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Development of a Freeze-Drying Strategy to Store Human Bone Marrow Mesenchymal Stem/Stromal Derived Extracellular Vesicles for Applications in StrokeDorus, Brian 25 January 2023 (has links)
Mesenchymal stem/stromal cells (MSCs) release Extracellular vesicles (EVs) that are believed to play a major role in nerve regeneration after stroke. However, a major complication when trying to transition MSC-EVs from a pre-clinical to clinical setting is the convenient long-term storage of MSC-EVs. Therefore, we developed a strategy to freeze dry MSC-EVs to store them for more practical clinical applications. We first determined the optimal trehalose concentration for freeze drying the MSC-EVs, and we subsequently investigated the optimal freezing conditions. It was determined that 100 mM of trehalose and freezing temperature at -20°C were the optimal conditions to freeze dry the EVs. The therapeutic capabilities of the freeze-dried MSC-EVs was tested via tube formation assay and co-culturing them with neural stem/progenitor cells (NSPCs). It was found that human vein umbilical endothelial cells (HUVECs) treated with rehydrated MSCEVs promoted tube formation suggesting the trophic factors in the MSC-EVs survived the freeze-drying process. As for the NSPC co-culture, all treatments involving rehydrated MSC-EVs protected by trehalose during the freeze-drying process promoted proliferation and did not affect their ability to differentiate into oligodendrocytes, astrocytes, or neurons. Determining the optimum freezing-drying conditions allows us to stockpile a large amount of MSC-EVs at room temperature for on-demand applications.
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Investigation of Porous Ceramic Structure by Freeze-CastingBakkar, Said Adnan 05 1900 (has links)
The design and fabrication of porous ceramic materials with anisotropic properties has, in recent years, gained popularity due to their potential application in various areas that include medical, energy, defense, space, and aerospace. Freeze-casting is an effective, low-cost, and safe method as a wet shaping technique to create these structures. To control the morphology of these materials, many critical factors were found to play an important role. In this dissertation, the processing parameters of the magnetic field-assisted freeze-casting method were optimized with a focus on comparing the structure obtained using vertical and horizontal magnetic fields and understanding the mechanisms that occur under different freezing modes. More specifically, this processing method was used to produce Al2O3 and B4C porous ceramics materials with unidirectionally-aligned pore channels. The effect of the vertical and horizontal magnetic field strength and direction, concentration of magnetic material (Fe3O4), cooling rate, and freezing time were examined. The resulting ceramics with highly aligned pore channels were infiltrated with molten metal to create metal matrix composites. The mechanical properties of these structures were measured and were subsequently correlated to their morphology and composition.
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Detection of Influenza A Viruses From Environmental Lake and Pond IceKoçer, Zeynep A. 09 July 2010 (has links)
No description available.
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Climatology of Freeze-Thaw Days in the Conterminous United States: 1982-2009Haley, Jason S. 12 April 2011 (has links)
No description available.
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Characterization and physiological regulation of glucose transporter 2 in the liver of the wood frog, <i>Rana sylvatica</i>: implications for freeze toleranceRosendale, Andrew J. 24 June 2014 (has links)
No description available.
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Geographical variation of freeze tolerance in the wood frog, <i>Rana sylvatica</i>: the role of hepatic glycogen metabolismdo Amaral, Maria Clara Figueirinhas 04 August 2014 (has links)
No description available.
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Utilization of a preclinical model for chemoprevention of esophageal cancer employing a food-based and single- agent approachAziz, Robeena M. 07 June 2004 (has links)
No description available.
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Scaffold design and characterisation for osteochondral tissue regenerationDeplaine ., Harmony 03 February 2012 (has links)
El objetivo principal de esta tesis doctoral es el diseño de un andamio polimérico bicapa macroporoso para la regeneración del complejo osteocondral. El material empleado para la fabricación del constructo ha sido el ácido poli(L-láctico), un polímero biodegradable de la familia de los poliésteres. Una de las capas del andamio ha sido diseñada para asistir la regeneración del cartílago articular. La otra capa sirve de anclaje al hueso subcondral, y se diferencia de la anterior en sus propiedades mecánicas y bioactividad. Este comportamiento ha sido logrado por combinación del ácido poli(L-láctico) con nanopartículas inorgánicas. Ambas capas están unidas entre sí por una fina capa de material no poroso que evita el flujo de células de una parte a otra del constructo.
Para lograr este objetivo se realizó un primer estudio de diseño variando la morfología de los andamios hasta obtener aquella arquitectura más adecuada para la regeneración de ambos tejidos. Se varió parámetros de síntesis tales como la concentración de polímero y el ratio entre polímero y porógeno. Los andamios fueron evaluados mecánica y fisicoquímicamente y se seleccionó los parámetros de síntesis del ácido poli(L-láctico) que dieron mejores resultados.
En la regeneración del tejido es esencial conocer cómo variarán las propiedades del material una vez sea implantado y comience su degradación. Por lo tanto, fue considerado oportuno realizar un estudio de degradación del material in vitro en diversas condiciones. El estudio de la degradación fue realizado en condiciones estáticas durante 6, 12, 18, 24 semanas y 1 año y en condiciones dinámicas durante 1, 2, 4 y 6 semanas. Se evaluó tanto las características mecánicas como las fisicoquímicas tras los diversos tiempos de la degradación.
Posteriormente, y para aumentar las características mecánicas y la bioactividad del anclaje óseo, se incorporó distintas cantidades de nanopartículas inorgánicas de hidroxiapatita y sílice a los andamios. / Deplaine ., H. (2012). Scaffold design and characterisation for osteochondral tissue regeneration [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/14638
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Use of Volumetric Heating to Improve Heat Transfer During Vial Freeze-DryingDolan, James Patrick Jr. 28 September 1998 (has links)
Freeze-drying (lyophilization) is a drying process which is used to remove water from heat sensitive products, usually for the purpose of preservation. By removing water, the product becomes more stable at room temperature. This is a common process in the pharmaceutical industry because freeze-drying offers the advantage of drying at low temperatures and producing very low residual moisture contents. Often the materials dried in this manner are heat sensitive and require the highest possible quality. However, freeze-drying is a very slow process, often requiring 24 to 48 hours. During the process, vacuum pumps and refrigeration systems run continuously, making freeze-drying a very expensive process.
The goal of this project was to show that volumetric heating can be used in pharmaceutical freeze-drying and that this mode of heating offers some advantages. There were two approaches taken to the work, one experimental and one analytical. The experimental approach was broken into two phases, one focused on comparing microwave and conventional freeze-drying and the other focused on demonstrating the advantages of volumetric heating. In the analytical approach, a mathematical model was used to confirm the trends observed in phase II of the experimental work.
Experiments were conducted in a conventional laboratory freeze-dryer and the drying rate results were compared to the results obtained with an experimental microwave freeze-drying apparatus. Experiments were also conducted with the vaccine strain <i>A. pleuropneumoniae</i>. A viability study was conducted, comparing the viability loss caused by each process. The viability study showed a slightly higher viability loss for the microwave process.
A comparison of drying curves showed that the microwave process resulted in a slight improvement in primary drying time: 2.5 hours for the microwave process compared to 3 hours for the conventional process. There was a significant difference in overall drying times: 4 hours for the microwave process compared to 11 hours for the conventional process. This result was caused by a lower residual moisture content at the start of secondary drying and a higher secondary drying temperature for the microwave process.
Experiments were also conducted to show that using lower chamber pressure results in higher drying rates. This is not the case in a conventional freeze-dryer since heating is dependent on the chamber pressure in the low pressure environment of freeze-drying. Thus, an advantage of volumetric heating was demonstrated. The results show that a modest increase in pressure, from 0.05 to 0.3 Torr, caused a one third reduction in primary drying time.
The mathematical model developed in the analytical work relied on the D'Arcy equation to describe the flow of vapor in the porous dried layer. The results of the model confirm trends seen in the measured temperature and weight profiles. Analyzing the effect of varying the chamber pressures shows that lowering the pressure in the range of 1 to 0.01 Torr results in a significant increase in drying rate giving as much as a two thirds reduction in drying time for the case studied. A model incorporating mass transport equations derived from the dusty gas model was also presented. This model offers the benefit of a more accurate prediction of mass transport through the porous dried layer.
NOTE: (09/2008) An updated copy of this ETD was added after there were patron reports of problems with the file. / Ph. D.
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Méthodes innovantes de séchage de suspensions de nanocristaux / Innovative methods to dry nanocrystalline suspensionsTouzet, Antoine 12 July 2018 (has links)
Les nouvelles molécules thérapeutiques présentent la plupart du temps une faible solubilité aqueuse, à l’origine d’une biodisponibilité restreinte lors de leur administration orale. La réduction de taille des cristaux de substance active à l’échelle submicronique (= production de nanocristaux) s’est imposée comme une voie majeure de formulation au cours de ces dernières années.Afin de pallier des problèmes d’instabilité et d’aboutir à des formes solides pratiques d’utilisation pour les patients, une étape de séchage est généralement effectuée après production des nanocristaux en suspension. L’objectif principal de cette étape est de générer un produit présentant à la fois des propriétés facilitant le déroulement d’étapes procédés avales mais aussi et surtout de prévenir l’agrégation des nanocristaux au risque de compromettre leur dissolution après administration.Plusieurs techniques sont aujourd’hui utilisées dans l’industrie pharmaceutique telle que l’enrobage/granulation en lit d’air fluidisé et le spray drying. Parallèlement à l’élaboration d’une « formulation standard » pour ces deux méthodes de référence, deux techniques innovantes de lyophilisation ont été investiguées dans ce travail. La stabilisation de nanocristaux de kétoconazole par cryopelletization et active freeze drying a été démontrée et les paramètres clefs identifiés. Les produits générés par ces deux techniques, respectivement sous forme de pellets ou de poudre fine, ont été comparés à ceux conçus par enrobage en lit d’air fluidisé et spray drying. En conclusion, ces travaux indiquent que la cryopelletization et l’active freeze drying se positionnent comme des procédés d’intérêt de seconde intention, permettant de traiter des cas particuliers (molécules sensibles au stress thermique, à l’humidité et/ou couteuses). / Newly active pharmaceutical ingredients very often suffer from low aqueous solubility, a fact that in many cases can lead to poor oral bioavailability. Nanosizing, referring to drug nanocrystals production by size reduction, has demonstrated over the past few years a great potential to overcome this major issue.Since solid oral dosage forms are generally preferred due to stability reasons and patient convenience, the production of nanocrystals in a liquid medium is usually followed by a drying step. The main objective of this drying step is to generate a product suitable for downstream processing operations while at the same time preventing nanocrystal aggregation which can adversely affect the dissolution performance of the dry product in vivo.Several drying techniques such as spray drying and coating/granulation in fluidized bed have been successfully implemented in the pharmaceutical industry. In this work, two innovative freeze drying techniques were investigated and compared to the two above mentioned reference methods. The suitability of cryopelletization and active freeze drying to stabilize ketoconazole nanocrystals has been demonstrated and the key process parameters identified. The formulations generated by these two innovative techniques in the form of pellets or fine powder particles, respectively, were compared to the previously manufactured by fluidized bed and spray drying. In conclusion, this work presents cryopelletization and active freeze drying as suitable second-line processes with potential to address the drying of formulations containing problematic molecules sensible to thermal stress, moisture and/or presenting high production costs.
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