Este estudo se propôs a analisar os efeitos das diferentes larguras temporais de pulso do laser de Er:YAG na rugosidade superficial, na perda de volume do material e na altura do degrau formado em uma Y-TZP (Yttrium-stabilized Tetragonal Zirconia Polycrystal) pré-sinterizada. Foram utilizados 5 blocos de Y-TZP pré-sinterizados destinados ao uso CAD/CAM que foram seccionados, obtendo-se 63 espécimes padronizados. Em seguida, esses espécimes foram lixados com uma sequência decrescente de lixas para padronização da superfície. Os grupos experimentais foram compostos a partir de 1 fator de variação - o tratamento de superfície. Os espécimes foram irradiados com o laser de Er:YAG (Fidelis III Fotona, Eslovênia) - 2,94 ?m, 100 mJ, 15,87J/cm2, 10 Hz, 1W, 7 mm, 60% água/40% ar. Assim formaram-se 7 grupos experimentais (n=9): G1 (50 ?s); G2 (100 ?s); G3 (300 ?s); G4 (600 ?s); G5 (1000 ?s); G6 (Abrasão triboquímica com partículas de 30 ?m); G7 (Sem tratamento). Após os tratamentos de superfície, os espécimes foram sinterizados em forno específico, de acordo com as recomendações do fabricante. Foram capturadas imagens em Microscópio Confocal a Laser 3D de cada espécime e avaliadas por meio de um software. A análise dos grupos irradiados revelou um aumento da rugosidade nos grupos G1, G2, G3 e G4 quando comparados aos grupos G6 e G7. O grupo G5 apresentou superfície totalmente plana e desfavorável para retenção. No que se refere à perda de volume e formação de degrau, os grupos G1, G2 e G3 demonstraram grande perda de volume e grande altura de degrau formado, o que pode levar a uma desadaptação da peça protética. No grupo G4 foram observados valores de rugosidade satisfatórios com pouca perda de volume e pequena altura de degrau formado, semelhante ao que foi notado para a abrasão trioboquímica (G6), podendo ser indicada para tratamento de superfície de Y-TZP pré-sinterizada. A irradiação com o laser de Er:YAG promoveu um padrão morfológico com muitas irregularidades, característico de ablação para os grupos G1, G2, G3 e G4 e, para todos os protocolos utilizados, não foi observada a presença de trincas ou carbonização. Sugere-se que a largura temporal de 600 ?s (G4) seja a mais indicada como alternativa para tratamento de superfície, objetivando a criação de micro-retenções superficiais, com pouca perda de volume de material e que, ao mesmo tempo, não provoque danos à estrutura da cerâmica policristalina. / This study evaluated the effects of different Er:YAG laser pulse width protocols on surface roughness, loss of volume of the material and the step height formed of pre-sintered Yttrium-stabilized tetragonal Zirconia Polycrystal (Y-TZP) by three-dimensional profilometric assessment. Blocks of pre-sintered Y-TZP were cut providing 63 standard 5mm thick samples which were divided by surface treatment, as follows (n=9): G1 (50?s); G2 (100?s); G3 (300?s); G4 (600?s); G5 (1000?s); G6 (tribochemical silica coating); G7 (Untreated). Laser settings: The Er:YAG laser (Fidelis III; Fotona, Ljubljana, Slovenia) 100mJ, 15,87J/cm2, 10Hz, 1W, (60%) and air (40%) cooling. After treated or not, samples were sintered according to the manufacturer\'s recommendations. Roughness, volume loss and step and were analyzed by 3D profilometric assessment with confocal laser microscopy. Irradiated groups showed an increased roughness in the groups G1, G2, G3 and G4 when compared to G6 and G7 groups. The G5 group showed a completely flat and unfavorable surface for retention. The groups G1, G2 and G3 shown great loss of volume and the step height formed, which can lead to a gap on the crowns. In G4 were observed satisfactory roughness with little loss of volume and the step height formed similar to G6. Irrespective of laser protocol, any of the specimens showed presence of cracks and carbonization. It is suggested that the pulse width 600?s (G4) is the most suitable pulse width protocol as an alternative surface treatment, promoting micro-retention, with little loss of volume of material, comparable to gold standard treatment.

Supramolecular structure of eight commercial types of iPP, its blends with poly(L-lactide) (PP-PLLA) and copolymers with ethylene-propylene rubber (PP-EPR) together with four types of polyethylene (LDPE, HDPE) was uncovered and observed in this work. The spherulitic structure of iPP and PE homopolymers and PP copolymers was uncovered by etching of six different solutions of mineral acids with KMnO4 and by dissolution in four selected solvents. The latter was found to be ineffective. The structure of PP-PLLA blends was uncovered only after recrystallization (the change of structure by annealing and cooling rate). The uncovered crystalline structure was observed directly by confocal laser scanning microscope (CLSM) and SEM. Crystallinity together with a size distribution of crystallites was determined by DSC. It was proved that spherulitic supramolecular structure was easy to uncover by chemical etching only for molded samples and for fracture surfaces of injected samples. The efficiency of etchants varied but, generally, the samples with high degree of crystallinity were etched earlier. The etchants containing nitric acid and high content of sulphuric acid uncovered fine details of shperulites with respect to mixtures containing orthophosporic acid. It was also observed that not only the etchant (especially with nitric acid) itself but also its vapors were effective. The structure was revealed later but the appearance was plastic (3D) and the structure was very fine. The supramolecular structure of PP-PLLA blends differed with amount of each component. The ethylene-rubber phase came forth with increasing time of etching while PP spherulites disappeared in PP-EPR samples. CLSM was shown to be very good tool for observing supramolecular structure of studied samples with respect to SEM, which was proved to be entirely unsuitable for polyolefines. DSC is recommended to be performe before uncovering supramolecular structure for basic information about proportion of amorphous/crystalline phase and size of crystallites.

Magister Chirurgiae Dentium (MChD) / Purpose: To evaluate and compare the in-vitro shear bond strength and micro leakage of two compomers with their adhesive systems and to examine the dentine-restorative interface under confocal scanning laser microscopy (CSLM). Matoiats and Methods: For shear bond strength (SBS) testing thirty non-carious human molars were used of which fifteen molars were restored with Dyract AP using Non-Rinse Conditioner (NRC) and Prime&Bond NT (PBNT) and fifteen were restored with F2000 and Scotchbond Multi-Purpose Plus (SBMP). For the microleakage evaluation cavity preparations were made on the facial surfaces of thirty non-carious premolars. These were then restored with the respective compomer system. The specimens were thermocycled, sectioned and examined for dye penetration. The dentine-restorative interface was examined through a confocal scanning laser microscope. The primers of the bonding agents were labelled with rhodamine B and the adhesive resins were labelled with fluorescein and examined under CSLM in fluorescent mode. Results: The mean SBS for PBNT and SBMP were 12.8 and 18.1 MPa, respectively. The microleakage scores showed Dyract with PBNT leaked on the dentine side in 13 of the 15 specimens examined. On the enamel side 2 of the 15 specimens showed microleakage. With F2000 and SBMP no micro leakage was observed on either enamel or dentine sides. The CSLM images show clear resin tag and hybrid layer formation for both the materials examined, although SBMP showed deeper penetration into the dentine with longer resin tags. The length of the resin tags and thickness of the hybrid layer for PBNT was found to be approximately 10 um and 2 um respectively. SBMP showed resin tags measuring about 100 um while the hybrid layer measured about 5 um. Conclusion: This study demonstrates that the acid-etch technique ofSBMP with F2000 produces higher bond strength and no micro leakage when compared to the self-etching/self-priming "non-rinse technique" of NRC with PBNT and Dyract.

The extracellular matrix (ECM) is a three-dimensional, acellular scaffold of living tissues. Incorporating the ECM into cell culture models is a goal of cell biology studies and requires biocompatible materials that can mimic the ECM. Among such materials are hydrogels: polymeric networks that derive most of their mass from water. With the tuning of their properties, these polymer networks can resemble living tissues. The microarchitectural properties of hydrogels, such as porosity, pore size, fiber length, and surface topology can determine cell plasticity. The adequate characterization of these parameters requires reliable and reproducible methods. However, most methods were historically standardized using other biological specimens, such as 2D cell cultures, biopsies, or even animal models. Therefore, their translation comes with technical limitations when applied to hydrogel-based cell culture systems. In our current work, we have reviewed the most common techniques employed in the characterization of hydrogel microarchitectures. Our review provides a concise description of the underlying principles of each method and summarizes the collective data obtained from cell-free and cell-loaded hydrogels. The advantages and limitations of each technique are discussed, and comparisons are made. The information presented in our current work will be of interest to researchers who employ hydrogels as platforms for cell culture, 3D bioprinting, and other fields within hydrogel-based research.

Plusieurs décennies de recherche ont permis de mieux comprendre les effets de l’athérosclérose sur le système cardiovasculaire, d’améliorer la prévention et de développer des traitements efficaces. Les effets de l’athéroslérose sur le cerveau demeurent toutefois mal compris même si le lien entre le fonctionnement cognitif et la santé du système vasculaire est maintenant bien établi. La venue de nouvelles méthodes d’imagerie telle la microscopie laser à 2-photons (TPLM) permet d’étudier l’impact de certaines maladies sur la microvasculature cérébrale en mesurant le flux sanguin dans des vaisseaux uniques situés dans des régions cérébrales millimétriques sous la surface. Les résultats des études in vitro peuvent dorénavant être corrélés à ceux obtenus in vivo. En premier lieu, ce mémoire revoit la théorie ayant permis le développement de la TPLM qui permet de prendre des mesures hémodynamiques in vivo dans des vaisseaux de très petits calibres tels des capillaires cérébraux de souris. Par la suite, son utilisation est décrite chez des souris anesthésiées afin de comparer les mesures d’hémodynamie cérébrale tels la vitesse des globules rouges, le flux de globules rouges, le flux sanguin cérébral, l’hématocrite sanguin et le diamètre des vaisseaux. Finalement, nous avons comparé les données hémodynamiques entre des souris de 3 mois normales (WT ; n=6) et des souris atteintes d’athérosclérose précoce (ATX ; n=6). Les résultats obtenus sur un nombre total de 209 capillaires (103 pour les souris WT et 106 pour les souris ATX) démontrent que les souris ATX possèdent une vitesse des globules rouges (+40%) plus grande, un flux de globule rouge plus grand (+12%) et un flux capillaire plus élevé (+14%) sans démontrer pour aucun de ces paramètres, une différence statistiquement significative. L’hématocrite moyen (35±4% vs 33±2% ; p=0.71) et le diamètre moyen des vaisseaux (4.88±0.22μm vs 4.86±0.20μm ; p=0.23) étaient également comparables. La vitesse des globules rouges a démontré une faible corrélation avec le diamètre des vaisseaux (r=0.39) et avec le flux de globules rouges/seconde (r=0.59). En conclusion, les travaux menés dans le cadre de ce mémoire de maîtrise permettent d'envisager, grâce aux nouvelles méthodes d’imagerie cérébrale telle la TPLM, une meilleure compréhension des mécanismes hémodynamiques sous-jacents à la microcirculation cérébrale. L’effet d’une pression pulsée augmentée, tel que proposée dans l’athérosclérose reste cependant à démontrer avec cette méthode d’imagerie. / Many decades of research have given us a better understanding of the effects of atherosclerosis on different organs. For example, works on the cardiovascular effects of atherosclerosis have improved the prevention, screening and treatment resulting in a better prognosis. The effects of atherosclerosis on cerebral vessels are misunderstood even if the link between brain function and cerebral perfusion is well known. With the improvements in brain imaging, there are more possibilities to better define the physiopathology of brain perfusion and the impact of disease on cerebral microvasculature. First, this work reviews the theory behind the development of two-photon laser microscopy (TPLM) and the measure of hemodynamics in small vessels such as cerebral capillaries in mice. We also describe how we measured cerebral hemodynamics in anesthetized mice: red blood cell speed, red blood cell flux, blood hematocrit, and vessel diameter. We compared results obtained between normal mice (WT ; n=6) and pathological mice (ATX ; n=6), all aged 3 months old. ATX mice are well recognized to develop early atherosclerosis and served as a model for high pulse pressure. We measured 209 cerebral capillaries (103 on WT mice and 106 on ATX mice). ATX mice tend to show a trend toward a higher red blood cell speed (+40%), higher red blood cell flux (+12%) and higher capillary flux (+14%) in ATX mice. Mean hematocrit (35±4% vs 33±2% ; p=0.71) and mean vessel diameter (4.88±0.22μm vs 4.86±0.20μm ; p=0.23) were not statistically different between both groups. Red blood cell speed showed a weak correlation with vessel diameter (r=0.39) and red blood cell flux (r=0.59). In conclusion, the TPLM should permit a better understanding of the effect of vascular disease in cerebral hemodynamics. However, the effect of high pulse pressure on cerebral microvasculature needs to be better defined.

Ce travail se situe à l’intersection des trois disciplines : méthodes numériques, techniques expérimentales et du machine learning, a pour but de proposer une famille de techniques d’identification par analyse inverse des lois de comportement en mécanique. Dans le domaine d’identification des matériaux, l’indentation instrumentée est particulièrement attractive, car elle permet de procéder à des essais non-destructifs sur l’échantillon ou sur une structure en service. L’essai d’indentation, similaire à un test de dureté, consiste à enfoncer la pointe de l’indenteur à une faible profondeur dans la matière tout en enregistrant le déplacement en fonction de la force appliquée. L’identification des propriétés élastoplastiques des matériaux est basée alors sur l’exploitation de la courbe force-déplacement (courbe P-h). Toutefois, le problème inverse est souvent mal posé et des problèmes d’unicité mènent à la notion de paires de "matériaux mystiques" produisant, dans des conditions d’essai donnés, des courbes P-h identiques, malgré des propriétés différentes. L’idée de notre travail est de compléter la procédure d’identification en faisant appel à des dispositifs expérimentaux récents, notamment à la microscopie laser, permettant de mesurer la carte 3D de l’empreinte résiduelle obtenue après le retrait de l’indenteur. Pour aborder la question de la richesse d’information de l’empreinte par rapport à la courbe P-h seule, nous proposons de construire, dans un espace affine réduit, la variété des formes d’empreinte admissibles au sens d’une loi de comportement et du modèle d’éléments finis de l’essai. La mesure de la dimension intrinsèque nous indique alors le nombre maximal de paramètres potentiellement identifiables. Cela nous permet de proposer et de valider numériquement des nouveaux procédés expérimentaux, plus représentatifs, à partir des données synthétiques, ainsi que des algorithmes d’identification associés. La prise en compte de l’erreur de modèle et de l’erreur de mesure, nous mène ensuite à proposer un ensemble d’algorithmes de projection d’empreintes expérimentales, réalisées en collaboration avec l’INSA de Rennes sur la variété synthétique. Nous abordons alors le problème d’identification des propriétés d’écrouissage de plusieurs matériaux de complexité croissante et départageons des "jumeaux mystiques" par des essais de multi-indentation, basés sur l’exploitation de l’empreinte seule ou en complément de la courbe P-h. / The thesis lies at the intersection of three disciplines : numerical methods, experimental techniques, and machine learning. The primary aim of this work is to develop a group of algorithms for characterization by inverse analysis of a material’s constitutive law. In the field of material characterization, indentation test is especially attractive since it is considered non-destructive, and may be performed even on a structure in service. The test, similar to a hardness test, consists in penetrating an indenter into the surface of the material. The force exerted on the indenter is recorded against the penetration depth over a series of time instants, leading to a force-displacement (P-h) curve, which is the most frequently used source of information for the identification of material properties. However, the inverse problem based solely on this curve tends to be ill-posed, leading to nonunique identification solution, i.e., the "mystical material pair", for whom the corresponding force-displacement curves are almost identical despite the very different material properties. The basic idea is then to complete the identification process with innovative experimental measurements, such as laser microscope, which allows measuring the 3D residual imprint after the withdrawal of the indenter. To address the advantage of this measurement over P-h curve, we propose to construct, within a reduced affine space, a manifold of shapes admissible to the postulated constitutive law, experimental and simulation setups, based on synthetic data. The intrinsic dimensionality of the manifold limits the number of identifiable parameters allowing to validate numerically experimental procedures. Considering both the model and measurement errors, we develop a series of local manifold learning algorithms to solve the inverse problem iteratively for experimental results obtained in cooperation with INSA de Rennes. This approach allows us to characterize diverse metallic materials of increasing complexity, based on actual experimental measurements. For example, for the Hollomon’s law, the mystical pair is alleviated in using a single imprint, while for the Voce law, a multi-depth experimental protocol is proposed to differentiate mystical siblings.

<p>Chromatography is a powerful technique, essential in chemical analyses and preparative separation in industry and research. Many different kinds of chromatographic material are needed, due to the large variety of applications. Detailed methods of characterisation are needed to design new chromatographic materials and understand their properties. In this thesis, confocal Raman spectroscopy and surface enhanced Raman spectroscopy (SERS) have been applied to micrometer-size chromatographic beads, for which these techniques have not been used earlier. New methodology, optimized for use with the chromatographic beads, has been developed and evaluated. </p><p>Confocal spectroscopy has been used to determine distributions of functional groups within single chromatographic beads. This distribution is of great importance in determining the chromatographic properties, since the material is porous and the solute molecules can diffuse inside the beads. Most of the confocal experiments have been performed with Raman spectroscopy; fluorescence spectroscopy, using Nd³⁺ ions or dye-labelled proteins as fluorescence probes, has been used for comparison. </p><p>The concentration of adsorbed analytes is very low within the beads. SERS was therefore used to enhance the Raman signal. SERS-active surfaces were prepared by incorporating gold nano-particles into the interior of the bead. TEM measurements showed that the gold nano-particles could be observed throughout, and it was possible to record analyte spectra from different positions within the bead. Enhanced spectra could be obtained both for small test molecules and for larger bio-molecules, although the spectra for the smaller analytes were much more intense.</p>

Chromatography is a powerful technique, essential in chemical analyses and preparative separation in industry and research. Many different kinds of chromatographic material are needed, due to the large variety of applications. Detailed methods of characterisation are needed to design new chromatographic materials and understand their properties. In this thesis, confocal Raman spectroscopy and surface enhanced Raman spectroscopy (SERS) have been applied to micrometer-size chromatographic beads, for which these techniques have not been used earlier. New methodology, optimized for use with the chromatographic beads, has been developed and evaluated. Confocal spectroscopy has been used to determine distributions of functional groups within single chromatographic beads. This distribution is of great importance in determining the chromatographic properties, since the material is porous and the solute molecules can diffuse inside the beads. Most of the confocal experiments have been performed with Raman spectroscopy; fluorescence spectroscopy, using Nd3+ ions or dye-labelled proteins as fluorescence probes, has been used for comparison. The concentration of adsorbed analytes is very low within the beads. SERS was therefore used to enhance the Raman signal. SERS-active surfaces were prepared by incorporating gold nano-particles into the interior of the bead. TEM measurements showed that the gold nano-particles could be observed throughout, and it was possible to record analyte spectra from different positions within the bead. Enhanced spectra could be obtained both for small test molecules and for larger bio-molecules, although the spectra for the smaller analytes were much more intense.

No description available.

The aim of submitted diploma thesis is the study of non-isothermal crystallization kinetics of polylactide (PLA) with selected agents (1 %) and observation of the emerging crystalline structure under polarizing optical microscope. The agents were talc, a mixture of organic salts with the addition of amorphous SiO2 (HPN 68L) and zinc stearate (HPN 20E) and LAK-301 (potassium salt of 5-dimethylsulfoisophtalate), which is a nucleating agent developer for PLA. The PLA matrix served as a reference. Non-isothermal crystallization took place on a differential scanning calorimeter at cooling rates () 0,3; 0,5; 0,7; 1; 1,5; 2 °C/min After non-isothermal crystallization, the crystalline fraction (Xc) od PLA was evaluated from X-ray diffraction analysis, and the supramolecular structure was observed after chemical degradative etching using confocal laser scanning microscope. The crystallization kinetics were evaluated by the methods of Jeziorny and Mo and the activation energy of the crystallization was determined according to the Friedmann method. All prepared materials were amorphous (Xc 40 % for up to 1,5 °C/min). However, for LAK-301, Xc decreased to 30 % already at the = 2 °C/min and it can be assumed that with increasing its nucleation activity will decrease. A spherulitic structure was observed in all samples, but the number and size of spherulites decreased with increasing and the appearance varied according to the type of agent. Both kinetic models proved to be unsuitable for materials with low Xc and the highest because the rate of crystallization did not change. With the Jeziorny method, it was possible to evaluate the kinetics only for the relative crystallinity Xt = 29–50 % and with the Mo method it was not possible to evaluate the data for the highest for PLA matrix and sample with HPN 68L. The samples with LAK-301 and HPN 68L showed the lowest activation energy.