In vitro, la formation de structures de type tubulaire avec des cellules endothéliales de veine ombilicale humaine (HUVEC) a été étudiée en combinant la fonctionnalisation de la chimie des matériaux et le développement de la géométrie tridimensionnelle. Le polycarbonate (PC) a été utilisé comme modèle pour le développement de l'échafaud. Le film de polysaccharide naturel, basé sur un dépôt alternatif couche par couche (LbL) d’acide hyaluronique (HA) et de chitosane (CHI), a d’abord été appliqué sur une surface PC et caractérisé en termes de croissance d’épaisseur microscopie à balayage lascar (CLSM). Cette première fonctionnalisation se traduit par un revêtement complet de la couche PC. Une biofonctionnalisation supplémentaire avec un peptide adhésif (RGD) et deux peptides angiogénétiques (SVV et QK) a été étudiée, immobilisant ces peptides sur le groupe carboxylique de HA précédemment déposé, en utilisant la chimie bien connue du carbodiimide. La version marquée de chaque peptide a été utilisée pour caractériser l’immobilisation et la pénétration des peptides dans les couches de polyélectrolytes, aboutissant à une greffe réussie avec une pénétration complète dans toute l’épaisseur du LbL. Des tests in vitro ont été effectués à l'aide de cellules HUVEC pour évaluer leur efficacité d'adhésion et leur activité métabolique sur la LbL avec et sans immobilisation de peptides, ce qui a permis d'améliorer l'activité préliminaire lorsque des combinaisons de peptides sont utilisées. Enfin, les micro-canaux PC (μCh) ont été développés et caractérisés pour la première fois, et les autres expériences ont été réalisées sur un micromètre de 25 μm de largeur, fonctionnalisé avec une architecture (HA / CHI) 12,5 (PC-LbL) avec des peptides RGD et QK -RGD + QK) ou avec des peptides RGD et SVV (PC-RGD + SVV). Notre première expérience de tubulogénèse a montré de manière surprenante la formation de structures de type tubulaire déjà après 2h d'incubation en utilisant la combinaison double-peptides, mais uniquement avec PC-RGD + QK. Les tubes étaient également présents après 3 et 4 heures de culture. L'expérience de co-culture avec des péricytes humains dérivés du placenta (hPC-PL) montre comment la stabilisation des tubes a été améliorée après 3 et 4 heures également pour l'échantillon de PC-RGD + SVV. Globalement, notre matériel bio-fonctionnel avec les peptides PC-RGD + QK et PC-RGD + SVV permet la formation d'une structure de type tubulaire à la fois dans une expérience de monoculture et de co-culture. / In vitro, tubular-like structures formation with human umbilical vein endothelial cells (HUVECs) was investigated by combining material chemistry functionalization and three-dimensional geometry development. Polycarbonate (PC) was used as a template for the development of the scaffold. Natural polysaccharide’s film based on alternate layer-by-layer (LbL) deposition of hyaluronic acid (HA) and chitosan (CHI), was first applied to PC surface and characterized in terms of thickness growth both, in dry conditions using ellipsometry, and confocal lascar scanning microscopy (CLSM). This first functionalization results in a complete coating of the PC layer. Further biofunctionalization with one adhesive peptide (RGD) and two angiogenetic peptides (SVV and QK) was investigated, immobilizing those peptides on the carboxylic group of HA previously deposited, using the well-known carbodiimide chemistry. The labeled version of each peptide was used to characterize the peptides’ immobilization and penetration into the polyelectrolytes layers, resulting in a successful grafting with complete penetration through the entire thickness of the LbL. In vitro tests were performed using HUVECs to assess their adhesion efficiency and their metabolic activity on the LbL with and without peptide immobilization, resulting in a preliminary improved activity when peptide-combinations is used. Finally, PC micro-channels (μCh) were first developed and characterized, and the rest of the experiments were performed on μCh of 25μm width, functionalized with (HA/CHI)12.5 architecture (PC-LbL) with RGD and QK peptides (PC-RGD+QK) or with RGD and SVV peptides (PC-RGD+SVV). Our first tubulogenesis experiment surprisingly showed the formation of tubular-like structures already after 2h of incubation using the double-peptides combination but only using PC-RGD+QK the tubes were present also after 3 and 4 hours of culture. The co-culture experiment with human pericytes derived from placenta (hPC-PL) demonstrates how the stabilization of the tubes was improved after 3 and 4 hours also for the PC-RGD+SVV sample. Globally our bio-functional material with PC-RGD+QK and PC-RGD+SVV peptides allow the formation of tubular-like structure in both mono and co-culture experiment.

Made available in DSpace on 2017-07-24T19:38:11Z (GMT). No. of bitstreams: 1 Jociani Ascari.pdf: 1347077 bytes, checksum: 5bf88ebdcff72a3eee1c4ed2152eb55c (MD5) Previous issue date: 2007-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Baccharis uncinella (Asteraceae), popularly known as vassoura, is little known from the chemical point of view, with only two earlier scientific works on their secondary metabolites - the volatile constituents. In the present study, the essential oil from the leaves of B. uncinella and nine extracts obtained in a Soxhlet apparatus with organic solvents starting from the leaves, barks and log were analyzed by CCD, UV and IV and submitted to activity tests against microorganisms; besides, the interactions between the aqueous extract of the leaves with human erytrocytary cells of the sanguine types O+, O-, A+ and B+ were also analyzed. In the antimicrobial tests, the microfungus Candida albicans, as well as the Gram-positive Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and S. saprophyticus and the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were used. The essential oil showed action against B. cereus, S. aureus and C. albicans, in the same way as all of the organic extracts, but the growth of S. saprophyticus was not inhibited by no one of the extracts. The extracts of the barks, when compared with those obtained from other parts of the plant, were active against the largest number of microorganisms and presented the more strong activities. The minimum inhibitory concentration of the essential oil was estimated as 100/mL against S. aureus as well as against C. albicans, wich could be considered a moderate antimicrobial activity. We have isolated small amounts of the three major phenolic components found in the leaves, and the analysis of their UV- and IR-spectra showed the structural features of hydroxylaryl-propenoic acids for these compounds that were found for the first time in this plant species. The analyses of GC-MS of the essential oil produced by hydrodistillation of the leaves collected at the Paraná State presented low monoterpene content (2.29%) and the following sesquiterpenes as the main components: spathulenol 17.20%, caryophyllene oxide 16.20%, transcaryophyllene 10.48%, eudesmol 7.55%, germacrene D 6.28% and cadinol 5.18%. Another oil sample was obtained using the same conditions, but starting with leaves collected in the highland area of Santa Catarina, also presented spathulenol as the main component (15.83%), but it contained 36% in monoterpenes, a very similar composition to those published for samples from the State of Rio Grande do Sul. The interactions among the aqueous extract of the leaves with human erytrocytes isolated from the sanguine types O+, O-, A+ and B+, being verified that the extract presents a non-lytic activity in the analyzed concentrations of 22, 110 and 220/mL. This non-hemolytic profile was observed as similar for all tested blood types. The serological types A+ and O- presented a similar pattern of free Hb oxidation in solution, and the same profile was observed in the new suspension of the precipitate. In the analysis of MEV we observed the qualitative aspects regarding the interaction of the aqueous extract with the architecture of the erythrocyte. In the assays of DPPH with low concentrations, 1.5 x 10-4 and 7.8 x 10-5 mg/mL, it was possible to quantify a partial reduction in the free radicals concentration of 20.10% and 9.25% respectively. / Baccharis uncinella (Asteraceae), popularmente conhecida como vassoura, é pouco conhecida do ponto de vista químico, havendo apenas dois trabalhos científicos anteriores sobre seus metabólitos secundários - os constituintes voláteis. No presente estudo, o óleo essencial das folhas de B.uncinella e nove extratos obtidos em Soxhlet com solventes orgânicos a partir das folhas, cascas e lenho foram analisados por CCD, UV e IV e submetidos a testes de atividade contra microrganismos; além disto, foram também analisadas as interações de extrato aquoso das folhas com células eritrocitárias humanas dos tipos sangüíneos O+, O-, A+ e B+. Nos testes antimicrobianos, foram usados o microfungo Candida albicans, as bactérias Gram-positivas Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus e S. saprophyticus e as bactérias Gram-negativas Escherichia coli, Klebsiella pneumoniae e Pseudomonas aeruginosa. O óleo essencial apresentou ação contra B. cereus, S. aureus e C. albicans, da mesma forma que todos os extratos orgânicos, mas o crescimento de S. saprophyticus não foi inibido por nenhum deles. Os extratos das cascas, quando comparados com os obtidos de outras partes da planta, foram ativos contra o maior número de microrganismos e apresentaram as mais fortes atividades. A concentração inibitória mínima do óleo essencial foi determinada como 50L/mL tanto contra S. aureus como contra C. albicans, podendo ser considerada uma atividade antimicrobiana moderada. Foram isoladas pequenas quantidades dos três compostos fenólicos majoritários, cujas análises indicam trata-se de derivados de ácidos hidroxiarilpropenóicos, encontrados pela primeira vez nesta espécie vegetal. As análises de CG-EM do óleo essencial produzido por hidrodestilação a partir de folhas coletadas no segundo planalto do Paraná, apresentaram baixo teor de monoterpenos (2,29%) e os seguintes sesquiterpenos como principais componentes: espatulenol 17,20%, cariofileno óxido 16,20%, trans-cariofileno 10,48%, eudesmol 7,55%, germacreno D 6,28% e cadinol 5,18%. Outra amostra de óleo obtida nas mesmas condições, mas a partir de folhas coletadas na região serrana de Santa Catarina, também apresentou espatulenol como principal componente (15,83%), mas quase 36% em monoterpenos, uma composição muito similar às publicadas sobre amostras da Serra Gaúcha. Foram analisadas as interações entre extrato aquoso das folhas com células eritrocitárias humanas dos tipos sangüíneos O+, O-, A+ e B+, sendo que o extrato apresenta uma atividade não-lítica nas concentrações analisadas de 22, 110 e 220 /mL, similar para todas tipagens analisadas. Os sorotipos A+ e O- testados apresentaram um padrão similar de oxidação da Hb livre em solução e, o mesmo perfil foi observado na ressuspensão dos precipitados. Na análise de MEV foram verificados aspectos qualitativos a respeito da interação do extrato aquoso com a arquitetura da célula eritrocitária. Baixas concentrações do extrato, 1,5 x 10-4 e 7,8 x 10-5 mg/mL, apresentaram atividade redutora em presença de DPPH, de 20,10% e 9,25% respectivamente.

Surgical repair of torn rotator cuff tendons have a high rate of failure and does not address the underlying pathophysiology. Tissue engineering strategies, employing the use of multipotent progenitor cells or growth factors, represent potential therapies to improve the outcome of rotator cuff surgery. The use of glycosaminoglycan-based biomaterials in these therapies may enhance the effectiveness of cell and growth factor delivery techniques. Furthermore, understanding the cellular and molecular mediators in tendon overuse can help elucidate the causes of tendon degeneration. Thus the overall goals of this dissertation were to 1) develop heparin-based biomaterials to enhance cell pre-culture and maintain growth factor bioactivity and 2) characterize the histological and enzymatic changes in a supraspinatus tendon overuse model. To investigate the use of heparin in enhancing dynamic signaling, mesenchymal stem cells (MSCs) were encapsulated in heparin-containing hydrogels and evaluated for differentiation markers when cocultured with a small population of differentiated cells. To probe the effect of sulfation of heparin on the interactions with protein, selectively desulfated heparin species were synthesized and evaluated for their ability to bind and protect proteins. Finally, to develop a tendon overuse model that can become a test bed for testing future targeted therapeutics, an animal model was evaluated for tissue damage and protease activity. Together these studies represent a multi-pronged approach to understanding how tendon tissues become degenerative and for developing technologies to improve the biological fixation of tendon to bone in order to reduce the need for revision surgeries.

L’arille de Momordica cochinchinensis (gac), un fruit de la famille des Cucurbitacées, est la source végétale la plus riche en lycopène et β-carotène. Ces deux composés ont, respectivement, un rôle de puissant antioxydant et de provitamine A, intéressant les compléments santé. Tout d'abord, un procédé d’extraction fractionnée douce a été développé pour extraire ces caroténoïdes naturels en gardant leur qualité originale. Puis, le lycopène et le β-carotène extraits ont été caractérisés et analysés. Au moins 95 % des extraits sont composés de l’isomère all-trans. Ils ne sont pas dégradés pendant le traitement thermique représentant les procédés de formulation. Leur stéréo-mutation thermique a été évaluée. Le lycopène est plus antioxydant et donc plus rapidement isomérisé que le β-carotène à haute température. L’isomérisation augmente leur activité antioxydante, qui a été évalué par test chimique TEAC et sur l’hémolyse des cellules sanguines (KRL) in vitro. Les deux caroténoïdes de l’arille de gac sont beaucoup plus antioxydants que le Trolox contre l’hémolyse. En comparant avec d’autres sources de β-carotène, les caroténoïdes extraits de gac dans ces conditions douces restent antioxydants même à des concentrations plus élevées contrairement à ceux extraits dans des conditions classiques qui deviennent prooxydants. Ces résultats permettent de discuter la bioactivité des caroténoïdes d'après leur qualité et de leur origine, c’est à dire leur source et leur procédé d’extraction. D’un point de vue applicatif, outre le procédé de fractionnement qui est industrialisable, le traitement thermique appliqué permet de contrôler la fonctionnalité des produits riches en caroténoïdes. / The aril of Momordica cochinchinensis (gac), plant from the Cucurbitaceae family, is the richest source of lycopene and β-carotene, which are a strong antioxidant and a pro-vitamin A, respectively, interesting for health-complements. First, a process of soft extraction-fractionation was developed for extracting effectively the natural carotenoids from gac without loss of their original quality. Then, the lycopene and β-carotene extracted from gac were analyzed and characterized. At least 95% of the extracts were composed of the all-trans isomer. They were not degraded during the heat-treatment mimicking formulation processing. Their thermal stereo-mutation was evaluated. Lycopene is more antioxidant, it is thus isomerized more rapidly than β-carotene at high temperature. The isomerization of carotenoids increases their antioxidant activity that was evaluated by the chemical test TEAC and through the hemolysis of red blood cells (KRL) in vitro. The lycopene and β-carotene from gac are notably more antioxidant than Trolox. By comparing with other sources of β-carotene, carotenoids extracted from gac in these soft conditions keep their antioxidant properties, even at high concentration, contrasting with extracts obtained in classical conditions that become prooxidant. From these results, the bioactivity of carotenoids is discussed from their quality and their origin that is their source and extraction process. For application, in addition to the fractionation process which is easily transferable to the industry scale, the heat-treatment used in this study is interesting for controlling products rich in functional carotenoids

Le Poly Vinyl Alcohol (PVA) a été associé aux verres élaborés dans un système quaternaire (BG) 46S6 par les procédés cités (fusion, sol-gel et sacffolds). Différents paramètres intervenant dans les synthèses des verres bioactifs ont été étudiés, nous citons à titre d’exemple : la température, le pH, la taille des particules, le rapport Polymère / verres, la microstructure, la porosité et la biodégradation. Les caractéristiques thermiques des verres élaborés ont été également déterminées après chaque synthèse par analyse thermique différentielle (DSC/TG, DTA/TG). Ainsi, la température de fusion, la température de transition vitreuse et la température de cristallisation ont été élucidées. Ces caractéristiques thermiques changent lorsque la composition chimique du verre est modifiée. A ce titre, les compositions chimiques ont été étudiées par Fluorescence (XRF) et Inductively Coupled Plasma-Opticale Emission Spectroscopy (ICP-OES) après chaque synthèse pour s’assurer de la pureté des verres bioactifs élaborés et destinés à des applications médicales. Plusieurs techniques physico chimiques d’analyses (DRX, MEB, MET, FT-IR, XRF, ICPOES) ont été mises en oeuvre pour déterminer les propriétés physico chimiques de nos verres bioactifs avant et après expérimentations « in vitro ». Le nano composite Polymère-Verres scaffolds que nous avons obtenu présente des particules de tailles comprises entre 40 et 61 nm et une porosité d’environ 85%. La biodégradation des verres scaffolds décroît lorsque la teneur en verre scaffolds dans le nano composite croît. Les expérimentations « in vitro » montrent qu’après immersion de ces nano composites dans un liquide physiologique synthétique (SBF), une couche d’apatite (phosphate de calcium) se forme à leur surface. L’épaisseur de la couche formée dépend clairement de la taille des particules et du rapport polymère / verre scaffolds. / The aim of the present work is the preparation of Bioactive Glass (BG) 46S6 by different techniques. Fabrication of composite scaffolds by using of Poly Vinyl Alcohol (PVA) and quaternary BG (two methods melting and sol-gel) with different ratios to the prepared scaffolds was carried out. Different factor affecting the final properties of the prepared composite scaffolds were investigated in this study, such as; temperature of treatment, BG particle size, polymer/glass ratio, microstructure, porosity, biodegradation, bioactivity, and drug release. The thermal behavior of the prepared bioactive glass by sol-gel and melting techniques were identified using Differential Scanning Calorimetric/Thermo Gravimetric (DSC/TG) or Differential Thermal Analysis/Thermo Gravimetric (DTA /TG). The elemental composition of the prepared bioactive glasses was determined by X-rays Fluorescence (XRF) to confirm that the prepared bioactive glasses have the same elemental compositions and high purity for biomedical applications. The particle size of the prepared bioactive glass was determined by Transmission Electron Microscopic (TEM). Nano-bioactive glass could be obtained by modified sol-gel and the obtained particle size ranged between 40 to 61 nm. The prepared bioactive glass by both applied methods has the same amorphous phase and all identified groups as well as. The porous scaffold has 85% porosity with a slight decrease by increasing the glass contents. The degradation rate decreased by increasing of glass content in the prepared scaffolds. The bioactivity of the prepared composite scaffolds was evaluated by XRD, FTIR, SEM coupled with EDX and Inductively Coupled Plasma-Optical Emission Spectroscopic (ICP-OES). It has been observed that after soaking in Simulated Body Fluid (SBF), there was an apatite layer formed on the surface of the prepared samples with different thickness depending on the glass particle size and polymer/glass ratio.

<p>The production of insects for protein is projected to reach a market share of 1.33 billion USD, a rapid increase from the estimated 144 million USD share of 2019 market. The isolation of insect protein produces by-products, including chitin. Currently chitin is extracted from aquaculture by-products, such as shrimp and crab shells, and used to produce chitosan for various applications in the supplement and food industry. With the insect market expected to continue its growth, the feasibility of sourcing commercial chitin and chitosan from reared crickets’, and the application properties of its counterpart, chitosan, was investigated in this dissertation. In the first part of this dissertation, chitin from two commonly reared crickets in the Unites States, <i>Acheta domesticus</i> and <i>Gryllodes sigillatus</i>, was successfully extracted, purified, and identified as a commercially viable option for chitin and chitosan. Extensive crustacean chitin studies served as the foundation of purification steps, however durations were adjusted to account for intrinsic differences between insects and crustacean exoskeletons. Furthermore, cricket chitosan was prepared and optimized with varying degrees of deacetylation. As expected, cricket chitosan had lower molecular but did not have a detectable effect on the bioactive properties tested. All cricket chitosan produced had similar lipid binding capacity <i>in vitro</i>. Additionally, the microbial inhibition of cricket chitosan and commercial chitosan (~70% DDA) were not significantly different when evaluated against <i>L. innocua</i> and <i>E. coli</i>. High DDA cricket chitosan showed greatest bacterial inhibition as expected. In the second part of this dissertation, cricket derived chitosan showed similar and improved food packaging properties, when evaluated against commercial shrimp chitosan. microstructure analysis provided by scanning electron microscopy showed greater compaction and agglomeration of cricket chitosan films. The change in microstructure may be attributed to the increased complexity generally attributed to insect chitosan materials, a result of remaining melanin and protein in close association with insect exoskeleton chitosan. As a result, cricket films had similar or increased tensile strengths but decreased elongation percentages when compared to shrimp films. Water vapor permeability of cricket films was decreased due to tortuosity. Residual melanin likely played an important role in increasing cricket film surface hydrophobicity and providing enhanced light barrier properties. Overall, this dissertation successfully shows the potential of crickets as insect derived chitin and chitosan, and its effectiveness as a lipid binding and antibacterial agent, as well as its potential use in biobased food packaging. </p>

This doctoral thesis was conducted in the framework of a co-supervised PhD between the Department of Agriculture, Forestry and Food Sciences of the University of Turin and the Unit of Pharmacognosy, Bioanalysis and Drug Discovery of the Faculty of Pharmacy at the Université libre de Bruxelles. The present manuscript was conceived as a thesis of articles and is composed of 9 different scientific publications. The general introduction of the work was issued from a published narrative review, while the result and discussion part is composed by eight chapters based on different scientific articles issued from the PhD project. The cocoa bean shell (CBS) is the external tegument that covers the cocoa bean, and is one of the major by-products in cocoa industry. It is normally discarded or underutilized, which could result in economical and environmental issues. However, CBS represents a notable source of polyphenols and methylxanthines (theobromine and caffeine) which can give it different biofunctionalities such as antioxidant and antidiabetic properties, among others. It also contains high amounts of dietary fiber (about 50% w/w), minerals, vitamins, and proteins. CBS has low-fat content, and interesting cocoa-aroma compounds. All this could make CBS useful as a food ingredient, and source of biofunctional compounds. The first part of the experimental work of this thesis is devoted to the chemical characterization of CBS and the establishment of its polyphenolic and volatile organic compound (aroma) profiles. The utilization of such profiles, determined by both complete characterization methods and screening methods, was also proposed for authentication purposes of CBS depending on its geographical origin and variety. In a second step, the utilization of CBS as a low-cost food ingredient for functional food production was envisaged. CBS-based beverages and biscuits were proposed as model foods. The influence of the CBS addition to the model foods was evaluated from both technological and nutritional points of view. Changes on the physicochemical characteristics of the model foods were assessed as well as their content in compounds of interest and potential biofunctionalities. Moreover, these studies served also to evaluate the effect of the different food matrices on the bioaccessibility and intestinal permeability of the bioactive compounds contained in CBS. In the third and last part of this work, a different utility was given to the study of the cocoa by-product. The antimicrobial potential of CBS was assessed against different bacterial and fungal strains and a metabolomic strategy was applied in order to individualize the putative active compounds against the Streptococcus mutans proliferation. This work was a contribution for the valorization of a high add-value product such as the CBS, and a step towards a zero-waste cocoa industry within the frame of sustainable circular economy. / Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie) / info:eu-repo/semantics/nonPublished

Les bioverres du système quaternaire SiO2-CaO-Na2O-P2O5 obtenus par fusion doivent être coulés à 1400°C, ce qui ne permet pas la mise en forme de matériaux complexes comme par exemple des composites macroporeux en biocéramiques (HA et TCP) recouverts d’une couche uniforme de bioverre. Pour contourner cette limitation, la voie sol-gel a été utilisée dans cette étude. Le but principal a été de synthétiser par le procédé sol-gel, deux compositions quaternaires du système SiO2-CaO-Na2O-P2O5 habituellement obtenues par fusion. Il s’agit des compositions 45S5® de L.L. Hench et 47Q de C. Duée. Ces verres sont inversés, c’est à dire qu’ils contiennent plus d’éléments modificateurs (calcium et sodium) que d’éléments formateurs (silicium et phosphore). Pour la synthèse sol-gel, du TEOS (TétraEthylOrthoSilicate) et du TEP (TriEthylPhosphate) ont été utilisés pour introduire les formateurs. En utilisant des précurseurs minéraux pour le calcium et le sodium, il existe des difficultés à maintenir le gel amorphe lors de son séchage. En effet, les précurseurs minéraux sont le siège de mécanismes de dissolution-précipitation liés entre autres aux solvants choisis, à la nature et à la concentration des anions dans le milieu. L’étude s’est donc orientée vers l’utilisation d’autres précurseurs des modificateurs afin de limiter la part des anions qui contribuent au phénomène de précipitation non souhaité. Deux procédés de synthèse originaux ont été alors mis au point avec des précurseurs alcoolates, acétates et du glycérol. Avec ces procédés, la nature amorphe des composés a été confirmée et leurs caractéristiques thermiques ont été ensuite étudiées. Tous les verres sol-gel réalisés sont bioactifs. Les compositions de base, 45S5® et 47Q, ont été ensuite dopées avec de l’argent afin de doter les bioverres d’une action antibactérienne. Des cellules L132 ont été utilisées pour évaluer la toxicité des poudres des deux procédés solgel. Le procédé le moins toxique a été conservé pour la suite de l’étude. Les bioverres dopés et non dopés ont été alors déposés à la surface d’échantillons plats et macroporeux en HA et TCP par une technique de trempage-retrait. Des tests de prolifération et de formation de biofilms par Pseudomonas aeruginosa ont été réalisés sur des pastilles de TCP recouvertes et ont mis en évidence un effet toxique des dépôts contenant de l’argent. Des mesures de prolifération et de vitalité sur des cellules humaines MG63 ont été également réalisées et ont permis d’observer également un effet toxique. Cet effet n’est pas souhaitable dans ce cas car il affecte la biocompatibilité de l’implant. Il devrait cependant être confirmé par d’autres tests réalisés avec d’autres lignées cellulaires. / Bioglasses of quaternary system SiO2-Na2O-CaO-P2O5 obtained by melting are cast at 1400 ° C, which does not allow the shape of complex materials, for example macroporous bioceramics composites (HA and TCP) coated with an uniform layer of bioglass. To overcome this limitation, the sol-gel process has been used in this study. The aim has been to synthesize by sol-gel process, two quaternary compositions usually obtained by melting in the SiO2- CaO-Na2O-P2O5 system. These are two compositions: 45S5® of L.L. Hench and 47Q made by C. Duée. These glasses are reversed, ie they contain more modifiers elements (calcium and sodium) as formers elements (silicon and phosphorus). For the sol-gel synthesis, TEOS (TetraEthylOrthoSilicate) and TEP (TriEthylPhosphate) have been used to introduce the formers. When minerals precursors are used for calcium and sodium, there are difficulties to maintain amorphous the gel during its drying. Indeed, minerals precursors have some dissolution-precipitation mechanisms linked to the selected solvents and the nature and concentration of anions in the medium. The use of other modifiers precursors has been made in the study to limit the proportion of anions contributing to the undesired precipitation phenomenon. Two original synthesis methods have been developed with alkoxide precursors, acetates and glycerol. With these processes, the amorphous nature of the compounds has been confirmed and their thermal characteristics have been studied. All sol-gel glasses are bioactives. The compositions, 45S5® and 47Q, have been doped with silver to add an antibacterial action to the bioglasses. L132 cells have been used to test the toxicity on the powders of the two sol-gel processes. The least toxic process is has been retained for the other tests. Doped and undoped bioglasses have been coated on the surface of flat and macroporous samples of HA and TCP by a dip-coating technique. Proliferation tests and biofilms formation by Pseudomonas aeruginosa have been made on coated TCP and show toxic effects of silver. Proliferation and vitality tests have been also made on MG63 human cells and have allowed to observe a toxic effect. This effect is not suitable in this case because it affects the biocompatibility of the implant. It would, however, be confirmed by other tests with other cell lines.

Les relations entre la structure des composés chimiques et leurs propriétés sont complexes et à haute dimension. Dans le processus de développement de médicaments, plusieurs proprié- tés d’un composé doivent souvent être optimisées simultanément, ce qui complique encore la tâche. Ce travail explore deux représentations des composés chimiques pour les tâches de prédiction des propriétés. L’objectif de ces représentations proposées est d’améliorer l’explicabilité afin de faciliter le processus d’optimisation des propriétés des composés. Pre- mièrement, nous décomposons l’algorithme ECFP (Extended connectivity Fingerprint) et le rendons plus simple pour la compréhension humaine. Nous remplaçons une fonction de hachage sujet aux collisions par une relation univoque de sous structure à bit. Nous consta- tons que ce changement ne se traduit pas par une meilleure performance prédictive d’un perceptron multicouche par rapport à l’ECFP. Toutefois, si la capacité du prédicteur est ra- menée à celle d’un prédicteur linéaire, ses performances sont meilleures que celles de l’ECFP. Deuxièmement, nous appliquons l’apprentissage automatique à l’analyse des paires molécu- laires appariées (MMPA), un paradigme de conception du développement de médicaments. La MMPA compare des paires de composés très similaires, dont la structure diffère par une modification sur un site. Nous formons des modèles de prédiction sur des paires de com- posés afin de prédire les différences d’activité. Nous utilisons des contraintes de similarité par paires comme MMPA, mais nous utilisons également des paires échantillonnées de façon aléatoire pour entraîner les modèles. Nous constatons que les modèles sont plus performants sur des paires choisies au hasard que sur des paires avec des contraintes de similarité strictes. Cependant, les meilleurs modèles par paires ne sont pas capables de battre les performances de prédiction du modèle simple de base. Ces deux études, RCFP et comparaisons par paires, visent à aborder la prédiction des propriétés d’une manière plus compréhensible. En utili- sant l’intuition et l’expérience des chimistes médicinaux dans le cadre de la modélisation prédictive, nous espérons encourager l’explicabilité en tant que composante nécessaire des modèles cheminformatiques prédictifs. / The relationships between the structure of chemical compounds and their properties are complex and high dimensional. In the drug development process, multiple properties of a compound often need to be optimized simultaneously, further complicating the task. This work explores two representations of chemical compounds for property prediction tasks. The goal of these suggested representations is improved explainability to better understand the compound property optimization process. First, we decompose the Extended Connectivity Fingerprint (ECFP) algorithm and make it more straightforward for human understanding. We replace a collision-prone hash function with a one-to-one substructure-to-bit relationship. We find that this change which does not translate to higher predictive performance of a multi- layer perceptron compared to ECFP. However, if the capacity of the predictor is lowered to that of a linear predictor, it does perform better than ECFP. Second, we apply machine learning to Matched Molecular Pair Analysis (MMPA), a drug development design paradigm. MMPA compares pairs of highly similar compounds, differing in structure by modification at one site. We train prediction models on pairs of compounds to predict differences in activity. We use pairwise similarity constraints like MMPA, but also use randomly sampled pairs to train the models. We find that models perform better on randomly chosen pairs than on pairs with strict similarity constraints. However, the best pairwise models are not able to beat the prediction performance of the simpler baseline single model. Both of these investigations, RCFP and pairwise comparisons, aim to approach property prediction in a more explainable way. By using intuition and experience of medicinal chemists within predictive modelling, we hope to encourage explainability as a necessary component of predictive cheminformatic models.

The aim of this work is to define relations between grain size and bioaktivity of oxide ceramics, specifically ZrO2, Al2O3 and HA. Ceramic materials with grain size from 100 nm up to 10 m, with various surface roughness, were tested for its bioactivity. Ceramography analysis was performed for all tested materials to precisely describe microstructures. Biological properties of the ceramic materials were tested via dilation tests directly in-vitro and by in-vitro extraction. Three cell culturing lines: osteoblast MG63, fibroblast L929, and epithelioid HeLa, were used for our testing. An influence of the grain size on the biological response was only found for the ceramic materials which had been thermally etched. The thermally etched nanocrystalline samples had larger areas covered by cells than ceramics with coarse grain microstructure. Biological tests on layered composites Al2O3×ZrO2 showed the cell selection determined by the type of material, where ZrO2 surfaces were preferably covered. Improved biological response of nanocrystalline ZrO2 was demonstrated on ceramic ZrO2, Al2O3 and SiO2 substrates with nanocrystalline coating of ZrO2. In this work a novel technological process for the formation of defect-free coatings was developed. Sintered coatings were tested using in-vitro technique with cell line HeLa, L929 and MG63 for up to 72 hours. The results of the biological tests of nanocrystalline coatings were consistent with results from the bulk nanocrystalline thermally etched ZrO2 ceramics.