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Plasmatic antiproteinase activity enhancement by insoluble functionalized polystyrene surfaces.

Antithrombogenic functional polymer surfaces have been obtained by grafting heparin or by substituting insoluble polystyrene with sulphonate and/or amino acid sulphamide groups. Their heparin-like properties have been related to their catalytic effects on the antithrombin III - thrombin complex formation. Amongst these antithrombogenic surfaces, this study demonstrates that some insoluble amino acid sulphamide derivatives of polystyrene strongly potentiate heparin cofactor II, in addition to antithrombin III. In contrast, an insoluble polystyrene sulphonate and, to a lesser extent, an insoluble heparin copolymer, are better catalysts of antithrombin III. It is hypothesized that such different behaviours result from different conformations of the species adsorbed onto the surfaces. The conclusions support the possible use of such amino acid sulphamide groups to prepare antithrombogenic surfaces in contact with blood.

Antithrombin III↗

Regulation by sulphonate groups of complement activation induced by hydroxymethyl groups on polystyrene surfaces.

Reducing the complement-activating capacity of a polymer surface is important in improving its blood compatibility. Polystyrene surfaces bearing hydroxymethyl (CH2OH) groups activate the alternative pathway of complement. This activation depends strongly on the density of the groups. Polystyrene surfaces bearing sulphonate (SO3-) groups adsorb proteins, resulting in an apparent activation. Polystyrene surfaces bearing both types of groups in close proportions are not activators in human serum, due to the adsorption of a protein of the alternative pathway, which has a protecting effect, not found when a polymer surface bearing hydroxyl groups is mixed in serum with another polymer surface bearing SO3- groups. In the presence of purified proteins of alternative pathway, C3 convertase activity can be created on each of these surfaces by deposition of C3b, but their susceptibility to inactivation by regulatory proteins H and I depends on the types of chemical groups present on the surface and whether the surfaces were passivated or not before C3b deposition.

Adsorption↗

Specific interactions of polystyrene biomaterials with factor D of human complement.

The contact of blood with some biomaterials results in complement activation, primarily by the alternative pathway (AP). Insoluble polystyrene derivatives bearing isolated sulphonate groups (PSSO3) deplete complement, whereas identical surfaces substituted with both sulphonate and hydroxymethyl groups (PSCH2OH-SO3) are non-activators. Polystyrene sulphonate derivatives possess high adsorptive properties, particularly for serine proteases of the coagulation cascade. Thus, we studied the interactions between polystyrene derivatives and factor D, an enzyme essential for AP activation. C3 was activated when normal human serum (NHS) was incubated with PSSO3, whereas PSCH2OH-SO3 did not induce any specific C3 activation. Both polymers adsorbed factor D from serum, as shown by the loss of haemolytic factor D from NHS incubated with the polymers and by the specific adsorption of radiolabelled factor D. When bound to the polymers, factor D was not functional. The disappearance of factor D was in contradiction to the observed complement activation induced by PSSO3. When other AP components were studied, it was evident that PSSO3 adsorbed factor H even more rapidly and efficiently than factor D. Thus, the net effect was an immediate deregulation of the AP resulting in C3 activation, followed by inhibition of the AP when factor D was finally depleted. Pre-exposure of PSSO3 to NHS prevented any complement activation because the polymer was saturated with factor H, but still adsorbed factor D. Such properties could be beneficial during haemodialysis with membranes for uremic patients who have increased levels of factor D in their serum.

Biocompatible Materials↗

Global degranulation of rat mast cells stimulated with DNP-polystyrene.

Mediator release was studied in rat peritoneal mast cells sensitized with a mouse monoclonal anti-DNP IgE antibody, and stimulated with DNP-ornithine covalently attached to radio-derivatized polystyrene petri dishes. Cells releasing serotonin at maximal rates were investigated by transmission electron microscopy. Generalized exocytosis of granules could be observed, suggesting non-directional release of mediators, and non-compartmentalized action of second messengers in mast cells stimulated with polystyrene-bound DNP. Stimulation of sensitized mast cells by DNP covalently bound to the rigid polystyrene surface is consistent with extrinsic mechanisms proposed for Fc(epsilon)RI receptor action, and suggests that internalization of Fc(epsilon)RI is not needed for triggering cell degranulation.

Animals↗

Some reactions of influenza viruses adsorbed to polystyrene for enzyme immunoassay.

Three influenza viruses adsorbed to the surface of polystyrene tubes exhibited the property of haemadsorption, but only two of the neuraminidases were still active. Reactions of the viruses with rabbit antisera indicated that at least some of the virus particles were ruptured by adsorption to the polystyrene and the exposed nucleoprotein could react with antibody. Enzyme immunoassays involving direct adsorption of influenza virus particles to polystyrene are unlikely, therefore, to differentiate between strains of influenza A.

Adsorption↗

Migration of styrene from polystyrene foam food-contact articles.

Polystyrene foam is used in many food-contact articles such as plates, cups, bowls, egg cartons, meat trays and hinged 'carryout' containers. In most of these applications the food is in contact with the article for a relatively short period of time at mild temperatures (up to 130 degrees F), or for longer periods of time at refrigerated temperatures (40 degrees F). The extent of migration of residual styrene from foam articles under these conditions is of interest in order to predict potential exposure of consumers to styrene from food-contact polymers. Studies of styrene migration from all polystyrene foam articles except egg cartons were completed using food oil as the simulant. Results showed that the amount of styrene migrating from the various food-contact articles made with thermoformed polystyrene sheet into food oil was proportional to the square root of time of exposure. The mean diffusion coefficients derived from these data, assuming the migration was Fickian in nature, showed a linear relationship between the log of the diffusion coefficient and the inverse of the absolute temperature of exposure from 70 to 150 degrees F (21-66 degrees C). The mean diffusion coefficients ranged from about 4.5E-11 cm2/s at 70 degrees F to 3.4E-9 cm2/s at 150 degrees F. Among the four thermoformed articles exposed to food oil, the diffusion coefficients varied by a factor of four or less at a given temperature. The migration of styrene from egg cartons was examined using conditions simulating 'typical' exposure temperature and time with 8% ethanol as the simulant (31 days at 40 degrees F (4 degrees C)). No migration of styrene was observed under this condition, with a detection limit of < 0.01 micrograms/cm2.

Diffusion↗

Release properties on gelatin-gum arabic microcapsules containing camphor oil with added polystyrene.

In this study, gelatin blended with arabic gum microcapsules containing camphor oil with added polystyrene were fabricated by a compound coacervation method. The parameters of oil/wall volume ratio, emulsification stirring speed, concentration of cross-linking agent, treated time and oil release properties were investigated. In order to improve the constant release effect of camphor oil, oil-soluble polystyrene (PS) was used as a sustained release agent. The camphor oil release curves were expressed by the exponential equation: psi(t)=C(eq)(1-e(-t/tau)), where psi(t) represent the variant of camphor oil concentration in the operation environment, C(eq) as the equilibrium state, t as the release time and tau as time constant. C(eq) and tau are significant factors pertaining to the camphor oil release properties. The results indicated that, for the microcapsules, the optimal oil/wall volume ratio was 0.75 to achieve the encapsulation efficiency of 99.6 wt.%. The average particle size were 294.7+/-14.2 microm, 167.2+/-11.2 microm, 85.7+/-8.7 microm at the homogenization stirring speed of 500, 1000, and 2000 rpm, respectively. The effect of sustained oil release will increase whereas the stirring speed decreases and the concentration of glutaraldehyde (GA) and treated time increases. Along with the increasing of quantity of polystyrene added, C(eq) decreased and tau increased, indicating that the sustained oil release amount and the release rate depend on the quantity of PS considerably.

Biocompatible Materials↗

Morphological studies of oligodeoxyribonucleotides probes covalently immobilized at polystyrene modified surfaces.

The immobilization of short ss-DNA (18- and 36-mer) and their hybridization were studied at gold and glassy carbon substrates modified with low molecular weight (approximately 12, 18 and 24 kg/mol) polystyrene thin films. Amino-modified DNA was attached to the surface by reaction with succinimide ester groups bound to the polystyrenes. A ferrocene modified DNA target was used to confirm the probe-target hybridization. Atomic force microscopy studies showed significant morphological changes after probe immobilization and hybridization compared to the featureless structure of the polystyrene film. Single-stranded DNA samples had a globular morphology with an average density of 3.8 and 2.2 (x 10(11)) globules/cm2 for the 18- and 36-mer, respectively. The formation of a porous structure with a 2.0 and 1.0 (x10(11)) average pore density corresponding to the 18- and 36-mer was observed after hybridization. A surface composition analysis was done by X-ray photoelectron spectroscopy to confirm and support the images interpretation. Ferrocene oxidation (+323 mV/18-mer, +367 mV/36-mer, versus Ag/AgCl) proved the presence of ds-DNA at the modified surfaces.

Coated Materials, Biocompatible↗

Protein-protein interaction analysis using an affinity peptide tag and hydrophilic polystyrene plate.

A sandwich ELISA method using peptide tags showing a specific affinity to a hydrophilic polystyrene surface (PS-tags), PS 19 composed of RAFIASRRIKRP and KPS19R10 of KRAFIASRRIRRP and a hydrophilic polystyrene (phi-PS) plate was used to analyze protein-protein interactions. An Escherichia coli cysteine synthase complex, in which serine acetyltransferase (SAT) interacts with O-acetylserine sulfhydrylase-A (OASS) was used as a model system. When the interaction was detected by the conventional sandwich ELISA method using a hydrophobic polystyrene (pho-PS) plate, for the exclusive use of ELISA, the signal intensity was barely detectable due to conformational change of the ligand protein, OASS in the adsorbed state. On the contrary, when OASS, genetically fused with PS19 (OASS-PS19) or chemically conjugated with KPS19R10 (OASS-KPS19R10), was immobilized on the phi-PS plate, a high signal intensity was detected. Furthermore, by applying the two-step sandwich ELISA, in which OASS-PS19 or OASS-KPS19R10 formed a complex with SAT in the blocking solution before immobilization on the phi-PS plate, the signal intensity was further increased with a much shorter operational time, because SAT in the blocking solution formed a complex with OASS-PS19 or OASS-KPS19R10 without any steric hindrance.

Enzyme-Linked Immunosorbent Assay↗

Improved cellular adhesion to acetone plasma modified polystyrene surfaces.

The plasma polymerization of acetone has been used to modify polystyrene substrates for the controlled growth of human fibroblast cells. The surface modified polystyrene was studied by X-ray photoelectron spectroscopy, water contact angle and atomic force microscopy. This showed the surface oxygen levels and wettability to increase rapidly with exposure to the acetone plasma. High-resolution XPS allowed the determination of the relative amounts of surface hydroxyl, carbonyl and carboxyl groups. This showed that there was little incorporation of carboxyl groups in the deposited films. AFM measurements revealed the films to be conformal with a surface roughness equivalent to that of the underlying polystyrene substrate with film growth rates of approximately 0.5 nm min(-1). High edge-definition patterns were produced with a simple masking procedure and allowed the confinement of cells to selected areas of the substrate. These chemically patterned surfaces allowed the study of cells confined to particular regions of the substrate as a function of incubation time.

Acetone↗

Molecular-scale observation of the surface of polystyrene particles by AFM.

The surface morphology of polystyrene particles prepared by soap-free emulsion polymerization was observed in situ using the tapping mode of an atomic force microscope (AFM). Here, the use of a cationic initiator enabled us to synthesize positively charged polystyrene, which then adsorbed electrostatically onto the negatively charged mica plate with molecular-scale smoothness. The following was found from AFM measurements in water. The surface of polystyrene particles changed from smooth to rough as the polymerization proceeded in the experimental condition, where the bulk generated tiny particles continuously, because of the heterocoagulation in the growth process between the newborn and tiny particles and the existing particles. However, it is easy to prepare the particles with smooth surfaces on the condition of rich monomers because the monomers in the bulk dissolve the polymers which form the surface morphology to be deformable and reduce the interfacial area.

Adsorption↗

Responses of peptide-specific T cells to stimulation with polystyrene beads carrying HLA class I molecules loaded with single peptides.

Cell-sized microbeads carrying single peptide-loaded HLA class I molecules were prepared for HLA-A2 and HLA-B7 by a simple procedure which transfers single peptide-loaded HLA class I molecules from cultured cells to polystyrene beads using anti-peptide antibodies directed to an intracellular segment of HLA-A alpha chains. The surface density of peptide-loaded HLA class I molecules on beads was comparable to that on the peptide-loaded cells. HLA-A2 beads loaded with an HCV peptide HCV1073 were tested for stimulation activity on an HCV1073-specific CD8+ T cell clone NS3-1. A substantial level of gamma-IFN production was induced. The stimulation was peptide-specific. The efficiency was dependent on the bead concentration and the surface HLA class I density on beads and enhanced significantly by co-coupling of anti-CD28 to peptide-loaded beads. The peptide-loading efficiency on HLA class I molecules and the transfer efficiency of HLA class I molecules to polystyrene beads were reasonably high for HLA-A2 and HLA-B7. Thus, polystyrene beads carrying these single peptide-loaded HLA class I molecules are potentially useful in further analysis of the co-stimulatory or inhibitory factors involved in CD8+ T cell responses and eventually in detection of cytotoxic T cells in PBLs.

Amino Acid Sequence↗

Vibrational spectroscopic encoding of polystyrene-based resin beads: converting the encoding peaks into barcodes.

A detailed approach is described for the vibrational spectroscopic encoding of polystyrene-based resin beads by converting the infrared absorption peaks suitable for encoding (encoding peaks) into barcodes. Based on combining the FT-IR measurements and the quantum-chemical computations, the vibrational characteristics of p-tert-butylstyrene monomer, polystyrene and poly(p-tert-butylstyrene) resin beads are analyzed, which are helpful for the selection of encoding peaks. The vibrational spectroscopic encoding of polystyrene-based resin beads could be obtained by converting the wavenumber, intensity and full width at half maximum (FWHM) of the encoding peaks into barcodes automatically through a computer program designed in our laboratory.

Electronic Data Processing↗

Laboratory and field comparisons of pyriproxyfen, polystyrene beads and other larvicidal methods against malaria vectors in Sri Lanka.

Hand-dug gem pits are important breeding sites for larvae of malaria vectors in Sri Lanka. Therefore, studies were carried out to help to select an effective, economic and convenient method that could be used to control malaria vector mosquito breeding in gem pits in a mining area. The effectiveness of four types of floating layers of polystyrene was compared in the laboratory and it was found that 2 mm expanded beads were the most effective for suffocating Anopheles larvae and pupae. The insect growth regulator, pyriproxyfen at dosages of 0.01 and 0.1 mg/l were tested in the laboratory and complete inhibition of emergence was found at both concentrations. A small-scale field trial was carried out for over a year to assess the efficacy of two concentrations of pyriproxyfen, 2 mm diameter expanded polystyrene beads, temephos, used engine oil and filling pits with soil. Pyriproxyfen only required re-application twice a year, whereas temephos or oil require 12 applications per year. Due to re-excavation by gem miners, polystyrene beads and filling of pits were not as permanent solutions as was expected. Calculations based on all available data showed that two annual treatments with pyriproxyfen at 0.01 mg/l would be the most cost-effective method with oil only slightly more expensive. However, the reduced required frequency for visiting every pit made the pyriproxyfen method the one of choice. The same low concentration of pyriproxyfen also effectively inhibited emergence of adults from river-bed pools.

Animals↗

Heparinization of gas plasma-modified polystyrene surfaces and the interactions of these surfaces with proteins studied with surface plasmon resonance.

Polystyrene surfaces obtained by spin-coating a solution of polystyrene in toluene on a gold layer were functionalized with carboxylic acid groups by preadsorption of the sodium salt of undecylenic acid, followed by an argon plasma treatment. A conjugate of albumin and heparin (alb-hep) was covalently immobilized onto the functionalized surface via preactivation of carboxylic acid groups with a water-soluble carbodiimide. The immobilization of alb-hep conjugate and the subsequent interactions of the heparinized surface with antithrombin III (ATIII, a heparin cofactor) and thrombin were monitored with surface plasmon resonance (SPR). The surface concentration of conjugate as determined with SPR deviated quantitatively from the results obtained with radiolabelled conjugate. The difference in surface concentrations of conjugate obtained with the two methods probably originates from the uncertainty of the refractive index of the alb-hep conjugate in the SPR technique. ATIII could be bound to the surface modified with alb-hep conjugate but not to a polystyrene surface modified with albumin. Rabbit anti-human ATIII did bind to the alb-hep surface previously exposed to ATIII, confirming the presence of surface bound ATIII. The alb-hep immobilized surface was able to bind much more thrombin than ATIII, which is probably due to the less specific heparin-thrombin interaction as compared to the heparin-ATIII interaction. This study shows that SPR is a technique that can be used to study, in real time, both the modification of polymer surfaces and the subsequent interactions of the modified surfaces with proteins.

Albumins↗

Biospecific interactions of vitamin K-dependent factors with phospholipid-like polystyrene derivatives. Part II: factor IX.

We previously demonstrated that phosphorylated polystyrene derivatives exhibit phospholipid-like behaviour and therefore are able to interact with factor II, one of the vitamin K-dependent coagulation factors. Under the same conditions as for factor II, we examined the interactions of factor IX with phosphorylated resins of various compositions in phosphate groups: these studies were carried out with or without albumin precoating of the polymers and either in the presence or absence of calcium ions. Adsorption experiments show that, in the absence of calcium ions, only one class of adsorption sites of factor IX can be evidenced with the interactions taking place through the formation of binary complexes, whereas in the presence of calcium ions, the affinity of factor IX for phosphorylated resins becomes very high and two types of adsorption sites have been evidenced with biospecific ternary complexes being formed. The domains of predominance of these complexes were determined. Moreover, the only functional groups borne by the phosphorylated polystyrene resins involved in factor IX-polymer interactions are phosphodiester groups. Comparison between factor II and factor IX adsorption onto the same polymers leads to the conclusion that the observed differences probably reflect the differences in the Gla domains of the vitamin K-dependent factors. Finally, this study demonstrates that phosphorylated polystyrene derivatives can be used as stationary phases for purification of factor IX by highly specific liquid biochromatography.

Adsorption↗

Prolonging the circulation time and modifying the body distribution of intravenously injected polystyrene nanospheres by prior intravenous administration of poloxamine-908. A 'hepatic-blockade' event or manipulation of nanosphere surface in vivo?

Intravenously injected uncoated small (60 nm) and large (250 nm) size model polystyrene particles (which are cleared rapidly from the blood by macrophages of the reticuloendothelial system) can be converted to long-circulatory and splenotropic particles in vivo, respectively, if such particles are injected shortly (up to 3 h) after an appropriate dose of the block polymer non-ionic surfactant, poloxamine-908. Evidence indicates that small and large size polystyrene beads can acquire a coating of poloxamine-908 and/or poloxamine-protein complexes in vivo. The adsorption of such complexes on to the bead surface could explain their altered body distribution since small and large size polystyrene beads that were precoated with poloxamine-908 exhibit similar biodistribution following i.v. injection.

Drug Carriers↗

Fractionation of functional polystyrenes, poly(ethylene oxide)s and poly(styrene)-b-poly(ethylene oxide) by liquid chromatography at the exclusion-adsorption transition point.

The paper reports the fractionation of functional polystyrenes (PSs) and poly(ethylene oxide)s (PEOs) as well as their block copolymers, by liquid chromatography at the exclusion adsorption transition point (EATP-LC), also called "critical conditions" mode. In this specific elution mode (EATP-LC), the fractionation is only governed by the nature and the number of functions attached to the polymer backbone, independent of the molar mass distribution of the whole sample. Functional polystyrenes (alpha- and/or alpha,omega-alcohol-, acetal-, aldehyde- and acidic-PS) could be readily separated from non-functional polystyrenes under various chromatographic conditions. The technique also allowed the fractionation of poly(ethylene oxide)s and PS-PEO block copolymers. In the latter cases, moderately polar columns (grafted silica) and water-based polar eluents were required to obtain a satisfactory fractionation.

Adsorption↗