Francisella tularensis is a Gram-negative, zoonotic bacterium that causes tularemia in animals and humans. The two subspecies tularensis (Type A) and holarctica (Type B) are considered Tier I Select Agents due to the bioweapon potential of these subspecies. Type A strains, considered the more virulent of the subspecies, are highly infective producing respiratory tularemia with inhalation of as few as 10 cells. Due to classification as a Select Agent, a vast amount of F. tularensis research has occurred in the last two decades after the September 11th terrorism attack and the use of Bacillus anthracis spores in a biological attack on the United States Postal Services in 2001. This research has uncovered many of the various virulence factors of F. tularensis including an intracellular nature, the unique lipopolysaccharide produced, and a genetic pathogenicity island. This dissertation aims to further characterize outer surface antigens of F. tularensis subspecies in regards to virulence, biofilm formation, and role in vaccine development. In addition, this dissertation will also investigate the use of a novel vaccine delivery vehicle, alginate microencapsulation, in increasing the efficacy of these mutant strains. F. novicida is a subspecies of F. tularensis and usually classified as being non-encapsulated. However, F. novicida has a similar capsule glycosylation locus as F. tularensis and could produce a similar capsule-like complex that has previously been described for the F. tularensis LVS strain. I was able to isolate and characterize this CLC of F. novicida, which contained a heterogenous mixture of proteins and possible glycosylated proteins. A mutant with a multi-gene interruption within the glycosylation locus (F. novicidaΔ1212-1218) produced significantly less carbohydrate than the parent strain, was attenuated in the mouse model, and was partially protective when used to immunize mice against a virulent challenge. Biofilms of F. novicida were also characterized in regards to biofilm formation in various growth media and biofilm formation of strains lacking the O-antigen of the lipopolysaccharide (LPS). In general, F. novicida produced the greatest amount of biofilm in a brain heart infusion (BHI) broth, compared to other media. Loss of the O-antigen led to increased biofilm production when grown in BHI and decreased or similar biofilm production as the wildtype when grown in other media. This highlights the need to carefully select the growth medium when assessing biofilm formation of Francisella strains in the future. A final study of this dissertation characterized the use of alginate microspheres as a vaccine vehicle for an attenuated F. tularensis type A O-antigen deficient strain. O-antigen deficient strains of F. tularensis are highly attenuated in vivo and would be a safe choice for a vaccine candidate. However, these strains produce less than ideal protection against virulent challenge when used to immunize mice, possibly due to a lack of persistence in the host. In an attempt to increase persistence, we encapsulated an O-antigen deficient strain within sodium alginate microspheres and used those microspheres to immunize mice. The immunized mice produced a higher level of antibody response than mice immunized with a non-encapsulated version. However, this immunization only partially protected mice from a virulent challenge and did not match the protection afforded by the former Live Vaccine Strain (LVS). In part the deficiency in protection appears to be due to a lack of a robust cellular immune response in mice immunized with the alginate microspheres. In summary, this dissertation focuses on the various extracellular polysaccharides of F. tularensis: the glycosylation of CLC, the O-antigen, and the biofilm. Each polysaccharide plays a role in the virulence and pathogenesis of F. tularensis. Glycosylation of the CLC and the O-antigen are important virulence factors in mammalian disease, and mutants lacking either (not type A strains) are attenuated in the mouse model. Both also appear to play a role in the formation of the F. tularensis biofilm in a manner dependent on the environment or culture medium used. Each of these extracellular polysaccharides contribute to the lifecycle of Francisella. / Ph.D. / Francisella tularensis is a highly infectious bacterial pathogen that can cause disease in a wide array of animals and in humans. F. tularensis is also considered a potential weapon of bioterrorism and the development of an effective vaccine is a critical area of research. One strategy of developing a tularemia vaccine includes mutating a strain of F. tularensis to reduce expression of extracellular components that include polysaccharides. Strains that cannot express these components are usually unable to produce clinical signs in the host and may provide protection against fully virulent F. tularensis strains. The work presented in this dissertation will focus on characterizing the polysaccharide extracellular components of F. tularensis and developing a novel vaccine vehicle to increase protection from strains that do not cause disease.

Yes / The aim of this work was to investigate the possibility of covalent cross-linker-free, polyelectrolyte complex formation at the nanoscale between alginic acid (as sodium alginate, ALG) and protamine (PROT). Optimisation of the self-assembly conditions was performed by varying the type of polymer used, pH of component solutions, mass mixing ratio of the components and the speed and order of component addition on the properties of complexes. Homogenous particles with nanometric sizes resulted when an aqueous dispersion of ALG was rapidly mixed with a solution of PROT. The polyelectrolyte complex between ALG and PROT was confirmed by infrared spectroscopy. To facilitate incorporation of drugs soluble at low pH, pH of ALG dispersion was decreased to 2; however, no nanoparticles (NPs) were formed upon complexation with PROT. Adjusting pH of PROT solution to 3 resulted in the formation of cationic or anionic NPs with a size range 70–300 nm. Colloidal stability of selected alginic acid low/PROT formulations was determined upon storage at room temperature and in liquid media at various pH. Physical stability of NPs correlated with the initial surface charge of particles and was time- and pH-dependent. Generally, better stability was observed for anionic NPs stored as native dispersions and in liquids covering a range of pH. / This study was funded by Merrion Pharmaceuticals Ireland. This work was also supported by the Synthesis and Solid State Pharmaceutical Centre funded by the Science Foundation Ireland under grant number 12/RC/2275.

Yes / Fly ash is a toxic industrial waste, mainly consisting of silica and alumina particles, that has been found discharged into the environment. It is proposed that alginate, a naturally occurring biopolymer, can bind to these minerals and thus play a role in water purification. The binding forces involved in this process consist of weak interactions, such as van der Waals forces and electrostatic interactions. Although the attachment of alginate to mineral surfaces is mainly governed by its carboxylate groups, hydroxyl moieties could play a role in the interaction between the polymer and minerals. This work aims to use the SiO2 and Al2O3 particles as models for fly ash and to show the use of alginate biopolymers (fluorescently labelled with an aminonaphthaline sulfonate fluorophore (AmNS)) to coagulate them. The addition of simple electrolytes like NaCl and CaCl2 encourages the coiling of the polymer chain at high pH values which has an effect on its capability to bind to the inorganic particles. A combination of fluorescence and ICP-MS demonstrated that alginate has a considerable adsorption affinity for Al2O3, whereas it attracts SiO2 weakly. The adsorption process is pH dependent: strong adsorption was observed at low pH values. The dependence of adsorption on the mineral (Al2O3 and SiO2) concentration was also examined under different pH conditions: the adsorption amount was observed to increase by increasing the solid concentration. Adsorption isotherms obtained at low and high mineral concentrations were found to be Henry in type.

Yes / While the continuous development of advanced bioprinting technologies is under fervent study, enhancing the regenerative potential of hydrogel-based constructs using external stimuli for wound dressing has yet to be tackled. Fibroblasts play a significant role in wound healing and tissue implants at different stages, including extracellular matrix production, collagen synthesis, and wound and tissue remodeling. This study explores the synergistic interplay between photothermal activity and nanomaterial-mediated cell proliferation. The use of different graphene-based materials (GBM) in the development of photoactive bioinks is investigated. In particular, we report the creation of a skin-inspired dressing for wound healing and regenerative medicine. Three distinct GBM, namely, graphene oxide (GO), reduced graphene oxide (rGO), and graphene platelets (GP), were rigorously characterized, and their photothermal capabilities were elucidated. Our investigations revealed that rGO exhibited the highest photothermal efficiency and antibacterial properties when irradiated, even at a concentration as low as 0.05 mg/mL, without compromising human fibroblast viability. Alginate-based bioinks alongside human fibroblasts were employed for the bioprinting with rGO. The scaffold did not affect the survival of fibroblasts for 3 days after bioprinting, as cell viability was not affected. Remarkably, the inclusion of rGO did not compromise the printability of the hydrogel, ensuring the successful fabrication of complex constructs. Furthermore, the presence of rGO in the final scaffold continued to provide the benefits of photothermal antimicrobial therapy without detrimentally affecting fibroblast growth. This outcome underscores the potential of rGO-enhanced hydrogels in tissue engineering and regenerative medicine applications. Our findings hold promise for developing game-changer strategies in 4D bioprinting to create smart and functional tissue constructs with high fibroblast proliferation and promising therapeutic capabilities in drug delivery and bactericidal skin-inspired dressings.

L'objectif de cette thèse était de développer de nouveaux « donneurs de NO » stables en liant du S-nitrosoglutathion (GSNO) à une structure polymérique. Dans une première étape, les polymères ont été liés au glutathion (GSH) : le chitosan-GSH et l'alginate-GSH ont ainsi été préparés par la « méthode des carbodiimides » et dans une deuxième étape, les polymères finaux [SNOC (S-nitrosoglutathione-oligosaccharide-chitosan) et SNA (S-nitrosoglutathione-alginate)] ont été préparés par nitrosation des deux conjugués précédent. La quantité de NO fixée a été déterminée par les méthodes Griess et Saville. L’aptitude des polymères à libérer du NO et à passer la barrière intestinale [SNOC et SNA] a été évaluée dans une chambre d’Ussing. Nous avons obtenu des polymères avec des quantités variables de NO en fonction de la méthode utilisée (159 µmol de NO/g à 525 µmol de NO/g pour le SNOC ; 174 µmol de NO/g à 468 µmol de NO/g pour le SNA). Le SNOC était stable pendant au moins 6h et le SNA pendant au moins 10h. Enfin, nous avons essayé de mettre au point des microparticules de GSH et GSNO par spray drying avec de l’Eudragit ® FS 30D gastro-résistant. La caractérisation des microparticules a été réalisée par microscopie électronique à balayage (SEM), par diffraction X (PXRD) et par spectroscopie infrarouge (FTIR). Les essais de libération in vitro ont été réalisés dans un tampon (pH 1,2, 3, 6, 6,8 et 7,4). Les microparticules étaient chargées négativement avec une taille moyenne allant de 5 à 7 µm. La formulation était stable à pH acide mais a montré une libération rapide à pH basique ; elle pourrait donc servir de système de délivrance du NO au niveau intestinal. / The aim of the thesis was to develop novel and stable NO-donors by linking S-nitrosoglutathione (GSNO) to a polymer backbone. In the first step, chitosan-GSH and alginate-GSH conjugates were prepared by a carbodiimide reaction and in the second step SNOC (S-nitrosoglutathione-oligosaccharide-chitosan) and SNA (S-nitrosoglutathione-alginate) were prepared by the nitrosation of both conjugates respectively. The amount of NO was determined by Griess and Saville methods. Stability and ex vivo experiments of SNOC and SNA were performed in an Ussing chamber through rat intestine. We obtained polymers with different amount of NO (i.e. 159 µmol of NO/g to 525 µmol of NO/g for SNOC; 174 µmol of NO/g to 468 µmol of NO/g for SNA) depending upon the procedure of nitrosation. SNOC was stable for at least 6h and SNA for at least 10h. Also, we aimed to develop spray dried microparticles of GSH and GSNO based on Eudragit® FS 30D polymer. The microparticles were characterized by scanning electron microscopy (SEM), X-ray diffraction (PXRD), infrared spectroscopy (FTIR) and in vitro release studies were performed in different pH conditions (pH 1.2, 3, 6, 6.8 and 7.4). The microparticles were negatively charged with mean particle size ranging from 5 to 7 µm. The formulation was stable and was resistant to acidic pH but showed rapid release in basic pH; hence, they can be used as colon specific drug delivery systems for the treatment of Crohn’s disease. We think that these formulations could be used in animal models in the treatment of Crohn’s disease.

Ce travail décrit la synthèse d'une série de matériaux à base de biomasse d'alginate et d'algues; ces matériaux ont été conçus sous différentes formes: billes et mousses. Des procédés spécifiques ont été mis au point pour l’utilisation directe de la biomasse algale (AB, sans ajout d’autres polymères) avec le souci de développer un processus de synthèse simple et respectueux de l’environnement (production réduite de sous-produits et sans additif supplémentaire). Ces matériaux ont été testés pour la décontamination d’effluents contenant des métaux lourds (Pb(II) et Cu(II)), mais également pour la valorisation des métaux (métaux du groupe platine, PGM: Pd(II) et Pt(IV)). Différents paramètres opératoires ont été testés afin d'évaluer les capacités de sorption et les étapes limitantes, mais également d'identifier des stratégies d’amélioration des performances d’adsorption. L’incorporation de poly(éthylèneimine) (PEI) est une méthode prometteuse pour augmenter la densité de groupes réactifs (fonctions amines). Différents procédés ont été testés: (a) l'incorporation de particules de PEI réticulées avec du glutaraldéhyde (billes hétérogènes: ABA/PEI), et (b) le greffage homogène de PEI sur de l'alginate (suivi de la réticulation par le glutaraldéhyde) (billes homogènes HABA/PEI). La spectroscopie FTIR et l'analyse MEB& MEB-EDX ont été utilisées pour interpréter les mécanismes de fixation ainsi que pour caractériser la structure des matériaux. Dans une deuxième étape, les matériaux sélectionnés ont été testés pour la catalyse supportée en utilisant la réaction d'hydrogénation du 3–nitrophénol (3-NP). Les résultats sont structurés en 3 parties développées successivement: (a) synthèse des billes d’alginate, AB et AB/PEI et étude de l’adsorption de métaux lourds et de PGMs, (b) comparaison des propriétés d’adsorption du Pd(II) par les billes composites AB/PEI préparées par les voies homogène et hétérogène (et leur application aux tests en catalyse supportée), et (c) la synthèse de mousses poreuses (préparées par réaction entre l’alginate et la PEI) appliquées à l’adsorption du Pd(II) et à la catalyse supportée en réacteur à lit fixe.Si la PEI a un effet limité sur la fixation des métaux lourds (interaction avec les groupes carboxyliques de la biomasse d'alginate ou d'algues), sa présence améliore l’adsorption des métaux dans le cas des PGMs (les groupes amine protonés ont une forte affinité pour les espèces chloro-anioniques du Pd(II)) en particulier pour les billes d'alginate et AB. Tous les adsorbants ont une préférence pour le Pb(II) par rapport au Cu(II) et pour le Pd(II) par rapport au Pt(IV); la présence de PEI limite la sélectivité du matériau pour le Pb(II) et le Pd(II). La capacité de fixation et la stabilité des billes composites d'alginate/PEI ont été améliorées en utilisant le mode de synthèse homogène (la PEI étant dispersée de manière homogène dans la bille avant la réticulation par le glutaraldéhyde). Les deux supports (billes hétérogènes ou homogènes) chargés en Pd(II) (ensuite réduit) ont permis des performances catalytiques comparables bien qu’inférieures à celles des catalyseurs classiques, mais le mode homogène améliore la stabilité à long terme du matériau. Le conditionnement du support catalytique sous forme de mousse a permis de tester la réaction catalytique dans un système à lit fixe : le conditionnement sous forme de mousse améliore les propriétés de transfert de masse par rapport aux billes et la constante de vitesse apparente n'est que légèrement réduite après 30 cycles de fonctionnement. / This work describes the synthesis of a series of materials based on alginate and algal biomass (AB); these materials have been designed under different shapes: beads and foams. Special processes have been developed for directly using the algal biomass (without adding other polymers) with the double objective of simple processing and environmentally-friendly manufacturing (reduced production of sub-products and without additional resources). These materials have been tested first for metal recovery for heavy metal decontamination (Pb(II) and Cu(II)) but also for the valorization of metals (platinum groups metals, PGMs: Pd(II) and Pt(IV)). These studies were performed investigating various operating conditions in order to evaluate sorption capacities and limiting steps but also to identify the processes to be used for improving sorption performance. The incorporation of poly(ethyleneimine), PEI, is a promising method for increasing the density of highly reactive groups (amine functions). Different processes have been tested: (a) the incorporation of particles of PEI crosslinked with glutaraldehyde (heterogeneous beads: ABA/PEI), and (b) the homogeneous grafting of PEI on alginate (followed by glutaraldehyde crosslinking) (HABA/PEI beads). Several techniques have been used for characterizing the sorption process and the structure of developed sorbents, including FTIR spectroscopy, SEM & SEM-EDX analysis. In a second step selected materials have been tested for supported catalysis using the simple reaction of hydrogenation of 3–nitrophenol (3-NP) as a test reaction. The results are structured in 3 parts successively developed: (a) synthesis of alginate, AB and AB/PEI beads and testing for sorption heavy metals and PGMs, (b) comparison of Pd(II) sorption properties of AB/PEI composite beads prepared by the homogeneous and the heterogeneous routes (and their application to supported catalytic tests), and (c) synthesis of highly porous foams (prepared by reaction of alginate with PEI) and the testing of Pd(II) sorption and Pd-supported catalysis (in fixed-bed reactor). While PEI hardly affects the sorption of heavy metals (due to direct interaction with carboxylic groups of alginate or algal biomass), the presence of PEI strongly improves metal binding in the case of PGMs (the protonated amine groups strongly bind chloro-anionic PGM species). All the sorbents have a preference for Pb(II) over Cu(II) and for Pd(II) over Pt(IV), especially for alginate and AB beads because the presence of PEI limits the selectivity of the material for Pb(II) and Pd(II). Both the sorption capacity and the stability of composite alginate/PEI beads were improved while using the homogeneous synthesis mode (the PEI polymer being homogeneously dispersed in the bead before glutaraldehyde crosslinking). The two supports (heterogeneous vs. homogeneous beads) loaded with Pd(II) and subsequently reduced gave comparable catalytic performance (lower than conventional catalysts) but the homogeneous mode improves the long-term stability. The conditioning of the catalytic support as a foam allows testing the catalytic reaction in fixed-bed system: the conditioning improves mass transfer properties compared to beads and the apparent rate constant is only slightly reduced after operating 30 cycles.

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.

La synthèse de glycomonomères à base d'oligoalginate (AlgiMERs) et leur polymérisations conventionnelles et RAFT en solution aqueuse ont été étudiés. Premièrement, l'oligoalginates de départ ont été transformés soit dans le glycosylamines correspondant ou en amino alditols (via une amination réductrice). A cette étape, l'optimisation des protocoles d'amination ont été identifiées par la réalisation d'une étude systématique sur un simple acide uronique (acide D-glucuronique). Deuxièmement, les sucres aminés ont été obtenus a réagi avec une électrophile portant un groupe vinyle polymérisable à céder AlgiMERs. Le glycomonomères résultant n'a pas homopolymérisé même en haute force ionique et pour temps de réaction longs, mais leur copolymérisations radicalaire conventionnelles avec N-(2-hydroxyéthyl)méthacrylamide (HEMAm) donne de glycopolymères avec de haute mass molaires (Mw ≈ 1.500.000 Da) contenant jusqu'à 50% en masse de oligoalginate. Une étude cinétique a confirmé que la consommation des deux monomères suivi une cinétique de premier ordre et que les AlgiMERs ont été intégrées tôt dans le processus de polymérisation. Basé sur ces résultats, l'enquête a été étendue à la copolymérisation radicalaire vivante en milieu aqueuse et glycopolymères gradient bien définies ont été obtenues (Mn = 12 000 Da - 90 000 Da; PDI ≤ 1,20). Enfin, j'ai pu prouver qu'un polymère synthétique portant des résidus d'oligo (1→4)-α-L-guluronan conduit des gels en présence d'ions Ca2+ et offre un hydrogel transparent et stable. / The synthesis of oligoalginate derived glycomonomers (AlgiMERs) and their conventional and Reversible Addition Fragmentation chain Transfer (RAFT) polymerizations in aqueous solution were investigated. Firstly, the starting oligoalginates were transformed either into the corresponding glycosylamines or into amino-alditols (via reductive amination). At this stage, optimized amination protocols were identified by carrying out a systematic study on a simpler uronic acid (D-glucuronic acid). Secondly, the obtained amino sugars were reacted with an electrophile bearing a polymerisable vinyl group to yield AlgiMERs. The resulting glycomonomers did not homopolymerize even in high ionic strength and for long reaction times, but their conventional free radical copolymerization with 2-hydroxyethyl methacrylamide (HEMAm) led to high molecular weight glycopolymers (Mw ≈ 1.500.000 Da) containing up to 50 % by mass of oligoalginate. A kinetic study confirmed that the consumption of both monomers followed a first order kinetic and that oligoalginate-derived monomers were incorporated early on in the polymerization process. Based on these results, the investigation was extended to the living radical copolymerization in aqueous solution and well defined gradient glycopolymers were obtained (Mn = 12 000 Da – 90 000 Da; PDI ≤ 1.20). Finally, I could prove that a synthetic polymer carrying oligo(1→4)-α-L-guluronan residues gels in the presence of Ca2+ ions and affords a transparent and stable hydrogel.

Cette thèse présente un travail sur l’obtention d’aérogels d’HDL par séchage en conditions CO2 supercritique et l’élaboration de nouveaux bionanocomposites formés par la coprécipitation d’hydroxydes doubles lamellaire (HDL) dans l'espace confiné des billes d'alginate. Grâce à la combinaison d’une synthèse par coprécipitation Flash et d’un séchage en conditions supercritiques au CO2, des aérogels d’HDL possédant des surfaces spécifiques élevées sont élaborés. Parallèlement, l’alginate est utilisé comme une matrice de confinement pour la précipitation inorganique d’HDL. D’une part, les billes d'alginate sont synthétisées par complexation des ions Ca2+ et la coprécipitation des phases HDL s’effectue en réalisant des imprégnations successives de réactifs. D’autre part, des billes d'alginate sont formées directement en présence des cations divalents (Mg2+, Ni2++, Co2+, ...) et des cations des métaux trivalents (Al3+), précurseurs des composés inorganiques. La coprécipitation des HDL se produit dans ce cas lors d'une étape d'imprégnation dans une solution d'hydroxyde de sodium. Tous les composés HDL, aérogels ou encore bionanocomposites sont caractérisés en détail par DRX, spectroscopie IR, MEB/MET, adsorption/désorption d’azote et ATG/DTG, pour obtenir un meilleur aperçu de la structure des particules, de leur taille et de leur morphologie. Des études menées sur l’adsorption de la trypsine pour les aérogels ou encore sur les performances d’électrodes modifiées HDL-alginate ont permis de montrer qu’il était possible d'améliorer les performances des HDL en augmenter leur porosité et en élaborant des bionanocomposites. / In this work, we investigated both the use of CO2 supercritical drying conditions and the use of biopolymer to modify the growth and aggregation of inorganic Layered Double Hydroxide (LDH) particles. Indeed, one possibility to enhance their performances is to increase the LDH porosity and to design them as nanostructured open structure. Thanks to the combinaison of fast coprecipitation and CO2 supercritical drying, highly porous LDH aerogels were obtained with enhanced textural properties. In parallel, the coprecipitation of Layered Double Hydroxides (LDH) in the confined space of alginate beads is reported. In our approach, Alginate acts as a template to support and confined the inorganic precipitation. In one hand, beads made of Alginate are synthesized by complexation of Ca2+ ions and LDH phases are coprecipitated using successive impregnations of reactants. In another hand, Alginate beads are formed directly in presence of the divalent (Mg2+, Ni2+, Co2+,…) and trivalent metal cations (Al3+), precursors of the inorganic compounds. LDH coprecipitation then occurs during a further impregnation step in a sodium hydroxide solution. All the LDH aerogels and LDH nanocomposites beads are deeply characterized using XRD, SEM/TEM, FTIR spectroscopic, adsorption/desorption of nitrogen and TGA/DTG to get better insight on particle structure, size and morphology Aerogels display enhanced adsorption behavior toward trypsine immobilization whereas a net improvement of the electrochemical response is noticed for the NiAl based bionanocomposites prepared by confined coprecipitation into Alginate.

Ce travail avait pour objectif la synthèse d’hydrogels d’alginates stables dans le temps et leur évaluation comme support de régénération des voies nerveuses lésées du Système Nerveux Central. Différents dérivés amphiphiles de l’alginate de sodium ont été préparés en fixant chimiquement des chaînes alkyles en C12 à différents taux sur le squelette polymère par l’intermédiaire, soit de liaisons ester (alginates esters), soit de liaisons amide (alginamides) moins sensibles à l’hydrolyse. Le comportement en solution des alginamides a été étudié en termes de stabilité dans le temps, de solubilité, de comportement rhéologique, et de taux de gonflement, puis comparé à celui des alginates esters. Les alginamides ont montré des propriétés en solution différentes de celles des alginates esters, en particulier des contraintes critiques faibles, qui sont les conséquences de la formation d’agrégats liée à l’existence d’une réaction secondaire de réticulation chimique lors de la synthèse. Néanmoins, à des concentrations et taux de greffage appropriés, il est possible d’obtenir un réseau polymère tridimensionnel stabilisé par des associations hydrophobes et pouvant servir de pont de régénération pour l’application visée. L’étude in vivo conclut à l’absence de signe de régénération des voies nerveuses chez les animaux lésées sur une durée d’un an. Toutefois, cette étude a permis de valider la stratégie d’implantation d’un gel physique rhéofluidifiant et thixotrope, de définir des méthodes d’analyse des tissus post mortem en présence d’hydrogel, d’examiner la possibilité d’encapsuler un traitement pharmacologique et de définir un cahier des charges élargi de ce biomatériau (pH, stabilité, gonflement …) / The aim of this work was to synthesise alginate hydrogels, stable in time, and to evaluate their potential use as scaffolds for the damaged nerve regeneration in central nervous system. Various amphiphilic derivatives of sodium alginate were prepared by covalent attachment of alkyl chains (12 carbons) onto the polysaccharide at different substitution ratio, either via ester (alginate ester) or amide (alginamide) linkages, these last ones being more stable toward hydrolysis. The properties in solution of the alginamide derivatives were studied in terms of solubility, stability as function of time, rheological behaviour and swelling ratio. Results were compared to those obtained with the alginate ester family and highlighted a different behaviour for the alginamide series in semi-dilute regime. In particular, alginamide hydrogels exhibited a low critical strain which has been attributed to the presence of aggregates in the solution. The formation of these aggregates was due to the occurrence of a secondary cross-linking reaction during the synthesis of polymers. Nevertheless, it was possible, by appropriate tuning of the substitution yield and of the solution concentration, to obtain a three-dimensional network stabilized by intermolecular hydrophobic interactions, which has been evaluated as regenerative support for the considered application. In vivo studies demonstrated the absence of nerve regeneration for the tested injured animals after one year. However, these studies allowed us to evaluate both the strategy for the implantation of a physical gel exhibiting a shear-thinning and thixotrope behavior and the possibility to encapsulate a pharmacological treatment. An enlarged project specification has also been defined for this biomaterial (pH, stability, swelling ratio…)