A.P. Piedade

A comparative study of carbon fiber-based microelectrodes for the measurement of nitric oxide in brain tissue.

The measurement of Nitric oxide (NO) in real-time has been a major concern due to the involvement of this ubiquitous free radical modulator in several physiological and pathological pathways in tissues. Here we performed a study aiming at evaluating different types of carbon fibers, namely Textron, Amoco, Courtaulds and carbon nanotubes (University of Kentucky) covered with Nafion/o-phenylenediamine (o-PD) for NO measurement in terms of sensitivity, LOD, response time and selectivity against major potential interferents in the brain (ascorbate, nitrite and dopamine). The results indicate that, as compared with the other carbon fibers and nanotubes, Textron carbon fiber microelectrodes coated with two layers of Nafion and o-PD exhibited better characteristics for NO measurement as they are highly selective against ascorbate (>30,000:1), nitrite (>2000:1) and dopamine (>80:1). These coated Textron microelectrodes showed an average sensitivity of 341+/-120pA/microM and a detection limit of 16+/-11nM. The better performance of the Textron fibers is likely related to a stronger adhesion or more uniform coating of the Nafion and o-PD polymers to the fiber surface. In addition, the background current of the Textron carbon fibers is low, contributing to the excellent signal-to-noise for detection of NO.

Covalent binding of urease on ammonium-selective potentiometric membranes☆

As part of the development of disposable urea bioselective probes, the covalent binding of urease on ammonium-selective potentiometric membranes has been assessed. Nonactin/bis(1-butylpentyl)adipate/poly(vinylchloride) (PVC) membranes, directly applied to an internal solid contact (conductive epoxy-graphite composite), has been used as a support for covalent immobilization of urease. Two types of all-solid-state construction process have been assayed: thin layers of cellulose acetate (CA) were coated on the PVC ammonium-selective membranes (type 1) and blends of PVC and CA at various ratios were used as ammonium-selective membrane matrices (type 2). Urease was covalently attached to CA via aldehyde groups. These groups were created on the polysaccharide with sodium periodate to which the enzyme was immobilized through a spacer (hexamethylenediamine). The viability of both types of probe for the determination of ammonium ions was assessed after each step of the activation process. Results indicated that type 2 potentiometric probes are altered after the treatment with sodium periodate. Good results were obtained with type 1 probes. Their dynamic concentration range of response to urea was from 2 x 10(-5) to 0.01 M with a sensibility of 50 mV/decade.

Influence of albumin on the tribological behavior of Ag-Ti (C, N) thin films for orthopedic implants

With the increase of elderly population and the health problems arising nowadays, such as cancer, knee and hip joint prostheses are widely used worldwide. It is estimated that 20% of hip replacement surgeries simply fail after 5years, due to wear loosening, instability and infection. In this paper it is reported the study of advanced materials with the ability to overcome some of these drawbacks. The development of ceramic coatings, based on carbonitrides of transition metals, such as TiCN, doped with silver, Ag, may represent an effective solution. Thin films of Ag-TiCN were produced by dc reactive magnetron sputtering with silver contents ranging from 4 to 8at.%. The physical, chemical, structural, morphological/topographical, mechanical and tribological properties were evaluated. The tribological tests were performed in a unidirectional wear simulator, pin on disk, being the antagonists of a ceramic Al2O3 ball, and using simulate body fluids as lubricant. Hanks Balanced Salt Solution (HBSS) and bovine serum albumin (BSA) in HBSS were chosen, in order to evaluate the lubrication ability of the solution containing the protein, albumin. The results revealed that the coatings with Ag content ranging from 4 to 8at.%, were the most promising, as the tribological properties were superior to the results reported by other authors, which also developed Ag-TiCN coatings containing similar Ag contents and using similar test conditions. The presence of albumin leads to a lower wear in all the test conditions, and this enhancement was higher in the hydrophobic surfaces.

Influence of surface features on the adhesion of Staphylococcus epidermidis to Ag–TiCN thin films

Abstract Staphylococcus epidermidis has emerged as one of the major nosocomial pathogens associated with infections of implanted medical devices. The initial adhesion of these organisms to the surface of biomaterials is assumed to be an important stage in their colonization. The main objective of this work is to assess the influence of surface features on the adhesion of S. epidermidis to Ag–TiCN coatings deposited by dc reactive magnetron sputtering. The structural results obtained by x-ray diffraction show that the coatings crystallize in a B1-NaCl crystal structure typical of TiC0.3N0.7. The increase of Ag content promoted the formation of Ag crystalline phases. According to the results obtained with atomic force microscopy, a decrease on the surface roughness of the films from 39 to 7 nm is observed as the Ag content increases from 0 to 15 at.%. Surface energy results show that the increase of Ag promotes an increase in hydrophobicity. Bacterial adhesion and biofilm formation on coatings were assessed by the enumeration of the number of viable cells. The results showed that the surface with lower roughness and higher hydrophobicity leads to greater bacterial adhesion and biofilm formation, highlighting that surface morphology and hydrophobicity rule the colonization of materials.

Aerobic uranium immobilization by Rhodanobacter A2-61 through formation of intracellular uranium-phosphate complexes.

Severe environmental problems arise from old uranium mines, which continue to discharge uranium (U) via acid mine drainage water, resulting in soil, subsoil and groundwater contamination. Bioremediation of U contaminated environments has been attempted, but most of the conceptual models propose U removal by cell suspensions of anaerobic bacteria. In this study, strain Rhodanobacter A2-61, isolated from Urgeiriça Mine, Portugal, was shown to resist up to 2 mM of U(vi). The conditions used (low nutrient content and pH 5) potentiated the interaction of the toxic uranyl ion with the tested strain. The strain was able to remove approximately 120 μM of U(vi) when grown aerobically in the presence of 500 μM U. Under these conditions, this strain was also able to lower the phosphate concentration in the medium and increased its capacity to take up inorganic phosphate, accumulating up to 0.52 μmol phosphate per optical density unit of the medium at 600 nm, after 24 hours, corresponding approximately to the late log phase of the bacterial culture. Microscopically dense intracellular structures with nanometer size were visible. The extent of U inside the cells was quantified by LS counting. EDS analysis of heated cells showed the presence of complexes composed of phosphate and uranium, suggesting the simultaneous precipitation of U and phosphate within the cells. XRD analysis of the cells containing the U-phosphate complexes suggested the presence of a meta-autunite-like mineral structure. SEM identified, in pyrolyzed cells, crystalline nanoparticles with shape in the tetragonal system characteristic of the meta-autunite-like mineral structures. U removal has been reported previously but mainly by cell suspensions and through release of phosphate. The innovative Rhodanobacter A2-61 can actively grow aerobically, in the presence of U, and can efficiently remove U(vi) from the environment, accumulating it in a structural form consistent with that of the mineral meta-autunite inside the cell, corresponding to effective metal immobilization. This work supports previous findings that U bioremediation could be achieved via the biomineralization of U(vi) in phosphate minerals.

Photoacoustic Measurement of Electron Injection Efficiencies and Energies from Excited Sensitizer Dyes into Nanocrystalline TiO2 Films

Time-resolved photoacoustic calorimetry is used to measure the energy released upon injection of an electron from an electronically excited dye adsorbed to nanocrystalline TiO2 into the conduction band of this material. More energy is released when the environment of the dye is made less polar, because the energy of the dye-oxidized state has a more pronounced solvent dependence than the edge of the conduction band of the TiO2 semiconductor. Such energy dependences should be considered in the design of more efficient dye-sensitized solar cells.

Thin films with chemically graded functionality based on fluorine polymers and stainless steel

Thin films of stainless steel and poly(tetrafluoroethylene) were co-deposited, by radiofrequency magnetron sputtering, in an inert atmosphere in order to produce a functionally graded material as a coating on a traditional biomaterial, where non-ferromagnetic characteristics and improved wettability must be ensured. These thin films are intended to modify the surface of SS316L used in stents, where the bulk/thin film couple should be regarded as a single material. This requires excellent adhesion of the coating to the substrate. All coatings were deposited with an average thickness of 500 nm. The chemical and phase characterization of the surface revealed that, with the increase in F content, the thin film evolves from a ferritic phase (alpha) to an amorphous phase with dispersion of a new crystalline ceramic phase (FeF(2)). For intermediate F content values, an austenitic (111) phase (gamma) was present. Bearing in mind the envisaged application, the best results were attained for thin films with a fluorine content between 10 and 20 at.%.

Silver activation on thin films of Ag–ZrCN coatings for antimicrobial activity

Nowadays, with the increase of elderly population and related health problems, knee and hip joint prosthesis are being widely used worldwide. However, failure of these invasive devices occurs in a high percentage thus demanding the revision of the chirurgical procedure. Within the reasons of failure, microbial infections, either hospital or subsequently-acquired, contribute in high number to the statistics. Staphylococcus epidermidis (S. epidermidis) has emerged as one of the major nosocomial pathogens associated with these infections. Silver has a historic performance in medicine due to its potent antimicrobial activity, with a broad-spectrum on the activity of different types of microorganisms. Consequently, the main goal of this work was to produce Ag-ZrCN coatings with antimicrobial activity, for the surface modification of hip prostheses. Thin films of ZrCN with several silver concentrations were deposited onto stainless steel 316 L, by DC reactive magnetron sputtering, using two targets, Zr and Zr with silver pellets (Zr+Ag target), in an atmosphere containing Ar, C2H2 and N2. The antimicrobial activity of the modified surfaces was tested against S. epidermidis and the influence of an activation step of silver was assessed by testing samples after immersion in a 5% (w/v) NaClO solution for 5 min. The activation procedure revealed to be essential for the antimicrobial activity, as observed by the presence of an inhibition halo on the surface with 11 at.% of Ag. The morphology analysis of the surface before and after the activation procedure revealed differences in silver distribution indicating segregation/diffusion of the metallic element to the films surface. Thus, the results indicate that the silver activation step is responsible for an antimicrobial effect of the coatings, due to silver oxidation and silver ion release.

In vitro behaviour of nanocrystalline silver-sputtered thin films

Silver thin films were deposited with different preferential orientations and special attention was paid to the bioreactivity of the surfaces. The study was essentially focused on the evaluation of the films by x-ray diffraction (XRD), atomic force microscopy (AFM), high-resolution transmission electron microscopy (HRTEM), electron probe microanalysis (EPMA) and contact angle measurements. The deposited thin films were characterized before and after immersion in S-enriched simulated human plasma in order to estimate the influence of the preferential crystallographic orientation on the in vitro behaviour. Silver thin films with and without (111) preferential crystallographic orientation were deposited by r.f. magnetron sputtering to yield nanocrystalline coatings, high compact structures, very hydrophobic surfaces and low roughness. These properties reduce the chemisorption of reactive species onto the film surface. The in vitro tests indicate that silver thin films can be used as coatings for biomaterials applications.

Zeta potential, contact angles, and AFM imaging of protein conformation adsorbed on hybrid nanocomposite surfaces.

The sputtering deposition of gold (Au) and poly(tetrafluoroethylene) (PTFE) was used to prepare a nanocomposite hybrid thin film suitable for protein adsorption while maintaining the native conformation of the biological material. The monolithic PTFE and the nanocomposite PTFE/Au thin films, with Au content up to 1 at %, were co-deposited by r.f. magnetron sputtering using argon as a discharge gas and deposited onto 316L stainless steel substrates, the most commonly used steel in biomaterials. The deposited thin films, before and after bovine serum albumin (BSA) adsorption, were thoroughly characterized with special emphasis on the surface properties/characteristics by atomic force microscopy (AFM), zeta potential, and static and dynamic contact angle measurements, in order to assess the relationship between structure and conformational changes. The influence of a pre-adsorbed peptide (RGD) was also evaluated. The nanotopographic and chemical changes induced by the presence of gold in the nanocomposite thin films enable RGD bonding, which is critical for the maintenance of the BSA native conformation after adsorption.