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Influence de l’agrégation érythrocytaire sur la migration axiale de microparticules simulant des plaquettes sanguinesGuilbert, Cyrille 06 1900 (has links)
Lors du phénomène d’hémostase primaire ou de thrombose vasculaire, les plaquettes sanguines doivent adhérer aux parois afin de remplir leur fonction réparatrice ou pathologique. Pour ce faire, certains facteurs rhéologiques et hémodynamiques tels que l’hématocrite, le taux de cisaillement local et les contraintes de cisaillement pariétal, entrent en jeu afin d’exclure les plaquettes sanguines de l’écoulement principal et de les transporter vers le site endommagé ou enflammé.
Cette exclusion pourrait aussi être influencée par l’agrégation de globules rouges qui est un phénomène naturel présent dans tout le système cardiovasculaire selon les conditions d’écoulement. La dérive de ces agrégats de globules rouges vers le centre des vaisseaux provoque la formation de réseaux d’agrégats dont la taille et la complexité varient en fonction de l’hématocrite et des conditions de cisaillement présentes. Il en résulte un écoulement bi-phasique avec un écoulement central composé d’agrégats de globules rouges avoisinés par une région moins dense en particules où l’on peut trouver des globules rouges singuliers, des petits rouleaux de globules rouges et une importante concentration en plaquettes et globules blancs. De ce fait, il est raisonnable de penser que plus la taille des agrégats qui occupent le centre du vaisseau augmente, plus il y aura de plaquettes expulsées vers les parois vasculaires. L'objectif du projet est de quantifier, in vitro, la migration des plaquettes sanguines en fonction du niveau d’agrégation érythrocytaire présent, en faisant varier l’hématocrite, le taux de cisaillement et en promouvant l’agrégation par l’ajout d’agents tels que le dextran à poids moléculaire élevé. Cependant, le comportement non Newtonien du sang dans un écoulement tubulaire peut être vu comme un facteur confondant à cause de son impact sur l’organisation spatiale des agrégats de globules rouges. De ce fait, les études ont été réalisées dans un appareil permettant de moduler, de façon homogène, la taille et la structure de ces agrégats et de quantifier ainsi leur effet sur la migration axiale des plaquettes. Du sang de porc anti coagulé a été ajusté à différents taux d’hématocrite et insérer dans un appareil à écoulement de Couette, à température ambiante. Les plaquettes sanguines, difficilement isolables in vitro sans en activer certains ligands membranaires, ont été remplacées par des fantômes en polystyrène ayant un revêtement de biotine. La quantification de la migration de ces fantômes de plaquettes a été réalisée grâce à l’utilisation de membranes biologiques fixées sur les parois internes de l’entrefer du rhéomètre de Couette. Ces membranes ont un revêtement de streptavidine assurant une très forte affinité d’adhésion avec les microparticules biotynilées.
À 40% d’hématocrite, à un cisaillement de 2 s-1, 566 ± 53 microparticules ont été comptées pour un protocole préétabli avec du sang non agrégeant, comparativement à 1077 ± 229 pour du sang normal et 1568 ± 131 pour du sang hyper agrégeant. Les résultats obtenus suggèrent une nette participation de l’agrégation érythrocytaire sur le transport des fantômes de plaquettes puisque l’adhésion de ces derniers à la paroi du rhéomètre de Couette augmente de façon quasi exponentielle selon le niveau d’agrégation présent. / During the primary hemostatis or thrombosis phenomenon, the human blood platelets must adhere to the vascular wall in order for them to perform their repairing or pathological function. To do so, certain rheological and hemodynamic factors such as the hematocrit, local shear rate and the wall shear stress, must come into play to exclude blood platelets from the main blood stream and transport them to the vicinity of the damaged or inflamed site.
This exclusion could also be influenced by red blood cell aggregation which is a natural process present throughout the entire cardiovascular system under certain flow conditions. The displacement of these rouleaux of red blood cells towards the centre of the vessel induces the formation of 3D networks of aggregates whose size and complexity vary as a function of the hematocrit and the shearing conditions present. It results in a two phase flow with an inner core composed of red blood cell aggregates surrounded by single red blood cells or small aggregates and large numbers of white blood cells and platelets. It is therefore reasonable to believe that the larger the inner core becomes, the more platelets will be expulsed towards the vascular wall. The objective of the study was to quantify, in vitro, the lateral migration of blood platelets as a function of the level of red blood cell aggregation present, by changing the hematocrit, the shear rate and by promoting red blood cell aggregation with the use of agents such as high molecular weight dextran. However, the non Newtonian behavior of blood in tube flow can be seen as a confounding factor to the understanding of the spatial organization of the red blood cell aggregates. In this study, whole blood was circulated in a simple shear flow apparatus, which allowed to homogeneously modulate the red blood cell aggregate sizes and structure, and quantify their effect on the axial migration of blood platelets. Anticoagulated porcine bloods were adjusted to different hematocrits and inserted into a Couette flow apparatus, at room temperature. Blood platelets, difficult to isolate in vitro without activating in a non reproducible manner specific membrane ligands, were replaced with biotin coated fluorescent polystyrene beads. The quantification of the migration of these platelet ghosts was conducted with the use of biological membranes fixed on the interior walls of the Couette apparatus. These streptavidin coated membranes ensure a strong adhesive affinity with the biotynilated beads.
At 40% hematocrit and at a shear of 2 s-1, 566 ± 53 micro particles were counted for non aggregated erythrocytes, 1077 ± 229 for aggregating red blood cells and 1568 ± 131 for hyper aggregating blood. The results obtained suggest a strong participation of the red blood cell aggregation on the transport of platelet ghosts since the number of ghost cells fixed on the wall of the Couette rheometer increases almost exponentially with the level of aggregation present.
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Rhéologie des polymères fondus à hauts taux de cisaillement : application à la microinjection / Polymer melts rheology at high shear rate : microinjection molding applicationMnekbi Djebali, Cheima 07 December 2012 (has links)
La rhéologie à hauts taux de cisaillement pour deux polymères, le PEHD semi-cristallin et le PMMA amorphe a été étudiée. Des outils de rhéométrie classique, un rhéomètre plan-plan en mode dynamique, et un rhéomètre capillaire, ont été utilisés dans des conditions extrêmes (avec des filières pour la rhéométrie capillaire de diamètres allant jusqu'à 0,3 mm) mais les dépouillements de ces résultats ont été fait suivant les hypothèses conventionnelles en négligeant les instabilités et les phénomènes physiques qui interviennent lors de ces écoulements.Nous avons par la suite développé un modèle mathématique de l'écoulement dans un capillaire pour rendre compte de l'importance des différents phénomènes physiques qui peuvent avoir lieu dans des écoulements extrêmes, à savoir l'échauffement et la piezodépendance de la viscosité, la compressibilité et le glissement à la paroi. Les résultats du modèle développé ont été comparés avec les résultats expérimentaux.Nous avons aidé au développement d'une presse de microinjection originale et nous l'avons testée avec un moule de plaque instrumenté d'épaisseur allant jusqu'à 0,2 mm. Nous avons montré qu'il était possible de réaliser des pièces de qualité ce qui est avéré par des mesures de pression, vitesse et de température bien reproductibles. Nous avons exploité les données rhéologiques expérimentales dans la modélisation de la phase de remplissage avec le logiciel de calcul Rem3D. Des corrélations entre les mesures expérimentales et les calculs ont été réalisées en comparant l'évolution des pressions dans le système d'alimentation et dans l'empreinte. / Rheology at high shear rate for both polymers, semi-crystalline HDPE and amorphous PMMA was studied. Classical rheometry tools, plane-plane dynamic mode rheometer and capillary rheometer, were used in extreme conditions (with channels diameters for capillary rheometry up to 0.3 mm). However, analyses of these results were made following conventional assumptions neglecting instabilities and physical phenomena involved in these flows.We then developed a mathematical model of a capillary flow in to reflect the importance of different physical phenomena that can occur in extreme flows, namely heating, pressure dependency of viscosity, compressibility and the wall slip. The results of the developed model were compared with experimental results.We helped develop a press microinjection original and we tested it with instrumented plate mold with thickness up to 0.2 mm. We have shown that it is possible to make quality parts which are proven by well reproducible pressure, speed and temperature measurements. We used the experimental rheological data in filling phase modeling with the calculation software Rem3D. Correlations between experimental measurements and calculations were carried out by comparing the pressure in the filling system and the cavity.
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Suitability of cellulose ester derivatives in hot melt extrusion : thermal, rheological and thermodynamic approaches used in the characterization of cellulose ester derivatives for their suitability in pharmaceutical hot melt extrusionKarandikar, Hrushikesh M. January 2015 (has links)
Applications of Hot Melt Extrusion (HME) in pharmaceuticals have become increasingly popular over the years but nonetheless a few obstacles still remain before wide scale implementation. In many instances these improvements are related to both processing and product performance. It is observed that HME process optimisation is majorly focused on the active pharmaceutical ingredient's (API) properties. Characterising polymeric properties for their suitability in HME should be equally studied since the impact of excipients on both product and process performance is just as vital. In this work, two well-established cellulose ester derivatives: Hydroxy Propyl Methyl Cellulose Acetate Succinate (HPMCAS) and Hydroxy Propyl Methyl Cellulose Phthalate (HPMCP) are studied for their HME suitability. Their thermal, thermodynamic, rheological, thermo-chemical and degradation kinetic properties were evaluated with model plasticisers and APIs. It was found the thermal properties of HPMCP are severely compromised whereas HPMCAS is more stable in the processing zone of 150 to 200 °C. Thermodynamic properties revealed that both polymers share an important solubility parameter range (20-30 MPa P1/2P) where the majority of plasticisers and BCS class II APIs lie. Thus, greater miscibility/solubility can be expected. Further, the processability of these two polymers investigated by rheometric measurements showed HPMCAS possesses better flow properties than HPMCP because HPMCP forms a weak network of chain interactions at a molecular level. However, adding plasticisers such as PEG and TEC the flow properties of HPMCP can be tailored. The study also showed that plasticisers have a major influence on thermo-chemical and kinetic properties of polymers. For instance, PEG reduced polymer degradation with reversal in kinetic parameters whereas blends of CA produced detrimental effects and increased polymer degradation with reduction in onset degradation temperatures. Further, both polymers are observed to be chemically reactive with the APIs containing free -OH, -SOR2RN- and -NH2 groups. Finally, these properties prove that suitability of HPMCP is highly debated for HME and demands great care in use while that of HPMCAS is relatively better than HPMCP in many instances.
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Influence de l’agrégation érythrocytaire sur la migration axiale de microparticules simulant des plaquettes sanguinesGuilbert, Cyrille 06 1900 (has links)
Lors du phénomène d’hémostase primaire ou de thrombose vasculaire, les plaquettes sanguines doivent adhérer aux parois afin de remplir leur fonction réparatrice ou pathologique. Pour ce faire, certains facteurs rhéologiques et hémodynamiques tels que l’hématocrite, le taux de cisaillement local et les contraintes de cisaillement pariétal, entrent en jeu afin d’exclure les plaquettes sanguines de l’écoulement principal et de les transporter vers le site endommagé ou enflammé.
Cette exclusion pourrait aussi être influencée par l’agrégation de globules rouges qui est un phénomène naturel présent dans tout le système cardiovasculaire selon les conditions d’écoulement. La dérive de ces agrégats de globules rouges vers le centre des vaisseaux provoque la formation de réseaux d’agrégats dont la taille et la complexité varient en fonction de l’hématocrite et des conditions de cisaillement présentes. Il en résulte un écoulement bi-phasique avec un écoulement central composé d’agrégats de globules rouges avoisinés par une région moins dense en particules où l’on peut trouver des globules rouges singuliers, des petits rouleaux de globules rouges et une importante concentration en plaquettes et globules blancs. De ce fait, il est raisonnable de penser que plus la taille des agrégats qui occupent le centre du vaisseau augmente, plus il y aura de plaquettes expulsées vers les parois vasculaires. L'objectif du projet est de quantifier, in vitro, la migration des plaquettes sanguines en fonction du niveau d’agrégation érythrocytaire présent, en faisant varier l’hématocrite, le taux de cisaillement et en promouvant l’agrégation par l’ajout d’agents tels que le dextran à poids moléculaire élevé. Cependant, le comportement non Newtonien du sang dans un écoulement tubulaire peut être vu comme un facteur confondant à cause de son impact sur l’organisation spatiale des agrégats de globules rouges. De ce fait, les études ont été réalisées dans un appareil permettant de moduler, de façon homogène, la taille et la structure de ces agrégats et de quantifier ainsi leur effet sur la migration axiale des plaquettes. Du sang de porc anti coagulé a été ajusté à différents taux d’hématocrite et insérer dans un appareil à écoulement de Couette, à température ambiante. Les plaquettes sanguines, difficilement isolables in vitro sans en activer certains ligands membranaires, ont été remplacées par des fantômes en polystyrène ayant un revêtement de biotine. La quantification de la migration de ces fantômes de plaquettes a été réalisée grâce à l’utilisation de membranes biologiques fixées sur les parois internes de l’entrefer du rhéomètre de Couette. Ces membranes ont un revêtement de streptavidine assurant une très forte affinité d’adhésion avec les microparticules biotynilées.
À 40% d’hématocrite, à un cisaillement de 2 s-1, 566 ± 53 microparticules ont été comptées pour un protocole préétabli avec du sang non agrégeant, comparativement à 1077 ± 229 pour du sang normal et 1568 ± 131 pour du sang hyper agrégeant. Les résultats obtenus suggèrent une nette participation de l’agrégation érythrocytaire sur le transport des fantômes de plaquettes puisque l’adhésion de ces derniers à la paroi du rhéomètre de Couette augmente de façon quasi exponentielle selon le niveau d’agrégation présent. / During the primary hemostatis or thrombosis phenomenon, the human blood platelets must adhere to the vascular wall in order for them to perform their repairing or pathological function. To do so, certain rheological and hemodynamic factors such as the hematocrit, local shear rate and the wall shear stress, must come into play to exclude blood platelets from the main blood stream and transport them to the vicinity of the damaged or inflamed site.
This exclusion could also be influenced by red blood cell aggregation which is a natural process present throughout the entire cardiovascular system under certain flow conditions. The displacement of these rouleaux of red blood cells towards the centre of the vessel induces the formation of 3D networks of aggregates whose size and complexity vary as a function of the hematocrit and the shearing conditions present. It results in a two phase flow with an inner core composed of red blood cell aggregates surrounded by single red blood cells or small aggregates and large numbers of white blood cells and platelets. It is therefore reasonable to believe that the larger the inner core becomes, the more platelets will be expulsed towards the vascular wall. The objective of the study was to quantify, in vitro, the lateral migration of blood platelets as a function of the level of red blood cell aggregation present, by changing the hematocrit, the shear rate and by promoting red blood cell aggregation with the use of agents such as high molecular weight dextran. However, the non Newtonian behavior of blood in tube flow can be seen as a confounding factor to the understanding of the spatial organization of the red blood cell aggregates. In this study, whole blood was circulated in a simple shear flow apparatus, which allowed to homogeneously modulate the red blood cell aggregate sizes and structure, and quantify their effect on the axial migration of blood platelets. Anticoagulated porcine bloods were adjusted to different hematocrits and inserted into a Couette flow apparatus, at room temperature. Blood platelets, difficult to isolate in vitro without activating in a non reproducible manner specific membrane ligands, were replaced with biotin coated fluorescent polystyrene beads. The quantification of the migration of these platelet ghosts was conducted with the use of biological membranes fixed on the interior walls of the Couette apparatus. These streptavidin coated membranes ensure a strong adhesive affinity with the biotynilated beads.
At 40% hematocrit and at a shear of 2 s-1, 566 ± 53 micro particles were counted for non aggregated erythrocytes, 1077 ± 229 for aggregating red blood cells and 1568 ± 131 for hyper aggregating blood. The results obtained suggest a strong participation of the red blood cell aggregation on the transport of platelet ghosts since the number of ghost cells fixed on the wall of the Couette rheometer increases almost exponentially with the level of aggregation present.
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Influência de aspectos geométricos na hidrodinâmica e transferência de oxigênio de biorreatores airlift de circulação internaEsperança, Mateus Nordi 28 February 2014 (has links)
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Previous issue date: 2014-02-28 / Agência Nacional de Petróleo / The performance of pneumatic bioreactors is highly related to their geometric characteristics, such as the bottom clearance, riser to downcomer cross sectional area ratio, liquid height and the gas-liquid separator design. Although new geometries have been proposed, it is still necessary deeper studies to obtain more adequate reactor projects for bioprocess applications. This study evaluated the influence of the gas-liquid separator design on the hydrodynamics and oxygen transfer of 10-L concentric-tube airlift bioreactors, using Newtonian and non- Newtonian fluids and in order to define the better set of geometric characteristics. To reach this aim, the gas-liquid separator openness angle (α) varied from 30° to 90° and the volume fraction of fluid present on the gas-liquid separator section (FVL,GLS) varied from 0.10 to 0.30. The results indicated that for both fluids (Newtonian and non-Newtonian), the overall volumetric oxygen transfer coefficient (kLa) increased with the increase in α and a decrease in FVL,GS. Meanwhile, this gas-liquid separator geometry caused low global gas hold-up (εG), suggesting the reduction of mean bubble diameter (dB) for this condition. Operating with the non-Newtonian fluid at 5.0 vvm, the best gas-liquid separator geometry (α=90°; FV L,GLS=0,10) exhibited kLa and εG of 0,017 s-1 and 0,11, respectively. Moreover, this set of geometric characteristics lead to a gas-liquid flow with intermediate values for the drag coefficient (CD), suggesting moderate shear conditions. For the best geometry, the average shear rate varied from 1500 to 2400 s-1, in a similar range when compared to other airlift bioreactors. These results indicate the feasibility to use this bioreactor geometry in applications with shear-sensitive microorganisms. / O desempenho de biorreatores pneumáticos depende fortemente das suas características geométricas, como o vão livre na base, a razão entre as áreas de escoamento, a altura de líquido e o design da região de mistura. Embora diferentes geometrias tenham sido propostas na literatura, ainda há necessidade de estudos mais aprofundados, com o intuito de se obter projetos de biorreatores mais adequados aos bioprocessos. O presente estudo avaliou a influência da geometria da região de mistura na hidrodinâmica e transferência de oxigênio em biorreatores airlift de cilindros concêntricos de 10 L, empregando-se fluidos Newtonianos e não- Newtonianos, a fim de se definir o melhor conjunto de características geométricas. Para isso, variou-se o ângulo da região de mistura (α) (ângulo entre a lateral da região de mistura e o eixo horizontal) de 30° a 90° e a fração volumétrica de líquido na região de mistura (FVL,RM) (razão entre o volume de líquido contido apenas na região de mistura e o volume de líquido total do biorreator) de 0,10 a 0,30. Os resultados mostraram que para ambos fluidos (Newtoniano e não-Newtoniano), o coeficiente volumétrico de transferência de oxigênio (kLa) aumentou com o incremento em α e a diminuição de FVL,RM. Entretanto, esta configuração de geometria da região de mistura proporcionou baixa retenção gasosa (εG), indicando baixos valores de diâmetro das bolhas (dB) nesta condição. Empregando-se a melhor geometria da região de mistura (α=90°; FV L,RM=0,10), para o fluido não- Newtoniano, na condição de 5,0 vvm, obteve-se valores de kLa e εG de 0,017 s-1 e 0,11, respectivamente. Além disso, verificou-se que esta combinação de parâmetros geométricos conduziu a um escoamento gás-líquido com valores intermediários de coeficiente de arrasto (CD), sugerindo condições amenas de cisalhamento. Através da estimativa da velocidade de cisalhamento média ( m & ) para a melhor geometria em termos de transferência de oxigênio, observou-se uma variação entre 1500 e 2400 s-1, sendo estes valores da mesma ordem de grandeza quando comparados a outros biorreatores airlift. Esses resultados reforçam a viabilidade de utilização desta geometria em aplicações com microrganismos sensíveis ao cisalhamento.
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Suitability of cellulose ester derivatives in hot melt extrusion.Thermal, rheological and thermodynamic approaches used in the characterization of cellulose ester derivatives for their suitability in pharmaceutical hot melt extrusionKarandikar, Hrushikesh M. January 2015 (has links)
Applications of Hot Melt Extrusion (HME) in pharmaceuticals have become increasingly popular over the years but nonetheless a few obstacles still remain before wide scale implementation. In many instances these improvements are related to both processing and product performance. It is observed that HME process optimisation is majorly focused on the active pharmaceutical ingredient's (API) properties. Characterising polymeric properties for their suitability in HME should be equally studied since the impact of excipients on both product and process performance is just as vital. In this work, two well-established cellulose ester derivatives: Hydroxy Propyl Methyl Cellulose Acetate Succinate (HPMCAS) and Hydroxy Propyl Methyl Cellulose Phthalate (HPMCP) are studied for their HME suitability. Their thermal, thermodynamic, rheological, thermo-chemical and degradation kinetic properties were evaluated with model plasticisers and APIs. It was found the thermal properties of HPMCP are severely compromised whereas HPMCAS is more stable in the processing zone of 150 to 200 °C. Thermodynamic properties revealed that both polymers share an important solubility parameter range (20-30 MPa P1/2P) where the majority of plasticisers and BCS class II APIs lie. Thus, greater miscibility/solubility can be expected. Further, the processability of these two polymers investigated by rheometric measurements showed HPMCAS possesses better flow properties than HPMCP because HPMCP forms a weak network of chain interactions at a molecular level. However, adding plasticisers such as PEG and TEC the flow properties of HPMCP can be tailored. The study also showed that plasticisers have a major influence on thermo-chemical and kinetic properties of polymers. For instance, PEG reduced polymer degradation with reversal in kinetic parameters whereas blends of CA produced detrimental effects and increased polymer degradation with reduction in onset degradation temperatures. Further, both polymers are observed to be chemically reactive with the APIs containing free -OH, -SOR2RN- and -NH2 groups. Finally, these properties prove that suitability of HPMCP is highly debated for HME and demands great care in use while that of HPMCAS is relatively better than HPMCP in many instances.
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Optimisation of methane production from anaerobically digested cow slurry using mixing regime and hydraulic retention timeHughes, Kevin Lewis William January 2015 (has links)
AD is regarded as a sustainable technology that could assist the UK Government meet internationally agreed GHG emission targets by 2050. However, the mature status of the technology is based on expensive systems that rely on high energy feedstock to be profitable. Meanwhile, the natural biodegradation of cow slurry is a recognised contributor to climate change despite having a relatively low CH4 potential because of the large volumes produced. Economic mixing is essential to the cost-effectiveness of farm AD but techniques applied are not always appropriate as slurry is a shear thinning thixotropic Herschel-Bulkley fluid and therefore challenging to mix. The apparent viscosity of slurry and the shear stress induced was most influenced by solids content (exponential change) followed by temperature (linear). Most shear thinning occurred before a rising shear rate of 20s-1 was achieved with the fluid acting near-Newtonian above. Thixotropic recovery occurred within 1 hour of resting. Rheological values were also much higher than previously reported. Highest CH4 production occurred in the first 10 days of the batch process using a range of mixing regimes with different shear rates and rest periods. During fed-batch operations, changing shear rate had a minimal effect on CH4 production using a 30-day HRT whereas shorter rest periods increased production. Specific CH4 production rate was highest when feeding and mixing coincided. However, when HRT was reduced (OLR increased) the CH4 produced by all mixed regimes significantly increased with highest values being achieved using high intensity mixing rested for short periods. Lower HRTs also requires smaller digesters. Parasitic mixing energy invariably had the most influence on net energy production. Signs of instability were evident after 20 days using the low HRT. Significant microbial adaptation was also observed as the experiments progressed. The research outcomes demonstrate that mixing regime and HRT can be managed to maximise net energy production whilst reducing capital expenditure.
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