A BACKING DEVICE BASED ON AN EMBEDDED STIFFENER AND RETRACTABLE INSERTION TOOL FOR THIN-FILM COCHLEAR ARRAYS

Intracochlear trauma from surgical insertion of bulky electrode arrays and inadequate pitch perception are areas of concern with current hand-assembled commercial cochlear implants. Parylene thin-film arrays with higher electrode densities and lower profiles are a potential solution, but lack rigidity and hence depend on manually fabricated permanently attached polyethylene terephthalate (PET) tubing based bulky backing devices. As a solution, we investigated a new backing device with two sub-systems. The first sub-system is a thin poly(lactic acid) (PLA) stiffener that will be embedded in the parylene array. The second sub-system is an attaching and detaching mechanism, utilizing a poly(N-vinylpyrrolidone)-block-poly(d,l-lactide) (PVP-b-PDLLA) copolymer-based biodegradable and water soluble adhesive, that will help to retract the PET insertion tool after implantation. As a proof-of-concept of sub-system one, a microfabrication process for patterning PLA stiffeners embedded in parylene has been developed. Conventional hotembossing, mechanical micromachining, and standard cleanroom processes were integrated for patterning fully released and discrete stiffeners coated with parylene. The released embedded stiffeners were thermoformed to demonstrate that imparting perimodiolar shapes to stiffener-embedded arrays will be possible. The developed process when integrated with the array fabrication process will allow fabrication of stiffener-embedded arrays in a single process. As a proof-of-concept of sub-system two, the feasibility of the attaching and detaching mechanism was demonstrated by adhering 1x and 1.5x scale PET tube-based insertion tools and PLA stiffeners embedded in parylene using the copolymer adhesive. The attached devices survived qualitative adhesion tests, thermoforming, and flexing. The viability of the detaching mechanism was tested by aging the assemblies in-vitro in phosphate buffer solution. The average detachment times, 2.6 minutes and 10 minutes for 1x and 1.5x scale devices respectively, were found to be clinically relevant with respect to the reported array insertion times during surgical implantation. Eventually, the stiffener-embedded arrays would not need to be permanently attached to current insertion tools which are left behind after implantation and congest the cochlear scala tympani chamber. Finally, a simulation-based approach for accelerated failure analysis of PLA stiffeners and characterization of PVP-b-PDLLA copolymer adhesive has been explored. The residual functional life of embedded PLA stiffeners exposed to body-fluid and thereby subjected to degradation and erosion has been estimated by simulating PLA stiffeners with different parylene coating failure types and different PLA types for a given parylene coating failure type. For characterizing the PVP-b-PDLLA copolymer adhesive, several formulations of the copolymer adhesive were simulated and compared based on the insertion tool detachment times that were predicted from the dissolution, degradation, and erosion behavior of the simulated adhesive formulations. Results indicate that the simulation-based approaches could be used to reduce the total number of time consuming and expensive in-vitro tests that must be conducted.

NEW MICROFLUIDIC SYSTEM TO INCREASE ROBUSTNESS OF ELECTRODE PERFORMANCE AND DEVELOP POINT-OF-CARE HEMATOCRIT DEVICE

The present dissertation aimed to develop a new microfluidic system for a point-of-care hematocrit device. Stabilization of microfluidic systems via surfactant additives and integration of semipermeable SnakeSkin® membranes was investigated. Both methods stabilized the microfluidic systems by controlling electrolysis bubbles. Surfactant additives, Triton X-100 and SDS stabilized promoted faster bubble detachment at electrode surfaces by lowering surface tension and decreased gas bubble formation by increasing gas solubility. The SnakeSkin® membranes blocked bubbles from entering the microchannel and thus less disturbance to the electric field by bubbles occurred in the microchannel. Platinum electrode performance was improved by carbonizing electrode surface using red blood cells. Irreversibly adsorbed RBCs lysed on platinum electrode surfaces and formed porous carbon layers while current response measurements. The formed carbon layers increase the platinum electrode surface area and thus electrode performance was improved by 140 %. The microfluidic system was simplified by employing DC field to use as a platform for a point-of-care hematocrit device. Feasibility of the microfluidic system for hematocrit determination was shown via current response measurements of red blood cell suspensions in phosphate buffered saline and plasma media. The linear trendline of current responses over red blood cell concentration was obtained in both phosphate buffered saline and plasma media. This research suggested that a new and simple microfluidic system could be a promising solution to develop an inexpensive and reliable point-of-care hematocrit device.

Functionalized Carbon Micro/Nanostructures for Biomolecular Detection

Advancements in the micro-and nano-scale fabrication techniques have opened up new avenues for the development of portable, scalable and easier-to-use biosensors. Over the last few years, electrodes made of carbon have been widely used as sensing units in biosensors due to their attractive physiochemical properties. The aim of this research is to investigate different strategies to develop functionalized high surface carbon micro/nano-structures for electrochemical and biosensing devices. High aspect ratio three-dimensional carbon microarrays were fabricated via carbon microelectromechanical systems (C-MEMS) technique, which is based on pyrolyzing pre-patterned organic photoresist polymers. To further increase the surface area of the carbon microstructures, surface porosity was introduced by two strategies, i.e. (i) using F127 as porogen and (ii) oxygen reactive ion etch (RIE) treatment. Electrochemical characterization showed that porous carbon thin film electrodes prepared by using F127 as porogen had an effective surface area (Aeff 185%) compared to the conventional carbon electrode. To achieve enhanced electrochemical sensitivity for C-MEMS based functional devices, graphene was conformally coated onto high aspect ratio three-dimensional (3D) carbon micropillar arrays using electrostatic spray deposition (ESD) technique. The amperometric response of graphene/carbon micropillar electrode arrays exhibited higher electrochemical activity, improved charge transfer and a linear response towards H2O2 detection between 250μM to 5.5mM. Furthermore, carbon structures with dimensions from 50 nano-to micrometer level have been fabricated by pyrolyzing photo-nanoimprint lithography patterned organic resist polymer. Microstructure, elemental composition and resistivity characterization of the carbon nanostructures produced by this process were very similar to conventional photoresist derived carbon. Surface functionalization of the carbon nanostructures was performed using direct amination technique. Considering the need for requisite functional groups to covalently attach bioreceptors on the carbon surface for biomolecule detection, different oxidation techniques were compared to study the types of carbon–oxygen groups formed on the surface and their percentages with respect to different oxidation pretreatment times. Finally, a label-free detection strategy using signaling aptamer/protein binding complex for platelet-derived growth factor oncoprotein detection on functionalized three-dimensional carbon microarrays platform was demonstrated. The sensor showed near linear relationship between the relative fluorescence difference and protein concentration even in the sub-nanomolar range with an excellent detection limit of 5 pmol.

Pelvic-Floor Rehabilitation, Part 1 : Comparison of Two Surface Electrode Placements During Stimulation of the Pelvic-Floor Musculature in Women Who Are Continent Using Bipolar Interferential Currents

Background and Purpose. Electrical stimulation of the pelvic floor is used as an adjunct in the conservative treatment of urinary incontinence. No consensus exists, however, regarding electrode placements for optimal stimulation of the pelvic-floor musculature. The purpose of this study was to compare two different bipolar electrode placements, one suggested by Laycock and Green (L2) the other by Dumoulin (D2), during electrical stimulation with interferential currents of the pelvic-floor musculature in continent women, using a two-group crossover design. Subjects. Ten continent female volunteers, ranging in age from 20 to 39 years (X̅=27.3, SD=5.6), were randomly assigned to one of two study groups. Methods. Each study group received neuromuscular electrical stimulation (NMES) of the pelvic-floor musculature using both electrode placements, the order of application being reversed for each group. Force of contraction was measured as pressure (in centimeters of water [cm H2O]) exerted on a vaginal pressure probe attached to a manometer. Data were analyzed using a two-way, mixed-model analysis of variance. Results. No difference in pressure was observed between the two electrode placements. Differences in current amplitude were observed, with the D2 electrode placement requiring less current amplitude to produce a maximum recorded pressure on the manometer. Subjective assessment by the subjects revealed a preference for the D2 electrode placement (7 of 10 subjects). Conclusion and Discussion. The lower current amplitudes required with the D2 placement to obtain recordings comparable to those obtained with the L2 technique suggest a more comfortable stimulation of the pelvic-floor muscles. The lower current amplitudes required also suggest that greater increases in pressure might be obtained with the D2 placement by increasing the current amplitude while remaining within the comfort threshold. These results will help to define treatment guidelines for a planned clinical study investigating the effects of NMES and exercise in the treatment of urinary stress incontinence in women postpartum.

The impact of storage conditions upon gentamicin coated antimicrobial implants

A systematic approach was developed to investigate the stability of gentamicin sulfate (GS) and GS/poly (lactic-co-glycolic acid) (PLGA) coatings on hydroxyapatite surfaces. The influence of environmental factors (light, humidity, oxidation and heat) upon degradation of the drug in the coatings was investigated using liquid chromatography with evaporative light scattering detection and mass spectrometry. GS coated rods were found to be stable across the range of environments assessed, with only an oxidizing atmosphere resulting in significant changes to the gentamicin composition. In contrast, rods coated with GS/PLGA were more sensitive to storage conditions with compositional changes being detected after storage at 60 °C, 75% relative humidity or exposure to light. The effect of γ-irradiation on the coated rods was also investigated and found to have no significant effect. Finally, liquid chromatography–mass spectrometry analysis revealed that known gentamines C1, C1a and C2 were the major degradants formed. Forced degradation of gentamicin coatings did not produce any unexpected degradants or impurities.

Pelvic-Floor Rehabilitation, Part 1 : Comparison of Two Surface Electrode Placements During Stimulation of the Pelvic-Floor Musculature in Women Who Are Continent Using Bipolar Interferential Currents

Background and Purpose. Electrical stimulation of the pelvic floor is used as an adjunct in the conservative treatment of urinary incontinence. No consensus exists, however, regarding electrode placements for optimal stimulation of the pelvic-floor musculature. The purpose of this study was to compare two different bipolar electrode placements, one suggested by Laycock and Green (L2) the other by Dumoulin (D2), during electrical stimulation with interferential currents of the pelvic-floor musculature in continent women, using a two-group crossover design. Subjects. Ten continent female volunteers, ranging in age from 20 to 39 years (X̅=27.3, SD=5.6), were randomly assigned to one of two study groups. Methods. Each study group received neuromuscular electrical stimulation (NMES) of the pelvic-floor musculature using both electrode placements, the order of application being reversed for each group. Force of contraction was measured as pressure (in centimeters of water [cm H2O]) exerted on a vaginal pressure probe attached to a manometer. Data were analyzed using a two-way, mixed-model analysis of variance. Results. No difference in pressure was observed between the two electrode placements. Differences in current amplitude were observed, with the D2 electrode placement requiring less current amplitude to produce a maximum recorded pressure on the manometer. Subjective assessment by the subjects revealed a preference for the D2 electrode placement (7 of 10 subjects). Conclusion and Discussion. The lower current amplitudes required with the D2 placement to obtain recordings comparable to those obtained with the L2 technique suggest a more comfortable stimulation of the pelvic-floor muscles. The lower current amplitudes required also suggest that greater increases in pressure might be obtained with the D2 placement by increasing the current amplitude while remaining within the comfort threshold. These results will help to define treatment guidelines for a planned clinical study investigating the effects of NMES and exercise in the treatment of urinary stress incontinence in women postpartum.

Desenvolvimento de métodos electroquímicos de quantificação de fármacos em amostras de água contaminadas

Este trabalho teve como objectivo, o desenvolvimento de um método electroquímico, para quantificação do fármaco carbamazepina (CBZ) em águas contaminadas. Neste trabalho foram utilizados quatro métodos voltamétricos: a voltametria cíclica, a voltametria de varrimento linear, a voltametria de onda quadrada e a voltametria de impulso diferencial. Os eléctrodos de trabalho utilizados foram, o eléctrodo de mercúrio de gota suspensa, o eléctrodo de carbono vítreo clássico e um eléctrodo de carbono vítreo modificado com um filme de nanotubos de carbono de paredes múltiplas (MWCNTs). O eléctrodo de mercúrio de gota suspensa permitiu o estudo da redução da CBZ numa região de potencial mais catódico, e os eléctrodos de carbono vítreo, com e sem modificação, permitiram o estudo da oxidação da CBZ numa região de potencial mais anódico. Nas condições experimentais estudadas, o eléctrodo de mercúrio de gota suspensa revelou ser um sensor voltamétrico pouco eficaz na determinação quantitativa da carbamazepina, em amostras com uma matriz complexa. Entre os eléctrodos de carbono vítreo, o eléctrodo de carbono vítreo modificado com os MWCNTs revelou ser o sensor voltamétrico mais eficaz e sensível, na detecção e determinação da carbamazepina. Modificado com um filme de nanotubos de carbono de paredes múltiplas, que previamente foram dispersos em dihexadecilhidrogenofosfato (DHP) e água, este novo eléctrodo permitiu obter uma resposta electroquímica da CBZ, consideravelmente superior ao eléctrodo não modificado. Utilizando a voltametria de varrimento linear e as condições experimentais consideradas óptimas, o eléctrodo nanoestruturado permitiu obter uma relação linear entre o sinal medido e a concentração da CBZ no intervalo 0.13- 1.60 M (30.7- 378 g -1), com os limites de detecção e quantificação mais baixos, até à data reportados com métodos electroquímicos (0.04 e 0.14M, respectivamente). O eléctrodo modificado foi aplicado na quantificação da CBZ, em formulações farmacêuticas, em águas naturais tratadas e em amostras de águas residuais, ambas dopadas, obtendo-se taxas de recuperação consideravelmente elevadas (100.6%, 98.0%,95.8%, respectivamente). Os resultados obtidos, na análise da CBZ em amostras ambientais, com o eléctrodo modificado, foram comparados com resultados obtidos por HPLC-UV e LC­ ESI-MS/MS, validando o método electroquímico desenvolvido neste trabalho. ABSTRACT: The aim of this work was to develop a new electrochemical method for the quantification of carbamazepine (CBZ) in contaminated waters. ln this study, four voltammetric methods were used: cyclic voltammetry, linear sweep voltammetry, square wave voltammetry and differential pulse voltammetry. the working electrodes used were the hanging mercury drop electrode (HMDE), the classical glassy carbon electrode (GCE), and a glassy carbon electrode modified with a film of multi-walled carbon nanotubes (MWCNls). Using HMDE, the reduction of CBZ was studied in the cathodic potential region. the CGE sensors, with or without modification, allowed the study of CBZ oxidation in the anodic potential region. ln the tested conditions, the results obtained for the quantification of CBZ using the HMDE sensor were not very satisfactory, especially when more complex samples were analysed. When the MWCNls-dihexadecyl hydrogen phosphate (DHP) film­ coated GCE was used for the voltammetric determination of CBZ, the results obtained showed that this modified electrode exhibits excellent enhancement effects on the electrochemical oxidation of CBZ. the oxidation peak current of CBZ at this film­ modified electrode increased significantly, when compared with that at a bare glassy carbon electrode. The enhanced electrooxidation and voltammetry of CBZ at the surface of MWCNTs-DHP film coated GCE in phosphate buffer solution (pH 6.71) was attributed to the unique properties of MWCNTs such as large specific surface area and strong adsorptive properties providing more reaction sites. The proposed method was applied to the quantification of CBZ in pharmaceutical formulations, drinking water and wastewater samples with good recoveries and low limits of detection and quantification (0.04 and 0.14 M, respectively), and was positively compared with chromatographic techniques usually used in the quantification of pharmaceutical compounds in environmental samples. HPLC-UV and LC-ESI-MS/MS were also used in the quantification of CBZ in pharmaceutical formulations and wastewater samples to prove the importance and accuracy of his voltammetric method.

Fertilizers with coated urea in upland rice production and nitrogen apparent recovery.

Urea is the most used N fertilizer for upland rice, however, a great percentage of N loss can occur with the use of this fertilizer. The use of products that provide reduction of N loss for urea fertilizers can contribute to increase N use efficiency. The objective of this study was to determine the effect of N rates applied in the form of coated urea in the content and accumulation of N in dry biomass, apparent recovery of nitrogen and grain yield of upland rice. The experimental design was a randomized complete blocks arranged in a 4 x 3 + 1 factorial scheme. The treatments consisted of four sources of N fertilizer [1. Common urea; 2. Polymer-coated urea for slow release of N (PCU); 3. urea with the urease inhibitor N-(n-Butyl) thiophosphoric triamide (NBPT); and 4. urea coated with copper sulfate and boric acid as urease inhibitors (UCCB)], with three fertilization rates (30, 60 and 90 kg ha-1 of N). In addition, we included a control treatment without N application. Coated urea did not provide increases in rice grain yield in relation to common urea. The increasing amount of N resulted in significant increases in rice grain yield (from 3217 to 5548 kg ha-1, 2010/11, and from 3392 to 4560 kg ha-1, 2011/12). The apparent nitrogen recovery rate decreased with the increase in N applied doses.

Synthesis and characterization of defective PBAs electrode material

Sodium manganese hexacyanoferrate (NaMnHCF) and its derivatives have been synthesized by simple co-precipitation method with addition of the citric and ascorbic acids respectively. The correspondent crystal structure, water content, chemical formula and a deep structural investigation of prepared samples have been performed by means of the combination of the laboratory and synchrotron techniques (PXRD, FT-IR, TGA, MP-AES and XAS). Electrochemical tests have been done using three-electrode system in sodium nitrate solution at different concentration. From cyclic voltammetry curves, Fe3+/2+ redox peak has been observed, whereas Mn3+/2+ peak was not always evident. Structural stability of the cycled samples has then been tested using 2D XRF imaging and Transmission X-ray microscopy (TXM) techniques. The intercalation of NaMnHCF after 20 cycles has been found by micro-XANES analysis of the highlighted spots which have been found in the XRF images. TXM has also confirmed the appearance of the intercalated particles after 50 cycles comparing the spectra between charged and discharged materials at three different edges (Mn, Fe and N). However, by comparison with lithium samples, it seems obvious that sodium samples are more homogeneous and intercalation is at the very beginning indicating the relative structural stability of sodium manganese hexacyanoferrate electrode material.

Graphene-based bioelectronic interfaces and devices and interpretative models targeting glial cells

Bioelectronic interfaces have significantly advanced in recent years, offering potential treatments for vision impairments, spinal cord injuries, and neurodegenerative diseases. However, the classical neurocentric vision drives the technological development toward neurons. Emerging evidence highlights the critical role of glial cells in the nervous system. Among them, astrocytes significantly influence neuronal networks throughout life and are implicated in several neuropathological states. Although they are incapable to fire action potentials, astrocytes communicate through diverse calcium (Ca2+) signalling pathways, crucial for cognitive functions and brain blood flow regulation. Current bioelectronic devices are primarily designed to interface neurons and are unsuitable for studying astrocytes. Graphene, with its unique electrical, mechanical and biocompatibility properties, has emerged as a promising neural interface material. However, its use as electrode interface to modulate astrocyte functionality remains unexplored. The aim of this PhD work was to exploit Graphene-oxide (GO) and reduced GO (rGO)-coated electrodes to control Ca2+ signalling in astrocytes by electrical stimulation. We discovered that distinct Ca2+dynamics in astrocytes can be evoked, in vitro and in brain slices, depending on the conductive/insulating properties of rGO/GO electrodes. Stimulation by rGO electrodes induces intracellular Ca2+ response with sharp peaks of oscillations (“P-type”), exclusively due to Ca2+ release from intracellular stores. Conversely, astrocytes stimulated by GO electrodes show slower and sustained Ca2+ response (“S-type”), largely mediated by external Ca2+ influx through specific ion channels. Astrocytes respond faster than neurons and activate distinct G-Protein Coupled Receptor intracellular signalling pathways. We propose a resistive/insulating model, hypothesizing that the different conductivity of the substrate influences the electric field at the cell/electrolyte or cell/material interfaces, favouring, respectively, the Ca2+ release from intracellular stores or the extracellular Ca2+ influx. This research provides a simple tool to selectively control distinct Ca2+ signals in brain astrocytes in neuroscience and bioelectronic medicine.

Characterization Of Morphology And Composition Of Inorganic Fillers In Dental Alginates.

Energy dispersive X-ray spectroscopy microanalysis (EDX), scanning electron microscopy (SEM), and Archimedes' Principle were used to determine the characteristics of inorganic filler particles in five dental alginates, including Cavex ColorChange (C), Hydrogum 5 (H5), Hydrogum (H), Orthoprint (O), and Jeltrate Plus (JP). The different alginate powders (0.5 mg) were fixed on plastic stubs (n = 5) and sputter coated with carbon for EDX analysis, then coated with gold, and observed using SEM. Volume fractions were determined by weighing a sample of each material in water before and after calcining at 450(°)C for 3 h. The alginate materials were mainly composed of silicon (Si) by weight (C-81.59%, H-79.89%, O-78.87%, H5-77.95%, JP-66.88%, wt). The filler fractions in volume (vt) were as follows: H5-84.85%, JP-74.76%, H-70.03%, O-68.31%, and C-56.10%. The tested materials demonstrated important differences in the inorganic elemental composition, filler fraction, and particle morphology.

Platinum Blue Staining Of Cells Grown In Electrospun Scaffolds.

Fibroblast cells grown in electrospun polymer scaffolds were stained with platinum blue, a heavy metal stain, and imaged using scanning electron microscopy. Good contrast on the cells was achieved compared with samples that were gold sputter coated. The cell morphology could be clearly observed, and the cells could be distinguished from the scaffold fibers. Here we optimized the required concentration of platinum blue for imaging cells grown in scaffolds and show that a higher concentration causes platinum aggregation. Overall, platinum blue is a useful stain for imaging cells because of its enhanced contrast using scanning electron microscopy (SEM). In the future it would be useful to investigate cell growth and morphology using three-dimensional imaging methods.