Made available in DSpace on 2014-07-29T15:12:48Z (GMT). No. of bitstreams: 1 Dissertacao - Joel Rocha da Silva.pdf: 557734 bytes, checksum: 97955f40a90305a3592aea224371f9ea (MD5) Previous issue date: 2008-05-27 / Magnetite nanoparticles were prepared by the coprecipitation of ions Fe2+ and Fe3+ using ammonia solution as precipitating agent. Maghemite nanoparticles were prepared by forced oxidation of magnetite nanoparticles in acidic medium using nitrate ions as oxidizing agent. The magnetic nanoparticles were used to the preparation of aqueous magnetic fluids samples by the functionalization of the nanoparticles with bilayers of laurate/laurate and laurate/Pluronic. Aliquots of the magnetic fluids were dried and the resultant powders were characterized by chemical analysis (the contents of ions Fe2+ and Fe3+), X-ray powder diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR). The contents of ions Fe2+ and Fe3+ in all the samples showed that the nanoparticles are not pure magnetite or maghemite phases. X-ray powder diffraction (XRD) indicated the existence of inverse cubic spinel phase, but didn't permit the distinction between magnetite and maghemite phases. Based on the results of chemical and XRD analyses, the nanoparticles could be better characterized as reduced maghemite, which mean maghemite phase containing ions Fe2+. The average sizes of the oxide nanoparticles estimated by XRD were around of 10 nm. FTIR analyses showed that the nanoparticles were functionalized with bilayers of laurate/laurate and laurate/Pluronic. FTIR analyses also were indicative of the maghemite phase. The hydrodynamic size of the functionalized nanoparticles measured by PCS were in the range of 70-90 nm for the samples based on laurate and in the range of 100-200 nm for the samples containing Pluronic. The measurements of zeta potential showed that the magnetic fluids based on laurate bilayers presented better colloidal stability than that one based on bilayers of laurate/Pluronic. On the other hand, the studies of colloidal stability in cell culture medium by hydrodynamic size measurements showed the lost of colloidal stability of all the samples, in all the concentrations of medium investigated. The fluids with laurate/laurate bilayers showed higher ratio of aggregation in the culture medium than the fluids with laurate/Pluronic. The association of amphotericin B with nanoparticles was carried out only using the samples with laurate bilayers. The results showed that the ratio of association depends on the concentration of amphotericin added to the magnetic fluid. It was observed 100% of amphotericin association in the range of nanoparticles/amphotericin concentrations used, which in turn are adequated for posterior in vivo studies / Nanopartículas de magnetita foram preparadas pela coprecipitação dos íons Fe2+ e Fe3+ usando uma solução de amônia como o agente de precipitação. As nanopartículas de maguemita foram preparadas por oxidação forçada de nanopartículas de magnetita usando íons nitrato como o agente de oxidação, em meio ácido. As nanopartículas magnéticas foram usadas para a preparação de fluidos magnéticos aquosos pela funcionalização das nanopartículas com bicamadas de laurato/laurato e laurato/Pluronic. As alíquotas dos fluidos magnéticos foram secas e o pó resultante foi caracterizado por análise química (teores dos íons Fe2+ e Fe3+), difração de raios X pelo método do pó (DRX) e espectroscopia na região do infravermelho com transformada de Fourier (FT-IR). Os teores dos íons Fe2+ e Fe3+ em todas as amostras mostram que as nanopartículas não são fases puras da magnetita ou de maguemita. A difração de raios X (DRX) pelo método do pó indicou a existência da fase cúbica de espinélio inverso, mas não permite a distinção entre a magnetita e as fases de maguemita. Baseado nos resultados de análises químicas e de DRX, as nanopartículas poderiam ser melhor caracterizadas como maguemita reduzida, a qual resulta em uma fase de maguemita contento íons Fe2+. Os tamanhos médios das nanopartículas dos óxidos estimados por DRX foram em torno de 10 nm. As analises de FT-IR mostram que as nanopartículas foram funcionalizadas com bicamadas de laurato/laurato e laurato/Pluronic. As análises de FT-IR também foram indicativos da fase maguemita. O tamanho hidrodinâmico das nanopartículas funcionalizadas medidas por PCS foi em torno de 70-90 nm para amostras a base de laurato e em torno de 100-200 nm para amostras contendo Pluronic. As medidas de potencial zeta mostraram que os fluidos magnéticos a base de bicamada de laurato apresentam melhor estabilidade coloidal do que os a base de bicamada de laurato/Pluronic. Por outro lado, o estudo da estabilidade coloidal em meio de cultura por medidas de tamanho hidrodinâmico mostra a perda da estabilidade coloidal de todas as amostras, em todas as concentrações do meio investigado. Os fluidos com bicamadas de laurato/laurato mostraram taxas mais altas de agregação no meio de cultura do que os fluidos com laurato/Pluronic. Os estudos de associação de anfotericina B com nanopartículas foram realizados somente usando as amostras com bicamadas laurato/laurato. Os resultados mostraram que a taxa de associação depende da concentração de anfotericina adicionada ao fluido magnético. Foi observado 100% de associação da anfotericina na faixa da concentração nanopartículas/anfotericina usadas, que por sua vez são adequadas para posteriores estudos in vivo

Les thérapies ciblées constituent une révolution médicale dans le traitement du cancer. Dans ce contexte, le pseudopeptide multivalent N6L, qui cible spécifiquement les cellules tumorales et induit leur mort, apparaît comme une molécule prometteuse. Le N6L cible en effet deux nucléoprotéines surexprimées à la surface des cellules cancéreuses qui sont la nucléoline et la nucléophosmine. L'étude du mécanisme d'action anti-métastatique du N6L, dans lequel l'implication du TIMP-3 a été soulignée, a permis d'identifier une nouvelle cible : les glycosaminoglycanes sulfatés (GAG).Dans le but de développer une approche multimodale pour le diagnostic et le traitement du cancer du sein, le N6L a été greffé à la surface de nanoparticules magnétiques (NPM-N6L). La propriété de ciblage tumorale des NPM-N6L a été démontrée in vitro et in vivo. Leur cible tumorale majeure fut les GAG.Par ailleurs, l'activité anti-tumorale du N6L dans le traitement du cancer de la prostate à différents stades de la maladie a été démontrée in vitro et in vivo. La cible tumorale mise en jeu fut la nucléophosmine. En outre, ce potentiel anti-tumoral implique une diminution de l'activité du récepteur aux androgènes et probablement une voie de signalisation impliquant une interaction entre la nucléophosmine phosphorylée et le récepteur aux androgènes. / Targeted therapies constitute a revolution in the medical treatment of cancer. In this context, the multivalent pseudopeptide N6L that specifically target tumor cells and induces their death seems a promising molecule. The N6L targets two nucleoproteins that are overexpressed on the surface of cancer cells: nucleolin and nucleophosmin. The study of the N6L anti-metastatic mechanism of action in which TIMP-3 was involved, has identified a new target: sulfated glycosaminoglycans (GAG).In order to develop a multimodal approach for diagnosis and treatment of breast cancer, the N6L was grafted to the surface of magnetic nanoparticles (MNP-N6L). Tumor targeting properties of MNP-N6L was demonstrated in vitro and in vivo. Their major tumor target was GAG.Furthermore, the N6L anti-tumor activity in the treatment of prostate cancer at different stages of the disease was demonstrated in vitro and in vivo. The target involved in this activity was nucleophosmin. Furthermore, this anti-tumor potential implies a decrease in the activity of the androgen receptor and probably a signaling pathway involving an interaction between phosphorylated nucleophosmin and the androgen receptor.

Les nanocolloïdes magnétiques sont des dispersions de nanostructures magnétiques dans un liquide porteur. Par la combinaison des propriétés du liquide et des particules magnétiques, ces dispersions peuvent être confinées, déplacées, déformées et contrôlées par l'application d'un champ magnétique externe et ont ainsi de nombreuses applications en nanosciences et les nanotechnologies. Nous avons étudié la dispersion de nanoparticules magnétiques (NPM) dans les solvants eutectiques profonds (DES). Ces solvants, constitués d'un mélange entre un sel d'ammonium (ici le chlorure de choline (Ch) et un donneur de liaison H (ici, l'ethyleneglycol (EG) ou l'urée (U)) ont des propriétés proches des liquides ioniques tout en étant biodégradables. L'un des verrous concernant ces dispersions est la nature des forces impliquées dans la stabilité colloïdale. En effet, on ne peut plus expliquer la stabilité des dispersions dans ces milieux par le modèle DLVO, classiquement utilisé dans l'eau, du fait de leur force ionique élevée. Nous avons dans en premier temps caractérisé soigneusement deux DES (ChEH (1:3) et ChU (1:2) en mol) du point de vue de la densité et viscosité pour des températures entre 20 et 45°C. Ceci nous a permis de montrer la forte association de ces liquides. Un protocole de dispersion de nanoparticules de maghémite (Fe2O3) ou de ferrite mixte (CoxZn1-xFe2O4) est ensuite proposé, et les dispersions sont étudiées par diffusion de rayonnement (lumière et SAXS). Il s'est avéré que les particules les plus petites étaient les mieux dispersées. Enfin, un test de synthèse de NPM dans des solutions d'argile a permis d'obtenir une polydispersité plus faible en sortie de synthèse. / Magnetic nanocolloids are dispersions of magnetic nanostructures in a carrier fluid. Thanks to the original properties of both the liquid and the magnetic particles, these dispersions can be confined, moved, deformed and controlled by applying an external magnetic field. Such dispersions thus have many applications in nanoscience and nanotechnologies.We studied the dispersion of magnetic nanoparticles in deep eutectic solvents (DES). These solvents (DES), obtained by mixing a quaternary ammonium salt (e.g., choline chloride Ch) and a hydrogen bond donor (e.g., ethyleneglycol EG or Urea U) have properties similar to ionic liquids, and are also biodegradable. One of the questions about these dispersions is the nature of the forces implied in colloidal stability, since the DLVO model classically used in water cannot be invoked here due to the very high ionic strength of the solvent.In a first step, we have carefully characterized two DES ((ChEG (1:3) and ChU (1:2) in mol), measuring the density and viscosity for temperatures between 20 and 45°C. We could thus show the high association in these liquids.A protocol to disperse nanoparticles of maghemite (Fe2O3) or mixed ferrite (CoxZn1-xFe2O4) is then proposed, and the obtained dispersions are studied by dynamic light scattering and SAXS. The size polydispersity was reduced by size sorting, and it reveals that the smallest particles are the most easy to disperse in the DES.Last, a synthesis of NMP in clay dispersion was tested and showed promising results with a reduced size polydispersity.

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The thesis has been focused on testing of micromethod of DNA isolation using magnetic particles from thermic-managed food products in a quality suitable for polymerase chain reaction (PCR). Currant jams were selected for the analysis. These were homogenized using plastic copist and stomacher in lysis buffer with cetyltrimethylammonium bromide (CTAB). The effect of chloroform-octanol and isopropanol in the preparation of homogenates was tested. Homogenates were used for DNA isolation by magnetic particles. Rough fraction of DNA was purified by binding on the magnetic particles after centrifugation of the CTAB complexes with proteins, polyphenols and polysaccharides. Two types of magnetic particles were tested: microparticles of poly(hydroxyethylmethacrylate-co-glycidylmethacrylate) - P(HEMA-co-GMA) and nanoparticles of iron oxides covered by poly(L-lysine) - PLL. Isolated DNA was analyzed spectrophotometrically - it was assessed its concentration and contamination by polyphenols and proteins. After that, amplification of the DNA was tested in PCR. Primers specific for plant ribosomal DNA were used. PCR products of expected length 700 bp were detected by agarose gel electrophoresis. It was shown that DNA isolated from currant jams using magnetic particles was in PCR-ready quality.

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In this study, the magnetically-induced vibrations of functionalized magnetic particle suspensions were probed for the development of a novel optical spectroscopy technique. Through this work (1) the frequency-dependence of the faraday rotation in ferrofluids and (2) the extension of this system to elucidating particle size and conformation as an alternative immunossay to costly and labor/time intensive Western Blotting and ELISA has been shown. With its sensitivity and specificity, this method has proven to be a promising multi-functional tool in biosensing, diagnostic, and therapeutic nanomedicine efforts. Due to its ubiquitous nature in all optically-transparent materials, the farady rotation, or circular birefringence, was developed as a robust and sensitive nanoscale biomolecule characterization technique through Brownian relaxation studies of particle suspensions. Current efforts have shown the applicability of this phenomenon in solid, pure liquid, and colloidal samples as well as simultaneous advancements of magnetic nanoparticle research in the magnetometric and magneto-optical regimes. By merging these two fields, a clinically relevant spectroscopy (fd-FRS, Frequency Domain Faraday Rotation Spectroscopy) was developed based on a newly revised model stemming from Debye relazation theory. Through this work, an optical bench with a variable permeability core electromagnet and a frequency-domain lock-in amplifier setup (DC to 20 kHz) have been used to distinguish between Fe3O4-core nanoparticles with functionalization layers of PEG4/PEG8 polymer with future applications involving the Anti-BSA/BSA antibody/antigen couple. Particle concentrations down to 500 nM (magnetic nanoparticles) and 0.01 Volume % (magnetic beads) were studied with diameters ranging from 200 nm to 1μm. currently, the characteristic peak corresponding to the out-of-phase relazation of the suspended particles has been elusive, despite a wide particle size distribution and the use of a balanced photodetector. Future work will involved highly monodisperse samples, faster scan times, and thermal characterization applications of fs-FRS.

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[pt] As nanopartículas magnéticas (NPMs) apresentam grande potencial em diversas aplicações tecnológicas e vem ganhando destaque na área da biomedicina devido suas propriedades superparamagnéticas. Para este trabalho foram sintetizadas nanopartículas de óxido de ferro (Fe3O4) pelo método de coprecipitação e recobertas com Pluronic F-127 (PL F-127), o qual se demonstrou, a partir de estudos anteriores, um surfactante com ótima estabilidade coloidal e alto grau de biocompatibilidade, sendo tais características relevantes para as aplicações na área da biomedicina. O método de produção se mostrou eficiente para produção de uma grande quantidade de amostra e baixo grau de oxidação, o que mantém a integridade dos resultados e do material para diversas análises no decorrer de longos períodos. A espectroscopia Raman e a difração de elétrons apontam para a composição majoritária de magnetita cristalina das amostras. As imagens obtidas através de microscopia de transmissão (MET) mostraram que o diâmetro médio das NPMs não é afetado pela concentração PL F-127 e está de acordo com os tamanhos obtidos pelas técnicas magnéticas. O MET também mostrou partículas monodispersas com formato esféricas. As técnicas de microscopia magnética de varredura (MMV), magnetômetro de amostra vibrante e de efeito Hall revelaram que o comportamento das NPMs é superparamagnético em temperatura ambiente e que a funcionalização não interferiu significativamente na magnetização de saturação. / [en] Magnetic nanoparticles (MNPs) have great potential in several technological applications and are gaining prominence in the biomedical area due to their superparamagnetic properties. For this work, iron oxide (Fe3O4) nanoparticles were synthesized by the coprecipitation method and coated with Pluronic F-127 (PL F127), which was demonstrated, from previus studies, a surfactante with excellent coloidal stability and high degree of biocompatibility, such characteristics being relevant for applications in the área of biomedicine. The production method proved to be efficient for producing a large amount of sample and a low degree of oxidation, which maintains the integrity of the results and material for several analyzes over long periods. Raman spectroscopy and eléctron diffraction indicate the samples are pure and crystalline magnetite. The images obtained through transmission eléctron microscopy (TEM) showed that the mean diameter of MNPs is not affected by the PL F-127 concentration and is in agreement with the sizes obtained by magnetic techniques. TEM also showed monodisperse particles with a spherical shape. Scanning magnetic microscopy (MMV), vibrating sample magnetometer and Hall effect techniques revealed that the behavior of NPMs is superparamagnetic at ambient temperature and that the functionalization did not significantly interfere in the saturation magnetization.