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Identification and Expression Analysis of Zebrafish Glypicans during Embryonic DevelopmentBrand, Michael, Gupta, Mansi 02 December 2015 (has links) (PDF)
Heparan sulfate Proteoglycans (HSPG) are ubiquitous molecules with indispensable functions in various biological processes. Glypicans are a family of HSPG’s, characterized by a Gpi-anchor which directs them to the cell surface and/or extracellular matrix where they regulate growth factor signaling during development and disease. We report the identification and expression pattern of glypican genes from zebrafish. The zebrafish genome contains 10 glypican homologs, as opposed to six in mammals, which are highly conserved and are phylogenetically related to the mammalian genes. Some of the fish glypicans like Gpc1a, Gpc3, Gpc4, Gpc6a and Gpc6b show conserved synteny with their mammalian cognate genes. Many glypicans are expressed during the gastrulation stage, but their expression becomes more tissue specific and defined during somitogenesis stages, particularly in the developing central nervous system. Existence of multiple glypican orthologs in fish with diverse expression pattern suggests highly specialized and/or redundant function of these genes during embryonic development.
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Interactions roches/saumures en contexte d'abandon d'exploitations souterraines de sel / Rocks/brines interactions in abandonned underground salt workingBoidin, Elie 06 February 2007 (has links)
La problématique de cette thèse est l’identification des phénomènes physico-chimiques se produisant entre les roches encaissantes du gisement de sel gemme de Lorraine et des saumures, puis l’étude de leurs effets sur le comportement mécanique. Une démarche multi-échelle a été entreprise : du minéral à l’échelle d’une exploitation en passant par celle des essais mécaniques. La confrontation de la géologie locale avec la géométrie des cavités de dissolution de sel (logiciel GOCAD) au travers de ces roches encaissantes rend compte d’un délitage relativement rapide lorsque les argilites sont au contact d’une saumure de cavité. A l’inverse, l’anhydrite et la dolomie peuvent constituer le toit de cavité pendant plusieurs années, avant de se rompre. Suite à une caractérisation minéralogique, et microtexturale des roches encaissantes (Marnes irisées inférieures et moyennes), une expérimentation de type batch a été mise en œuvre afin de comprendre ces différences: les faciès lithologiques qualifiés de majeurs ont été immergés dans des saumures pendant plus d’un an. Des analyses chimiques et microscopiques ont permis de suivre les modifications minéralogiques et microtexturales. Au contact de saumures saturées en chlorure de sodium, les faciès argileux se délitent en raison de l’hydratation en gypse du minéral anhydrite. Au niveau de l’anhydrite massive, cette hydratation n’affecte qu’une frange superficielle des échantillons, en raison d’une porosité connectée quasi-nulle. En présence d’une saumure de cavité, l’hydratation est promue par la présence de potassium et de strontium en solution. Ces résultats permettent d’expliquer qualitativement la dilatation voire la rupture des éprouvettes d’argilite lors d’essais de fluage en saumures. Le comportement en flexion de l’anhydrite massive ne semble pas être affecté par la présence de saumure, en raison d’une porosité trop faible pour permettre l’accès de la saumure au site réactionnel que sont les cristaux d’anhydrite / The aim of this study is the understanding of the physico-chemical interactions between saturated brine and the rocks (Marnes Irisées inférieures) enclosing the underground salt workings in Lorraine (eastern France), and also the study of their effect on the mechanical behaviour. A multi-scale study was undertaken, from the mineral scale to the one of a salt working. Whereas anhydrite-rich argillites flake quickly with the presence of saturated brine at the border of solutions cavities, the dolomudstone and massive anhydrite don’t and can constitute the top of cavities for several years. In order to explain this difference, these three lithologies were analysed in terms of mineralogy, micro-texture and porous media. Then, samples of argillites and massive anhydrite were immersed in saturated brines for more than one year. Chemical and microscopic analysis testified the hydration of anhydrite crystals into gypsum. This transformation occurs in a superficial way on massive anhydrite; to the contrary, it is located inside the anhydrite-rich argilites. As this transformation induces a volume increasing of 63%, the anhydrite crystals swell. In saturated brine, the water activity is low enough to prevent the swelling of clays such as smectites. Thus, anhydrite swelling might be the responsible of the argillites splitting in a saturated brine environment. The superficial anhydrite hydration on massive anhydrite can be explained by the low values of connected porosity (less than 1%) for this lithology. This results can explain, in a qualitative way, the dilatant behaviour of argilites samples during creep tests with brine. The bending behaviour of massive anhydrite don’t seem to be affected by the presence of brine for one year or less, probably because of the too low porosity of this lithology
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Le rôle des sulfates de sodium dans l’altération des pierres du patrimoine bâti : méthodes indirectes d'identification pour l'approche expérimentale / The role of sodium sulfate in the stones weathering of heritage building : indirect methods of identification for the experimental approachDenecker, Mélanie 20 June 2014 (has links)
Les sulfates de sodium sont admis comme étant les sels les plus destructeurs pour les pierres naturelles et autre matériaux poreux. Les processus de cristallisation de ces sels demeurent encore mal compris, plus particulièrement les relations entre l'heptahydrate (SS7, Na2SO4-7H2O) et la mirabilite (SS10, Na2SO4-10H2O). L'heptahydrate, métastable, a longtemps été négligée dans les travaux scientifiques portant sur l'altération saline jusqu'à récemment, et son rôle dans l'altération des roches n'est toujours pas établi à ce jour. Afin de mieux comprendre le rôle des différentes phases dans l'altération, il est important de pouvoir identifier quelle phase cristallise sous des conditions environnementales spécifiques. La cristallisation du sel se produit le plus souvent dans les matériaux poreux où les processus de cristallisation ne peuvent pas être observés directement in situ. Dans cette étude, nous utilisons le suivi de la température pour détecter et identifier la cristallisation des hydrates au cours de cycles de refroidissement/chauffage d'une solution de sulfate de sodium et de l'échantillon de calcaire saturé de cette même solution. En parallèle, d'autres approches expérimentales consistent à mesurer la réponse électrique et élastique des processus de cristallisation / dissolution se produisant dans le milieu poreux. Ces méthodes permettent d'identifier les différentes transitions de phases se produisant au cours de cycles de température et peuvent être utilisées dans des études expérimentales de vieillissement accéléré, afin d'estimer le rôle respectif des différents hydrates de sulfate de sodium dans l'altération des pierres par les sels. / Sodium sulfates are well known to be the most damaging salts in building materials and rocks. The crystallization processes of sodium sulfates are not completely understood, and more specifically the transition between the heptahydrate (SS7, Na2SO4-7H2O) and the mirabilite (SS10, Na2SO4-10H2O). The metastable heptahydrate has long been neglected in scientific works on salt damage until recently, and its role in rock weathering has not been established so far. In order to better understand the role of the different phases in damaging process, it is important to be able to identify which phase crystallizes under specific environmental conditions. Unfortunately, salt crystallization occurs very often within porous material and crystallization processes cannot be observed directly in situ. In this study, we use temperature monitoring to detect and identify the hydrates crystallization during cooling/heating cycles a sodium sulfate solution and limestone samples saturated with the same solution. In parallel, other experimental approaches consist to measure the electrical and elastic responses of the crystallization/dissolution processes occurring in the porous media. These methods enable to characterize the different phases occurring during temperature cycle and can be used in further experimental studies about ageing test in order to estimate the role of the different sodium sulfate hydrates involved in the salt weathering of rocks.
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Medida do Perfil de Dose no Alumínio Irradiado com Raios Gama de Captura de Nêutrons Térmicos Utilizando o Dosímetro Termoluminescente CaSO4:Dy / Measurement of dose profile in aluminum, irradiated wtth thermal neutron capture gamma rays, by using the termoluminescent dosimeter CaSO4:DyFederico, Claudio Antonio 10 May 2002 (has links)
Radiação gama com energias maiores que 1,25 MeV são usualmente encontradas em ambientes de reatores, aceleradores de partículas e campos de radiação de origem cósmica. Nestas energias, a resposta de um dosímetro varia fortemente com a espessura de material absorvedor, até atingir um valor máximo conhecido como espessura de equibl^rio de partículas carregadas. O objetivo principal do presente trabalho foi a determinação experimental do perfil de dose absorvida por uma amostra de alumínio, para várias energias de feixes de raios gama, de forma a obter uma relação entre a energia média do campo de radiação gama e a espessura de equilíbrio de partículas carregadas para o alumínio. Os dosímetros foram irradiados com radiação gama produzida pela captura de neutrons térmicos em 23 materiais alvos no arranjo experimental montado no canal tangencial do reator do IPEN-CNEN/SP. Na determinação da camada de equilíbrio de partículas carregadas, foi ajustada aos dados experimentais uma função semi-empirica que permitiu obter a espessura de ETPC (Equilíbrio Transitório de Partículas Carregadas) para cada espectro de radiação gama utilizado neste trabalho, com energia média variando no intervalo de 3,26 a 7,85 MeV. Os resultados experimentais do presente trabalho permitiram obter uma relação entre a energia média do campo de radiação gama e a espessura de ETPC, que apresenta uma excelente concordância com o alcance corrigido dos elétrons gerados naquela energia. / Gamma radiation with energies greater than 1.25 MeV are usually produced in reactor environments, particle accelerators and in cosmic radiation fields. For these energies, the response of a dosimeter heavily vary with the absorber material thickness, up to attain a maximum value named as charged particle equilibrium thickness. The main gol of this paper was the experimental determination of the absorbed dose profile in an aluminum sample for several energies of gamma ray beam, in order to obtain a relationship between the average energy of the gamma radiation field and the charged particle equilibrium thickness. The dosimeters were irradiated with gamma radiation produced by thermal neutron capture in 23 target materials in the experimental arrangement mounted at the tangential beam hole of the BPEN-CNEN/SP reactor. For the determination of the charged particle equilibrium thickness, it was fitted to the experimental data a semi-empirical fimction which allowed to obtain the thickness of CTPE ( Charged Particle Transient Equilibrium ) for each gamma radiation spectrum used in this work, with average energy varying in the interval from 3.26 to 7.85 MeV. The experimental results of the present paper allowed to obtain a relationship between average energy of the radiation field and the CTPE thickness, which presents an excellent agreement with the corrected range for electrons produced in that energy.
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Detecção de bactérias redutoras de sulfato em efluente e sedimento de mina de urânioSheila Kênia de Almeida 17 August 2005 (has links)
Um dos graves problemas ambientais oriundos da indústria de mineração é a drenagem ácida de mina que ocorre quando a pirita e outros minerais sulfetados são oxidados devido à presença de oxigênio e água, produzindo ácido sulfúrico que solubiliza metais presentes no solo/rocha. Em uma planta das Indústrias Nucleares do Brasil INB, na Unidade de Tratamento de Minérios (UTM) este problema tem se pronunciado de forma preocupante levando a dissolução de espécies radioativas e metálicas presentes. O tratamento da água ácida usando bactérias redutoras de sulfato, proporciona decréscimo da acidez que é decorrente da redução do sulfato a sulfito e precipitação dos metais como sulfetos. Nesse contexto, o presente estudo foi realizado visando a caracterizar sazonalmente e espacialmente populações de bactérias redutoras de sulfato (BRS) nos efluentes líquidos e amostras de sedimento da cava da mina (CM) e nos bota-foras 4 e 8 (BF4 e BF8) coletadas na UTM. Tais informações poderão permitir posteriormente, estudar mecanismos de biomanipulação a fim de remediar situações impactantes. Menores valores de pH , abaixo de 3,5, foram medidos no período de março a abril em amostras de água da cava da mina e a maior população de BRS (2,8 NMP. mL-1) foi observada em fevereiro em amostras do BF8. Os valores encontrados para matéria orgânica na água foram menores do que aqueles encontrados no sedimento. A concentração de oxigênio dissolvido na água variou de 0,65 g/L a 13,3 g/L. As amostras de sedimento apresentaram maiores valores de BRS (10,2 NMP/mL), quando comparadas com amostras de água (0,63 NMP/mL). Tais resultados eram esperados uma vez que, nas amostras de água coletadas próximas ao sedimento foram observadas menores concentrações de oxigênio dissolvido (7,10g/L) e maiores concentrações de matéria orgânica disponível (17,0 mg/L) quando os resultados foram comparados com aqueles obtidos para amostras de água coletadas na superfície (0,004mg/L). Os resultados mostram, portanto, que o efluente ácido gerado apresenta altos teores de metais estáveis e radioativos, sulfato, baixo pH e presença de bactérias redutoras de sulfato. / One of the most serious environmental problems created by the mining industry is acid mine drainage. In one plant of Nuclear Industries of Brazil - INB, this problem is a matter of concern. The presence of iron sulfites, such as pyrite, generates water with acidity above the levels allowed by the legislation and therefore, inappropriate for releasing straight into the environment. The industry maintain a high cost treatment in acid water from mines and waste disposal which consists in neutralizing and precipitating heavy metals. The treatment of acid water using SRB (sulfate-reducing bacteria) has been used in other countries with quite good technical results as well as economical advantages and thus, the object of this research. The use Sulfate Reducing Bacteria takes to a decreasing of the acidity by reducing sulfate to sulfite and precipitating the stable metals as sulfides. A seasonal study was carried out on the sulfate-reducing bacteria present in the liquid effluent discharged from two wastes disposal of the uranium mine, in phase of decommission, in Caldas/MG. This study shows the presence of SRB in the analyzed environmental, as well as some factors that are related with the amount of SRB presents, such as: dissolved oxygen, pH and organic matter. SRB was presented in water samples with high concentrations of heavy metals and low pH values, as well as in samples with high oxygen levels. The sediment samples were the preferential place for SRB occurrence and site BF8 presented the highest values of SRB.
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Medida do Perfil de Dose no Alumínio Irradiado com Raios Gama de Captura de Nêutrons Térmicos Utilizando o Dosímetro Termoluminescente CaSO4:Dy / Measurement of dose profile in aluminum, irradiated wtth thermal neutron capture gamma rays, by using the termoluminescent dosimeter CaSO4:DyClaudio Antonio Federico 10 May 2002 (has links)
Radiação gama com energias maiores que 1,25 MeV são usualmente encontradas em ambientes de reatores, aceleradores de partículas e campos de radiação de origem cósmica. Nestas energias, a resposta de um dosímetro varia fortemente com a espessura de material absorvedor, até atingir um valor máximo conhecido como espessura de equibl^rio de partículas carregadas. O objetivo principal do presente trabalho foi a determinação experimental do perfil de dose absorvida por uma amostra de alumínio, para várias energias de feixes de raios gama, de forma a obter uma relação entre a energia média do campo de radiação gama e a espessura de equilíbrio de partículas carregadas para o alumínio. Os dosímetros foram irradiados com radiação gama produzida pela captura de neutrons térmicos em 23 materiais alvos no arranjo experimental montado no canal tangencial do reator do IPEN-CNEN/SP. Na determinação da camada de equilíbrio de partículas carregadas, foi ajustada aos dados experimentais uma função semi-empirica que permitiu obter a espessura de ETPC (Equilíbrio Transitório de Partículas Carregadas) para cada espectro de radiação gama utilizado neste trabalho, com energia média variando no intervalo de 3,26 a 7,85 MeV. Os resultados experimentais do presente trabalho permitiram obter uma relação entre a energia média do campo de radiação gama e a espessura de ETPC, que apresenta uma excelente concordância com o alcance corrigido dos elétrons gerados naquela energia. / Gamma radiation with energies greater than 1.25 MeV are usually produced in reactor environments, particle accelerators and in cosmic radiation fields. For these energies, the response of a dosimeter heavily vary with the absorber material thickness, up to attain a maximum value named as charged particle equilibrium thickness. The main gol of this paper was the experimental determination of the absorbed dose profile in an aluminum sample for several energies of gamma ray beam, in order to obtain a relationship between the average energy of the gamma radiation field and the charged particle equilibrium thickness. The dosimeters were irradiated with gamma radiation produced by thermal neutron capture in 23 target materials in the experimental arrangement mounted at the tangential beam hole of the BPEN-CNEN/SP reactor. For the determination of the charged particle equilibrium thickness, it was fitted to the experimental data a semi-empirical fimction which allowed to obtain the thickness of CTPE ( Charged Particle Transient Equilibrium ) for each gamma radiation spectrum used in this work, with average energy varying in the interval from 3.26 to 7.85 MeV. The experimental results of the present paper allowed to obtain a relationship between average energy of the radiation field and the CTPE thickness, which presents an excellent agreement with the corrected range for electrons produced in that energy.
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Efeito da reação álcali-agregado (RAA) associada ao ataque interno por sulfato (isa) na absorção e porosidade de compósito de matriz cimentíciaPereira, Elias 23 February 2018 (has links)
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Previous issue date: 2018-02-23 / A degradação da argamassa ocasionada pelo ataque por sulfatos e reação álcali-agregado, ocorre pela presença de contaminantes nos agregados que reagem com os produtos da hidratação do cimento, sendo formados produtos expansivos, provocando efeitos deletérios ao material. Este trabalho tem por objetivo a análise isolada e combinada da reação álcali-agregado pela presença de quartzo deformado nos agregados e do ataque interno por sulfato devido à presença de pirita, também nos agregados, em argamassas de cimento Portland, em diferentes idades e meios de cura, assim como a investigação das mudanças microestruturais, com foco na absorção e na mudança de estrutura porosa do material. O programa experimental foi divido em quatro etapas: (i) caracterização dos agregados; (ii) ensaios acelerados e envelhecimento natural em prismas de argamassa; (iii) investigação do comportamento, em ambiente natural, de corpos de prova de concretos com pirita; (iv) análise da absorção e porosidade. As análises petrográficas, variações dimensionais de barras de argamassa, resistências à compressão e resistência à tração por compressão diametral, foram bons indicativos para a ocorrência da RAA e ISA de forma isolada e combinada, onde as curas parcialmente e totalmente imersa em água mantida a 40 °C intensificaram as reações pela incidência de umidade e oxigenação constantes. As porosidades, absorções de água por imersão e por capilaridade dos corpos de prova se mostraram indicativos da intensidade das reações álcali-agregado e ataque por sulfatos, sendo que para agregados contendo sulfetos as variações puderam ser atribuídas a fissuração produzida pela formação de cristais expansivos de sulfatos, e para os agregados reativos as variações puderam ser atribuídas a presença de intercrescimento mirmequíticos e extinção ondulante dos silicatos, culminando na formação de géis expansíveis. Em ambas as manifestações patológicas, a demasiada formação de fases expansivas promoveu a fissuração da matriz e aumento da proporção de macroporos nas idades finais. As análises de diâmetro de poros por intrusão de mercúrio demonstram que a RAA ocorre primeiramente nos poros próximos a 50 nm, devido ao seu caráter de borda de reação, e a ISA promove o refinamento e colmatação dos poros de toda a gama de diâmetros pela formação de sais de sulfatos, além de ambos os processos gerarem tensões internas e fissuração da matriz. Para os agregados com efeito combinado, estes processos ocorrem concomitantemente, tendo influência sobre as capacidades de absorção de água por imersão e capilaridade, além das resistências mecânicas e variações dimensionais. / The degradation of the mortar caused by the attack by sulfates and alkali-aggregate reaction occurs by the presence of contaminants in the aggregates that react with the products of the cement hydration, being formed expansive products, causing deleterious effects to the material. This work aims at the isolated and combined analysis of the alkali-aggregate reaction by the presence of deformed quartz in the aggregate and the internal attack by sulphate due to the presence of pyrite, also in the aggregate, in Portland cement mortars, at different ages and mortar cure means, as well as the investigation of the microstructural changes, focusing on the absorption and the change in the porous structure of the material. The experimental program was divided into four stages: (i) characterization of the aggregates; (ii) accelerated tests and natural aging on mortar prisms; (iii) investigation of the behavior, in the natural environment, of concrete specimens with pyrite; (iv) absorption and porosity analysis. The petrographic analyzes, dimensional variations of mortar bars, compressive strengths and tensile strength by diametrical compression were good indicative for the occurrence of RAA and ISA in an isolated and combined manner, where the cures were partially and totally immersed in water maintained at 40 ° C intensified the reactions by the incidence of constant moisture and oxygenation. The porosities, water absorptions by immersion and by capillarity of the test specimens were indicative of the intensity of the alkali-aggregate and sulfate attack reactions, and being that for aggregates containing sulfides the variations could be attributed to the cracking produced by the formation of expansive crystals of sulfates, and for the reactive aggregates the variations could be attributed to the presence of myrhythmic intergrowth and undulating extinction of the silicates, culminating in the formation of expandable gels. In both pathological manifestations, the excessive formation of expansive phases promoted matrix cracking and increased proportion of macropores in the final ages. The analysis of pore diameter by mercury intrusion shows that the RAA occurs first in pores close to 50 nm, due to its character of reaction border, and ISA promotes the refinement and filling of the pores of the whole range of diameters by the formation of sulfate salts, in addition to both processes generating internal stresses and cracking of the matrix. For aggregates with combined effect, these processes occur concomitantly, having influence on the water absorption capacities by immersion and capillarity, besides the mechanical resistances and dimensional variations.
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Relations sulfates/carbonates pendant la diagenèse des sédiments marins : étude sédimentologique du delta éocene du Sobrarbe (Espagne) et modélisation thermodynamique des données ODP /IODPHoareau, Guilhem 30 November 2009 (has links) (PDF)
Ce travail a pour but l'étude de la stabilité des minéraux sulfatés et carbonatés dans les sédiments marins au cours de la diagenèse précoce (températures inférieures à 70°C et enfouissement maximal de 2 kilomètres). La première partie présente l'étude pétrographique et géochimique de concrétions dolomitiques contenant de la calcite et des sulfates (célestine et barytine), dans le delta éocène du Sobrarbe (Espagne). Nous déterminons l'ordre d'apparition des minéraux ainsi que leurs conditions de formation dans un contexte tectonique actif. Dans la deuxième partie, l'état de saturation des eaux interstitielles des sédiments marins par rapport aux carbonates et sulfates est calculé en utilisant l'intégralité de la base de données ODP/IODP (14000 échantillons). Pour les sulfates (célestine, gypse), la saturation est fréquente et caractérisée par l'équilibre chimique. Ce travail montre comment la modélisation des interactions eau/roche peu expliquer la présence de minéraux diagénétiques
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Delivery of thermostabilized chondroitinase ABC enhances axonal sprouting and functional recovery after spinal cord injuryLee, Hyun-Jung 10 November 2009 (has links)
Chondroitin sulfate proteoglycans (CSPGs) are one major class of axon growth inhibitors that are upregulated and accumulated around the lesion site after spinal cord injury (SCI), and result in regenerative failure. To overcome CSPG-mediated inhibition, digestion of CSPGs with chondroitinase ABC (chABC) has been explored and it has shown promising results. chABC digests glycosaminoglycan chains on CSPGs and can thereby enhance axonal regeneration and promote functional recovery when delivered at the site of injury. However, chABC has a crucial limitation; it is thermally unstable and loses its enzymatic activity rapidly at 37 ºC. Therefore, it necessitates the use of repeated injections or local infusions with a pump for days to weeks to provide fresh chABC to retain its enzymatic activity. Maintaining these infusion systems is invasive and clinically problematic.
In this dissertation, three studies are reported that demonstrate our strategy to overcome current limitations of using chABC and develop a delivery system for facilitating chABC treatment after SCI: First, we enhanced the thermostability of chABC by adding trehalose, a protein stabilizer, and developed a system for its sustained local delivery in vivo. Enzymatic activity was assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and dimethylmethylene blue (DMMB), and conformational change of the enzyme was measured via circular dichroism (CD) with and without trehalose. When stabilized with trehalose, chABC remained enzymatically active at 37 ºC for up to 4 weeks in vitro. We developed a lipid microtube-agarose hydrogel delivery system for a sustained release and showed that chABC released from the delivery system is still functionally active and slowly released over 2 weeks in vitro. Second, the hydrogel-microtube system was used to locally deliver chABC over two weeks at the lesion site following a dorsal over hemisection injury at T10. The scaffold consisting of hydrogel and chABC loaded lipid microtubes was implanted at the top of the lesion site immediately following injury. To determine effectiveness of topical delivery of thermostabilized chABC, animal groups treated with single injection or gel scaffold implantation of chABC and penicillinase (P'ase) were included as controls. Two weeks after surgery, the functionality of released chABC and the cellular responses were examined by immunohistological analysis with 3B3, CS-56, GFAP and Wisteria floribunda agglutinin (WFA). The results demonstrated that thermostabilized chABC was successfully delivered slowly and locally without the need for an indwelling catheter by using the hydrogel-microtube delivery system in vivo. The results demonstrated that released chABC from the gel scaffold effectively digested CSPGs, and therefore, there were significant differences in CSPG digestion at the lesion site between groups treated with chABC loaded microtube-hydrogel scaffolds and controls. Third, a long term in vivo study (45 days) was conducted to examine axonal sprouting/regeneration and functional recovery with both a single treatment each of microtube loaded chABC or Neurotrophin-3 (NT-3), and a combination of them by using the hydrogel-microtube delivery system. Over the long term study period, the treated animals showed significant improvement in locomotor function and more sprouting of cholera toxin B subunit (CTB)-positive ascending dorsal column fibers and 5-HT serotonergic fibers around the lesion site.
We demonstrated that this significant improvement of chABC thermostability facilitates the development of a minimally invasive method for sustained, local delivery of chABC that is potentially a useful and effective approach for treating SCI. In addition to that, we demonstrated that combinatorial therapy with chABC and neurotrophic factors could provide a synergistic effect on axonal regrowth and functional recovery after SCI.
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The development of glycosaminoglycan-based materials to promote chondrogenic differentiation of mesenchymal stem cellsLim, Jeremy James 03 July 2012 (has links)
Tissue engineering strategies represent exciting potential therapies to repair cartilage injuries; however, difficulty regenerating the complex extracellular matrix (ECM) organization of native cartilage remains a significant challenge. Cartilaginous ECM molecules, specifically chondroitin sulfate (CS) glycosaminoglycan, may possess the ability to promote and direct MSC differentiation down a chondrogenic lineage. CS may interact with the stem cell microenvironment through its highly negative charge, generation of osmotic pressure, and sequestration of growth factors; however, the role of CS in directing differentiation down a chondrogenic lineage remains unclear. The overall goal of this dissertation was to develop versatile biomaterial platforms to control CS presentation to mesenchymal stem cells (MSCs) in order to improve understanding of the interactions with CS that promote chondrogenic differentiation.
To investigate chondrogenic response to a diverse set of CS materials, progenitor cells were cultured in the presence of CS proteoglycans and CS chains in a variety of 2D and 3D material systems. Surfaces were coated with aggrecan proteoglycan to alter cell morphology, CS-based nano- and microspheres were developed as small particle carriers for growth factor delivery, and desulfated chondroitin hydrogels were synthesized to examine electrostatic interactions with growth factors and the role of sulfation in the chondrogenic differentiation of MSCs. Together these studies provided valuable insight into the unique ability of CS-based materials to control cellular microenvironments via morphological and material cues to promote chondrogenic differentiation in the development of tissue engineering strategies for cartilage regeneration and repair.
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