Resumo: O terpinen-4-ol é o principal componente da M. alternifólia, apresenta atividade antiinflamatória, antibacteriana e antifúngica; em estudos recentes in vitro a atividade antineoplásica para linhagens celulares de melanoma (M14) foi demonstrada. O objetivo específico deste trabalho foi verificar a atividade farmacológica in vitro dos isômeros óticos (-) terpinen-4-ol e (+) terpinen-4-ol e do óleo de Melaleuca alternifolia em nove linhagens diferentes de células tumorais, incluindo a célula UACC-62 do melanoma que ainda não havia sido estudada, e a concentração eficaz para o desenvolvimento de formulação que possa ter ação contra o melanoma. Após realização dos ensaios farmacológicos in vitro verificou-se que o (-)terpinen-4-ol apresentou melhor atividade nas linhagens celulares de melanoma, sendo o princípio ativo de escolha para as duas formulações que foram desenvolvidas, testadas e comprovadas a qualidade e estabilidade. Para caracterização das formulações foram efetuados os ensaios reológicos, sendo comprovado que as emulsões desenvolvidas são adequadas para o uso tópico no que diz respeito às suas características reológicas, sendo que as duas apresentaram comportamento de fluxo semelhante, diferindo apenas na área de histerese. Posteriormente foram feitos testes de permeação e retenção cutânea in vitro, para verificar qual formulação permite que o princípio ativo atinja em maior concentração a camada de interesse para atividade contra o melanoma. O método proposto para quantificação do (-)-terpinen-4-ol foi validado e utilizado nos ensaios de estabilidade acelerada, liberação, permeação e retenção cutânea in vitro. Os ensaios de liberação demonstraram que as duas formulações apresentam cinética de zero ordem, com fluxo de 14,62 µg/cm2 /h para formulação 1 e de 6,70 µg/cm2 /h para formulação F2. Na retenção cutânea ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The terpinen-4-ol is the major component of M. alternifolia, has anti-inflammatory, antibacterial and antifungal, in recent studies in vitro anticancer activity for melanoma cell lines (M14) was demonstrated. The specific objective of this study was to assess the in vitro pharmacological activity of optical isomers (-) terpinen-4-ol and (+) terpinen-4-ol and oil of Melaleuca alternifolia in nine different tumor cell lines, including cell UACC -62 melanoma that had not yet been studied, and effective concentration for the formulation development that may have action against melanoma. After performing the in vitro pharmacological tests showed that the (-) terpinen-4-ol showed better activity in melanoma cell lines, with the active ingredient of choice for the two formulations were developed, tested and proven quality and stability. For characterization of the formulations were made the rheological tests and confirmed that the developed emulsions are suitable for topical use in relation to their rheological characteristics, and the two had similar flow behavior, differing only in the area of hysteresis. Subsequent tests were performed permeation and skin retention in vitro, to determine which formulation allows the active ingredient reaches higher concentration in the layer of interest for activity against melanoma. The proposed method for quantification of (-)-terpinen-4-ol was validated and used in accelerated stability testing, release, skin permeation and retention in vitro. The release assays demonstrated that both formulations have zero order kinetics at a rate of 14.62 g/cm2/h for formulation 1 and 6.70 g/cm2/h for formulation F2. In vitro skin retention was found that the F1 showed retention of terpinen-4-ol smaller in the stratum corneum, compared to F2. Conversely, the retention in the epidermis over the dermis of terpinen-4-ol was higher in F1. It was found that formulation F1 ... (Complete abstract click electronic access below) / Orientador: Maria Virgínia Scarpa / Coorientador: Marcos Antonio Corrêa / Banca: Leila Aparecida Chiavacci / Banca: Renata Fonseca Vianna Lopez / Mestre

ADHD pharmacotherapy uses methylphenidate (MPH), D-amphetamine (D- amph), two psychostimulants targeting dopamine transporters, or atomoxetine (ATX), specifically targeting norepinephrine transporters. We have assessed the pharmacological mechanisms of these three drugs on the in vitro efflux of neurotransmitters in rat prefrontal cortex (PFC) and striatal slices as well as on the in vivo electrical activities of PFC pyramidal neurons, striatal medium spiny neurons, ventral tegmental area dopamine neurons or dorsal raphe nucleus serotonin neurons, using single cell extracellular electrophysiological recording techniques. We have also tested whether chronic methylphenidate treatment, during either adolescence or adulthood, could have long-lasting consequences on body growth, depression and neuronal functions. Release experiments showed that all ADHD drugs induce dose-dependent dopamine efflux in both the PFC and striatum, with different efficacies, while only D- amph induced cortical norepinephrine efflux. Atomoxetine induced an unexpected massive dopamine outflow in striatal regions, by mechanisms that depend on physiological parameters. Our electrophysiological studies indicate that all three drugs equally stimulate the excitability of PFC pyramidal neurons, in basal and NMDA-evoked conditions, when administered acutely (3 mg/kg). While the electrophysiological effects elicited by psychostimulants may be dependent on D1 receptor activation, those induced by atomoxetine relied on different mechanisms. In the ventral tegmental area (VTA), methylphenidate (2 mg/kg), but not atomoxetine, induced firing and burst activity reductions, through dopamine D2 autoreceptor activation. Reversal of such effects (eticlopride 0.2 mg/kg) revealed an excitatory effect of methylphenidate on midbrain dopamine neurons that appear to be dependent on glutamate pathways and the combination of D1 and alpha-1 receptors. Finally, acute intraperitoneal psychostimulant injections increased vertical locomotor activity as well as NMDA2B protein expression in the striatum. Some animals chronically treated with intraperitoneal administrations (methylphenidate 4 mg/kg/day or saline 1.2 ml/kg/day) showed decreased body weight gain. Voluntary oral methylphenidate intake induces desensitisation to subsequent intravenous methylphenidate challenges, without altering dopamine D2 receptor plasticity. Significant decreases in striatal NMDA2B protein expression were observed in animals chronically treated. After adolescent MPH treatment, midbrain dopaminergic neurons do not display either desensitisation or sensitisation to intravenous methylphenidate re-challenges. However, partial dopamine D2 receptor desensitisation was observed in midbrain dopamine neurons. Using behavioural experiments, cross-sensitisation between adolescent methylphenidate exposure and later-life D-amphetamine challenge was observed. Significant decreases in striatal NMDA2B protein expression were observed in animals chronically treated, while striatal medium spiny neurons showed decreased sensitivities to locally applied NMDA and dopamine. While caffeine is devoid of action on baseline spike generation and burst activity of dopamine neurons, nicotine induces either firing rate enhancement, firing rate reduction, or has no consequences. Adolescent methylphenidate treatment leads to decreased neuronal sensitivities to the combination of nicotine, MPH and eticlopride, compared to controls. Finally, nicotine partially prevented D-amphetamine-induced increase of rearing activities. Our results show that increases in the excitability of PFC neurons in basal conditions and via NMDA receptor activation may be involved in the therapeutic response to ADHD drugs. Long-term consequences were observed after psychostimulant exposure. Such novel findings strengthen the mixed hypothesis in ADHD, whereby both dopamine and glutamate neurotransmissions are dysregulated. Therefore, ADHD therapy may now focus on adequate balancing between glutamate and dopamine.

Les copolymères à blocs amphiphiles peuvent s’auto-assembler sous forme de vésicules,appelées polymersomes. Ces vésicules ont été développées et étudiées depuis de nombreusesannées notamment pour l’encapsulation et la délivrance contrôlée de médicaments. Depuisquelques temps, elles connaissent des applications dans le domaine du biomimétisme cellulaire.Plus robustes que leurs analogues lipidiques (liposomes), les avantages à utiliser lespolymersomes comme mimes synthétiques de cellules biologiques ne sont plus à démontrer.Ainsi, des structures compartimentées à base de polymères ont été développés comme mimesstructurels de cellules. Ces systèmes ont été utilisés comme bioréacteurs, avec la réalisation deréactions chimiques ou enzymatiques en cascade en milieu confiné. Toutefois, l’un desobstacles qu’il reste à franchir est de trouver des moyens simples et efficaces pour déclencherla réaction au sein de ces systèmes. C’est dans ce contexte que s’inscrivent les travaux de cettethèse. Une membrane synthétique asymétrique à base de lipide et polymère a été développée etla méthode d’émulsion-centrifugation a été utilisée pour produire des systèmes compartimentésbiomimétiques. De plus, deux approches différentes ont été suivies pour provoquer la libérationcontrôlée d’espèces encapsulées, l’une utilisant la température et l’autre la lumière. Enfin, desétudes de co-encapsulation de cellules synthétiques (polymersomes) et biologiques au sein demilieux 3D ont été réalisées dans le but d’évaluer leur compatibilité et la possibilité de les cocultiver. / Amphiphilic block copolymers can self-assemble into vesicles, also called polymersomes.These vesicles have been developed and studied for many years especially in the field of drugloading and controlled release. More recently, their use as cell mimics have attracted a lot ofattention, mainly because polymersomes exhibit many advantages in contrast to their lipidicanalogues (liposomes). In such, compartmentalized polymer systems have especially beendeveloped as structural mimics of cells. These systems have found applications as bioreactorsthat can confine cascade chemical or enzymatic reactions. However, a major goal that stillremains to achieve is to find ways to trigger the beginning of these chemical reactions insidethe compartmentalized structures. The work carried out during this PhD thesis was actually totackle this challenge. A synthetic asymmetric lipid – polymer membrane, that mimics themembrane of biological cells was developed and the emulsion-centrifugation protocol wasfollowed to prepare biomimetic compartmentalized structures. In addition, two different waysto control the independent release of multiple species from individual compartments weredeveloped, based on temperature or light activation. Lastly, co-encapsulation of synthetic cells(polymersomes) and biological cells were performed in 3D media with the aim to study theircompatibility for co-culture experiments.

Les arthrogryposes multiples congénitales (AMC), limitations articulaires multiples survenant au cours du développement et présentes à la naissance, sont un ensemble hétérogène de maladies dont le dénominateur commun est une diminution des mouvements fœtaux. Dans la majorité des cas, l'AMC est liée à un mécanisme impliquant le système neuromusculaire. Les causes sont dans un grand nombre de cas d'origine génétique. La connaissance de cette cause permet de proposer un conseil génétique avec une évaluation précise du risque de récurrence et éventuellement un diagnostic anténatal. La connaissance du mécanisme sous-jacent participe à l'évaluation du pronostic et permet d'élargir les connaissances sur la mise en place du système neuromusculaire. Mes travaux de thèse ont porté sur ces deux aspects. Dans un premier temps, j'ai étudié l'expression du récepteur de la ryanodine RyR1 dans le muscle squelettique fœtal humain. Les mutations de RYR1 sont responsables de myopathies congénitales. Dans les formes sévères précoces, une AMC peut être occasionnellement associée. L'expression de RyR1 est détectée dès 14 semaines d'aménorrhée dans le muscle fœtal humain. Ce résultat confirme l'expression précoce de RyR1 chez l'homme et permet d'expliquer la possibilité de limitations articulaires. Dans un second temps, j'ai étudié une famille consanguine avec trois enfants atteints d'arthrogrypose distale à la recherche de la cause génétique sous-jacente. L'analyse pangénomique m'a permis de lier pour la première fois le gène ECEL1, codant une endopeptidase, à une pathologie humaine. La recherche de mutations d'ECEL1 dans une cohorte de 20 patients a permis d'identifier cinq autres familles. Toutes les mutations sont transmises sur un mode autosomique récessif et conduisent à une perte de fonction de la protéine. L'ensemble des patients présentent un phénotype clinique et IRM semblable et distinct des autres tableaux d'arthrogrypose distale. Au total, ces travaux confirment l'expression précoce de RyR1 chez l'homme et identifient le gène ECEL1 comme une cause récurrente d'un type particulier d'arthrogrypose distale autosomique récessive. / Arthrogryposis multiplex congenita (AMC) is a heterogeneous group of disorders characterised by multiple joint contractures at birth due to diminished foetal movements. In most cases, the underlying mechanism involves the neuromuscular system. Genetic causes are frequent. Identifying the genetic cause is paramount for precise recurrent risk assessment, genetic counselling and prenatal diagnosis. Elucidating the underlying mechanism allows for prognostic evaluation and expands our knowledge on the development of the human neuromuscular system. My thesis focused on these two aspects. First I studied the expression of the ryanodine receptor 1 in human foetal skeletal muscle. RYR1 mutations cause congenital myopathies. AMC can occasionally be associated with severe forms of RYR1 related congenital myopathies. RyR1 is expressed at 14 weeks of gestational age in human skeletal muscle. Thus it confirms the early expression of RyR1 in human and can account for the occurrence of joint contractures. Second, in order to identify an underlying genetic cause, I studied a consanguineous family with three affected children showing a distal arthrogryposis phenotype. The genome wide linkage study allowed me to link for the first time the endopeptidase coding gene ECEL1 to a human disease. Five other families were shown to carry ECEL1 mutations after screening a cohort of 20 families with distal arthrogryposis. All mutations were recessive and predicted to lead to a loss of function of the protein. All patients showed a recognisable clinical and MRI phenotype that differed to that of currently known distal arthrogryposes. Altogether, these results confirm the early expression of RyR1 in human and identify ECEL1 as a recurrent cause of a distinct type of distal arthrogryposis.

Les nanogels hybrides constitués de polymères thermosensibles et de nanoparticules inorganiques stimulables telles que des nanoparticules magnétiques (NPMs) ou des nanobatônnets d’or (AuNRs) sont extrêmement intéressants pour des applications biomédicales. Leur matrice en polymère permet d’encapsuler et de libérer de grandes quantités de molécules actives, alors que les nanoparticules peuvent générer de la chaleur lorsqu’elles sont exposées à un champ magnétique alternatif (AMF) pour les NPMs, et à une irradiation proche infrarouge (NIR-L) pour les AuNRs. Ce manuscrit de thèse porte sur la synthèse et la caractérisation de nanogels biocompatibles, pH- et thermosensibles, à base de monomères en oligo (éthylène glycol) méthyl éther méthacrylate (OEGMAs), d’acide méthacrylique (MAA) et encapsulant des NPMs et/ou des AuNRs pour déclencher de manière contrôlée, par hyperthermie magnétique ou par photothermie, la libération d’une molécule anticancéreuse, la doxorubicine (DOX). Des nanogels hybrides magnétiques, plasmoniques et magnéto-plasmoniques ont été synthétisés. Ces nanogels ont un diamètre hydrodynamique entre 200 et 500 nm et une température de transition de phase volumique comprise entre 30 et 54 °C. Le comportement de gonflement-dégonflement des nanogels peut être induit par plusieurs stimuli (température, pH, AMF, NIR-L). Ces résultats démontrent que les MagNanoGels sont d’excellents nanovecteurs pour accroître l’internalisation cellulaire en augmentant la cytotoxicité de la DOX et qu’il est possible de déclencher à distance la libération intracellulaire de DOX sous AMF dans des conditions athermiques. Par ailleurs, les PlasMagNanoGels peuvent générer efficacement de la chaleur par photothermie pour une thermothérapie. En outre, les propriétés intrinsèques des NPMs, pour le ciblage magnétique et en tant qu’agents de contraste pour l’imagerie par résonance magnétique (MRI), font de ces nanogels des candidats idéaux pour une nouvelle approche thérapeutique (diagnostique et traitement) contre le cancer. / Hybrid nanogels, composed of thermoresponsive polymers and inorganic responsive nanoparticles, such as magnetic nanoparticles (NPMs) and gold nanorods (AuNRs) are highly interesting for biomedical applications. Their polymeric matrix makes them able to uptake and release high quantities of drugs, whereas nanoparticles can generate heat when exposed to an alternating magnetic field (AMF) for NPMs, and to a near-infrared light for AuNRs. This thesis manuscript focuses on the synthesis and the characterization of biocompatible, pH- and thermoresponsive nanogels, based on oligo(ethylene glycol) monomers (OEGMAs), methacrylic acid (MAA) and encapsulating NPMs and/or AuNR for remotely triggered doxorubicin (DOX, anticancer drug) release, by magnetic hyperthermia or phothothermia. Hybrid magnetic, plasmonic and magneto-plasmonic nanogels were synthesized. Theses nanogels have a hydrodynamic diameter between 200 and 500 nm and a volume phase transition temperature (VPTT) from 30 to 54°C. The nanogels’ swelling-deswelling behavior can be induced by several stimuli (temperature, pH, AMF, NIR-L). These results demonstrate that MagNanoGels are excellent nanocarriers for enhancing cellular internalization enhancing DOX cytotoxicity and that DOX release was significantly enhanced upon exposure to AMF in athermic conditions. In addition, PlasMagNanoGels can efficiently generate heat by photothermy for thermotherapy. Therefore, the intrinsic properties of NPMs for magnetic targeting and as contrast agents for Magnetic Resonance Imaging (MRI), make these nanogels ideal candidates for a new therapeutic approach (diagnosis and treatment) against cancer.

This report revolves around a project for Boliden which is a thesis work for a master in engineering within industrial design engineering. This project had a primary focus of developing a new mount for fire extinguishers that will be suspended on vehicles in Boliden's mines. The foundation of the project was an accident in Boliden's mine Aitik where a fire extinguisher exploded due to damages caused by the vehicle mount in combination with vibrations. The objective of the project is to identify the main problem with the vehicle mounts used today in Boliden's mines and solve it. The process used in this project is a fairly standard process that is separated into four phases; Context immersion, Ideation, Conceptual design and Final design. The last two stages focused on prototyping and testing to finalize the design, whereas the beginning of the process was trying to get a grasp of the problem. This included understanding the unique environment of the mines as well as getting to the bottom of the problem with the vehicle mount. Interviews and observations were conducted during a visit to Boliden's mine, Renström, in Skellefteå to investigate the problem. The conclusions from the Context immersion was that the main problem with the vehicle mounts was that they were not adapted for the mining conditions at all. A large number of vibrations and a lack of vibration damping for the mount was the leading cause for the damages on the fire extinguisher. This caused the vehicle mount to fail and in the process also damage the fire extinguisher. The solution to this problem turned out to be a more robust product using a stronger material with higher damping to suspend the fire extinguisher. This made the vehicle mount cope with the vibrations in a better way. A quick release mechanism was also implemented that assists with the vibration durability as well as make the fire extinguisher easy and fast to release. The fast release, as well as the ease of use, makes the vehicle mount adapted for emergencies as well.  The durability and the use of the new vehicle mount were reassured by a vibration analysis made in NX 12, as well as a short usability test. / Den här rapporten behandlar ett projekt för Boliden vilket också är ett examensarbete inom civilingenjör teknisk design med inriktning mot produktutveckling. Projektet har fokuserats på att utveckla en ny fordonshållare för brandsläckare som kommer användas på fordon i Bolidens gruvor. Grunden för projektet var en olycka i Bolidens gruva Aitik där en brandsläckare exploderade då den hade blivit skadad av fordonshållaren samt vibrationer. Syftet med projektet är att identifiera den underliggande problemen som finns med fordonhållarna samt ta fram en bättre lösning. Processen som användes i det här projektet är baserad på en normal designprocess som består av fyra faser. De sista två faserna i processen fokuserade på att prototypa och testa för att färdigställa designen, medan början av processen försökte problemet förstås. Det innebar att gruvmiljön som är unik var tvungen att utforskas samt att gå till botten med problemet med hållarna. För att göra detta gjordes främst ett besök till Renströms gruvan i Skellefteå där intervjuer och observationer utfördes. I slutet av Kontextfasen fastställdes det att huvudproblemet var att hållaren inte var tillräckligt anpassad för gruvmiljön. Den stora mängden vibrationer från miljön i kombination med en brist av vibrations dämpning för hållaren var orsaken till att skador uppstår på brandsläckare. Lösningen på problemet var en ny hållare som var mer robust samt att materialet som används för att hålla brandsläckaren var starkare samt hade högre dämpningsgrad. Ett snabblås lösning var också implementerad som också bidrar till ökad vibrationstålighet, den gör också att brandsläckaren går snabbt och enkelt att ta loss i nödsituationer. Hållbarheten samt användbarheten säkerhetsställs genom en vibrationsanalys utförd i NX 12 samt ett kort användartest.

Doctor Legum - LLD / In South Africa, prison authorities are not primarily concerned with the health of the prison population. This is evidenced by inter alia: the vast number of complaints regarding health care received by the Judicial Inspectorate of Correctional Centres; natural deaths in prisons reported annually; litigation regarding health care and treatment in prisons; and the notoriously poor conditions of detention which inevitably have a negative impact on prisoners' health. There is as a result a noticeable difference between state provided health care to the public and health care in prisons. This thesis is therefore aimed at unpacking what the right to health means in respect of terminally ill prisoners. This question has been overshadowed by issues regarding medical parole in South Africa and intermittently by calls for palliative care in prisons. Whilst these issues are relevant to their plight, there is a need to articulate the scope of the right to health of terminally ill prisoners. This is imperative as not all prisoners who are terminally ill are eligible for medical parole and there are instances where the granting of such parole may be impractical. An analysis of the right to health in relation to terminally ill prisoners will provide legal certainty as to the legal entitlements regarding health care for one of the most vulnerable groups in society. They will know what they may legally claim and what they cannot insist upon in terms of the law.

The purpose of this rapport/paper is to summarize and compare three different reparation methods, which can be used in repairing damaged concrete dams, which are one of the most important parts of electricity production in Sweden. Most of big dams in Sweden built between1900- 1960 are in a great need of restoration. Time, harsh winter conditions and constant presence of water have caused great damages to these dams. There are numerous restoration methods, which can be used in dam reconstruction.  However, choosing the right method plays a crucial role in terms of economy and quality for the power plant companies. Due to negatively impacting global warming, carbon dioxide emissions can also be another concern for power plant companies when selecting a reparation method. Sometimes determining and choosing the “right” method is rather difficult when facing different reparation methods.  When deciding which method to choose, comparisons are made in terms of economy, quality, and carbon dioxide emissions causing global warming. The methods compared in this rapport are applying waterjet mechanical chiseling, and Chesterton filler.  Accordingly, some boundaries are set and some, but not all, factors are taken into account.  For example, winter expenses and transportation are not included in this rapport. The method which has been used in this study is both qualitative and quantitative. In the qualitative method, interviews, literature studies and experience of experts are gathered and used. The quantitative method used for calculation of costs and CO2 (e) emissions to the air. The study shows that in a minor spalled concrete damage, the waterjet method and Chesterton filler are appropriate with regards to quality.  Mechanical chiseling is the cheapest method, and the Chesterton filler method releases the least amount of carbon dioxide to the air of all methods. Hopefully, the comparison in this rapport can aid those who want to choose a reparation method based on the criterias for the outcome/method. / Dammkonstruktioner är en av de viktigaste delarna av elproduktionen i Sverige. De flesta av de stora betongdammarna i Sverige som är byggda mellan 1900 och 1960 är i behov av reparation. Åldern, svåra vinterförhållanden och ständig närvaron av vatten påverkar dammarna negativt och orsakar skador i betongen. Det finns gott om olika reparationsmetoder som kan användas vid dammreparationer. Att välja rätt metod vid reparationer kan spela en avgörande roll när det gäller ekonomi och kvalitet för kraftverksbolagen. Då de olika metoderna resulterar i olika mycket koldioxidutsläpp till luften och därmed effekt på den globala uppvärmningen är just koldioxidutsläppen en annan faktor som kan beaktas av elbolagen när de vill välja en reparationsmetod. Valet av reparationsmetod är inte lätt. Ofta går det att välja mellan flera olika metoder. Syftet med denna rapport att upprätta sammanfattningar för tre olika reparationsmetoder vid reparation av betongdammar, samt en jämförelse mellan dessa metoder. Jämförelser har gjorts i förhållande till ekonomi, kvalitet och koldioxidutsläpp till luften. De reparationsmetoder som har jämförts i denna rapport är vattenbilning, mekanisk bilning och Chesterton spackel. För att möjliggöra jämförelsen i denna studie har några avgränsningar gjorts i för de faktorer som jämförts, bland annat har vinterkostnader och transporter inte inkluderats i jämförelsen.   Metoden i denna uppsats bygger på kvalitativ och kvantitativ studie. Den kvalitativa studien består av litteraturstudier och intervjuer genom samtal och email, och den kvantitativa studien består av beräkningar för kostnader och CO2 (e)-utsläpp med hjälp av grundvärden och faktorer för 1 m² betongyta. Studien visar att vid en enkel frostskada så är vattenbilningsmetoden och Chestertonspackel lämpligast med avseende på kvalitet. Mekanisk bilning är billigast och Chestertonspackel släpper ut minst koldioxid till luften. Förhoppningsvis kan jämförelsen i denna rapport hjälpa de som vill välja reperationsmetod baserat på önskade kriterier för slutresultatet. / <p>2017-06-27</p>

La formulation de médicaments regroupe des technologies innovantes où l'usage de matières premières naturelles émerge du fait de contraintes environnementales grandissantes. Les huiles végétales par exemple sont plébiscitées grâce à leurs propriétés remarquables en terme de solubilisation de principes actifs, de biocompatibilité et de biodégradabilité. Afin d'élargir leurs applications, notamment pour la libération prolongée de principes actifs, la solidification de ces huiles par réticulation a été envisagée à l'aide d'une chimie douce, c'est-à-dire respectueuse des principes de la chimie verte. La silice a été choisie pour réaliser la réticulation par réaction sol-gel d'huiles végétales et ainsi obtenir des matériaux hybrides inorganique/organique d'origine naturelle. L'étude a consisté à fonctionnaliser des huiles végétales à l'aide de précurseurs silylés en utilisant une chimie sans solvant ni catalyseur, pour obtenir des huiles réticulables par voie sol-gel. Deux voies chimiques de fonctionnalisation ont été employées. La première, difficilement contrôlable, été basée sur une réaction époxy-amine où des huiles de lin ou de soja époxydées ont été utilisées. La seconde reposait sur la valorisation d'huile de ricin porteuse d'un groupement hydroxyle via une réaction hydroxy-isocyanate. Les excellents résultats obtenus ont permis de poursuivre l'étude par la formulation de l'huile de ricin silylée grâce à un nouveau procédé d'émulsion huile/eau thermo-stabilisée. Simple et robuste, ce procédé a rendu possible de façon concomitante la mise en forme et la solidification de microparticules hybrides. Les microparticules présentent des distributions de taille homogènes (20 à 200 µm), sont sphériques et capables de piéger des molécules lipophiles. Ainsi une molécule modèle, a été encapsulé avec des rendements très satisfaisants et sa libération a été totale après 8h en milieu physiologiques simulé. En modifiant la composition des microparticules (ratio inorganique / organique), il a été possible de prolonger les cinétiques de libération et de réduire significativement « l’effet burst ». La biocompatibilité de ces microparticules a été démontrée in vitro. Enfin dans l’optique de correctement caractériser la réaction sol-gel, différentes techniques analytiques ont été explorées pour l’étude in situ de la réticulation et l’identification d’un catalyseur biocompatible. Ces travaux ont permis de détailler les mécanismes des réactions d’hydrolyse et de polycondensation et ont ouvert la voie à un meilleur contrôle des cinétiques de réticulation des microparticules. De plus, il est apparu que la maîtrise de cette réaction semble être indispensable afin d'obtenir des objets "stabilisés " aux propriétés identiques et aux cinétiques de libération reproductibles. En conclusion, il a été démontré que la chimie sol-gel appliquée aux huiles naturelles offre des possibilités d’innovation en formulation galénique tout en respectant les contraintes environnementales et de santé et a permis le développement de matériaux originaux aux propriétés modulables. / Drug formulation is gathering innovative technologies where the use of natural products for the preparation of drug delivery systems is getting more and more considerations because of environmental concerns. For instance, vegetable oils get increasingly used because of their outstanding properties in terms of drug solubilization, biocompatibility and biodegradability. For expanding their application, to drug sustained-release for instance, oils have to be hardened under mild conditions with respect to green chemistry principals. The silica condensation was therefore chosen as cross-linking reaction by the mean the sol-gel reaction which when applied to vegetable oils led to biosourced hybrid organic/inorganic materials. To do so, oils were functionalized with alkoxysilanes precursors without solvent nor catalyst in order to obtain cross-linkable systems. Two chemical paths were studied. The first one, based on an epoxy-amine reaction between epoxydized linseed or soybean oils and the alkoxysilane precursor ended up with an uncontrolled reaction and a triglyceride disruption. The second, used castor oil as an hydroxylated raw material and was based on the hydroxy-isocyanate reaction. Valuable results were obtained and this silylated oil was formulated thanks to a new oil/water thermo-stabilized emulsion process. Simple and robust, this process allowed to simultaneously shape and harden hybrid microparticles. Ranging between 20 and 200 µm in diameter, hybrid microparticles were spherical, homogeneously distributed and were capable of entrapping and releasing lipophilic molecules. As a model, ibuprofen was efficiently encapsulated and was fully released over 8 hours in a simulated buffer. Furthermore, by changing the composition of hybrid microparticles (inorganic/organic ratio), it was also possible to extend release kinetics and significantly reduce the burst effect. Then, biocompatibility of those hybrid microparticles was demonstrated in vitro and an innovative study of the cross-linking reaction was performed. This study aimed to properly understand hydrolysis and polycondensation mechanisms and took the form of an in situ the sol-gel reaction monitoring. It allowed to identify an alternative biocompatible catalysts and gave an insight on how those reactions can be controlled to reach “stabilized" hybrids with constant properties and exhibiting robust and reproducible drug sustained-releases. Finally, it has been demonstrated that the sol-gel chemistry applied to vegetable oils for the synthesis of original and tunable hybrid materials with concerns to environmental and health issues, opened the gate towards innovations in drug formulation.

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-09-12T20:11:13Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) regina_inez_souza.pdf: 2322505 bytes, checksum: fb22e286692f3f78b323a1aa5767c86b (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-09-18T13:03:41Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) regina_inez_souza.pdf: 2322505 bytes, checksum: fb22e286692f3f78b323a1aa5767c86b (MD5) / Made available in DSpace on 2017-09-18T13:03:41Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) regina_inez_souza.pdf: 2322505 bytes, checksum: fb22e286692f3f78b323a1aa5767c86b (MD5) Previous issue date: 2017 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Na ind?stria farmac?utica, os pol?meros s?o bastante utilizados em sistemas para libera??o controlada de f?rmacos, por serem capazes de liber?-los efetivamente no local alvo, aumentando os benef?cios terap?uticos e minimizando os efeitos colaterais. Entre os pol?meros utilizados para essa finalidade, tem-se o poli(3-hidroxibutirato) (PHB) pertencente a classe dos poli(hidroxialcanoatos), sendo um pol?mero de origem microbiana, que destacase por ser biorreabsorv?vel e biocompat?vel. Quando utilizado em misturas f?sicas com outros pol?meros, as denominadas blendas, modifica-se propriedades f?sicas, mec?nicas e biol?gicas do PHB para aumentar sua aplicabilidade. Nesse sentido, o propilenoglicol (PPG) foi utilizado para prepara??o de blendas PHBPPG (90/10). O objetivo do trabalho foi preparar e estudar filmes polim?ricos constitu?dos por PHB e PHBPPG (90/10) contendo 1% e 5% de Snitrosoglutationa (GSNO), um doador de ?xido n?trico (NO) bem como sintetizar o f?rmaco e avaliar sua estabilidade ap?s armazenamento a 23?C, 4?C e -18?C durante 90 dias por an?lise t?rmica e por espectroscopia na regi?o do infravermelho. Os filmes obtidos contendo GSNO foram caracterizados por an?lise t?rmica, espectroscopia no infravermelho e avaliou-se o perfil de libera??o in vitro dos mesmos. A citotoxicidade dos extratos das matrizes constitu?das somente por PHB e PHBPPG (90/10) foi avaliada sobre c?lulas mononucleares do sangue perif?rico (PBMC) e leuc?citos totais pelo m?todo de exclus?o com azul de Tripan e sobre hem?cias pela avalia??o da atividade hemol?tica. Os resultados da an?lise t?rmica e infravermelho mostraram que a melhor forma de armazenamento de GSNO ? em 4?C e -18?C. As matrizes polim?ricas de PHB e da blenda PHBPPG (90/10) contendo 1% e 5% de GSNO, avaliadas por an?lise t?rmica foram capazes de liberar NO e apresentaram-se mais est?veis termicamente quando comparados aos filmes de PHB e da blenda sem o f?rmaco. O perfil de libera??o dos filmes de PHB e da blenda PHBPPG (90/10) mostrou que h? uma libera??o r?pida inicial de GSNO nas 24 horas iniciais seguido por um padr?o de libera??o controlada no per?odo de 1 a 8 dias para os filmes contendo 5% de GSNO, sendo o filme da blenda PHBPPG (90/10) contendo 5% de f?rmaco, o que apresentou maior porcentagem de libera??o no per?odo avaliado. Extratos provenientes dos filmes de PHB e da blenda PHBPPG (90/10) n?o apresentaram car?ter citot?xico por n?o induzirem hem?lise das hem?cias e n?o reduzirem a viabilidade celular de PBMCs e leuc?citos totais. As blendas desenvolvidas neste trabalho possuem a capacidade de libera??o controlada e in situ de ?xido n?trico, e possuem aplica??o potencial, por exemplo, no revestimento de stents coronarianos para preven??o da restenose p?s-angioplastia. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. / Polymers are widely used for controlled drug release systems improving the therapeutic benefits and minimizing side effects by the pharmaceutical industry due their capability of releasing it effectively at the target site. Among the polymers used for this purpose there is poly(3-hydroxybutyrate, PHB), a polymer from microbial origin of the poly(hydroxyalkanoate) class that is both bioresorbable and biocompatible. When used as a physical blends with other polymers several properties of PHB (physical, mechanical and biological) could be modified to increase their applicability. To produce blends with PHB the propyleneglycol (PPG) was used to prepare PHBPPG blends (90/10). The aim of this work was to prepare and study polymeric films composed by PHB and PHBPPG (90/10) containing 1% and 5% S-nitrosoglutathione (GSNO), a nitric oxide (NO) donor. The GSNO was synthesized and had its stability evaluated after storage at 23 ? C, 4 ? C and -18 ? C for 90 days by thermal analysis and infrared spectroscopy. The polymer materials containing GSNO were characterized by thermal analysis, infrared spectroscopy and also their drug release capability was evaluated. The cytotoxicity of matrices constituted by PHB and PHBPPG (90/10) was evaluated on peripheral blood mononuclear cells (PBMC) and total leukocytes by the exclusion method with Tripan blue and on red blood cells by the evaluation of hemolytic activity. The thermal and infrared analysis showed that the best form of GSNO storage is at 4 ? C and -18 ? C. Polymeric matrices of PHB and PHBPPG (90/10) containing 1% and 5% of GSNO, were able to release NO and were more thermally stable when compared to PHB and blends without the drug. The release profile of the PHB and PHBPPG blends (90/10) showed an initial fast release of GSNO at the initial 24 hours followed by a controlled release pattern in the period of 1 to 8 days for materials containing 5% of GSNO. The PHBPPG (90/10) blends containing 5% of drug presented the highest percentage of release in the evaluated period. The PHB and PHBPPG (90/10) blends did not present cytotoxic effects. The studied materials did not induced hemolysis of the red blood cells and did not reduced the cellular viability of PBMCs and total leukocytes. The developed blends are capable to exert the in situ controlled-release of nitric oxide and can potentially be used for example to coat coronary stents and thus help to prevent post-angioplasty restenosis.