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Electrogenerated chemiluminescence from polymer-bound ortho-metallated iridium(III) systems.

Three ortho-metallated iridium complexes whose emission maxima fall in different regions of the electromagnetic spectrum were bound in either Nafion or poly(9-vinylcarbazole) and their electrogenerated chemiluminescence (ECL) reported. The reaction of F(Ir)pic [bis(3,5-difluoro-2-(2-pyridyl)phenyl-(2-carboxypyridyl)-iridium III] with the oxidative-reductive co-reactant tri-n-propylamine (TPrA) resulted in ECL when the iridium complex was bound in Nafion. No significant ECL was observed for (btp)(2)Ir(acac) (bis[2,(2'-benzothienyl)-pyridinato-N,C3'](acetylacetonate)Ir(III)), and Ir(ppy)(3) (where ppy = 2-phenylpyridine) under these conditions. However, all three compounds displayed ECL with TPrA when bound in poly(9-vinylcarbazole).

Cations↗

Solubilization of carbon nanotubes by Nafion toward the preparation of amperometric biosensors.

The ability to solubilize single-wall and multiwall carbon nanotubes (CNT) in the presence of the perfluorinated polymer Nafion is described. Such use of Nafion as a solubilizing agent for CNT overcomes a major obstacle for creating CNT-based biosensing devices. Their association with Nafion does not impair the electrocatalytic properties of CNT. The resulting CNT/Nafion modified glassy-carbon electrodes exhibit a strong and stable electrocatalytic response toward hydrogen peroxide. The marked acceleration of the hydrogen peroxide redox process is very attractive for the operation of oxidase-based amperometric biosensors, as illustrated for the highly selective low-potential (-0.05 V vs Ag/AgCl) biosensing of glucose. These findings open the door for using CNT in a wide range of chemical sensors and nanoscale electronic devices.

Biosensing Techniques↗

Microfilters for biomedicine obtained by swift ion processing.

Swift ions with 50 MeV energy are used to irradiate bio-polymeric films. Lithium, carbon and oxygen beams, with fluencies ranging between 10(8) and 10(13) ions/cm2, are swept on large surfaces of hydrogenated and fluorated polymers. The electronic stopping power of the ion induces high rates of ionization, excitation and radical formation along the ion path with production of chain-scission and cross-linking processes which can be investigated by the molecular emission from the irradiated polymer. The radiation damage increases the chemical solubility of the material around the ion track up to distances of the order of 100 nm. A suitable chemical etching removes the damaged material leaving micrometric holes in the polymeric film. This process gives rise to micro-perforated films which can be used as microfilters, with sub-micrometric pore dimension, high selectivity and controllable porosity. Their use is very interesting for special applications in ultra- and microfiltration of biological liquids.

Biocompatible Materials↗

Identification of a protein-tyrosine phosphatase from human platelet membranes by an immobilon-based solid phase assay.

The present investigations show that a 53-kDa platelet-membrane protein is a protein tyrosine phosphatase. Identification involved a novel methodology in which membrane proteins are resolved by polyacrylamide gel electrophoresis and then transblotted to a polyvinylidene difluoride membrane surface-labeled with [32P-Tyr](Glu4.Tyr1)n. Phosphatase activity appears as clear areas in the autoradiograph of the renatured 32P Western blot. Studies using this new solid-phase system indicate that a 53-kDa platelet-membrane protein dephosphorylates [32P-Tyr](Glu4.Tyr1)n. Dephosphorylation is both time- and dose-dependent. The protein-tyrosine phosphatase antagonists orthovanadate and molybdate block dephosphorylation in a concentration-dependent manner. Inhibitors of protein serine/threonine phosphatases and of acid and alkaline phosphatases do not significantly affect enzymatic activity. The enzyme is active toward phosphotyrosyl proteins but not phosphoseryl proteins. Results from the solid-phase phosphatase assays correlate with data from standard liquid-phase studies. A Lineweaver-Burk plot gives an apparent Km of 4.3 microM and an apparent Vmax = 168 nmol P(i) transferred/min/mg for the membrane enzyme towards the 32P-labeled polymer in a liquid system. Platelet lysate was shown by the solid-phase assay to possess a protein-tyrosine phosphatase of approximately equal to 50 kDa. The tyrosine phosphatase activities associated with a placental preparation, pure recombinant protein-tyrosine phosphatase 1B, and YOPR, the product of the YOP51 gene of Yersinia enterocolitica, which contains the C235R mutation, were also visualized by this technique. However, the catalytic activity of domain 1 of the transmembrane leukocyte antigen-related protein-tyrosine phosphatase was not detected using the solid-phase assay.

Amino Acid Sequence↗

Cell responses to biomaterials. II: Endothelial cell adhesion and growth on perfluorosulfonic acid.

We report here the use of perfluorosulfonic acid (Nafion) as a substratum for the growth of bovine aortal endothelial cells. This support which can be generated in a number of forms is at least as efficient in maintaining the growth of endothelial and other cell types as tissue culture grade polystyrene (TCP) and represents an advance in this regard over polytetrafluoroethylene (Teflon). The mechanism underlying the different cell attachment capacities of these three polymers is not readily related to their different protein binding patterns. While Nafion adsorbs more total protein from serum than Teflon or TCP, it adsorbs relatively less of the major cell adhesive proteins, vitronectin and fibronectin, than does Teflon. Both Nafion and Teflon had comparable but low thrombogenic potential by in vitro tests. Teflon or expanded Teflon (Gore-tex) coated with a thin film of Nafion assumes the cell supportive characteristics of Nafion and hence the modification of these surfaces by the induction of a stable bond between Teflon (in various forms) and Nafion may provide a composite vascular graft material which has all the desirable qualities of both materials.

Animals↗

Frequency response of PVDF needle-type hydrophones.

This paper examines the factors governing the frequency response of ultrasonic polyvinylidene fluoride (PVDF) polymer needle-type hydrophones, in particular the sensitivity variations in the lower frequency range of 1-6 MHz. A theoretical model was used to analyze the influence of the hydrophone's diameter, the metal electrodes, thickness, PVDF material properties, the adhesive layer acoustical characteristics and the backing material, on the frequency response of the hydrophone. The results of the theoretical modelling differ by less than +/- 0.5 dB from those obtained experimentally from the reciprocity calibration in the frequency range 1-20 MHz. It is shown that the needle hydrophone's diameter and backing material are the main reasons for the sensitivity variations observed in the frequency range below 6 MHz.

Acoustics↗

Metal ion reduction and resultant deposition on viologen-functionalized LDPE films and viologen-containing microporous membranes.

Photo-induced reduction of gold and platinum metal salt solutions was carried out using viologen graft copolymerized on low-density polyethylene (LDPE) films and viologen-containing poly(vinylidene fluoride) (PVDF-PVBV) microporous membranes. The effects of the UV irradiation time and concentration of the metal salt solutions on the metal ion reduction process and the resultant metal deposition on the polymeric substrates were investigated. The metal-polymer composites were characterized using X-ray photoelectron spectroscopy (XPS), scanning and transmission electron microscopy (SEM and TEM), atomic force microscopy (AFM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and UV-visible absorption spectroscopy. The amount of metal uptake, the state of the metal, and the size of the metal particles were found to be strongly dependent on the UV irradiation time and the type and concentration of the metal salt solution. The microporous structure and the high viologen content of the PVDF-PVBV membrane constitute an effective matrix for metal ion reduction and preparation of metal nanoparticles.

Gold↗

Human monocyte adhesion and activation on crystalline polymers with different morphology and wettability in vitro.

This study evaluated the effects of crystalline polyamide (Nylon-66), poly(ethylene-co-vinyl alcohol) (PEVA), and poly(vinylidene fluoride) (PVDF) polymers with nonporous and porous morphologies on the ability of monocytes to adhere and subsequently activate to produce IL-1beta, IL-6, and tumor necrosis factor alpha. The results indicated monocyte adhesion and activation on a material might differ to a great extent, depending on the surface morphology and wettability. As the polymer wettability increases, the ability of monocytes to adhere increases but the ability to produce cytokines decreases. Similarly, these polymers, when prepared with porous surfaces, enhance monocyte adhesion but suppress monocyte release of cytokines. Therefore, the hydrophobic PVDF with a nonporous surface stimulates the most activity in adherent monocytes but shows the greatest inhibition of monocyte adhesion when compared with all of the other membranes. In contrast, the hydrophilic Nylon-66, which has a porous surface, is a relatively better substrate for this work. Therefore, monocyte behavior on a biomaterial may be influenced by a specific surface property. Based on this result, we propose that monocyte adhesion is regulated by a different mechanism than monocyte activation. Consequently, the generation of cytokines by monocytes is not proportional to the number of cells adherent to the surface.

Adult↗

In vitro and short-term in vivo characteristics of a Kel-F thin film modified glucose sensor.

A new outer layer composition, consisting of polytetrafluoroethylene (PTFE), Kel-F oil, and Nafion, is suggested to minimize the detrimental effect of dissolved oxygen and to extend the linear response range of a glucose oxidase(GOx)-based sensor using nonconducting polymer. The morphology of Kel-F/PTFE/Kel-F/Nafion polymeric laminate was followed during fabrication by SEM. When Kel-F film was formed on the PTFE outer layer, the linear response was extended to 21 mM, at a sensitivity of 2.8 +/- 0.8 nA/mM mm2. We demonstrate that a sensor without Kel-F/PTFE/Kel-F/Nafion outer layer is relatively oxygen dependent, whereas by comparison a sensor with Kel-F/PTFE/Kel-F/Nafion outer layer is oxygen independent. The current of such a glucose sensor implanted in the subcutaneous tissue stabilized within 60 min, and the lag between blood glucose changes and sensor output was within 1 min. The in vivo characteristics of the glucose sensor described show great promise for one-point in vivo calibration.

Animals↗

Removal of methyl tert-butyl ether (MTBE) with Nafion.

A solid organic polymer, Nafion, is tested for the removal of methyl tert-butyl ether (MTBE) in water. Nafion with perfluorosulfonic acid backbone and terminal sulfonic acid groups has a surface acidity similar to 100% sulfuric acid, and has been commonly used as a strong-acid catalyst in many organic reactions. Sorption and subsequent transformation of MTBE were observed in batch experiments. The transformation of MTBE by porous nanocomposite Nafion SAC-13 to tert-butyl alcohol (TBA), acetone, isobutene and probably methanol was found. Subsequent transformation of TBA to acetone was also observed. Results suggest that transformational pathways may include hydrolysis, dehydrogenation and oxidation. Dissolved oxygen is needed for the oxidation of isobutene to acetone. As Nafion is insoluble in water, chemically stable, and regenerable, its use in packed-bed reactors for MTBE removal looks promising.

Acetone↗

Narrowband impedance matching layer for high efficiency thickness mode ultrasonic transducers.

A new matching layer design concept has been proposed for narrowband continuous wave (CW) devices. Analysis has shown that the mechanical impedance of a resonant-type transducer in thickness mode CW operation does not equal its acoustic impedance rhoVs but roughly equals rhoVs/Q, where p is density, Vs is acoustic velocity, and Q is the mechanical quality factor. The value of rhoVs/Q is much lower than the acoustic impedance of water for any transducer material, including lead zirconium titanate (PZT), single crystals, or polyvinylidene fluoride (PVDF). With this new approach, the impedance of the matching layer must also be between water and pVs/Q, but there are few such practical low impedance materials. To realize equivalent low impedance structure, a novel double layer design is presented: a relatively low impedance material (such as polyethylene or polyurethane) on the inside and a relatively high impedance material (such as polyester or metal) on the outside. A high power CW transducer structure was designed and fabricated with PVDF-TrFE (polyvinylidene fluoride trifluoroethylene) to operate at 1.4 MHz. The basic quarter wavelength resonator structure is 0.7-mm alumina/0.2-mm piezo-polymer/0.25-mm polyester, and the matching section is 0.2-mm polyurethane and 0.25-mm polyester. A maximum power output of 6 to 9 W/cm2 with conversion efficiency of 30 to 35% was observed. For the transducer without matching section, the observed power was 3 to 4 W/cm2. Mason's model analyses (1) predict that the traditional matching layer is for broadband purposes and reduces output power both for PZT and PVDF-TrFE (2); this new matching scheme can be applied to PZT high power transducer. This high efficiency technique has application in various CW systems, such as Doppler sensors, interferometry, phase-sensitive imaging, or high energy focused beam systems.

Aluminum↗

A thin-layer electrochemical detector coated with nafion film for liquid chromatography.

The selectivity and analytical application of a thin-layer electrochemical detector comprised of glassy carbon electrode coated with Nafion film were investigated. As a result of the ion-exchange characteristics of the Nafion polymer, the selectivity and stability were improved greatly. The coated electrode has a good response only for cations with the same sensitivity as an uncoated (bare) electrode, but not for anions, and the response for neutral molecules is decreased three-fold. The diffusion of electroactive compounds in Nafion film is discussed based on the results of flow injection experiments. The peak current at the coated electrode was independent of the flow-rate of the mobile phase. Electrode poisoning due to protein adsorption was minimized. The use of 30% methanol or 10% acetonitrile in the mobile phase did not affect the performance of the coated electrode. Various analytes having three kinds of charge state, i.e., anionic, cationic and neutral, were tested. Liquid chromatography with electrochemical detection of ascorbic acid, norepinephrine, epinephrine, dopamine and uric acid was demonstrated.

Animals↗

Cast thin film biosensor design based on a Nafion backbone, a multiwalled carbon nanotube conduit, and a glucose oxidase function.

Novel electroanalytical sensing nanobiocomposite materials are reported. These materials are prepared by mixing multiwalled carbon nanotubes (MWNTs), a Nafion cation exchanger, and glucose oxidase (GOD) in appropriate amounts. The MWNTs are cylindrical with a diameter in the range 40-60 nm and with a length of up to several micrometers, and they provide electrical conductivity. Nafion acts as a polymer backbone to give stable and homogeneous cast thin films. Both MWNTs and Nafion provide negative functionalities to bind to positively charged redox enzymes such as glucose oxidase. The resulting biosensing composite material is inexpensive, reliable, and easy to use. The homogeneity of the MWNT-Nafion-GOD nanobiocomposite films was characterized by atomic force microscopy (AFM). Amperometric transducers fabricated with these materials were characterized electrochemically using cyclic voltammetry and amperometry in the presence of hydrogen peroxide and in the presence of glucose. Their linear response to hydrogen peroxide was demonstrated. The glucose biosensor sensitivity was strongly influenced by the glucose oxidase concentration within the nanobiocomposite film. The optimized glucose biosensor (2.5 mg/mL GOD) displayed a sensitivity of 330 nA/mM, a linear range of up to 2 mM, a detection limit of 4 microM, and a response time of <3 s.

Biosensing Techniques↗

Piezoelectric immunoassay for complement C4 based on a Nafion-modified interface for antibody immobilization.

A piezoelectric immunosensor based on a Nafion membrane interface with a simple immobilization procedure has been developed for the determination of complement C4 in human serum. The polyanionic perfluorosulfonated Nafion polymer was used to modify the electrode surface of quartz-crystal microbalance (QCM) as a platform for the immobilization of complement C4 antibodies. The surface morphology of QCM modified with Nafion membrane was investigated using scanning electron microscopy. The system was optimized with regard to the parameters involved in the preparation of the immunosensor and the assay process. The proposed immunosensor responds well to C4 in the range of 0.08-1.6 microg/ml with a relative standard deviation of <5.3%. Moreover, the proposed immunosensor has advantages in terms of the speed and simplicity of the immobilization procedure as well as the simple and advantageous regeneration process. The experimental results obtained with regard to nonspecific adsorption and recovery indicate that the proposed immunosensor offers a promising alternative tool for the clinical diagnosis of complement C4.

Antibodies↗

Enantioselective screen-printed amperometric biosensor for the determination of D-amino acids.

D-amino acids are generally considered to be important markers of bacterial contamination of food products. A screen-printed amperometric biosensor for the detection of D-amino acids has been constructed by the immobilization of D-amino acid oxidase on a graphite working electrode of a screen-printed strip modified with Prussian Blue and Nafion layers. Enzyme immobilization was then carried out by cross-linking of a mixture of the enzyme and bovine serum albumin with glutaraldehyde. As a result of the mediator addition and because of the multi-layer construction of the biosensor, including a polymer layer to avoid the interferences, the limit of the detection of the developed biosensor was two orders of magnitude improved in comparison to other screen-printed biosensors, as far as the determination of amino acids is concerned. Additional modification of the graphite electrode with carbon nanotubes led to a significant enhancement of the signal magnitude. A fast linear response of the developed biosensor was subsequently observed in static measurements for D-alanine in the concentration range from 5 to 200 microM. Excellent enantioselectivity towards D-amino acids was discovered. During the experiment, D-amino acids were detected in fruit juices and some milk samples. The complex matrix of natural milk samples had no influence on the response of the biosensor. The results were in good agreement with those obtained by capillary electrophoresis measurements.

Alanine↗

Prevention of the rapid degradation of subcutaneously implanted Ag/AgCl reference electrodes using polymer coatings.

To assess the effect of the biological response to implanted Ag/AgCl reference electrodes on the electrode stability, uncoated and polymer-coated Ag/AgCl electrodes were implanted subcutaneously in rats. After 1 week of implantation, uncoated Ag/AgCl electrode potentials, measured in 0.1 M KCl, shifted by about -180 mV, and both voltammetry and electron microscopy showed that all the AgCl was removed. The electrodes could be significantly protected by coating with polyurethane or a perfluorinated ionomer (Nafion) cured at 120 degrees C for 1 h. Electron micrographs showed the 120 degrees C cured Nafion and polyurethane coatings remained intact over 2 weeks of implantation. Following 2 weeks of implantation the cured, Nafion-coated electrodes' potentials were shifted by -15 +/- 7 mV relative to the initial values. Voltammetry showed that they were still not polarizable. The current densities obtained with the coated reference electrodes are sufficient for their use as counter/pseudoreference electrodes with implantable two-electrode glucose sensor systems. The tissue response to coated electrodes was minimal in comparison to the response to uncoated reference electrodes.

Animals↗

Improving the environment for immobilized dehydrogenase enzymes by modifying Nafion with tetraalkylammonium bromides.

Recent research in our group has shown that mixture-casting Nafion with quaternary ammonium bromides can increase the electrochemical flux of redox couples through the membrane and allow for larger redox species to diffuse to the electrode surface. The research has also suggested that when these salts are cast with Nafion micellar pore size is changing. Therefore, it was proposed that the quaternary ammonium salts could be employed to tailor the structure of the Nafion membrane for immobilizing enzymes in the polymer. For cations with a high affinity for the sulfonic acid groups of Nafion, the modified structure of Nafion can also help to stabilize the enzyme and increase activity by providing a protective outer shell and an ideal chemical environment that resists a decrease in pH within the pore structure. This research examines the ability to immobilize dehydrogenase enzymes in Nafion that has been modified with quaternary ammonium bromides. Fluorescence assays, fluorescence microscopy, and cyclic voltammetric studies were employed to analyze the ability to immobilize an enzyme within the membrane, to determine the activity of the immobilized enzyme and to examine the transport of coenzyme within the membrane. Dehydrogenase enzymes immobilized in tetrabutylammonium bromide/Nafion membranes have shown high catalytic activity and enzyme active lifetimes of greater than 45 days. A variety of dehydrogenase enzymes have been successfully immobilized in the membrane, including: alcohol dehydrogenase, aldehyde dehydrogenase, glucose dehydrogenase, and lactic dehydrogenase.

Alcohol Dehydrogenase↗

Why make monofilament sutures out of polyvinylidene fluoride?

In recent years some clinical reports have associated suture failures with polypropylene monofilaments. Therefore there is interest in developing an alternative suture material that is less thrombogenic than polyester and similar in handling characteristics but less prone to mechanical failure than polypropylene. To this end, Peters Laboratoire Pharmaceutique has developed a new monofilament suture material from polyvinylidene fluoride (PVDF), which has been subjected to a special treatment to modify its crystalline form and level of crystallinity. The purpose of this study was to evaluate its mechanical, chemical, and biologic properties and to compare its performance, in a peripheral vascular application, to that of a polypropylene control. A series of in vitro tests were performed to study the morphology, tensile properties, creep, surface chemistry, thermal characteristics, and resistance to iatrogenic trauma. In addition, an in vivo trial was undertaken in which vascular prostheses anastomosed with either PVDF or polypropylene sutures were implanted as a thoracoabdominal bypass for 6 months in the dog. Histologic and degradation analyses were performed on the explants. The results from the mechanical tests on 4-0, 5-0, and 6-0 PVDF and polypropylene sutures demonstrated that although both materials have similar breaking strengths, the PVDF has a higher extension at break, has less delayed extension when under tensile creep testing, and suffers less trauma than the polypropylene when compressed by a standard needle holder. While chemical analyses found evidence of surface oxidation on both types of sutures, thermal analysis confirmed that the level of crystallinity of the PVDF polymer is higher than that of the polypropylene control. During the pilot study in animals, PVDF sutures were found to have good handling and frictional characteristics that facilitated the tying of knots. Histologic analysis of the explants found no inflammatory cells in the tissue surrounding either the PVDF or polypropylene sutures, and scanning electron microscopic examination of the cleaned suture surfaces found no evidence of degradation during 6 months in vivo. Though preliminary in nature, these findings indicate that monofilament sutures made from PVDF provide an attractive alternative to those made from polypropylene for use in cardiovascular surgery. In addition to providing acceptable in vivo behavior and being easy to manipulate and more resistant to iatrogenic injury, PVDF materials can be sterilized by beta or gamma radiation and so can reduce dependence upon ethylene oxide and chlorofluorohydrocarbons.

Animals↗