, nonadhesive and mobile PEO, and the high hydrophilicity and enhanced blood compatibility. Therefore, PU-PEO1000-SO3 is promising as biostable and calcification-resistant biomaterial.

Surface structure and inert surface characteristics of perfluorodecanoic acid‐grafted polyurethane

AbstractThe surface compositions and characteristics of perfluorodecanoic acid (PFDA)‐grafted polyurethane (PU) were investigated to understand its enhanced blood compatibility. The grafting of PFDA was identified by attenuated total reflectance–Fourier transform infrared (ATR–FTIR). The results from angular‐dependent electron spectroscopy for chemical analysis (ADESCA) demonstrated the fluorocarbon enrichment at the outermost layer in PFDA‐grafted PU. Static secondary ion mass spectroscopy (SIMS) experiments supported the results drawn from ATR–FTIR and ADESCA data about the surface compositions. Energy‐dispersive X‐ray analysis (EDXA) data by fluorine mapping also indicated a considerable coverage with fluoroalkyl groups on the PU–PFDA surface. The critical surface tension, γc, of the highly hydrophobic PU–PFDA surface revealed an extremely low value of 6.9 dyn/cm due to the optimal orientation of CF3 groups to the uppermost surface. Therefore, such an inert low‐energy surface may contribute to improve blood compatibility of PFDA‐grafted PU. © 1993 John Wiley &amp; Sons, Inc.

Preparation and surface properties of PEO-sulfonate grafted polyurethanes for enhanced blood compatibility

Preparation and surface properties of PEO-sulfonate grafted polyurethanes for enhanced blood compatibility

Preparation and surface properties of PEO-sulfonate grafted polyurethanes for enhanced blood compatibility.

In order to improve the thromboresistance of the commercial polyurethane(PU), its surface modification was accomplished by three new different methods and their surface characteristics were investigated using ATR-FTIR, ESCA, SEM, and dynamic contact angle measurements. Sulfonations using propane sultone were performed directly onto PU or onto hydrophobic dodecanediol (DDO) grafted PU or onto hydrophilic poly(ethyleneoxide) (PEO) grafted PU. ESCA data coincided well with ATR-IR results, as more 0 at. % for PEO grafted PUs and the presence of S for the sulfonated PUs were revealed. At SEM observation the surfaces of PU-DDO and PU-PEO were relatively smooth, whereas all the sulfonated PU surfaces showed excellent smoothness and homogeneity. The hydrophilicity of the surfaces was considerably increased after PEO grafting or sulfonation. In addition, all the sulfonated PU surfaces, particularly PU-PEO-SO3, which has further hydrophilicity, exhibited complete wetting behavior due to the negatively charged SO3 groups.

Surface characteristics and blood compatibility of polyurethanes grafted by perfluoroalkyl chains.

Polyurethane (PU) surface was chemically modified by grafting of perfluorodecanoic acid (PFDA) to produce a highly hydrophobic surface to compare the blood compatability with hydrophilic poly(ethylene oxide) (PEO) grafted PUs. The advancing contact angle of modified PU-PFDA was increased up to 115 deg, while that of untreated PU was 86 deg. The PFDA grafted PU exhibited less adhesion and shape change of platelets than untreated PU, and the activated partial thromboplastin time (APTT) of PU-PFDA was considerably extended. The ex vivo occlusion time of untreated PU was only 50 min, but that of PFDA grafted PU was extended to 130 min, indicating that this hydrophobic surface is significantly blood compatible. It is interesting to find that the enhanced blood compatibility of very hydrophobic PU-PFDA was equivalent to hydrophilic PU-PEO.

Titanium-Protein Interaction: Changes with Oxide Layer Thickness

Since titanium is being used for various biomedical applications requiring enhanced blood compatibility, which may be partly due to its extremely stable oxide layer, an attempt is made here to understand the effect of oxide layer thickness on protein adsorption. Different thickness of oxide layers have been coated on titanium foil using anodizing method and thickness of oxide layers deposited have been measured by an ellipsometer. Studies of competitive adsorption of proteins, using I125 labelled protein from a mixture of 25 mg% albumin, 15 mg% gamma-globulin and 7.5 mg% fibrinogen indicate an increased adsorption of proteins onto the oxide layer coated surfaces compared to the bare surface.