Actuellement les traitements thérapeutiques pour soigner la schizophrénie, par voie intraveineuse ou orale, ne sont qu’en partie efficaces et associés généralement à des effets extrapyramidaux souvent dangereux et très gênants pour les patients. Afin d’augmenter l’efficacité du traitement toute en neutralisant les effets indésirables, ce travail a eu comme objectif de concevoir des capsules composites (PLGA-PEG / alginate) destinées à être administrées par voie orale et capables de libérer localement, de façon spécifique et contrôlée, le neuroleptique halopéridol dans le cerveau. L’optimisation du protocole de synthèse a permis d’obtenir de façon reproductible des nanocapsules de PLGA poreuses monodisperses et peu agrégées, possédant un diamètre hydrodynamique moyen inférieur à 80 nm et une bonne stabilité en solution aqueuse. Une fois fonctionnalisées avec le Poly (éthylène glycol) diamine, des études in vitro ont montré la faible toxicité de ces nanoparticules furtives ainsi que leur capacité à encapsuler une quantité satisfaisante d’halopéridol et de libérer ce principe actif sur une durée d’un mois avec un faible effet « burst ». L’incorporation des nanoparticules pégylées dans des matrices préparées à haute concentration d’alginate et de 100 % CaCl2 a permis d’obtenir des billes nanocomposites possédants une meilleure stabilité à la sortie du milieu gastrique simulé et persistent environ 30 minutes en milieu intestinal simulé. Enfin des études in vivo préliminaires sur des souris adultes utilisant des nanoparticules injectées et des billes nanocomposites ingérées ont démontré l’efficacité de ces systèmes à délivrer l’halopéridol au cerveau. / Currently therapeutic treatments for schizophrenia, intravenously or orally, are only partially effective and generally associated with extrapyramidal effects often dangerous and very troublesome for patients. In order, to increase the treatment efficiency by neutralizing any side effects the aim of this work was to design composite capsules (PLGA-PEG / alginate) intended to be administered by way oral and able to release locally, in a specific and controlled way, the neuroleptic “haloperidol” in the brain. The optimization of the protocol of synthesis allowed to obtain in a reproducible way of the nanocapsules of monodisperse and not very aggregate porous PLGA, having an average hydrodynamic diameter lower than 80 Nm and a good stability in aqueous solution. Once functionalized with Poly (ethylene glycol) diamine, in vitro studies showed the low toxicity of these furtive nanoparticles as well as their ability to encapsulate a satisfactory amount of haloperidol and release this active principle over a period of one month with a low burst effect. The incorporation of the PEGylated nanoparticles in matrices prepared with a high concentration of alginate and 100% CaCl2 made it possible to obtain nanocomposite beads having a better stability at the exit from the simulated gastric medium and persist approximately 30 minutes in simulated intestinal medium. Finally, preliminary in vivo studies on adult mice using injected nanoparticles and ingested nanocomposite balls showed the effectiveness of these systems to deliver haloperidol in the brain.

Neste trabalho foram desenvolvidos sistemas microfluídicos para aplicações na produção de micro e nanopartículas. Os dispositivos microfluídicos foram microfabricados em vidros do tipo borosilicato e em cerâmica verde LTCC (Low Temperature Co-fired Ceramic). Para os dispositivos em vidro foram utilizadas técnicas de fotolitografia, corrosão úmida e soldagem por cola UV. Com estas técnicas foram produzidos sistemas planares com diversas geometrias, sistemas com dispositivos em paralelo com duas e três camadas de vidros. Além disso, dois polímeros o Benzociclobuteno (BCB) e o Parylene-C foram apresentados como ferramenta para a modificação da superfície do vidro de hidrofílica para hidrofóbica. A cerâmica LTCC foi utilizada para a produção de um sistema microfluídico para focalização hidrodinâmica em 3 dimensões. Os dispositivos microfabricados foram utilizados para estudos sobre a produção de emulsões simples e duplas, observando a influência de parâmetros como taxa de fluxo, razão entre fluxos e diferentes tipos de emulsificantes no tamanho das gotas e no tipo de corte obtido em cada situação. Observou-se que o tamanho máximo das gotas obtidas fica restringido ao tamanho do canal microfabricado, cerca de 50m e o tamanho mínimo obtido foi de 15m. Além da produção de emulsões, foi estudada a produção de nanosuspensões pela técnica de nanoprecipitação anti-solvente. Para este estudo, além das geometrias planares, foram testadas as geometrias 3D e também sistemas para o aumento de escala de produção, onde o sistema integrado possuía 4 dispositivos para nanoprecipitação. Os resultados obtidos indicaram que os sistemas microfluídicos permitem a produção de nanopartículas amorfas, na faixa de 100 a 1000 nm, com baixa polidispersão, sendo ainda reprodutíveis em sistema de maior escala. O desenvolvimento deste trabalho mostrou que a microfluídica oferece ferramentas importantes na obtenção de micro e nanopartículas. / In this work microfluidic systems were developed for applications in micro and nanoparticles production. Microfluidic devices were microfabricated in borosilicate glasses substrates and LTCC ceramic (Low Temperature Co-fired Ceramic). For glass devices were used techniques such as photolithography, wet etching and UV glue for sealing glass wafers. With these techniques were manufactured planar systems with various geometries, systems with devices in parallel with two and three glass layers. In addition, two polymers, BCB and Parylene-C, were presented as a tool for glass surface modification, from hydrophilic to hydrophobic. The ceramic LTCC was used for the production of a microfluidic system for hydrodynamic focusing in three dimensions. The devices were used for studies on the single and double emulsions production, observing the influence of parameters such as flow rate, ratio between flows and different types of surfactants at the droplet size and droplet cut type obtained in each situation. It was observed that the maximum size of the droplets obtained is restricted by the channel size, the maximum was about 50m and the minimum size of 15m. Besides, the devices were used to produce nanoparticles using anti-solvent nanoprecipitation technique. For these studies, besides the planar geometries, 3D geometries were tested and also systems for increasing scale production, where 4 devices were integrated in one system for nanoprecipitation. The results indicated that the microfluidic systems allow the production of amorphous nanoparticles in the range of 100 to 1000 nm with low polydispersity, being also reproducible in a larger scale system. The development of this work has shown that microfluidics offers valuable tools in obtaining micro-and nanoparticles.

Neste trabalho foram desenvolvidos sistemas microfluídicos para aplicações na produção de micro e nanopartículas. Os dispositivos microfluídicos foram microfabricados em vidros do tipo borosilicato e em cerâmica verde LTCC (Low Temperature Co-fired Ceramic). Para os dispositivos em vidro foram utilizadas técnicas de fotolitografia, corrosão úmida e soldagem por cola UV. Com estas técnicas foram produzidos sistemas planares com diversas geometrias, sistemas com dispositivos em paralelo com duas e três camadas de vidros. Além disso, dois polímeros o Benzociclobuteno (BCB) e o Parylene-C foram apresentados como ferramenta para a modificação da superfície do vidro de hidrofílica para hidrofóbica. A cerâmica LTCC foi utilizada para a produção de um sistema microfluídico para focalização hidrodinâmica em 3 dimensões. Os dispositivos microfabricados foram utilizados para estudos sobre a produção de emulsões simples e duplas, observando a influência de parâmetros como taxa de fluxo, razão entre fluxos e diferentes tipos de emulsificantes no tamanho das gotas e no tipo de corte obtido em cada situação. Observou-se que o tamanho máximo das gotas obtidas fica restringido ao tamanho do canal microfabricado, cerca de 50m e o tamanho mínimo obtido foi de 15m. Além da produção de emulsões, foi estudada a produção de nanosuspensões pela técnica de nanoprecipitação anti-solvente. Para este estudo, além das geometrias planares, foram testadas as geometrias 3D e também sistemas para o aumento de escala de produção, onde o sistema integrado possuía 4 dispositivos para nanoprecipitação. Os resultados obtidos indicaram que os sistemas microfluídicos permitem a produção de nanopartículas amorfas, na faixa de 100 a 1000 nm, com baixa polidispersão, sendo ainda reprodutíveis em sistema de maior escala. O desenvolvimento deste trabalho mostrou que a microfluídica oferece ferramentas importantes na obtenção de micro e nanopartículas. / In this work microfluidic systems were developed for applications in micro and nanoparticles production. Microfluidic devices were microfabricated in borosilicate glasses substrates and LTCC ceramic (Low Temperature Co-fired Ceramic). For glass devices were used techniques such as photolithography, wet etching and UV glue for sealing glass wafers. With these techniques were manufactured planar systems with various geometries, systems with devices in parallel with two and three glass layers. In addition, two polymers, BCB and Parylene-C, were presented as a tool for glass surface modification, from hydrophilic to hydrophobic. The ceramic LTCC was used for the production of a microfluidic system for hydrodynamic focusing in three dimensions. The devices were used for studies on the single and double emulsions production, observing the influence of parameters such as flow rate, ratio between flows and different types of surfactants at the droplet size and droplet cut type obtained in each situation. It was observed that the maximum size of the droplets obtained is restricted by the channel size, the maximum was about 50m and the minimum size of 15m. Besides, the devices were used to produce nanoparticles using anti-solvent nanoprecipitation technique. For these studies, besides the planar geometries, 3D geometries were tested and also systems for increasing scale production, where 4 devices were integrated in one system for nanoprecipitation. The results indicated that the microfluidic systems allow the production of amorphous nanoparticles in the range of 100 to 1000 nm with low polydispersity, being also reproducible in a larger scale system. The development of this work has shown that microfluidics offers valuable tools in obtaining micro-and nanoparticles.

This work deals with the fabrication process of low density foam shells and the sharp control of their shape (diameter, thickness, density, sphericity, non-concentricity). During this PhD we focused on the non-concentricity criterion which has to be lower than 1%. The shells are synthesized using a microencapsulation process leading to a double emulsion and followed by a thermal polymerization at 60°C. According to the literature, three major parameters, the density of the three phases, the deformations of the shells along the process and the kinetics of the polymerization have a direct influence on the shells non-concentricity. The results obtained showed that when the density gap between the internal water phase and the organic phase increases, the TMPTMA shells non-concentricity improves. A density gap of 0.078 g.cm-3 at 60°C, leads to an average non-concentricity of 2.4% with a yield of shells of 58%. It was also shown that the synthesis process can be considered as reproducible. While using the same internal water phase, equivalent non-concentricity results are obtained using either a straight tube, a tube with areas of constriction or a short wound tube. The time required to fix the shell's shape is at least 20 minutes with thermal polymerization. So, it seems that the time spent by the shells inside the rotating flask allows the centering of the internal water phase inside the organic phase, whatever the circulation process used. In order to get higher polymerization rates and to avoid destabilization phenomena, we then focused our study on photopolymerization. When the synthesis is performed using a UV lamp with an efficient light intensity, the shells have a slightly higher thickness than the shells synthesized by thermal polymerization. Moreover, a really higher yield, around 80%, is achieved with UV polymerization. However, the average non-concentricity of the shells synthesized lays around 20%, which is really high compared to the 2.4% average non-concentricity obtained with thermal polymerization. It would be interesting to expose the shells to UV light at different times after collection in order to study the influence of the agitation time on the shells non-concentricity.

Ce travail porte sur le processus de fabrication de microballons en mousse basse densité et le contrôle fin de leur forme (diamètre, épaisseur, densité, sphéricité, non-concentricité). Durant cette thèse nous nous sommes concentrés sur le critère de non-concentricité qui doit être inférieure à 1%. Les microballons sont synthétisés en utilisant un procédé de microencapsulation conduisant à une émulsion double, suivie d'une polymérisation thermique à 60°C. Selon la littérature, trois paramètres majeurs, la densité des trois phases, les déformations du microballon pendant le procédé et la cinétique de polymérisation ont une influence directe sur la non-concentricité des microballons. Les résultats obtenus ont montré que lorsque l'écart de densité entre la phase aqueuse interne et la phase organique augmente, la non-concentricité des microballons TMPTMA s'améliore. Un écart de densité de 0,078 g.cm-3 à 60°C conduit à une non-concentricité moyenne de 2,4% avec un rendement en microballons de 58%. Il a également été montré que la synthèse peut être considérée comme reproductible. Pour une même phase aqueuse interne, les résultats de non-concentricité sont équivalents en utilisant soit un tube droit, un tube à étranglement ou un serpentin court. Le temps requis pour fixer la forme des microballons est d'au moins 20 minutes avec la polymérisation thermique. Ainsi, il semble que le temps passé par les microballons à l'intérieur des bouteilles de réception permet le centrage de la phase aqueuse interne à l'intérieur de la phase organique, quel que soit le processus de circulation précédemment utilisé. Afin d'obtenir des vitesses de polymérisation plus élevées et d'éviter les phénomènes de déstabilisation, nous avons alors concentré notre étude sur la photopolymérisation. Lorsque la synthèse est effectuée en utilisant une lampe UV avec une intensité lumineuse efficace, les microballons ont une épaisseur légèrement supérieure à celle des microballons synthétisés par voie thermique. Par ailleurs, un rendement plus élevé, environ 80%, est obtenu avec la polymérisation UV. Toutefois, la non-concentricité moyenne des microballons synthétisés est environ de 20%, ce qui est vraiment élevé par rapport à la non-concentricité moyenne de 2,4% obtenue par polymérisation thermique. Il serait intéressant d'exposer les microballons à la lumière UV, à différents moments après la collecte afin d'étudier l'influence du temps d'agitation sur la non-concentricité des microballons. / This work deals with the fabrication process of low density foam shells and the sharp control of their shape (diameter, thickness, density, sphericity, non-concentricity). During this PhD we focused on the non-concentricity criterion which has to be lower than 1%. The shells are synthesized using a microencapsulation process leading to a double emulsion and followed by a thermal polymerization at 60°C. According to the literature, three major parameters, the density of the three phases, the deformations of the shells along the process and the kinetics of the polymerization have a direct influence on the shells non-concentricity. The results obtained showed that when the density gap between the internal water phase and the organic phase increases, the TMPTMA shells non-concentricity improves. A density gap of 0.078 g.cm-3 at 60°C, leads to an average non-concentricity of 2.4% with a yield of shells of 58%. It was also shown that the synthesis process can be considered as reproducible. While using the same internal water phase, equivalent non-concentricity results are obtained using either a straight tube, a tube with areas of constriction or a short wound tube. The time required to fix the shell’s shape is at least 20 minutes with thermal polymerization. So, it seems that the time spent by the shells inside the rotating flask allows the centering of the internal water phase inside the organic phase, whatever the circulation process used. In order to get higher polymerization rates and to avoid destabilization phenomena, we then focused our study on photopolymerization. When the synthesis is performed using a UV lamp with an efficient light intensity, the shells have a slightly higher thickness than the shells synthesized by thermal polymerization. Moreover, a really higher yield, around 80%, is achieved with UV polymerization. However, the average non-concentricity of the shells synthesized lays around 20%, which is really high compared to the 2.4% average non-concentricity obtained with thermal polymerization. It would be interesting to expose the shells to UV light at different times after collection in order to study the influence of the agitation time on the shells non-concentricity.

L'objectif de cette étude était de préparer et de caractériser les particules submicroniques multifonctionnelles utilisables simultanément pour le diagnostic et le traitement de plusieurs maladies mortelles telles que le cancer. Pour ce faire, une étude systématique a été réalisée afin de comprendre les mécanismes impliqués et d'optimiser les paramètres du procédé de double émulsion-évaporation de solvant pour la préparation de ces particules. Pour l’imagerie in vitro, des nanoparticules polymériques fluorescentes (FluoSpheres®) ont été encapsulées dans une matrice polycaprolactone dégradable en utilisant le procédé de l’émulsion double-évaporation de solvant. Pour l’imagerie invivo, des nanoparticules d'or colloïdal ont été préparées et encapsulées via le même procédé et parfaitement caractérisées. Enfin, pour application theranostic, les nanoparticules d'or (comme agent de contraste) et un actif moléculaire (hydrophile Nefopam et hydrophobe benzoate de benzyle) ont été encapsulés simultanément dans des particules de polycaprolactone. Ces particules multifonctionnelles ont été caractérisées et évaluées in vitro comme model de pénétration cutané / The objective of this study was to prepare and characterize multifunctional submicron particles that can be used for diagnosis and therapy of several fatal diseases including cancer (i.e theranostic). For this purpose, a systematic study was performed in order to optimize the process parameters for preparation of polymeric particle that can be used as a platform for effective delivery of drugs and imaging labels. The imaging agent (FluoSpheres®) was encapsulated via double emulsion solvent evaporation technique to be used fluorescent contrast agent and their in vitro evaluation was performed. Then, gold nanoparticles were prepared by using NaBH4 reduction method, characterized and encapsulated by polycaprolactone polymer for in vitro applications. Finally, the gold nanoparticle were loaded into polycaprolactone particle along with a hydrophilic drug (Nefopam) and a hydrophobic drug (benzyl benzoate) simultaneously. The prepared particles were then characterized physicochemically and in vitro skin penetration study was performed

Des biomolécules telles que les protéines et les acides nucléiques sont d'un grand intérêt dans l'immunothérapie, la thérapie génique tels que des cancers, le traitement de SIDA, les troubles auto-immuns liés à la déficience immunitaire sévère et combinés à autres maladies. Afin de cibler ces objectifs, l'encapsulation et le ciblage sont incontestablement nécessaires pour protéger les biomolécules contre la dégradation rapide et aussi pour éviter les effets secondaires. Ainsi, dans le cadre de cette thèse, l'encapsulation de protéines et de l'ADN en utilisant le procédé basé sur la double émulsion évaporation de solvant a été étudiée. Mais avant de conduire l’étude expérimentale, l'état de l'art des procédés d'encapsulation a été effectué et une attention particulière a été consacrée à l’encapsulation de l'ADN et les protéines et aux procédés d’encapsulations basés sur la formulation des polymères biodégradables. En ce qui concerne l’étude expérimentale, une étude systématique des paramètres pertinents sur les propriétés physico-chimique et colloïdales des particules a été examinée. Cette étude a permis de mettre en évidence les facteurs clés capables d’affecter non seulement le procédé mais aussi les propriétés colloïdales des particules élaborées (taille, distribution en taille, propriétés électrocinétiques et stabilité colloïdale…). En second lieu, la formulation optimisée permettant l’obtention de tailles submicroniques a été utilisée pour l’encapsulation de l'ADN et les protéines. L'encapsulation de l'ADN a été étudiée en fonction de la concentration de l'ADN (sperme de saumon) et une encapsulation totale d'ADN a été observée sans modification ou altération des propriétés colloïdales des particules finales. L'encapsulation de protéines a été également examinée en utilisant HAS et une autre protéine fluorescente comme modèles. L'efficacité de l'encapsulation a été déterminée et trouvée de l’ordre de 100%. L'utilisation de la protéine fluorescente indique l'encapsulation totale avec une bonne répartition dans les cavités de la matrice polymère. En conclusion, le procédé de la double émulsion – évaporation de solvant est efficace pour l’encapsulation de l'ADN et des protéines sans modification des propriétés colloïdales des particules finales qui restent bien submicroniques / Biomolecules such as proteins and nucleic acids are of great interest in immunotherapy and gene therapy such as cancers, AIDS, autoimmune disorders, X-linked severe combined immune deficiency and many other diseases. In order to target such objectives, the encapsulation and the targeting are incontestably necessary principally to protect biomolecules against rapid degradation and also to avoid side effects. Then, in this thesis, the encapsulation of proteins and DNA using double emulsion solvent evaporation technique was performed. But before the encapsulation, the state of the art of the encapsulation processes was performed and special attention has been dedicated to DNA and proteins encapsulation. Firstly, systematic study of double emulsion solvent evaporation process was investigated in order to point out the key parameters controlling the particles size and the colloidal stability. Secondly, the optimized formulation conditions leading to submicron colloidal was used for DNA and Proteins encapsulation. The DNA encapsulation was investigated as a function of DNA concentration and it was found total encapsulation of DNA without any modification of the colloidal properties. The protein encapsulation was examined using HAS and fluorescent proteins as model. The encapsulation efficiency was found to be high and reaches 100%. The use of fluorescent protein shows the total encapsulation and good distribution of proteins in polymer matrix. As a general conclusion, the high encapsulation efficiency of DNA and protein, reveled the compatibility of the used process to encapsulated hydrophilic biomolecules even in submicron size biodegradable colloidal submicron particles