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The effect of chitosan and PVDF substrates on the behavior of embryonic rat cerebral cortical stem cells.

In this study, the behavior of neural stem cells from embryonic rat cerebral cortex were compared on the chitosan and poly(vinylidene fluoride) (PVDF) substrates at single-cell and neurosphere level. It was found that chitosan and PVDF substrates inhibited the proliferation and differentiation of single neural stem cells. It seemed that single-cell cultures on both substrates show cells remained dormant. However, neurospheres could exhibit different or similar behavior on these two substrates, which is dependent on the presence or absence of serum. More cells migrated outside from the neurospheres and longer processes extended from differentiated cells on chitosan than on PVDF when neurospheres were cultured in the serum-free medium. On the contrary, when serum was added to the culture system, chitosan and PVDF could induce the neurosphere-forming cells into an extensive cellular substratum of protoplasmic cells upon which process-bearing cells spread. In addition, based on the immunocytochemical analysis, the percentages of differentiated cell phenotypes of neurospheres cultured on chitosan and PVDF substrates became similar in the presence of serum. Therefore, it is reasonable to suggest that biomaterials may stimulate or inhibit the proliferation and differentiation of neural stem cells according to the complex environmental conditions. The information presented here should be useful for the development of biomaterials to regulate the preservation, proliferation, and differentiation of neural stem cells.

Animals↗

Urea biosensor based on PANi(urease)-Nafion/Au composite electrode.

The polyaniline (PANi)-Nafion composite film was prepared onto the ceramic plate by the cyclic voltammetry (CV) method with the various cycle numbers. When the PANi-Nafion/Au/ceramic plate with the preparing cycle number of 5 was as working electrode, the cathodic peak current was achieved as 84.0 microA in 60 mg dl(-1) NH4Cl buffer solution. On the other hand, the small cathodic peak currents for buffer solution in the presence of 60 mg dl(-1) LiOH, NaCl and KCl, respectively, were found with the same composite electrode as working electrode. The cathodic peak current decreased from 84.0 to 16.3 microA in the 60 mg dl(-1) NH4Cl buffer solution when the cycle number for preparing PANi-Nafion/Au/ceramic plate composite electrode with the CV method increased from 5 to 15. The enzyme of urease was immobilized onto the PANi-Nafion/Au/ceramic plate composite film by the electrochemical immobilization and the casting methods and used as sensing electrode to detect the concentration of urea in the buffer solution. The sensitivity of composite electrode immobilized with the casting method was greater than that of electrochemical immobilization method. The sensitivity and the detecting limit of the urea sensor were found to be 0.7 and 5.27 microA (mg dl(-1))(-1)cm(-2), as well as 6 and 0.3 mg dl(-1), respectively, when urease was immobilized by glutaraldehyde (GA) cross-linker and Nafion network, respectively.

Biosensing Techniques↗

A glucose biosensor based on electrodeposition of palladium nanoparticles and glucose oxidase onto Nafion-solubilized carbon nanotube electrode.

Electrodeposition was used for the co-deposition of glucose oxidase (GOx) enzymes and palladium nanoparticles onto a Nafion-solubilized carbon nanotube (CNT) film. The co-deposited Pd-GOx-Nafion CNT bioelectrode retains its biocatalytic activity and offers an efficient oxidation and reduction of the enzymatically liberated H2O2, allowing for fast and sensitive glucose quantification. The combination of Pd-GOx electrodeposition with Nafion-solubilized CNTs enhances the storage time and performance of the sensor. An extra Nafion coating was used to eliminate common interferents such as uric and ascorbic acids. The fabricated Pd-GOx-Nafion CNT glucose biosensor exhibits a linear response up to 12 mM glucose and a detection limit of 0.15 mM (S/N = 3).

Biosensing Techniques↗

A novel nitric oxide biosensor based on electropolymerization poly(toluidine blue) film electrode and its application to nitric oxide released in liver homogenate.

A novel poly(toluidine blue)-modified electrode has been constructed for the determination of nitric oxide in biological sample. The electrochemical behavior of poly(toluidine blue) film electrode and its electrocatalytic activity toward NO were studied in detail by cyclic voltammetry. Possible interferences were tested and evaluated after further coated with Nafion. The poly(toluidine blue) and Nafion composite film-modified electrode exhibits a good linear relationship over a NO concentration of 1.8 x 10(-7) to 8.6 x 10(-5)mol/L, and the detection limit is 1.8 x 10(-8)mol/L (S/N=3). NO release from the rat liver homogenate stimulated by l-arginine was studied, and the responses were decreased by the nitric oxide synthase inhibitor N(omega)-nitro-l-arginine.

Animals↗

Formaldehyde assay by capacitance versus voltage and impedance measurements using bi-layer bio-recognition membrane.

A novel formaldehyde sensitive biosensor based on bacterial formaldehyde dehydrogenase (FDH) as a bio-recognition element has been developed. The bio-recognition membrane had bi-layer architecture and consisted of FDH, cross-linked with albumin, and of the cofactor NAD at a high concentration level (first layer). The second layer was a negatively charged Nafion membrane, which prevented a leakage of negatively charged NAD molecules from the bio-membrane. As transducers, gold electrodes SiO(2)/Si/SiO(2)/Ti/Au and electrolyte-insulator-semiconductor Si/SiO(2) (EIS) structures have been used. Changes in capacitance and impedance properties of the bio-recognition membrane have been used for monitoring formaldehyde concentration in a bulk solution. It has been shown that formaldehyde can be detected within a concentration range from 1 microM to 20mM depending on the type of transduction used, with a detection limit of 1 and 100 microM for gold-based and EIS-based transducers, respectively.

Aldehyde Oxidoreductases↗

Comparison of amperometric biosensors fabricated by palladium sputtering, palladium electrodeposition and Nafion/carbon nanotube casting on screen-printed carbon electrodes.

Different strategies, including palladium electrodeposition (Pd(CV)), Pd sputtering (Pd(S)) and Nafion-solubilized carbon nanotube casting (Nafion/CNT), were used to modify screen-printed carbon electrodes (SPCEs) for the fabrication of amperometric enzyme biosensors. The electrochemical properties of the bare and modified SPCEs and the optimal conditions for surface modification were determined. The electrochemical response of the bare SPCE to H(2)O(2) under the potential of 0.3 V could be improved about 100-fold by Pd modification by electrodeposition or sputtering. By contrast, the electrochemical response of the bare SPCE was enhanced by only about 11-fold by Nafion/CNT casting. Moreover, the Pd(CV)-SPCEs exhibited better reproducibility of electrochemical response (a relative standard deviation (R.S.D.)<6.0%) than freshly prepared Pd(S)-SPCEs (R.S.D.>10%). The glucose biosensor fabricated from Pd-modified electrodes could be stored for up to 108 days without loosing significant activity. The Pd(CV)-SPCE also showed very reliable signal characteristics upon 50 consecutively repeated measurements of ascorbic acid. The electrocatalytic detection of the Pd-SPCE was combined with additional advantages of resistance to surface fouling and hence good stability. In conclusion, this study demonstrated that deposition of Pd thin film on SPCEs by electrodeposition or sputtering provided superior enhancement of electrochemical properties compared to Nafion/CNT-SPCEs. Despite their high electrochemical response, Pd(S)-SPCEs required an activation process to improve stability and Pd(CV)-SPCEs suffered from poor between electrode reproducibility.

Biosensing Techniques↗

Development of a micro-planar amperometric bile acid biosensor for urinalysis.

The determination of bile acid concentration in urine is useful for the screening and diagnosis of various hepatobiliary diseases. Currently, there is no concise method to determine bile acid concentration in urine. This study describes a bile acid biosensor fabricated by electrochemical technique for urinalysis. The micro-planar electrodes employed for the study consisted of a working electrode (platinum), a counter electrode (platinum) and a reference electrode (silver/silver chloride (Ag/AgCl)). The sensor chip was coated with Nafion using a spin-coater in order to both eliminate many interference species in urine and achieve long-term stability of the reference electrode. Nafion coating allowed the sensor chip to prevent the electrode reaction from interference species in urine, because it is charged negative strongly (Nafion contains sulfonic acid group). Three enzymes (bile acid sulfate sulfatase: BSS, beta-hydroxysteroid dehydrogenase: beta-HSD, and NADH oxidase: NHO) were immobilized by glutaraldehyde (GA: cross-linker) onto the sensor chip, because the immobilization of enzymes by GA is simple and commonly carried out. The sensor chip was able to detect bile acid in buffer solution. The optimum enzyme ratio immobilized onto the sensor chip was BSS:beta-HSD:NHO=4:4:20 U/1 chip. There was a relationship between the concentration of bile acid and the response current value. The dynamic range of the sensor chip was 2-100 microM for bile acid. Additionally, bile acid in the urine specimen could be detected using this bile acid biosensor. We present a simple and rapid bile acid biosensor with high sensitivity and high reproducibility.

Bile Acids and Salts↗

Inhibitory effect of PVDF tubes on biofilm formation in dental unit waterlines.

OBJECTIVES: It has been reported that dental unit waterlines (DUWLs) are contaminated with bacterial biofilm, and that water discharged from a DUWL contains bacteria that might be opportunistic pathogens. This study aimed to investigate the ability of polyvinylidene fluoride (PVDF) tubing to inhibit bacterial contamination in DUWLs. METHODS: Newly installed dental units were equipped with either a conventional polyurethane tube (unit A) or a PVDF tube (unit B), and the numbers of bacteria discharged from high- and low-speed handpiece lines were counted using R2A agar plates. Bacterial attachment on surfaces was observed with a scanning electron microscope (SEM) up to 185 days. Bacterial outflow during 1-day clinical service from a DUWL after 1-year usage was also examined. The surface free energy of each tube was determined based on the measurement of contact angles. RESULTS: The number of bacteria discharged from unit B was lower than from unit A at 80 days and thereafter. SEM examination demonstrated that the unit A tube was covered by biofilm constituting rods and filaments after 94 days, while no biofilm was observed in the unit B tube even after 185 days. After 1-year of usage, the unit B released significantly less bacteria than the unit A at every sampling period of 1-day clinic work. Surface free energies, calculated from contact angles measured, of PVDF and polyurethane tubes were 37.7 and 77.8, respectively. SIGNIFICANCE: The present results indicate that PVDF tubes, which have lower surface free energy than the conventional tubes, were effective in inhibiting biofilm formation and reducing bacterial outflow from DUWLs.

Bacterial Adhesion↗

Fluoropolymer-coated dacron versus PTFE grafts for femorofemoral crossover bypass: randomised trial.

OBJECTIVES: To investigate whether patency of a thin walled 8 mm fluoropassivated Dacron graft was similar to that of a standard 8mm PTFE graft for femorofemoral crossover bypass surgery. DESIGN: A randomised multicentre clinical trial comparing two vascular grafts with participation of 10 departments of vascular surgery in Denmark, Sweden and Norway. PATIENTS AND METHODS: 198 patients were randomised to PTFE (n=107) or fluoropolymer-coated Dacron grafts (n=91), 63% underwent surgery for claudication, 27% for ischaemic rest pain and 10% for tissue loss. The median follow-up time was 24 months (IQR 19-26 months). RESULTS: The primary patency rate of the two grafts was similar (log rank test: p=0.35). The primary patency rates (95% CI) for coated Dacron and PTFE grafts were 92% (86-98) and 94% (89-99) at 12 months and 87% (74-95) and 93% (87-99) at 24 months, respectively. CONCLUSION: In patients with unilateral iliac artery disease not amenable to angioplasty, the femoral-femoral bypass is durable and effective. No difference in patency was found between the two graft materials (fluoropolymer coated Dacron and PTFE).

Aged↗

Nafion-coated bismuth film and nafion-coated mercury film electrodes for anodic stripping voltammetry combined on-line with ICP-mass spectrometry.

Nafion-coated bismuth film electrodes (NCBFEs) and Nafion-coated mercury film electrodes (NCMFEs) were used to electrochemically preconcentrate metal analytes for subsequent analysis by inductively coupled plasma-mass spectrometry (ICP-MS). Either type of electrodes is part of a thin-layer electrochemical flow cell that is positioned upstream of a microconcentric nebulizer for the ICP-MS. Performances of these electrodes were compared in terms of the analytical "figures of merit" (e.g., dynamic ranges, reproducibility, hydrodynamic stability, and elimination of matrix effects detrimental to ICP-MS). The coupled technique (ASV-ICP-MS) is found to possess a wide dynamic range (at least 4 to 5 orders of magnitude) and to be reproducible. Both electrodes are much more stable than the thin mercury film electrode (TMFE) traditionally used for ASV-ICP-MS, with the lifetime of the NCBFE exceeding 8 h. Adopting these electrodes for ASV-ICP-MS overcomes the problems associated with a TMFE, the erosion of which decreases the sample throughput, affects the analysis precision, and contaminates conventional glass nebulizers and spray chambers of the spectrometer. The medium exchange procedure inherent in ASV is successfully implemented with a two-valve flow injection system for the accumulation of trace Cd2+ into the electrode from a certified seawater sample, followed by stripping Cd into a solution that is compatible to the ICP-MS operation.

Adsorption↗

Novel potentiometry immunoassay with amplified sensitivity for diphtheria antigen based on Nafion, colloidal Ag and polyvinyl butyral as matrixes.

A novel potentiometry immunoassay with amplified sensitivity has been developed for the detection of diphtheria antigen (Diph) via immobilizing diphtheria antibody (anti-Diph) on a platinum electrode based on Nafion, colloidal Ag (Ag), and polyvinyl butyral (PVB) as matrixes in this study. The modified procedure was further characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The influence and factors influencing the performance of resulting immunosensor were studied in detail. The resulting immunosensor exhibited sigmoid curve with log Diph concentrations, high sensitivity (51.4 mV/decade), wide linear range from 8 to 800 ng ml(-1) with a detection limit of 1.5 ng ml(-1), rapid potentiometric response (<3 min) and long-term stability (>6 months). Analytical results of clinical samples show that the developed immunoassay is comparable with the enzyme-linked immunosorbent assays (ELISAs) method, implying a promising alternative approach for detecting diphtheria antigen in the clinical diagnosis.

Antigens↗

Ion-exchange fibers and drugs: a novel device for the screening of iontophoretic systems.

The objective of this study was to theoretically model and experimentally measure the extent of drug release from ion-exchange fibers. The release was measured as a function of current density and NaCl concentration using a novel iontophoretic cell. The fibers tested contained weak carboxylate (-COOH) ion-exchange groups. The cationic model drugs tacrine and metoprolol were chosen on the basis of previous research, where tacrine had the lowest release rate and metoprolol the highest release rate. An in-house designed three compartment test cell was developed to test the suitability of drugs for iontophoretic drug delivery. In this cell, the anode and the drug containing ion-exchange fiber compartments were separated with a Nafion ion-selective membrane, while the fiber and the return electrode compartments were separated with a porous membrane. Tacrine proved to be a good drug candidate for this system as the release of the tacrine from the device was controllable with salt concentration and current density. Metoprolol release from the device was, however, not controllable.

Chromatography, Ion Exchange↗

Intraocular lens haptic fracturing with the neodymium:YAG laser In vitro study.

PURPOSE: To determine the neodymium:YAG (Nd:YAG) laser energy threshold to fracture the haptic of various currently marketed intraocular lenses. SETTING: Dokuz Eylul University, Ophthalmology Department, Izmir, Turkey. METHODS: Four haptic materials-poly(methyl methacrylate) (PMMA), hydrophobic acrylic, hydrophilic acrylic, and polyvinylidene fluoride-were tested. The intraocular lenses (IOLs) were grouped according to haptic material. Each group comprised 9 IOLs. Beginning laser energy was 4 mJ. If no crack was noted, energy was increased by 0.5 mJ at every 20 shots and the procedure continued until a fracture was noted. If the fracture could not be developed despite reaching an energy level of 10 mJ, the procedure was terminated. Statistical analysis was performed to determine the differences within groups. RESULTS: Hydrophobic acrylic and polyvinylidene fluoride haptics required higher energy and more laser shots to create a fracture than hydrophilic acrylic and PMMA haptics. CONCLUSIONS: Neodymium:YAG laser haptic fracturing can be a viable option to cut the fibrosed haptic before explanting any IOL. It was demonstrated in vitro that it is easier to fracture hydrophilic acrylic and PMMA haptics than hydrophobic acrylic and polyvinylidene fluoride haptics.

Acrylic Resins↗

Compensating for pulse imperfections in REDOR.

Rotational-echo double resonance (REDOR) is a magic-angle spinning technique for measuring heteronuclear dipolar couplings. Rotor-synchronized pi pulses recouple the dipolar interaction. The accuracy of a REDOR determination of distance or orientation depends totally on the quality of the dephased (recoupled) and full-echo spectra. We present a scheme for measuring and compensating for the effects of pulse imperfections in REDOR experiments. No assumptions are made about the quality of the pi pulses, and no pulses are added or taken away in implementing the compensation for incomplete REDOR dephasing by imperfect pi pulses.

Artifacts↗

A novel ELISA using PVDF microplates.

Here we describe the development of a novel specific, rapid ELISA system, which is performed on modified microplates where polyvinylidine fluoride (PVDF) forms the base of each well. The use of microplates with PVDF membranes as the solid phase allows for a greater binding capacity of protein in comparison to the solid phases of traditional ELISAs. The increased binding capacity of the solid phase provides for the direct binding of antigens, which can subsequently be assayed using a single, specific and well-characterized antibody. This direct assay system eliminates the need for two distinct antibodies that are often necessary in conventional two site ELISA systems. The system is able to specifically detect purified proteins as well as antigens in crude preparations of tissue homogenates. The PVDF-based ELISA performs with similar sensitivity and reproducibility as conventional two site ELISAs in tissue homogenates. The intra- and inter-assay coefficients of variation for the measurement of actin in crude rat brain homogenate were 2.36 and 5.15%, respectively.

Actins↗

Prospective study of carotid endarterectomy with modified polytetrafluoroethylene (ACUSEAL) patching: early and late results.

OBJECTIVE: Carotid endarterectomies (CEAs) with standard polytetrafluoroethylene (PTFE) patching have been shown to have results comparable with those of autogenous vein patching; however, prolonged bleeding through needle holes in PTFE is a commonly recognized problem. This is the first study of CEA using a new hemostatic modified PTFE patch (GORE-TEX) analyzing the early and late outcomes. METHODS: Two hundred consecutive CEAs were entered into this protocol. All patients had an immediate postoperative carotid duplex ultrasound scan that was repeated at 1 month and every 6 to 12 months thereafter. A Kaplan-Meier analysis was used to estimate the stroke-free survival and the risk of restenosis. The mean follow-up was 21 months (range, 1 to 48 months). RESULTS: The perioperative stroke rate was 1.5% (1% ipsilateral and 0.5% contralateral, two minor strokes and one major stroke) with no perioperative mortality or perioperative carotid thrombosis. The incidence of perioperative transient ischemic attacks was 3.5% (2.5% ipsilateral and 1% contralateral). The mean hemostasis time after completion of the patching was 3 minutes, in contrast to 14 minutes for conventional PTFE (in a previous study). The rates of freedom from ipsilateral strokes at 1, 2, 3, and 4 years were 99%, 99%, 99%, and 99%, respectively. The cumulative stroke-free survival rates at 1, 2, 3, and 4 years were 98%, 96%, 93%, and 93%, respectively. The rates of freedom from > or =70% restenosis at 1, 2, 3, and 4 years were 97%, 97%, 94%, and 94%, respectively. CONCLUSIONS: CEAs with a new modified PTFE patch are safe, have low perioperative stroke rates, are durable, and have an acceptable hemostasis time.

Adult↗

Biomedical sensor using thick film technology for transcutaneous oxygen measurement.

Transcutaneous blood gas monitoring is a non-invasive measurement technique for obtaining fast and relatively accurate responses to determine the respiratory conditions of patients. In this investigation, a screen-printed, disposable, transcutaneous oxygen sensor based on the working principle of amperometry, incorporates an integral heating element to enhance transcutaneous diffusion of blood gases typically at 44 degrees C. A Clark cell configuration is employed with gold working and counter electrodes and a silver/silver chloride reference electrode. Two different types of electrolytes namely potassium nitrate gel and polyelectrolyte (Nafion) have been studied under laboratory test conditions. A fully computer-controlled gas testing rig has been constructed to automate the varying of oxygen levels. Linear relationships have been established with an averaged sensitivity level of 0.029 microA/mmHg. In addition, a brief pilot clinical trial involving a fully grown human subject has been carried out alongside a commercial transcutaneous blood gas analyser. The investigations have shown that although the measured signals are weaker than those obtained from the laboratory test, the thick film sensor displays a repeatable and linear relationship when correlating with the commercial system. This study has greatly contributed towards the understanding for the suitability of the materials in achieving a viable, low-cost biomedical sensor.

Biosensing Techniques↗