Une approche synthétique a été développée pour préparer les phtalocyanines dimériques de fer azote ponté substituées par les substituants alkylesulfoniles attracteurs d’électrons. Six nouvelles phtalocyanines avec des petits substituants (méthylesulfonile, éthylesulfonile et hexylesulfonile) et des gros substituants (t-butylesulfonile, adamantylesulfonile et cyclohexylesulfonile) ont été préparées et caractérisées par ionisation électrospray (ESI-MS), UV-vis, FT-IR et RPE. Deux complexes (avec substituants hexylesulfoniles et t-butylesulfoniles) ont été caractérisés par spectroscopie Mössbauer, spectroscopie photoélectronique de rayons X (XPS) et par spectroscopie d’absorption des rayons X (XANES, EXAFS, spectroscopie d’émission Kβ haute résolution). Il a été montré que l’état électronique de fer de ces complexes dépend de la taille de substituants. Les phtalocyanines de fer à l’azote ponté avec les gros substituants (t-butylesulfonile, adamantylesulfonile et cyclohexylesulfonile) sont cationiques (PcFeIVNFeIVPc)+N3-, tandis que les complexes avec les substituants plus gros (t-butylesulfonyles, adamantylesulfonyles et cyclohexylesulfonyles) sont non-chargés, formellement PcFeIIINFeIVPc. Les propriétés catalytiques des six complexes ont été étudiées en utilisant du butyle hydroperoxyde comme oxydant en oxydation de toluène, de p-xylène et des alcools. Ce travail montre l’efficacité des phtalocyanines dimériques de fer azote ponté substituées par les groupements alkylesulfoniles attracteurs d’électrons comme catalyseurs pour oxydation dans les conditions favorables pour les applications industrielles propres / The synthetic approach was developed for the preparation of N-bridged diiron phthalocyanines substituted by different electron-withdrawing alkylsulfonyl substituents. Six novel phthalocyanines bearing small (methylsulfonyl, ethylsulfonyl and hexylsulfonyl) and bulky (t-butylsulfonyl, adamantylsulfonyl and cyclohexylsulfonyl) substituents have been prepared and characterized by electrospray ionization mass spectrometry (ESI-MS), UV-Vis, FT-IR and EPR. Two complexes (with hexylsulfonyl and t-butylsulfonyl substituents) were characterized in addition by Mössbauer techniques, X-ray photoelectron spectroscopy (XPS) and Fe K-edge X-ray absorption spectroscopy (XANES, EXAFS, high resolution Kβ emission spectroscopy). It has been evidenced that the electronic state of iron in these complexes depends on the size of the substituents. While N-bridged diiron phthalocyanines having methylsulfonyl, ethylsulfonyl and hexylsulfonyl substituents are cationic (PcFeIVNFeIVPc)+N3- complexes, N-bridged diiron phthalocyanines with bulkier t-butylsulfonyl, adamantylsulfonyl and cyclohexylsulfonyl substituents are formally neutral PcFeIIINFeIVPc species. The catalytic properties of six N-bridged diiron phthalocyanines have been studied, using tert-butyl hydroperoxide (tBuOOH) as the oxidant in the oxidation of toluene, p-xylene as well as in the oxidation of various alcohols. This thesis demonstrates the efficiency of N-bridged diiron phthalocyanines substituted by electron-withdrawing alkylsulfonyl groups as oxidation catalysts, in conditions required by environmental and industrial preoccupations

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2016-08-05T10:23:01Z No. of bitstreams: 2 Dissertação - Taise Maria dos Anjos Oliveira - 2016.pdf: 1420716 bytes, checksum: 75628b01c19c9db89c0e4ed2e9469284 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-05T15:03:14Z (GMT) No. of bitstreams: 2 Dissertação - Taise Maria dos Anjos Oliveira - 2016.pdf: 1420716 bytes, checksum: 75628b01c19c9db89c0e4ed2e9469284 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-05T15:03:14Z (GMT). No. of bitstreams: 2 Dissertação - Taise Maria dos Anjos Oliveira - 2016.pdf: 1420716 bytes, checksum: 75628b01c19c9db89c0e4ed2e9469284 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-02-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Photodynamic inactivation is a technique based on the synergistic combination of a photosensitizer, light and oxygen, resulting in the generation of reactive oxygen species, which are cytotoxic and cause cell death by apoptosis or necrosis. Bovine herpesvirus 1 is responsible for diseases of the respiratory and reproductive systems, it is estimated to occur an economic loss of a billion dollars a year, due to the action of the virus in the herd. In order to develop new alternatives and /or therapeutic approaches that are effective in viral clearance, we evaluated two photosensitizers, zinc phthalocyanine and aluminum phthalocyanine, the photoinactivation bovine herpesvirus 1. As a control we used irradiated virus without the presence of photosensitizer (light control) viruses incubated with photosensitizers without radiating (photosensitizer control) and viruses without any treatment. The light control showed that the use of light without the presence of photosensitizer does not have phototoxic effect on the virus; control showed that the photosensitizer zinc and aluminum phthalocyanines have no cytotoxicity in the absence of light. Both controls were compared to the virus that received no treatment. To better evaluate the irradiation time and the optimal concentration of the two photosensitizers, viral suspension aliquots containing 105.75 TCID50/mL were used, incubated with photosensitizers at concentrations of 5 and 10 μM and irradiated at 0, 15, 30, 45, 60, 75, and 90 minutes. After irradiation, the sample was added in permissive cell cultures virus, which was analyzed for the presence of cytopathic effect and the results were expressed as viral titers. The zinc phthalocyanine showed better efficiency in photoinactivation bovine herpesvirus 1, wherein the concentration of 10 μM with 30 minutes of irradiation was the most effective. The phthalocyanine aluminum inactivated virus completely after 60 minutes of irradiation, but there was no significant difference between concentrations. These results indicate that both phthalocyanines have a good applicability in photodynamic viral inactivation, but the zinc phthalocyanine has a better efficiency. / A inativação fotodinâmica é uma técnica baseada na combinação sinérgica de um fotossensibilizador, oxigênio e luz, que resulta na geração de espécies reativas do oxigênio, que são citotóxicas e causam a morte celular por apoptose ou necrose. O herpesvírus bovino 1 é responsável por enfermidades no sistema respiratório e reprodutivo, estima-se que ocorra uma perda econômica de um bilhões de dólares por ano, devido à ação do vírus no rebanho. Visando novas alternativas e/ou abordagens terapêuticas que sejam eficazes na eliminação viral, avaliamos dois fotossensibilizadores, ftalocianina zinco e ftalocianina alumínio, na fotoinativação do herpesvírus bovino 1. Como controle foram utilizados vírus irradiado sem a presença do fotossensibilizador (controle da luz), vírus incubado com os fotossensibilizadores sem irradiar (controle do fotossensibilizador) e vírus sem nenhum tratamento. O controle da luz mostrou que o emprego da luz sem a presença do fotossensibilizador não possui efeito fototóxico sobre o vírus, o controle do fotossensibilizador mostrou que as ftalocianinas zinco e alumínio não possuem citotoxidade na ausência da luz. Ambos os controles foram comparados ao vírus que não recebeu nenhum tratamento. Para avaliar o melhor tempo de irradiação e a melhor concentração dos dois fotossensibilizadores, foram utilizadas alíquotas de suspensão viral contendo 105,75 TCID50/mL, incubadas com os fotossensibilizadores nas concentrações de 5 e 10 μM e irradiadas nos tempos 0, 15, 30, 45, 60, 75 e 90 minutos. Após a irradiação a amostra foi adicionada em culturas de células permissíveis ao vírus, onde foi analisada a presença de efeito citopático e os resultados foram expressos em títulos virais. A ftalocianina zinco apresentou melhor eficiência na fotoinativação do herpesvírus bovino 1, sendo a concentração de 10 μM com 30 minutos de irradiação a mais eficaz. A ftalocianina alumínio inativou o vírus totalmente após 60 minutos de irradiação, mas não apresentou diferença significativa entre as concentrações. Estes resultados indicam que ambas ftalocianinas possuem uma boa aplicabilidade na inativação fotodinâmica viral, porém a ftalocianina zinco possui uma melhor eficiência.

Made available in DSpace on 2014-06-11T19:32:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-06-26Bitstream added on 2014-06-13T19:43:38Z : No. of bitstreams: 1 zanfolim_aa_dr_bauru.pdf: 2078904 bytes, checksum: 89fecc6406c0fe251fffb9f3101d20dd (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Neste trabalho foram preparados filmes finos de ftalocianinas de zinco (ZnPc) e de níquel (NiPc) através da técnica de evaporação térmica à vácuo (PVD - physical vapor deposition) em diversas espessuras e em escala nanométrica com o objetivo de determinar a arquitetura molecular destes filmes bem como suas propriedades ópticas e elétricas. Em última análise buscam-se gerar subsídios para as possíveis aplicações, especialmente dispositivos eletrônicos a base de semicondutores orgânicos e sensores de gás. A ZnPc e a NiPc em pó foram caracterizadas utilizando-se as técnicas de termogravimetria (TG), calorimetria exploratória diferencial (DSC) e os filmes PVD por espectroscopias de absorção no ultravioleta-visível (UV-vis), no infravermelho com transformada de Fourier (FTIR), espalhamento Raman, difração de raios-X, microscopias óptica e de força atômica (AFM) e caracterização elétrica cc (tensão x corrente). Os resultados mostraram que é possível a fabricação de filmes PVD de ZnPc e NiPc, uma vez que estas moléculas não são termicamente degradadas durante o processo de evaporação térmica a vácuo, e que o crescimento dos filmes pode ser controlado em escala nanométrica para ambos os materiais. Em termos estruturais, os filmes PVD de ZnPc e NiPc são cristalinos (forma α) e possuem as moléculas arranjado-se na forma de agregados e monômetros e ordenadas com o anel macrociclo inclinado em relação à superfície do substrato. Tais agregados podem ser vistos em escala nanométrica, porém, em escala micrométrica os filmes apresentam-se morfologicamente homogêneos. Em relação às propriedades ópticas e elétricas, observou-se que ambos os filmes PVD absorvem na região do visível com a ZnPc apresentando fotoluminescência quando irradiado com laser 785 nm. A condutividade elétrica é de 1,2x'10 POT. -10' S/m para a ZnPc e de 72x'10 POT... / In this work thin films of phthalocyanines of zinc (ZnPc) and nickel (NiPc) were fabricated through the vacuum thermal evaporation technique (PVD - physical vapor deposition) for different thicknesses at nanometric scale with the objective of determining the molecular architecture of these films as well as their optical and electrical properties. The final idea is to generate subsidies for applications of these films in electronic devices based on organic semiconductors and gas sensors. The PVD films were characterized using thermogravimetry (TG), differential scanning calorimetry (DSC), ultraviolet-visible (UV-vis) and Fourier transform infrared (FTIR) absorption spectroscopies, Raman scattering, X-ray diffraction, optical and atomic force (AFM) microscopies, and electrical characterization (tension x current dc). The results showed that the fabrication of ZnPc and NiPc films is possible since these molecules are not thermally degraded during the process of vacuum thermal evaporation and that the growth of the films can be controlled at nanometric scale for both materials. Structurally, the PVD films of ZnPc and NiPc possess the molecules organized with the macrocycle ring tilted in relation to the substrate surface. They are crystalline (α form) and possess molecular aggregates in the form of dimmers or higher order of aggregates and monomers. Such aggregates can be seen at nanometric scale, however, at micrometric scale the films are morphologically homogeneous. In relation to the optical and electrical properties, it was observed that boh PVD films absorb in the visible region with the ZnPc presenting photoluminescence when irradiated with the 785 nm laser line. The electric conductivity at 1,2x'10 POT. -10' S/m for ZnPc and 72x'10 POT. -10' S/m for NiPc. They also presented photoconductivity with the ZnPc more photoconductor than NiPc. Finally, after thermal treatment... (Complete abstract click electronic access below)

Cette thèse s’intéresse à la fabrication de jonctions magnétiques à effet tunnel organiques. Les MTJ organiques remplacent la barrière par une molécule. Il a fallu d’abord vaincre les problèmes liés à la fabrication de ces MTJ. En effet, ce type de jonction est très fragile du point de vue de sa fabrication, car incompatible avec les solvants. Un nouveau procédé de fabrication a été mis au point. Ce procédé fait appel à de petites « billes » nanométriques dispersées à la surface d’un échantillon. Ce procédé a été utilisé avec succès. Nous avons obtenu une réponse magnétique des échantillons. Des mesures XAS et de magnéto-transport ont été menées sur des jonctions MgO. Une approche in operando innovante a été utilisée. Ces mesures ont démontré que la présence d’oxyde de fer aux interfaces limite la TMR. Pour finir, des mesures SR-PES ont été menées dans le but d’étudier la polarisation d’interface de Cu/MnPc dans le système Cu(100)//Co/Cu/MnPc. Ces mesures ont révélé que cette interface est très fortement polarisée en spin. Les structures qui apparaissent dans les spectres ne peuvent être expliquées par une simple atténuation du signal du cobalt due à la couverture de molécules. / This thesis concerns the fabrication of organic magnetics tunnel junctions. Organic MTJs replace the barrier with a molecule. First, we had to overcome the problems of MTJ manufacturing. Indeed, this type of junction is very fragile from the point of view of its manufacturing because they are incompatible with solvents. A new manufacturing process has been developed. This process uses small nanometric "beads" scattered on the surface of a sample. This method has been used successfully and we obtained a magnetic response of the samples. XAS and magneto-transport measurements were conducted on MgO junctions. An innovative in operando approach was used. These measurements revealed that the presence of oxide at the interfaces limits the TMR. Finally, SR-PES measurements were carried out in order to study the Cu/MnPc interface polarization in the Cu (100)//Co/Cu/MnPc system. These measurements revealed that this interface is strongly spin polarized. Structures appearing in the spectra cannot be explained by a simple attenuation of the cobalt signal due to molecule coverage.

L'avenir de la spintronique repose sur le développement de matériaux ayant des propriétés magnétiques remarquables. L'objectif de cette thèse est de comprendre la physique des deux matériaux fonctionnels proposés pour des applications spintroniques qui utilisent des simulations de la densité fonctionnelle.Nous nous sommes intéressés dans une première partie au ferrite de gallium pour lequel il a été montré que les propriétés dépendaient de la concentration de fer.Les spectres optiques ont été calculés et comparés aux spectres expérimentaux suggérant des niveaux élevés de désordre. Dans la deuxième partie, nous avons montré une polarisation de spin à l’interface hybride formée entre la phthalocyanine de manganèse et la surface de cobalt,en accord avec les expériences de photoémission.La formation de la spinterface a été expliquée par différents mécanismes d'hybridation dans chaque canal de spin.Cette polarisation de spin est coordonnée avec des moments magnétiques induits sur les sites moléculaires. / The future of the spintronics technology requires developing functional materials with remarkable magnetic properties. The aim of this thesis is to understand the physics of functional materials proposed for spintronic applications using ab-initio density functional simulations. We investigated the properties of two different functional materials. We first studied the magnetoelectric gallium ferrite GFO. The dependence of the different properties on the iron concentration has been demonstrated and discussed. The optical spectra were calculated and compared to the experimental once suggesting high levels of iron disorder. In the second part, we demonstrated a highly spin polarized hybrid interface formed between manganese phthalocyanine and cobalt surface in agreement with photoemission experiments. The formation of this spinterface was described by different hybridization mechanisms in each spin channel. This high spin polarization is coordinated with induced magnetic moments on the molecular sites.

L'oxydation de liaisons C-H fortes et la transformation de liaisons carbone-halogène dans des conditions douces sont particulièrement difficiles et représentent un challenge pour les chimistes. Ces réactions sont réalisées dans la nature par l'intermédiaire des métalloenzymes. Dans le but de reproduire cette chimie efficace, nous développons au laboratoire des catalyseurs inspirés de deux des plus puissantes monooxygenases : les cytochromes P450 et la méthane monooxygenase soluble. Il s'agit de complexes binucléaires où deux atomes de fer sont pontés par un atome et stabilisés par des ligands macrocycliques. En particulier, les dimères de phthalocyanine de fer pontés par un atome d'azote (μ-nitrido) sont capables d'activer les peroxydes comme H2O2 afin oxyder, dans des conditions douces, les substrats les plus inertes (méthane, benzène, dichlorométhane). Ces travaux de thèse ont consisté en l'utilisation de ces systèmes bio-inspirés pour décrire la première transformation oxydante des liaisons C-F aromatiques. Cette nouvelle approche peut être utilisée pour la minéralisation de polluants notoires : les composés aromatiques halogénés. La démarche générale de cette thèse a été de (i) décrire la réactivité des complexes de phthalocyanines et porphyrines et (ii) en élucider le mécanisme afin de (iii) comprendre les paramètres clés gouvernants l'efficacité catalytique. Nous avons ainsi décrit le premier système capable de minéraliser les aromatiques perfluorés en conditions douces par transformation oxydante de la liaison C-F en liaison C-OH. Un mécanisme d'époxydation du système-π du substrat fluoré par l'intermédiaire d'une espèce fer-oxo a été proposé et argumenté. Nous avons déterminé l'effet de la nature de l'atome pontant sur la structure du catalyseur et expliqué l'impact crucial de la structure Fe(μ-N)Fe sur l'efficacité catalytique grâce aux caractérisations EXAFS, XANES et des calculs DFT. La synthèse, la caractérisation, et la comparaison de dimères à ligands porphyrazine, porphyrine ou phthalocyanine nous a enfin permis de décrire l'influence du ligand macrocyclique sur la géométrie moléculaire et la structure électronique des complexes / Oxidations of strong C-H and transformation of carbon-halogen bonds under mild conditions are especially difficult and challenging reactions. This reactivity can be achieved in Nature through iron metalloenzymes. In order to reproduce the efficient chemistry found in Nature, we develop bio-inspired diiron macrocyclic catalysts combining the structural features of two the most powerful monooxygenases: cytochrome P450 and soluble methane monooxygenase. Diiron catalysts where two iron atoms are bridged by μ-nitrido group and stabilized by macrocyclic supporting ligands can activate peroxides such as H2O2 for the most challenging oxidations reactions (methane, benzene, dichloromethane) under mild and practical conditions. This thesis is focused on the use of these bio inspired systems for the previously unknown oxidative transformation of the aromatic C-F bonds. This novel approach to the activation of C-F bonds can be used for the mineralization of notorious pollutants: the halogenated aromatics. We (i) described the reactivity of phthalocyanine and porphyrin complexes and (ii) studied the mechanism to (iii) understand and explain the key parameters which gouverns the catalytic efficiency. We had reported the first system able to transform aromatic C-F bonds onto C-OH bonds under mild conditions. Based on mechanistic studies, an epoxidation mechanism mediated by diiron-oxo species has been proposed. We have studied the influence of the bridging atom on the structure and the catalytic efficiency of the catalysts using both spectroscopic and theoretical methods. Finally, the effect of the macrocyclic ligand on molecular geometry and electronic structure of the complexes have been described through the synthesis, the characterization and the comparison of diiron complexes based on porphyrazine, porphyrin or phthalocyanine ligands

Small molecule organic semiconductors such as phthalocyanines and their derivatives represent a very interesting alternative to inorganic semiconductor materials for the development of flexible electronic devices such as organic thin field effect transistors, organic Light Emitting Diodes and photo-voltaic cells. Phthalocyanine molecules can easily accommodate a variety of metal atoms as well in the central core of the molecule, resulting in wide range of magnetic properties. Exploration of optical properties of organic crystalline semiconductors thin films is challenging due to sub-micron grain sizes and the presence of numerous structural defects, disorder and grain boundaries. However, this can be overcome by fabricating macroscopically ordered semiconductor films by solution processing. Presence of fewer grain boundaries and defects while modifying the pi orbital overlap by controlling the molecular stacking by forming macroscopic long range ordered grains, was essential in understanding the delocalization and diffusion length and its role in magnetic exchange mechanisms in this system. The origins of magnetic exchange mechanism between delocalized ligand electrons and spins in organic semiconductors has been of key interest as it underlies many complex optical and transport properties and is investigated in this thesis. The interaction between magnetic ions in organic magnetic semiconductors is quite challenging to interpret due to competing exchange mechanisms present in crystalline thin films. Better understanding of these exchange mechanisms is essential for tuning of magnetic exchange interaction to suit the need of practical magnetic storage and spintronic devices. Optical techniques such as linear dichroism, magnetic circular dichroism and magneto-photoluminescence are used and provided key understanding about the relation between excitons, spin exchange mechanisms and collective magnetic behavior of delocalized electrons in organic semiconductors. An enhancement in the collective magnetization of the crystalline thin films with strong exchange coupling between the delocalized ligand electrons and d-electrons is observed. The electronic states responsible for magnetic exchange are identified utilizing magnetic field and temperature dependent studies. Furthermore, soluble organics allowed engineering of organic analogues to diluted magnetic semiconductors (DMS) by creation of metal/metal-free Pc alloys. Optical studies provided crucial information about delocalization and diffusion lengths in these systems allowing fine tuning of this delocalization length scale and metal-metal distance. The exploration of magnetic behavior in metal/metal-free Pc alloys opens an avenue for tuning magnetic properties through an exchange similar to Ruderman-Kittel-Kasuya Yosida (RKKY) type interaction in organic magnetic semiconductors.

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department: Department of Biophysics and Physical Chemistry Candidate: Pavel Cermak Supervisor: Assoc. Prof. Veronika Novakova, PhD. Title of Thesis: Preparation and photophysical evaluation of tetra-3,4- pyridoporphyrazines suitable for the photodynamic therapy Tetra-3,4-pyridoporphyrazines (TPyPz) are aza-analogues of phthalocyanines. Their large system of conjugated bonds enables them to absorb light in the red part of the absorption spectrum. Due to their ability to produce singlet oxygen, they can be potentially used as photosensitizers in photodynamic therapy (PDT). Its mechanism is based on co-functioning of three elements - photosensitizer, light and oxygen. Photosensitizer excited by light absorption transfers its energy into tissue oxygen, thus, creating cytotoxic singlet oxygen. This method is beneficial for its high selectivity, low toxicity, minimal invasion and fast effect. The aim of this work was to synthetize and study water-soluble TPyPz suitable for PDT. Water solubility was achieved by quarternized amines, forming of salts or using suitable delivery systems (hydrophilic emulsion). Hydrophilicity was also increased by introduction of hydrophilic non-charged substituents (OH). At first, appropriate precursors for...

O processo de cicatrização cutânea pode ser favorecido pela aplicação de laser de baixa potência, sendo a matriz de colágeno e elastina um substituto dérmico no tratamento de feridas descrita como um sistema adequado na engenharia tecidual em sistemas tridimensionais da pele. Com este intuito, foi elaborado um estudo utilizando o sistema nanoemulsão contendo cloro alumínio ftalocianina (ClAlPc) como o agente fotossensibilizante em biópsias de explants de pele de pacientes saudáveis irradiadas por luz proveniente de laser de baixa potência, visando estabelecer a melhor dose de luz para a bioestimulação de colágeno tipo I e elastina neste tecido. O sistema de liberação de fármacos foi sintetizado utilizando protocolos já conhecidos, cujas propriedades fotoquímicas e fotofísicas foram confirmadas por espectrofotometria de absorção e fluorescência, além de estudos de estabilidade medindo-se o tamanho das partículas, potencial Zeta e índice de polidispersão. Uma vez caracterizado este sistema e conhecendo-se a sua faixa de absorção, elaborou-se um protocolo para avaliar a ação do fármaco nos principais componentes da matriz extracelular, como colágeno e elastina, e a sua ação combinada com luz laser de baixa potência em três doses conhecidas distintas: 70, 140 e 700 mJ/cm2. Foi avaliada também a ação somente da irradiação sobre as biópsias, nas mesmas doses, podendo-se assim observar os seus efeitos. As estruturas morfológicas da pele foram estudadas por histologia, e posteriormente foram comparadas quantitativamente as porções de colágeno e elastina da pele tratada e irradiada com as amostras do controle, as quais não receberam nenhum tipo de tratamento. Tanto a análise para colágeno tipo I quanto a análise para elastina apontaram um aumento de quase 20% em relação às amostras não tratadas, utilizando a dose intermediária de 140 mJ/cm2 nas amostras tratadas com o fármaco de ftalocianina, durante um período de 14 dias após a irradiação. Este efeito foi bastante significativo, ao ser comparado com a ação somente da irradiação, que apresentou desempenho inferior. Outra técnica explorada neste trabalho e utilizada na detecção da expressão das enzimas MMP-2 e MMP-9, participantes do processo cicatricial, foi a zimografia de gelatina, utilizando o meio de cultivo de cada amostra. As bandas relacionadas à degradação da gelatina para cada amostra no zimograma foram quantificadas e os níveis de expressão de MMP-2 e MMP-9 comparados. Os resultados obtidos confirmaram a análise histológica, apontando um maior nível de expressão dessas enzimas nos grupos tratados com o fármaco fotossensibilizante e luz na dose intermediária (140 mJ/cm2) sendo esta combinação a mais promissora para um estudo mais aprofundado. / Wound healing process can be favored by Low Level Laser Therapy, with the collagen and elastin matrix a dermic substitute described as an appropriate system in tissue engineering and in tridimensional skin systems. With this purpose, a study was elaborated using the system nanoemulsion of aluminium-chloride phthalocyanine (ClAlPc) as photosensitizer in skin biopsies obtained after plastic surgery and irradiated by a low level laser, to establish the most appropriate dose of light for biostimulation of type I collagen and elastin fibers. The drug delivering system was synthesized using a well-known protocol and its photophysical and photochemical properties were confirmed by absorption and fluorescence spectrophotometry, besides stability studies measuring particle size, zeta potential and polidispersion index. Once characterized this system and known its absorption range in the UV-Vis region of light spectrum, a protocol was elaborated for evaluating the effect of the photosensitizer direct into extracellular matrix components, collagen and elastin, and the combined effect with low level laser therapy using three different doses: 70, 140 and 700 mJ/cm2. Also, only the effect of irradiation was evaluated using the same doses. Morphological structures of the skin were analyzed by histology, and portions of collagen and elastin of skin biopsies after the photosensitizer and light treatment were quantified and compared to the non-treated samples. The analysis for type I collagen and elastin pointed an increase of more than 20% compared to the non-treated samples, for the samples treated with the combination phosensitizer/light140 mJ/cm2 after 14 days of treatment. This effect was significant when compared to the effect of the irradiation only. Another technique was used in this work to detect the expression of MMP-2 and MMP-9, participant enzymes in various processes including tumoral progression and wound healing. Samples of biopsies culture medium were collected and analyzed by gelatin zymography, the bands related to gelatin degradation were quantified and MMP-2 and MMP-9 expression levels compared. The obtained results confirmed histological analysis, pointing a higher expression level of these enzymes for the group treated with the photosensitizer and the intermediate dose of light (140 mJ/cm2), leading to a promising combination of treatment for future studies.

A Terapia Fotodinâmica (TFD) contra o melanoma cutâneo tem encontrado várias limitações devido à interferência de cromóforos endógenos (melanina) na irradiação deste tipo de cancêr de pele. Agentes fotossensibilizantes que absorverem em comprimentos de onda superiores a 650 nm evitam a competição com a melanina (absorção máxima 530 nm). Neste cenário, surge a ftalocianina de cloroalumínio com forte absorção entre 670-680 nm. Entretanto, esta molécula é lipofílica, o que impede sua aplicação na terapêutica. Para superar este problema, esforços têm sido direcionados para o desenvolvimento de sistemas de veiculação de fármacos hidrofóbicos. Portanto, o objetivo deste trabalho foi desenvolver, caracterizar e avaliar o efeito fotodinâmico de ClAlPc encapsulada em nanocápsulas e nanopartículas de organogel. As nanocápsulas contendo ClAlPc foram obtidas pelo método de nanoprecipitação por meio de um planejamento fatorial 23 e as nanopartículas de organogel contendo ClAlPc foram preparadas pela dispersão a quente do organogel em solução aquosa. Ambas as formulações coloidais foram caracterizadas com relação ao diâmetro médio, índice de polidispersão (IPd), potencial zeta, eficiência de encapsulação (E.E.) e estabilidade física. Os ensaios in vitro de toxicidade e fotocitotoxicidade de ClAlPc nanoencapsulada e/ou livre foram realizados sobre as linhagens melanocíticas WM1552C, WM278, WM1617 e B16-F10. O método de nanoprecipitação foi capaz de produzir nanocápsulas em tamanho nanométrico (233,0 nm ±2,00), com uma distribuição de tamanho homogênea e monodispersa (0,309 ±0,0038), bem como potencial zeta significantemente negativo de 29,6 mV (±3,91). A formulação de nanocápsulas apresentou uma boa E.E. de 63,7% para a ClAlPc. O conteúdo de ClAlPc presente nas formulações de nanocápsulas foi determinado pelos métodos analíticos espectrofotométrico e espectrofluorimétrico validados, os quais foram capazes de quantificar a ClAlPc com precisão e exatidão. O estudo de estabilidade física das formulações de nanocápsulas revelou o caráter estável desta formulação por um período de até 12 meses. As nanopartículas de organogel contendo ClAlPc apresentaram diâmetro médio de 282,7 nm (±2,99), IPd de 0,343 (±0,0280) e potencial de superfície de + 49,3 mV (±1,84). A formulação de nanopartículas de organogel apresentou uma E.E.de ClAlPc de 60%. O estudo de estabilidade física também revelou um comportamento estável das nanopartículas de organogel por um período de 6 meses. Os estudos de toxicidade na ausência de luz demonstraram o caráter biocompatível das nanocápsulas desenvolvidas e foi possível comprovar o excelente efeito fototóxico da ClAlPc nanoencapsulada sobre todas as linhagens melanocíticas com combinação de 0,30 µg.mL-1 de ClAlPc nanoencapsulada e doses de luz de 150 ou 500 mJ.cm-2, nestas condições a fração de morte celular foi superior a 90%. Portanto, estes resultados confirmam o potencial de nanocarreadores contendo ClAlPc como sistema de veiculação de compostos hidrofóbicos aplicados à TFD. / Photodynamic Therapy (PDT) against melanoma has found several limitations due to interference from endogenous chromophores (melanin) in the irradiation of this skin cancer. Photosensitizer agents which absorb at wavelengths above 650 nm may avoid melanin competition. In this context, the chloroaluminum phthalocyanine is a promissor photosensitizer with strong absorption between 670-680 nm. However, this molecule is lipophilic difficulting its application in therapy. In order to overcome such problem, efforts have focused on development of drug delivery systems containing hydrophobic photosensitizers. Therefore, the aim of this study was to develop, characterize, and evaluate the photodynamic effect of ClAlPc encapsulated into nanocapsules and organogel nanoparticles. The nanocapsules containing ClAlPc were obtained by nanoprecipitation method using a factorial design 23, and organogel nanoparticles containing ClAlPc were prepared hot dispersion of organogel in aqueous solution. Both colloidal formulations were characterized with respect to average diameter, polydispersity index (PdI), zeta potential, encapsulation efficiency (EE), and physical stability. The in vitro tests for toxicity and photocytotoxicity of nanoencapsulated and free ClAlPc were performed on melanocytic cell lines (WM1552C, WM278, WM1617, and B16-F10). The nanoprecipitation method was able to produce nanocapsules with nanometer-size (233.0 nm ± 2.00), PdI of 0.309 (± 0.0038) indicating homogeneous and monodisperse formulations, as well as significantly negative zeta potential of 29.6 mV (± 3.91). The formulation of nanocapsules showed a good EE of 63.7% for ClAlPc. The ClAlPc content present in nanocapsules was determined by validated analytical spectrophotometric and spectrofluorimetric methods, which were reliably able to determine ClAlPc in nanocapsules. The study of physical stability for nanocapsules showed the stable character of this formulation for a period of 12 months. The nanoparticles of organogel containing ClAlPc had an average diameter of 282.7 nm (± 2.99), IPD of 0.343 (± 0.0280), and surface potential of + 49.3 mV (± 1.84). The formulation of organogel nanoparticles showed a ClAlPc EE of 60%. The physical stability study also revealed a behavior stable organogel nanoparticles for a period of 6 months. Toxicity studies in darkness conditions have been confirmed biocompatibility of nanocapsules. Besides that, the excellent photodynamic effect of ClAlPc nanoencapsulated was reached on all melanocytic cell lines using an arrangement of 0.30 g.mL-1 of ClAlPc encapsulated, and light doses of 150 or 500 mJ.cm-2, under these conditions the cell death fraction was more than 90%. Therefore, these results confirm the potential of nanocarriers containing ClAlPc as delivery system for hydrophobic photosensitizers applied to PDT.