Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

This Diploma Thesis presents results obtained by solution of two partial projects: a) Preparation of monomers from renewable sources using metathesis and tandem hydrogenation catalyzed with ruthenium compounds - project solved during my Erasmus stay at the Université de Rennes 1 in France; b) Preparation and properties of ,-bis(tpy)quarterthiophene oligomers carrying ionic side groups as oligomonomers for polyelectrolyte conjugated dynamers - project solved at the Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University in Prague. Project a): Self-metathesis of 1,2-epoxyhex-5-ene (but-3-enyloxirane) and its cross-metathesis with methyl acrylate and acrylonitrile catalyzed with ruthenium compounds as well as tandem design of these metatheses and consecutive hydrogenation of their products by gaseous hydrogen have been optimized. The following influences have been studied and tuned: (i) type of the catalyst (Grubbs, Hoveyda, Zhan) and its concentration and method of dosing, (ii) concentration of reactants and additives, (iii) type of solvent, and (iv) reaction temperature. Reactions were monitored by the GC, GC and MS methods and the products were characterized by the NMR method. Methyl 6,7-epoxyheptanoate (methyl 5-oxiranylpentanoate) obtained by the tandem...

Cette thèse concerne la fonctionnalisation des biomatériaux, le titane et l'alliage de titane Ti6Al4V ainsi que le PEEK (Poly-Éther-Éther-Ketone) pour l'application de ces biomatériaux dans les domaines de l'implantologie dentaire et de la reconstruction maxillo-faciale. Au cours de la première partie de ce travail nous avons synthétisé quatre peptides bi-fonctionnels avec une grande affinité pour la surface implantaire de Ti et de Ti6Al4V et également pour les kératinocytes oraux. La spectroscopie de force de la cellule unique (Single Cell Force Spectroscopy - AFM) a été utilisée pour étudier l'adhésion d'une cellule sur les surfaces fonctionnalisées par les quatre peptides bi-fonctionnels. A l'aide du test colorimétrique de para-nitrophenyl phosphate (pNPP) on a démontré l'adhésion des kératynocytes après 4 heures d'incubation sur les surfaces fonctionnalisées avec les peptides bi-fonctionnels. Les résultats de ces études ont démontré la présence d'un peptide bi-fonctionnel qui augmente l'adhésion cellulaire instantanée et l'adhésion 4h après incubation des kératinocytes oraux sur les métaux. Ce peptide résiste à l'influence de facteurs externes tels que l'adsorption d'albumine et représente une proposition prometteuse pour la fonctionnalisation de l'implant de Ti et de Ti6Al4V. La deuxième partie de notre mémoire de thèse décrit une méthode de dépôt par spray des films des multicouches de PLL/PGA (poly-l-lysine/acide polyglutamique), des protéines et des cellules souches pulpaires. Les outils de spray d'usage unique ont été adaptés aux exigences de la bonne pratique de fabrication (GMP - good manufacturing practice). Les premiers tests de prolifération cellulaire ont démontré qu'elle n'a pas été affectée par le spray. L'étude en AFM a démontré que la rugosité et l'épaisseur du film augmentaient exponentiellement avec l'augmentation du nombre des couches déposées. Le traitement physique par UV rayonnement ainsi que le traitement par la déshydratation et la réhydratation des films ont provoqué des changements dans l'épaisseur, l'élasticité et la dureté des films. Le dépôt des protéines sur les MPEs a aussi augmenté l'épaisseur et a influencé la dureté de la surface. Les changements chimiques de la structure des MPEs après le traitement physique et après le dépôt des protéines naturelles ont été étudiés par la spectroscopie Raman. La prolifération cellulaire au 1er, 3e et 8e jours après leur spray sur des films de MPEs et des protéines a été ensuite évaluée. Les MPEs traitées par UV combiné avec déshydratation/réhydratation ainsi que les PEMs couvertes par protéine - CaP ont entrainé une meilleure adhésion et prolifération cellulaires que les MPEs non-traitées. Finalement la méthode de dépôt par spray des MPEs, protéines et aussi des cellules souches pulpaires a été appliquée sur la surface de PEEK. La meilleure prolifération cellulaire au 8e jour après le spray était sur la surface couverte par le film de (PLL-PGA)5-protéine/CaP. Une étude in vitro sur le degré de minéralisation au 21e jour après l'incubation des cellules pulpaire dans un milieu ostéconductuer sur des différentes (MPEs couvertes-PEEK) surfaces a été effectuée. Les résultats de la microscopie électronique à balayage (MEB), microscopie électronique à transmission (MET), combiné avec des études histologiques ont confirmé la formation d'une matrice extracellulaire minéralisée des autour des cellules pulpaires sur la surface de PEEK. Conclusions: Dans ce travail nous avons décrit une nouvelle approche de fonctionnalisation des surfaces de Ti and Ti6Al4V par des peptides bi-fonctionnels en proposant une séquence prometteuse qui augmente l'adhésion des kératinocytes oraux. Ensuite nous avons développé une méthode de fonctionnalisation de la surface de PEEK par des multicouches de polyeléctrolytes, des protéines naturelles et des cellules souches de la pulpe dentaire. La différentiation ostéoblastique in vitro a été finalement évaluée. / The objective of this thesis was to develop new techniques of surface functionalization of titanium; titanium alloy (Ti6Al4V) and PEEK (poly-ether-ether ketone) surfaces for their application on dental implantology and maxillofacial surgery. During the first part of the thesis we have synthesized four metal binding-cell specific peptides (MCSPs) with high affinity to titanium surface and to oral keratinocytes cells. Single Cell Force Spectroscopy (AFM) was used to study the instantaneous cell adhesion force of keratinicyte cell on MCSPs functionalized surfaces. The colorimetric para-nitrophenyl phosphate (pNPP) essay demonstrates the surface cell adhesion four hours after incubation. The results demonstrate the presence of one bi-functional peptide (MCSP-2) who increases both: the instantaneous and the 4 h cell adhesions. MCSP-2 resist to the influence of external factors like BSA adsorption and could be an interesting candidate for implant surface functionalisation. In the second part of the thesis we have developed a spray deposition method of polyelectrolyte multilayer (PEM) films build. PLL/PGA (poly-l-lysine/poly-glutamic acid) and proteins were used for surface coating. Consequently dental pulp stem cells (DPSC) are sprayed on the PEM films. The aims were to improve a spray -method for cells and the polyelectrolytes deposition on the PEEK implant. The entire spray device was designed for single use, which correspond to good manufacturing practice (GMP) conditions. Cell proliferation in 24 hours after spraying was not disturbed by the spray. Physicochemical properties of PEMs deposited by spray on glass surfaces were performed by Atomic Force Microscopy (AFM) and by contact angle measurements. The results have demonstrated the changes in films thickness and films roughness with increasing numbers of layers corresponding to exponential growth of the films. Physical treatment by UV irradiation and drying-wetting process affects the film thickness and the film elasticity and increases the stiffness of the film. The deposition of protein-CaP and collagen coatings on PEM films increased the layer thickness and influenced the hardness of the surface. Chemical changes in the polyelectrolyte structure during the physical treatment and after proteins deposition were studied by Raman spectroscopy. Cell proliferation of the pulp cells at 1st, 3th and 8th days after pulp cells deposition on PEMs coated glass surfaces, was then evaluated. The results demonstrated that 10 UV/ dray/wetted films and the natural proteins coated films enhanced cell adhesion and cell proliferation. The protein-CaP PEMs covered surface creates the best microenvironment to ensure the cell behavior. Finally PEM films coatings are applied on PEEK implant surface. The AFM study shows the changes of homogeneity and roughness of this surface after PEM film deposition. Contact angle measurement demonstrates decreases of surface hydrophobicity. Cell proliferation of dental pulp stem cells (DPSC) on (PLL-PGA)5-protein/CaP coated PEEK surface was highest compared to other functionalized surfaces. In vitro study of DPSCs osteogenic differentiation was evaluated by the degree of mineralization of the extracellular matrix on the PEEK surface at 21st day after cell incubation. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) combined with histological studies improved the formation of mineralized extracellular matrix and confirmed the osteogenic differentiation of DPSCs on the PEMs coated PEEK surface. Conclusions: In this work we validated the method of Ti and Ti6Al4V functionalization with bi-functional peptides. One peptide that could increase the epithelial cell adhesion to the surface was proposed. Spray deposition technics of PEMs, protein and pulp stem cells were applied for PEEK implant functionalization. Finally we evaluated the differentiation of dental pulp stem cells on the PEEK surface in vitro.

Notre thèse vise la conception d'un biocapteur ultra-sensible à base d'Arthrospira platensis pour la détection des substances toxiques (métaux lourds et pesticides) pour un contrôle environnemental. Cette étude concerne, dans un premier lieu, le développement de systèmes impédimétriques et à ondes acoustiques de Love, associés à des puces en Polydimethylsiloxane (PDMS) et fonctionnalisés par des cellules d'Arthrospira platensis immobilisées sur une multicouche de polyélectrolytes déposés par la méthode « Layer by Layer ». L'étude fondamentale des interactions physiques-chimiques-biologiques mises en jeu lors des étapes de fonctionnalisation du capteur, et son application à la détection de cadmium et de mercure, est également abordée, à travers une étude acoustique (suivi en temps réel), électrochimique et microscopique (microscopie à force atomique (AFM) en milieu sec ou liquide et microscopique électronique à balayage (MEB)). Dans un deuxième lieu, cette étude présente les systèmes conductimétriques basés sur l'inhibition des activités enzymatiques de la cyanobactérie par les polluants. Différents modes d'immobilisation sur des microcapteurs interdigités (IDF) en or, ont été étudiés : immobilisation via les monocouches auto-assemblées « Self Assembled Monolayers », immobilisation via les nanoparticules d'or revêtues par du PAH et co-réticulées à l'albumine à l'albumine de sérum bovin sous vapeur saturée de glutaraldéhyde. Ce principe de détection consiste à inhiber l'Activité Phophatase Alcaline (APA) par les métaux lourds et l'Activité Estérase (AChE) par les pesticides. Des limites de détection excellente (10-20 M) ont été obtenues. Un biocapteur bi-enzymatique a été élaboré avec le même biorécepteur (Arthrospira platensis) pour une multi-détection des polluants. Ce capteur a été testé sur des échantillons d'eaux prélevés sur des effluents hospitaliers et urbains (eaux brutes et traitées) / This thesis focuses on the design of an ultra-sensitive biosensor based on Arthrospira platensis for the detection of toxic substances (heavy metals and pesticides) for environmental monitoring. This study deals with, first, the development of impedimetric and Love acoustic wave systems, associated with Polydimethylsiloxane (PDMS) chips and functionalized with Arthrospira platensis cells immobilized on polyelectrolyte multilayers deposited by the “Layer and Layer” method. Electrochemical, acoustic and microscopic interactions with cadmium and mercury (atomic force microscopy (AFM) in dry or liquid medium and studies of scanning electron microscopy (SEM)) have been performed. This work focuses also on conductometric system based on the inhibition of enzymatic activities of cyanobacteria by pollutants. Different modes of immobilization on gold interdigitated microsensors (IDTs) were studied: immobilization via self-assembled monolayers, immobilization with gold nanoparticles coated with PAH and co-reticulated with bovine serum albumin under saturated vapor of glutaraldehyde. They were based on the inhibition of Alkaline Phosphatase activity (APA) by heavy metals Esterase Activity (AChE) by pesticides. Excellent detection limits (10-20 M) were obtained. A bi-enzymatic biosensor was developed with the same bioreceptor (Arthrospira platensis) for polluants multi-detection. This sensor has been tested on water samples of urban and hospitals effluents (before and after treatment)

Die Elektrorotation von fixierten Erythrozyten wurde im Frequenzbereich von 16 Hz bis 33 MHz untersucht. Zwischen 16 Hz und 1 kHz zeigen die fixierten Erythrozyten eine Rotation parallel zur Feldrichtung mit einer maximalen Rotationsgeschwindigkeit zwischen 30 Hz und 70 Hz. Es wurde sowohl die Abhängigkeit von der äußeren Leitfähigkeit untersucht als auch von der Oberflächenladung. Die experimentellen Resultate erwiesen sich als konsistent mit einer erst kürzlich entwickelten Theorie zur Elektrorotation im niederfrequenten Bereich (LFER). Sie zeigen, daß die Elektrorotation im niederfrequenten Bereich von der Oberflächenladung und -leitfähigkeit entscheidend mitbestimmt werden kann. Fixierte Thrombozyten wurden mittels Elektrorotation im Frequenzbereich von 16 Hz bis 33 MHz untersucht. Zur Interpretation der Daten wurde ein theoretisches Modell weiterentwickelt, welches die innere vesikuläre Struktur der Thrombozyten berücksichtigt, und mit dem niederfrequenten Modell zur Elektrorotation superponiert. In Lösungen mit Dextran unterschiedlicher Molekulargewichte und Konzentrationen wurde sowohl die elektrophoretische Mobilität als auch Elektrorotation von fixierten Erythrozyten untersucht. Es konnte gezeigt werden, daß sich im niederfrequenten Bereich Depletionschichten an Hand von Elektrorotationsspektren erfassen lassen. Diese Daten bestätigen auch die Theorie zur LFER. Messungen der elektrophoretischen Mobilität von nativen Erythrozyten wurden in Lösungen mit dem Polyelektrolyten Polystyrensulfonat in Anhängigkeit vom Molekulargewicht und der Salzkonzentration durchgeführt. Es zeigte sich, daß das Polymer zum einen reversibel adsorbiert und zum anderen einen deutlichen Depletioneffekt herbeiführt. Im letzten Teil der Arbeit wurde ein Verfahren entwickelt, welches die Herstellung von Polyelektrolyt-Kapseln auf der Grundlage von biologischen Zellen ermöglicht, welche in Form und Größe identisch mit den verwendeten biologischen Templaten sind. / Electrorotation of fixed red blood cells (RBC) has been investigated in a frequency range between 16 Hz and 33 Mhz. Between 16 Hz and 1 kHz fixed red blood cells undergo co-field rotation with a maximum of rotation betwen 30 and 70 Hz. The rotation was studied as a function of electrolyte conductivity and surface charge density. These observations are consistent with a recently developed theory of the low frequency electrorotation (LFER) and demonstrate that the surface charge and the surface conductivity can play a significant role in this frequency range. Fixed platalets were investigated by means of electrorotation in the frequency range from 16 Hz to 33 Mhz. For the interpretation of the data a model which takes into account the inner structure of the platalets was developed and added to the theory which describes the rotation in the low frequency range. In solutions with Dextran and fixed platalets the electrophoretic mobility as well as the electrorotation was measured. It was shown that in the low frequency range depletion layers are detectable. Furthermore this results verify the LFER theory. Measurements of the electrophoretic mobility of native RBC were carried out in solutions of the polyelectrolyte Polystyrenesulfonate in dependence on the molecular weight and the ionic strength. It was shown that the polymer adsorbs reversible and forms a significant depletion effect. In the last part of this work a method was developed, which allows the construction of polyelectrolyte caspules with biological cells as template, which are identical in size and shape with the templates used.

In dieser Arbeit wurden Gold- und PlatinNanopartikel in sphärischen Polyelektolyt-Bürsten (SPB) synthetisiert. Diese wurden zu mechanistischen Untersuchungen der p-Nitrophenol-Reduktion mittels Natriumborhydrid herangezogen. Dabei konnte der Mechanismus der Reaktion auf der Oberfläche der Nanopartikel aufgeklärt werden. Die Reaktion folgt einem Langmuir Hinshelwood (LH) Mechanismus. Hierbei adsorbieren beide Edukte auf die Oberfläche, bevor sie im zu p-Aminophenol umgesetzt werden. Nach der Reaktion desorbiert das Reaktionsprodukt. Mittels des LH Modells konnten für verschiedene Temperaturen die intrinsische Geschwindigkeitskonstante, sowie die Adsorptionskonstanten der Edukte bestimmt werden. Mit diesen Daten konnten dann die Enthalpie und Entropie der Adsorption der Edukte und die Aktivierungsenergie berechnet werden. Neben dem Reaktionsmechanismus wurde die Induktionszeit der p-Nitrophenol Reduktion untersucht. Hierbei konnte gezeigt werden, dass diese Totzeit der Reaktion wahrscheinlich auf eine Restrukturierung der Nanopartikeloberfläche zurückzuführen ist. Sie ist unabhängig von den eingesetzten Konzentrationen des Borhydrids, hingegen abhängig von der Konzentration an p-Nitrophenol auf der Oberfläche der Nanopartikel, was auf Restrukturierung der Nanopartikel durch p-Nitrophenol hindeutet. Zudem wurden Hinweise auf eine spontane Rekonstruktion der Nanopartikel gefunden, die unabhängig von der Konzentration des p-Nitrophenols ist. Des Weiteren wurde die katalytische Oxidation von Morin mit Manganoxid Nanopartikeln untersucht. Diese sind in der Polyelektrolytschale der SPB immobilisiert. Analysen der Reaktionskinetik der Morin Oxidation ergaben, dass auch in diesem Fall der LH Mechanismus vorliegt. Hierbei konnten die Adsorptionskonstanten und Geschwindigkeitskonstanten für verschiedene Temperaturen ermittelt werden und somit die Aktivierungsenergie der Oxidation sowie die Adsorptionsenthalpie und Entropie der Edukte. / In this work, gold and platinum nanoparticles were synthesized into spherical polyelectrolyte brushes (SPB) in order to apply them as catalysts for kinetic studies of the reduction of p-nitrophenol by sodium borohydride. It was found that the reaction follows the Langmuir-Hinshelwood (LH) mechanism where both educts must adsorb onto the surface of the catalyst in order to react. Thereby, the rate determining step is the surface reaction of both educts. After the reaction, the product desorbs from the surface and a free active site is formed. With this model the intrinsic reaction rate and the adsorption constants for both educts could be determined. The measurements at different temperatures allowed the calculation of the activation energy and the adsorption enthalpy and entropy of the educts. Besides the reaction mechanism, the induction time of the reaction was analyzed. Here, it was shown that the reason of this delay time is a restructuring of the nanoparticle surface. The induction time is solely dependent on the concentration of p-nitrophenol on the surface of the nanoparticles and independent of the applied concentrations of borohydride. Moreover, hints for a spontaneous reconstruction of the nanoparticles without p-nitrophenol were found. In the second part, the catalytic oxidation of morin by manganese oxide has been studied. These nanoparticles were embedded inside the polyelectrolyte layer of the SPB. These nanoparticles were used for systematic studies of the oxidation of morin with hydrogen peroxide. It was shown that in this case the reaction followed a LH kinetics as well. Here, the intrinsic rate constants and the adsorption constants could be obtained for different temperatures. The activation energy and the adsorption enthalpy and entropy could be determined accordingly. The adsorption enthalpy is exothermic in both cases.

Made available in DSpace on 2017-07-20T12:40:21Z (GMT). No. of bitstreams: 1 Cliciane Guadalupe de Jesus.pdf: 1994532 bytes, checksum: 0f732ce8dacf9081816c07b36ec9e75d (MD5) Previous issue date: 2013-06-21 / In this thesis, it is described the preparation, characterization and application of LbL films based on the 3- n - propylpyridinium silsesquioxane polymer (SiPy+Cl-) and copper (II) tetrasulfophthalocyanine (CuTsPc), denoted as (SiPy+Cl-/CuTsPc)n, for construction of sensors and biosensors. These films (SiPy+Cl-/CuTsPc)n, were applied as a sensor for promethazine hydrochloride (PMZ) and hydrogen peroxide (H2O2). In addition, taking advantage of the property to the film in electrocatalyzing the reduction of H2O2, an enzymatic amperometric biosensor was constructed by the immobilization of the glucose oxidase enzyme (GOx) on the film for detection of glucose, based on the detection of H2O2 produced in the enzymatic reaction. The UV-Vis spectroscopic technique showed that the adsorption of phthalocyanine occurred linearly indicating that the approximately the same amount of the species is immobilized at each bilayer. The interaction between the sulfonic group and the pyridinium ring is responsible for the support of the films due to the shift of the vibration associated with sulfonic groups. The film (SiPy+Cl-/CuTsPc)5 was highly electrocatalytic and sensitive determination of PMZ. The oxidation process of PMZ molecule presented two peaks at 0.48 and 0.79 V. After optimization of the square wave voltammetry parameters (f = 100 s-1, a = 40 mV and ΔEs = 2 mV), the peak at 0.79 V was used for quantification. Accordingly, the modified electrode obtained a linear response of 3.98 x 10-7 to 3.85 x 10-6 mol L-1 with limits of detection (LOD) and quantification (LOQ) equal to 8.71 x 10-9 mol L-1 (2.79 g L-1) and 9.31 x 10-8 mol L-1 (9.31 g L-1), respectively. The recovery in pharmaceutical samples showed high stability, specificity and accuracy of the method in complex samples without any pre-treatment. The film (SiPy+Cl-/CuTsPc)2 showed excellent electrocatalytic activity for the reduction of H2O2 with a wide linear range (4.97 x 10-6 a 4.76 x 10-5 mol L-1) and low detection limit (0.33 x 10-6 mol L-1). This electrode was modified with the glucose oxidase enzyme (GOx) and coated with Nafion® (Nf) film, which was used to prevent leaching of GOx. The Fourier transform infrared, fluorescence and circular dichroism spectroscopics, showed that the GOx enzyme retains its native secondary structure when immobilized on the LbL film. The resulting biosensor (SiPy+Cl-/CuTsPc)2(SiPy+Cl-/GOx/Nf) exhibited a current response which varied linearly with the concentration of glucose in the range of 1-10 mmol L-1, with LOD = 0.17 mmol L-1, sensitivity of 1.397 μA x 10-7/mmol L-1 and a rapid response time, less than 5 seconds. The Michaelis-Menten constant apparent ( appmK ) was 10 mmol L-1. Furthermore, the biosensor also showed good reproducibility, excellent stability and a long life, without loss of enzymatic activity by a one-week period and remained active for more than three weeks. The use of the polymer Nf decreased the effect of interferents on the response of the biosensor. / Esta tese descreve a preparação, a caracterização e a aplicação de filmes automontados entre o polímero cloreto de 3- n -propilpiridínio silsesquioxano (SiPy+Cl-) e a ftalocianina tetrassulfonada de cobre (II) (CuTsPc), designados como (SiPy+Cl-/CuTsPc)n, para a construção de sensores e biossensores. Tais filmes, (SiPy+Cl-/CuTsPc)n, se mostraram promissores como sensor para detecção de cloridrato de prometazina (PMZ) e peróxido de hidrogênio (H2O2). Adicionalmente, tirando-se proveito da propriedade do filme em eletrocatalisar a redução do H2O2, construiu-se um biossensor enzimático amperométrico, a partir da imobilização da enzima glicose oxidase (GOx) sobre o filme, para a detecção de glicose, baseado na detecção do H2O2 produzido na reação enzimática. A técnica de espectroscopia de UV-Vis revelou que a adsorção de ftalocianina ocorre de maneira linear indicando que a cada bicamada depositada aproximadamente a mesma quantidade de material é imobilizado. A interação entre os grupos sulfônicos e o anel piridínio é a responsável pela sustentação dos filmes, devido ao deslocamento da banda associada à vibração dos grupos sulfônicos. O filme (SiPy+Cl-/CuTsPc)5 mostrou propriedades eletrocatalíticas na determinação de PMZ. Este apresentou dois processos de oxidação em 0,48 e 0,79 V. Após a otimização dos parâmetros da voltametria de onda quadrada (f = 100 s-1, a = 40 mV e ΔEs = 2 mV), o pico a 0,79 V foi utilizado para sua quantificação. Nestas condições, o eletrodo modificado obteve uma resposta linear de 3,98 x10-7 a 3,85 x 10-6 mol L-1 com limites de detecção (LD) e quantificação (LQ) iguais a 8,71 x 10-9 mol L-1 (2,79 μg L-1) e 9,31 x 10-8 mol L-1 (9,31 μg L-1), respectivamente. A recuperação em amostras farmacêuticas revelou alta estabilidade, especificidade e precisão do método, sem qualquer pré-tratamento. O filme (SiPy+Cl-/CuTsPc)2 demonstrou excelente atividade eletrocatalítica para redução de H2O2 com uma ampla resposta linear (4,97 x 10-6 a 4,76 x 10-5 mol L-1) e baixo limite de detecção (0,33 x 10-6 mol L-1). Este eletrodo foi modificado com a enzima glicose oxidase (GOx) e recoberto com filme de Nafion® (Nf), que foi usado para prevenir o lixiviamento da GOx. As técnicas espectroscópicas de infravermelho, fluorescência e dicroísmo circular mostraram que a enzima GOx mantém a sua estrutura nativa secundária quando imobilizado sobre o filme LbL. O biossensor resultante (SiPy+Cl-/CuTsPc)2(SiPy+Cl-/GOx/Nf) exibiu uma resposta de corrente que variou linearmente com a concentração de glicose na faixa de 1 a 10 mmol L-1, com LD = 0,17 mmol L-1, sensibilidade de 1,397 x 10-7 μA/mmol L-1 e um rápido tempo de resposta, inferior a 5 s. A constante de Michaelis-Menten aparente ( appmK ) foi de 10 mmol L-1. Além disso, o biossensor também apresentou boa reprodutibilidade, excelente estabilidade e longo tempo de vida, sem diminuição da atividade enzimática por um período de uma semana e permaneceu ativo por mais de três semanas. A utilização do polímero Nf possibilitou a diminuição do efeito de interferentes na resposta do biossensor. Palavras chave: Filmes LbL, polieletrólito Silsesquioxano, Biossensor Amperométrico, Glicose Oxidase.

Made available in DSpace on 2014-12-17T15:42:27Z (GMT). No. of bitstreams: 1 MariaSPL.pdf: 2188057 bytes, checksum: 3142bea2af0c353bf960b7b878efd707 (MD5) Previous issue date: 2006-11-17 / The aim of this study was to generate an asymmetric biocompactible and biodegradable chitosan membrane modified by the contact with a poly(acrylic acid) solution at one of its sides at room temperature and 60◦C. The pure chitosan membrane, as well as the ones treated with poly(acrylic acid) were characterized by infrared spectroscopy (FTIRATR) at angles of 39◦, 45◦ and 60◦ , swelling capacity in water, thermal analysis (TG/DTG), scanning electronic microscopy (SEM) and permeation experiments using metronidazole at 0,1% and 0,2% as a model drug. The results confirmed the presence of ionic interaction between chitosan and poly(acrylic acid) by means of a polyelectrolyte complex (PEC) formation. They also showed that such interactions were more effective at 60◦C since this temperature is above the chitosan glass transition temperature wich makes the diffusion of poly(acrylic acid) easier, and that the two treated membranes were asymmetrics, more thermically stable and less permeable in relation to metronidazole than the pure chitosan membrane / O prop?sito deste estudo foi produzir uma membrana assim?trica biocompat?vel e biodegrad?vel de quitosana modificada pelo contato com uma solu??o de poli(?cido acr?lico) em uma de suas superf?cies ? temperatura ambiente e a 60◦C. As membranas de quitosana pura, quitosana com poli(?cido acr?lico) a 25◦C e quitosana com poli(?cido acr?lico) a 60◦C foram caracterizadas por: espectroscopia no infravermelho (FTIRATR) em ?ngulos de incid?ncia de 39◦, 45◦ e 60◦, ganho de massa em ?gua, an?lise t?rmica (TG/ DTG), microscopia eletr?nica de varredura (MEV) e, ainda, atrav?s dos ensaios de permea??o in vitro utilizando como f?rmaco modelo o metronidazol em solu??o aquosa nas concentra??es de 0,1 e 0,2%. Os resultados obtidos comprovaram a exist?ncia de intera??es i?nicas entre os dois pol?meros, atrav?s da forma??o dos chamados complexos polieletrol?ticos. Tamb?m mostraram que a reticula??o foi mais efetiva a 60◦C , uma vez que essa temperatura est? acima da temperatura de transi??o v?trea da quitosana, o que facilita a difus?o do poli(?cido acr?lico) e que as membranas resultantes s?o assim?tricas, mais est?veis termicamente e menos perme?veis ao metronidazol do que a membrana de quitosana pura

Made available in DSpace on 2017-05-12T14:47:16Z (GMT). No. of bitstreams: 1 Juliana Cecchet.pdf: 2090330 bytes, checksum: 58b3febaf50310456ede4d0b8648a7f0 (MD5) Previous issue date: 2007-07-06 / The aim of this work was to value the performance of three types of coagulants, two of them of polichlorate of alum class and one of them was organic, combined to three polyelectrolytes class (anionic, cationic and no ionic) applying the dissolved air flotation (DAF), to viability their use on treatment of effluents of soybean oil refinery. The study was conducted on laboratory scale where first was tested the coagulants on several dosages combined to several bands of pH and three ascension velocities (Va), finding a optimum band of this combination for one of each three batteries. For this, was fixed parameters how time and fast mixed gradient (1 min / 150 s-1), time and slow mixed gradient (10 min / 40 s-1 5 min / 20 s-1), pressure on saturation chamber (450 kPa) and recirculation ratio (30%). Finding the optimum concentration of coagulant (Cc ótima), optimum coagulation pH (pHcoag ótimo) and optimum ascension velocities (Vaótima), fix those parameters and then goes for a variation of polyelectrolyte, testing several dosages and finding a optimum concentrate of polyelectrolyte (Cf ótimo). Finally, with junction of all optimized parameters, was varied de recirculation rate (R) (10, 20 e 30%), choosing the best R and collecting samples for the determination of levels of turbidity, total suspended solids (TSS), volatile suspended solids (VSS), oil and greases (O&G) and chemical oxygen demand (COD) remaining on treated effluent presenting a percentage of remotion COD of 95,4%, 93,7% and 95,1% and O&G of 100%, 94,2% e 99,8% for the batteries 1, 2 e 3 respectively. Despite of being excellent efficiencies of remotion, the treated effluent was not present characteristics for the reuse on the cooling water towers, because its organics load on the raw effluent its too much higher and even with so good levels of remotion, still was necessary a pos treatment to think on reuse. / Este trabalho foi desenvolvido com o objetivo de avaliar o desempenho de três tipos de coagulantes, dois da classe dos PACs (cloreto de polialumínio) e um orgânico, combinados a três classes de polieletrólitos (aniônico, catiônico e não iônico), aplicando o uso de Flotação por Ar Dissolvido (FAD), para viabilizar seu uso em tratamento de efluentes de refinaria de óleo de soja. O experimento foi conduzido em escala de laboratório e primeiramente foram testados os coagulantes em diversas dosagens combinados a diversas faixas de pH e três velocidades ascensionais (Va), encontrando uma faixa ótima dessa combinação para cada uma das três baterias. Para tanto foram fixados parâmetros como tempo e gradiente de mistura rápida (1 min / 150 s-1) tempo e gradiente de mistura lenta (10 min / 40 s-1 5 min / 20 s-1), pressão na câmara de saturação (450 kPa) e razão de recirculação (30%). Encontrando-se a concentração de coagulante ótima (Cc ótima), pH de coagulação ótimo (pHcoag ótimo) e velocidade ascensional ótima (Vaótima). Fixados esses parâmetros foi verificada a variação do polieletrólito, testando-se em diversas dosagens e encontrando uma concentração de floculante ótima (Cf ótimo). Finalmente, com a junção de todos os parâmetros otimizados, efetua-se a variação da razão de recirculação (R) (10, 20 e 30%), escolhendo a melhor R e coletando-se amostras para a determinação dos níveis de turbidez, sólidos suspensos totais (SST), sólidos suspensos voláteis (SSV), óleos e graxas (O&G) e demanda química de oxigênio (DQO) remanescentes no efluente tratado apresentando um grau de remoção de DQO de 95,4%, 93,7% e 95,1% e de O&G de 100%, 94,2% e 99,8% para as baterias 1, 2 e 3 respectivamente. Apesar de serem eficiências excelentes de remoção, o efluente tratado não apresentou características de reuso em torres de resfriamento, pois sua carga orgânica no efluente bruto é muito alta e mesmo com níveis tão bons de remoção ainda seria necessário um pós-tratamento para se avaliar a possibilidade de reuso.

La régénération des défauts osseux de taille critique reste un défi majeur pour la santé. Les limitations des greffes de tissus communes nous ont incités à développer une alternative synthétique basée sur la construction d’un biomatériau, des facteurs ostéoinductifs et des cellules souches. Pour la construction du biomatériau, nous nous sommes concentrés sur les microporteurs polymères poreux, car ils supportent une expansion cellulaire à grande échelle et un assemblage modulaire des tissus, contournant deux goulots d'étranglement importants pour la traduction clinique. Pour assurer l'approvisionnement industriel et l'approbation réglementaire, nous avons conçu une méthode de fabrication sans solvant organique basée sur la cristallisation sphérulitique du poly(L-lactide) (PLLA) dans ses mélanges avec du polyéthylène glycol (PEG). Les sphérulites de PLLA ont été facilement récupérées sous forme de microporteurs en éliminant par rinçage le PEG soluble dans l'eau. Leur taille et leur porosité pourraient être contrôlées indépendamment en ajustant le rapport PLLA / PEG et la température de cristallisation. La biocompatibilité et l'ostéoconductivité des microporteurs à PLLA ont été confirmées par l'expansion et la différenciation ostéogénique des cellules souches adipeuses humaines (hASC). Comme cette dernière fonction hASC est sensible à différents paramètres de culture, nous avons ensuite utilisé l'approche de conception d'expériences pour leur dépistage rapide. En combinaison avec l'analyse à haut débit, nous avons identifié plusieurs paramètres ayant une influence marquée sur leur différenciation ostéogénique. Enfin, pour la délivrance de facteurs ostéoinducteurs, nous avons élaboré des multicouches de polyélectrolytes (PEM) à base de poly (L-ornithine) et d'acide hyaluronique biocompatibles. Ces PEM ont été caractérisées en termes de croissance, de morphologie, d'aptitude à incorporer des protéines morphogénétiques osseuses (BMP) et à fonctionner en tant que revêtements sur des microporteurs à PLLA. Nos résultats préliminaires ont montré que l’incorporation de BMP dans les PEM avait un effet important sur l’adhérence des hASC. Bien que des études supplémentaires soient nécessaires, les microporteurs à PLLA recouverts de PEM chargés de BMP et ensemencés avec hASC pourraient être un implant synthétique prometteur pour une régénération osseuse améliorée. / The regeneration of critical-sized bone defects remains a major healthcare challenge. The limitations of common tissue grafts prompted us to develop a synthetic alternative based on a biomaterial construct, osteoinductive factors and stem cells. For biomaterial construct, we focused on porous polymeric microcarriers as they support large-scale cell expansion and modular tissue assembly, circumventing two important bottlenecks for clinical translation. To insure industrial supply and regulatory approval, we designed an organic-solvent-free method for their fabrication based on the spherulitic crystallization of poly(L-lactide) (PLLA) in its blends with polyethylene glycol (PEG). The PLLA spherulites were easily recovered as microcarriers by rinsing away the water-soluble PEG. Their size and porosity could be independently controlled by tuning the PLLA/PEG ratio and crystallization temperature. The biocompatibility and osteoconductivity of PLLA microcarriers were confirmed through the expansion and osteogenic differentiation of human adipose stem cells (hASCs). Because the latter hASC function is sensitive to different culture parameters, we then used the Design of Experiments approach for their rapid screening. In combination with high-throughput analysis, we identified several parameters that had a pronounced influence on their osteogenic differentiation. Finally, for the delivery of osteoinductive factors, we elaborated polyelectrolyte multilayers (PEM) based on biocompatible poly(L-ornithine) and hyaluronic acid. These PEMs were characterized in terms of their growth, morphology, the ability to incorporate bone morphogenetic proteins (BMP) and to function as coatings on PLLA microcarriers. Our preliminary results showed that the incorporation of BMPs inside PEMs had a strong effect on hASC adhesion. While further studies are needed, hASC-seeded PLLA microcarriers coated with BMP-loaded PEMs could be a promising synthetic implant for improved bone regeneration.