Preparation of non-thrombogenic materials using 2-methacryloyloxyethyl phosphorylcholine.

This review addresses the non-thrombogenic characteristics of copolymers based on 2-methacryloyloxyethyl phosphorylcholine (MPC), originally developed by Nakabayashi and colleagues. The hypothesis underlying these developments was that such materials would adsorb phospholipids from blood, yielding surfaces with good natural blood compatibility. Methacrylates were found to have excellent properties for this copolymerisation. The characteristics of the MPC copolymers relevant to the improved blood compatibility were minimisation of protein adsorption through an increase in the amount of free water in the MPC hydrogels, which prevents protein conformational change and increased protein stability in solution. Non-thrombogenicity has been evaluated by in vitro, ex vivo and in vivo procedures. Non-thrombogenic dialysis membranes and a durable glucose biosensor have been developed using this MPC copolymer.

Synthesis of phospholipid polymers having a urethane bond in the side chain as coating material on segmented polyurethane and their platelet adhesion-resistant properties.

Surface modification of segmented polyurethanes (SPUs) was carried out using new blood compatible polymers having both phospholipid polar groups and urethane bonds in the side chains. The polymers were composed of 2-methacryloyloxyethyl phosphorylcholine (MPC), n-butyl methacrylate (BMA) and methacrylate with a urethane bond (MU). The MPC copolymers were soluble in ethanol. The SPU membranes were immersed in an ethanol solution of MPC copolymers and dried in vacuo for coating. The surface formed was completely covered with the MPC copolymer which was confirmed by X-ray photoelectron spectroscopic analysis. The polymer coatings were hardly detached in water, ethanol and 40% aqueous solution of ethanol compared with poly(MPC-co-BMA) which did not have the MU moieties. Therefore, the MU moieties had affinity for the SPU. The surface modification of the SPUs suppressed platelet adhesion effectively after contact with platelet-rich plasma for 180 min.

Improvement of blood compatibility on cellulose dialysis membrane. I. Grafting of 2-methacryloyloxyethyl phosphorylcholine on to a cellulose membrane surface.

A methacrylate with a phospholipid polar group, 2-methacryloyloxyethyl phosphorylcholine (MPC), was grafted on cellulose membrane for haemodialysis in an aqueous medium using cerium ion (Ce4+) as an initiator. The effects of the concentrations of MPC and Ce4+, and degassing of feed solution on the grafting of MPC on the surface and the membrane properties such as permeability and mechanical strength were examined. The grafted MPC composition depended on the concentrations of both the monomer and initiator in the feed solution. When the grafted MPC distribution was controlled by the monomer concentration, the permeability of the membrane decreased with an increase in grafted MPC distribution. On the other hand, the permeability was not changed from the original membrane's value when the MPC distribution was regulated by Ce4+ concentration. The tensile strength of the membrane did not change during the grafting of MPC and this indicated that the grafting had taken place in the amorphous region of the cellulose. These results suggested that this method is a promising way to improve the blood compatibility of a cellulose membrane without having an adverse effect on the haemodialysis membrane.

Interaction between phospholipids and biocompatible polymers containing a phosphorylcholine moiety.

Random and block copolymers containing a phospholipid polar group in their side chain were synthesized by the copolymerization between 2-methacryloyloxyethyl phosphorylcholine and styrene. These copolymers showed amphiphilic character, especially poly(methacryloyloxyethyl phosphorylcholine-block-styrene) formed stable polymer micelles in water. The interaction between natural phospholipid, dipalmitoylphosphatidylcholine and methacryloyloxyethyl phosphorylcholine copolymers was investigated. The amount absorbed of dipalmitoylphosphatidylcholine from its liposomal solution on to the poly(methacryloyloxyethyl phosphorylcholine-co-styrene) surface increased with increase of methacryloyloxyethyl phosphorylcholine composition. Moreover, when poly(methacryloyloxyethyl phosphorylcholine-block-styrene) was added to dipalmitoylphosphatidylcholine solution, organization of dipalmitoylphosphatidylcholine molecules and stabilization of bilayer structure of dipalmitoylphosphatidylcholine liposome were found. This means that methacryloyloxyethyl phosphorylcholine moieties in the copolymer have a strong affinity to dipalmitoylphosphatidylcholine molecules. The blood compatibility of methacryloyloxyethyl phosphorylcholine copolymers was also investigated with particular attention to the aggregation ability of platelets after contacting methacryloyloxyethyl phosphorylcholine copolymers; this ability decreased when platelets were put in contact with polymers without a methacryloyloxyethyl phosphorylcholine moiety. On the other hand, aggregation ability remained at almost the same level to that of original platelets after contact with methacryloyloxyethyl phosphorylcholine copolymers. From these findings, we concluded that methacryloyloxyethyl phosphorylcholine copolymers show excellent blood compatibility due to adsorption of lipids from plasma and the formation of an organized adsorption layer of lipids on the surface of the methacryloyloxyethyl phosphorylcholine copolymers.

Chemical modification of silk fibroin with 2-methacryloyloxyethyl phosphorylcholine. II. Graft-polymerization onto fabric through 2-methacryloyloxyethyl isocyanate and interaction between fabric and platelets.

2-Methacryloyloxyethyl phosphorylcholine (MPC) was grafted onto silk fabric in a two-step heterogeneous system through the vinyl bonds of 2-methacryloyloxyethyl isocyanate (MOI) modified on the fabric. First, habutae silk fabric was modified with the MOI monomer in anhydrous dimethyl sulfoxide using di-n-butyltin (IV) dilaurate and hydroquinone at 35 degrees C. The saturated weight gain of modified MOI monomer on the fabric was 7.3 wt% versus the original silk. Second, graft polymerization with MPC onto the MOI modified silk was conducted using 2,2'-azo bis[2-(2-imidazolin-2-yl)propane dihydrochloride] (VA-044) as an azo polymerization initiator. The weight of the grafted MPC eventually gained was about 26.0 wt%. The MOI-modified and MPC-grafted silk fabrics were analyzed by Fourier transform infrared (FT-IR) spectroscopy. To confirm the improved biocompatibility of the silk fabric, platelet adhesion was preliminarily tested measuring lactate dehydrogenase. The number of platelets adhering to polyMPC-grafted silk fabric decreased by about one tenth compared to original and MOI-modified silk after 60 min of contact with human platelet-rich plasma (1.0 x 10(6) platelets cm(-2)).

Performance of adhesive bone cement containing hydroxyapatite particles.

A new acrylic bone cement which can adhere to both bone and prostheses was developed based on a methyl methacrylate (MMA) monomer containing 4-methacryloyloxyethyl trimellitate anhydride (4-META) as adhesion promoting agent. Moreover, hydroxyapatite (HA) particles were introduced into the 4-META cement as a bone compatible filler. The mechanical strengths of an acrylic bone cement without 4-META decreased drastically with an increase in the percentage of HA particles in the cement. However, the mechanical strengths of the HA-containing 4-META cement did not change in the same way as that of the 4-META cement without HA due to adhesion between the cement HA particles and matrix. The HA particles did not affect the adhesion of the 4-META cement to bone and metals. Implantation of the 4-META cement and the HA-containing 4-META cement in animals demonstrated that these cements did not disturb bone ingrowth and the new bone was able to contact the cement directly. The 4-META cements, with and without HA particles, could adhere to bone in vivo.

Modification of polysulfone with phospholipid polymer for improvement of the blood compatibility. Part 1. Surface characterization.

To improve the surface blood compatibility of polysulfone (PSf) membranes, we prepared novel polymeric additives which have suitable blood compatibility. They were polymers with a phosphorylcholine group, a 2-methacryloyloxyethyl phosphorylcholine (MPC) unit. The MPC polymer could be blended with polysulfone by a solvent evaporation method during membrane processing, and a transparent membrane could be obtained. The mechanical properties of the blend membrane were similar to that of the original PSf membrane. Surface analysis of the blend membrane by X-ray photoelectron spectroscopy and dynamic contact angle measurement revealed that the MPC unit in the polymeric additive was concentrated on the surface of the membrane. The blend membrane significantly reduced plasma protein adsorption compared with that of the PSf membrane.

Modification of polysulfone with phospholipid polymer for improvement of the blood compatibility. Part 2. Protein adsorption and platelet adhesion.

Protein adsorption and platelet adhesion from human plasma on polysulfone (PSf) membranes modified with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer were studied. The modification was carried out by blending of the MPC polymer in the PSf. The amount of protein adsorbed on the PSf/MPC polymer blend membrane was significantly decreased with an increase in the composition of the blended MPC polymer. The distribution of the specific proteins adsorbed on the membrane surface was also determined by a gold-colloid immunoassay. Albumin, gamma-globulin and fibrinogen were observed on every membrane surface after contact with plasma. However, in the case of the blended membrane, the density of the adsorbed proteins decreased compared with that of original PSf membrane. That is, the MPC polymer blended in the membrane could function as a protein-adsorption-resistant additive. The number of platelets adhered on the PSf membrane was reduced, and change in the morphology of adherent platelets was also suppressed by the modification with the MPC polymer. Therefore, the PSf/MPC polymer blend membrane had improved blood compatibility compared with the PSf membrane.

Improvement of blood compatibility on cellulose dialysis membrane. 2. Blood compatibility of phospholipid polymer grafted cellulose membrane.

The blood compatibility of a cellulose haemodialysis membrane whose surface was grafted with a methacrylate having a phospholipid polar group, 2-methacryloyloxyethyl phosphorylcholine, was evaluated with attention to platelet adhesion to the membrane surface and complement activation induced by the membrane. When the original cellulose membrane came in contact with platelet-rich plasma for 30 min, numerous platelets adhered to the surface and aggregated. On the other hand, the membrane grafted with 2-methacryloyloxyethyl phosphorylcholine effectively suppressed platelet adhesion and activation. This effect became more pronounced with increasing surface distribution. Especially, the 2-methacryloyloxyethyl phosphorylcholine grafted membranes, whose distribution exceeded 0.27, completely inhibited platelet adhesion, even when the contact time was 180 min. Moreover, the complement activation was also reduced with increased 2-methacryloyloxyethyl phosphorylcholine distribution on the surface of the membrane.

Preparation of nanoparticles composed with bioinspired 2-methacryloyloxyethyl phosphorylcholine polymer.

The poly(L-lactic acid) nanoparticles immobilized with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which has excellent blood compatibility, were prepared by a solvent evaporation technique using the water-soluble amphiphilic MPC polymer as an emulsifier and a surface modifier. The diameter and zeta-potential of the obtained nanoparticles strongly depended on the concentration of the MPC polymer. When the nanoparticles were prepared in 1.0 mg/ml of an MPC polymer aqueous solution, the diameter was 221 nm which was determined by atomic force microscopy and dynamic light scattering measurements. The X-ray photoelectron spectroscopic analysis indicated that the phosphorylcholine groups of the MPC unit were located at the surface of the nanoparticles, that is, the MPC polymer was immobilized on the PLA particles and the surface zeta-potential was -2.5 mV. Various hydrophobic fluorescence probes could permeate through the MPC polymer layer and adsorb on the PLA surface. The amount of bovine serum albumin adsorbed on the nanoparticles was significantly smaller compared with that on the conventional polystyrene nanoparticles. It is suggested that the nanoparticles immobilized with the MPC polymer have the potential for use as both a novel drug carrier and diagnostic reagent which can come in contact with blood components.

Fabrication and characterization of dipalmitoylphosphatidylcholine-attracting elastomeric material for joint replacements.

Wear debris associated with the polyethylene components of total joint replacements has been shown to induce bone resorption which contributes to implant loosening. In an effort to promote lubrication and reduce wear in artificial hip joints, the use of the cushion bearing concept has been proposed previously; however, an elastomeric material tested as a cushion bearing has been shown to have poor tribological properties during initiation of motion from rest. The goal of this project was to fabricate and characterize an elastomer that has the ability to attract to its surface naturally occurring boundary lubricants from an aqueous solution. The test elastomer and appropriate controls were characterized using fluorescence, electron spin resonance and X-ray photoelectron spectroscopy. The test elastomer was found to have an enhanced ability to attract dipalmitoylphosphatidylcholine, a known physiological boundary lubricant. A cushion bearing that also encourages boundary lubrication represents a potential improvement over currently existing orthopaedic implant-bearing materials.

The effect of the chemical structure of the phospholipid polymer on fibronectin adsorption and fibroblast adhesion on the gradient phospholipid surface.

The interaction between biocomponents and the polyethylene (PE) surface modified with poly[omega-methacryloyloxyalkyl phosphorylcholine (MAPC)] was considered taking into account the surface characteristics, i.e., density, mobility, and orientation of the poly(MAPC). The PE surface, grafted gradually with the poly(MAPC) was prepared by corona irradiation method. The amount of peroxide produced on the PE surface which was determined with 1,1-diphenyl-2-picryl-hydrazyl, increased with an increase in the energy of the corona. The surface density of the poly(MAPC) was increased with an increase in the amount of the peroxides produced by the corona irradiation. The orientation and mobility of the poly(MAPC) grafted on the PE surface was evaluated with 1,6-diphenyl-1,3,5-hexatriene. The orientation of the poly[6-methacryloyloxyhexyl phosphorylcholine (MHPC)] which has six methylene chains between the phospholipid polar group and the backbone was higher than that of other poly(MAPC)s. The mobility of the poly(MAPC) decreased with an increase in the methylene chain length in the MAPC unit. The fibronectin adsorption on the gradient PE sheet grafted with poly(MAPC) was determined with enzyme-labeled immunoassay. The amount of adsorbed fibronectin on the PE grafted with poly[2-methacryloyloxyethyl phospohorylcholine(MPC)] and poly(MHPC) decreased with an increase in their surface density. Especially, the PE sheet grafted with the poly(MHPC) was effectively reduced compared with other poly(MAPC)s. On the poly[10-methacryloyloxydecyl phosphorylcholine (MDPC)], there is a minimum amount of adsorbed fibronectin. The fibronectin adsorption pattern on the PE sheet grafted with poly(MAPC) was quite different from the chemical structure of the MAPC unit. The human normal diploid fibroblasts (WI-38 cells) were cultured on the gradient PE sheet grafted with poly(MAPC) changing the concentration of seeded WI-38 cells. The adhesion behavior of the WI-38 cells was different depending on the concentration of the seeded WI-38 cells. When the concentration was low, the number of the adherent WI-38 cells had the same tendency as fibronectin adsorption. The gradient PE sheet grafted with the poly(MHPC) effectively reduced WI-38 cells adhesion even when the concentration of the WI-38 cells was high. The biocompatibility of polymer surfaces can be improved by highly oriented phosphorylcholine group.

Adhesive 4-META/MMA-TBB opaque resin with poly(methyl methacrylate)-coated titanium dioxide.

An adhesive opaque resin for veneering on cast metal was developed with 4-methacryloxyethyl trimellitate anhydride and poly (methyl methacrylate)-coated titanium dioxide prepared by aqueous phase polymerization. The opaque resin was a modified 4-META/MMA-TBB resin. The powder consisted of 20% of the encapsulated material and 80% PMMA instead of pure PMMA powder. This resin hides the metal color when the thickness of the resin is as thin as 50 microns. The opaque resin bonded strongly to both cobalt-chromium alloy and visible-light-cured veneering resin. This self-curing opaque resin is applicable not only for bonding veneering resin to an alloy surface but also for bonding fixed partial dentures to enamel surfaces.

Effect of 2-(methacryloxy)ethyl phenyl hydrogen phosphate on adhesion to dentin.

A variety of methacrylate-based materials has been developed with the capacity of adhering to dentin. This study investigated the effectiveness of 2-(methacryloxy)ethyl phenyl hydrogenphosphate (phenyl-P) for bonding 5% phenyl-P in methyl methacrylate (MMA) to dentinal surfaces. Polymerization of the phenyl-P/MMA monomer was initiated by partially oxidized tri-n-butyl borane catalyst (TBB). The mean tensile bond strength of 5% phenyl-P in MMA to dentin that was pre-treated with an aqueous solution of 10% citric acid/3% ferric chloride, abbreviated as 10-3, was found to be 10.5 MPa. Scanning electron microscope examination demonstrated the formation of a transitional or "hybrid" layer of resin-reinforced dentin, created by the intermingling and entanglement of polymerized resin with collagen bundles exposed by dentin pre-treatment with 10-3, an effective remover of the dentinal smear layer. This "hybrid" layer or zone was essential for high tensile bond strength to be attained. Phenyl-P was found to be effective in promoting monomer diffusion and impregnating monomer into demineralized dentinal surfaces. The formation of the "hybrid" layer of resin-reinforced dentin followed in situ resin polymerization initiated by partially oxidized tri-n-butyl borane (TBB). Ferric (Fe3+) ions deposited on dentinal surfaces from the 10-3 solution also acted to improve monomer diffusion and entanglement with demineralized dentin, and facilitated the formation of the "hybrid" layer/zone.

Bonding to intact dentin.

It has been reported that the presence of a smear layer on dentinal substrates can compromise bonding. Typically, smear layers are removed by acidic agents that selectively extract calcium salts from dentin surfaces to leave a collagen-rich substrate. Acid-conditioned dentin (i.e., demineralized) is then primed and an adhesive agent applied. In the present study, we removed smear layers by "polishing" dentin specimens with a hydroxyapatite paste and ultrasonication. Bonding procedures were carried out by means of an aqueous solution of 20% 2-methacryloyloxyethyl phenyl phosphoric acid (phenyl-P) and 30% 2-hydroxyethyl methacrylate, referred to as 2OP-30H, a "self-etching primer". The 20P-30H solution was applied to "intact" dentin (i.e., non-demineralized) for either 30 or 60 s. Control samples received no application (O s) of the self-etching primer. Mean tensile bond strengths (10 MPa) were similar in both the 30-second- and 60-second-primed groups. The widths of formed hybrid layers varied from 0.3 +/- 0.2 micron at O s application (control) to 2.1 +/- 0.3 micron for the 30-second group and 4.1 +/- 0.2 micron for the 60-second group. SEM and TEM observations revealed that the 20P-30H self-etching primer created diffusion channels into "intact" calcium-rich dentin which permitted monomer to infiltrate dentin substrates. Hybrid layers identified under microscopic examination demonstrated resistance to both HCI and NaOCI treatments, suggesting that the hybrid layer was not defective, and that bonding was stable.

Bonding to ground dentin by a phenyl-P self-etching primer.

Increasingly higher concentrations of Phenyl-P in 30% HEMA were used as dentin conditioners to improve the bonding of adhesive resins to smear layers. The maximum bond strength (10.4 MPa) was obtained at a concentration of 20% Phenyl-P. Transmission electron microscopy of fractured, lightly smeared, and compactly smeared dentin demonstrated that 20% Phenyl-P in 30% HEMA demineralized the dentin surface by partially dissolving mineral crystals from around collagen. When applied to smear layers, this resin system demineralized the smear layer and incorporated it into the applied resin which penetrated a short distance into the underlying dentin, thereby creating a hybrid layer that contained the original smear layer. This single-step conditioner/primer offers several advantages over previous bonding systems by permitting a single solution to serve as both a conditioner and a primer.

A white band and a transparent zone observed at an interface between a polymerizing polyfunctional monomer and a poly(methyl methacrylate) rod, I.

Macro- and microscopic studies were made of an interfacial microstructure which developed between a bifunctional monomer and a PMMA rod (or beads) during polymerization. The microstructure consisted of a white band and a blue transparent zone.

4-META opaque resin--a new resin strongly adhesive to nickel-chromium alloy.

1) A new adhesive opaque resin containing a reactive monomer, 4-methacryloxy-ethyl trimellitate anhydride (4-META), was prepared, and its application to thermosetting acrylic resin veneer crowns was studied. 2) The 4-META opaque resin was applied to a variety of nickel-chromium dental alloy specimens which had undergone different treatment, and endurance tests were conducted to evaluate the durability of adhesion. 3) Stable adhesion against water penetration was achieved with metal surfaces first etched with HCl and then oxidized with HNO3. A bond strength of 250 kg/cm2 was maintained even after immersion in water at 37 degrees C for 30 wk or at 80 degrees C for ten wk. Furthermore, this value did not decrease even after the specimens were subjected to 500 thermal cycles. 4) The 4-META opaque resin studied can eliminate the necessity for retention devices on metal castings. 5) The smooth 4-META opaque resin should have no adverse effects on gingivae.

Influence of dentinal polyelectrolytes on wet demineralized dentin, a bonding substrate.

The objective of this study was to show the influence of dissolved dentinal polyelectrolytes on the characteristics of dentin (bonding substrate) demineralized by citric acid in the absence or presence of ferric chloride. The demineralizing agent was an aqueous mixture of 0, 1, 3, or 10% ferric chloride in 10% citric acid (10-0, 10-1, 10-3, 10-10, respectively). The hypothesis was that the concentration of dissolved dentinal noncollagenous substances, mainly polyelectrolytes soluble in water, must be decreased by their aggregation with ferric ions, which changes the characteristics of demineralized dentin, the rates of demineralization, and dehydration. Cervical bovine dentin was prepared in 3 x 2 x 2-mm blocks, each weighing 20.0 +/- 0.5 mg. The rate of demineralization was investigated by measuring the weight loss resulting from demineralization by immersion in 10 mL of conditioner at 2-h intervals. The dehydration rate of wet demineralized dentin was determined using two methods: (1) weight loss in a desiccator under 263 Pa pressure and (2) differential scanning calorimetry (DSC). Twenty, 12, 8, and 4 h were required to complete demineralization of the blocks with the 10-0, 10-1, 10-3, and 10-10 solutions, respectively. The 10-10 wet demineralized dentin showed the highest rate of dehydration, followed in descending order by the 10-3, 10-1, and 10-0 specimens. Ferric chloride in dentin conditioners provided both a higher rate of dentin demineralization and a higher dehydration rate of wet demineralized dentin. These results suggest that in the presence of ferric chloride, a decreasing amount of dissolved polyelectrolytes aggregated with ferric ions in the substrates may increase the permeability of dentin to water and citric acid. Improvement of monomer permeability is essential to the preparation of good hybridized dentin, providing a more stable and reliable bonding and also protecting the dentin and pulp from infection. A further study of bonding substrates is required in order to understand the role of hybridized dentin in improved dental treatment.

Effect of HEMA on bonding to dentin.

The present study investigated the effectiveness of treating dentin with 2-hydroxyethyl methacrylate (HEMA) prior to application of an adhesive resin. The adhesive resin was 5% 4-methacryloxyethyl trimellitate anhydride (4-META) in methyl methacrylate (MMA) combined with poly-MMA powder. Polymerization of this resin was initiated by tri-n-butyl borane (TBB). Bovine dentin samples were ground with 600-grit Carbimet paper discs, and demineralized with either an aqueous solution of 10% citric acid/3% ferric chloride (10-3) or an aqueous solution of 10% citric acid (10-0). Improved bond strengths were achieved with HEMA treatment of bovine dentin samples, and improvement of bond strengths was dependent upon the time period of HEMA application. Scanning electron microscope (SEM) examination revealed the formation of a transitional zone of resin-reinforced dentin, termed the "hybrid" layer, in those specimens receiving 10-3 pre-treatment. The adhesive resin impregnated the exposed collagen bundles with which it entangled to create the "hybrid", essential in the attainment of high tensile bond strengths. Specimens pre-treated with 10-0 did not readily form "hybrid" layers. However, if HEMA application followed the 10-0 pre-treatment, "hybrids" were demonstrated on SEM, and bond strength increased to 13 MPa. The ferric ions in the 10-3 effectively improved the diffusivity of dentinal substrates, as did HEMA. This study indicates that HEMA applied to dentinal substrates enhances monomer diffusion and entanglement with dentinal components, and facilitates the formation of a "hybrid" layer.