Submitted by Simone Souza (simonecgsouza@hotmail.com) on 2017-11-01T14:56:26Z No. of bitstreams: 1 DISS_2014_Jonas Mangoni Rambo.pdf: 9222881 bytes, checksum: 541ad16fc560f2a2f8e066273f9c5d87 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2017-11-08T15:34:43Z (GMT) No. of bitstreams: 1 DISS_2014_Jonas Mangoni Rambo.pdf: 9222881 bytes, checksum: 541ad16fc560f2a2f8e066273f9c5d87 (MD5) / Made available in DSpace on 2017-11-08T15:34:43Z (GMT). No. of bitstreams: 1 DISS_2014_Jonas Mangoni Rambo.pdf: 9222881 bytes, checksum: 541ad16fc560f2a2f8e066273f9c5d87 (MD5) Previous issue date: 2014-04-15 / A Província Aurífera Alta Floresta (PAAF) localiza-se no norte do estado de Mato Grosso, delimitada ao norte pelo Gráben do Cachimbo e ao sul pelo Gráben dos Caiabís. Se enquadra na porção sul do Cráton Amazônico, entre as províncias geocronológicas Ventuari – Tapajós e Rio Negro – Juruena. Na porção leste da PAAF foram selecionados 5 alvos para elaboração deste trabalho: (i) Mina do Edu; (ii) Garimpo do Peteca; (iii) Filão do Laje; (iv) Filão do Cocheira e (v) Filão do Vinte. O corpo mineralizado do Edu tem atitudes de N55ºE/subvertical, e é classificado como estruturas do tipo T. Essas estruturas são fraturas extensionais que ocorreram com a mesma atitude do esforço principal e, portanto, indicam a direção do esforço compressivo principal e são preenchidas por veios de quartzo. A Mina do Edu apresenta fluídos carbônicos e aquosa-carbônico. O Garimpo do Peteca e o Filão do Laje, com atitudes de E-W/subvertical, são classificados como estruturas do tipo R. Os Filões do Cocheira e do Vinte, com atitudes N-S/subvertical, são classificados como estruturas do tipo X, que são zonas confinadas de cisalhamento transcorrente geradas a partir da nucleação de fraturas de cisalhamento dextral. O Garimpo do Peteca, o Filão do Laje, Filão do Cocheira e Filão do Vinte apresentam fluídos estritamente aquosos, variando de alta a média salinidade. Esses fluídos distintos sugerem mineralizações provenientes de uma mesma fonte magmática, no entanto, devido à distância de algumas ocorrências da fonte, o fluído foi interagindo com outros fluídos, se distinguindo assim do fluído primário. / The Alta Floresta Gold Province (PAAF) is located in the northern state of Mato Grosso, bounded on the north by the Gráben do Cachimbo and on the south by Gráben dos Caiabis. The PAAF fits in the southern portion of the Amazon Craton, between the Ventuari - Tapajós and Rio Negro - Juruena geochronological provinces. In the eastern portion of PAAF 5 targets were selected for the present work: (i) Mina do Edu (ii) Garimpo do Peteca (iii) Filão do Laje , (iv) Filão do Cocheira (v) Filão do Vinte. The mineralized body of the Mina do Edu has attitude of N55ºE/subvertical, and is classified as structures of the T type. These structures are extensional fractures that occurred with the same attitude as the main effort, and therefore indicate the direction of the main compressive stress and are filled by quartz veins. The Mina do Edu presents carbonic and aqueous-carbonic fluid inclusions. The Garimpo do Peteca and Filão do Laje, with attitudes of EW/subvertical, are classified as structures of the R type. The Filão do Cocheira and Filão do Vinte, with attitudes NS/subvertical, are classified as structures of the X type, which are enclosures transcurrent shear generated from the nucleation of dextral shear fractures. The Garimpo do Peteca, Filão do Laje, Filão do Cocheira and Filão do Vinte presents strictly aqueous fluid inclusions, ranging from high to medium salinity. These different fluids suggest mineralization from the same magmatic source, however, due to the distance of the source from some instances, the fluid was interacting with other fluids, thus distinguishing from the primary fluid.

Background: Collagens of marine origin are applied increasingly as alternatives to mammalian collagens in tissue engineering. The aim of the present study was to develop a biphasic scaffold from exclusively marine collagens supporting both osteogenic and chondrogenic differentiation and to find a suitable setup for in vitro chondrogenic and osteogenic differentiation of human mesenchymal stroma cells (hMSC). Methods: Biphasic scaffolds from biomimetically mineralized salmon collagen and fibrillized jellyfish collagen were fabricated by joint freeze-drying and crosslinking. Different experiments were performed to analyze the influence of cell density and TGF-β on osteogenic differentiation of the cells in the scaffolds. Gene expression analysis and analysis of cartilage extracellular matrix components were performed and activity of alkaline phosphatase was determined. Furthermore, histological sections of differentiated cells in the biphasic scaffolds were analyzed. Results: Stable biphasic scaffolds from two different marine collagens were prepared. An in vitro setup for osteochondral differentiation was developed involving (1) different seeding densities in the phases; (2) additional application of alginate hydrogel in the chondral part; (3) pre-differentiation and sequential seeding of the scaffolds and (4) osteochondral medium. Spatially separated osteogenic and chondrogenic differentiation of hMSC was achieved in this setup, while osteochondral medium in combination with the biphasic scaffolds alone was not sufficient to reach this ambition. Conclusions: Biphasic, but monolithic scaffolds from exclusively marine collagens are suitable for the development of osteochondral constructs.

Les alliages Mg-Ca sont des alliages biocompatibles et biodégradables largement utilisés pour des applications biomédicales comme prothèses bio-résorbables. Ils sont très utilisés car leurs taux de corrosion et de dégradation dans l'organisme peuvent être contrôlés par le taux de calcium et par l'influence du milieu d'implantation. Ces solutions ont inspirées le Groupe SEB qui cherche à mettre au point un procédé fiable de minéralisation d'une eau par dissolution d'alliages Mg-Ca. L'objectif de ce travail de thèse est double, à savoir, la conception des alliages Mg-Ca pour ce procédé, en choisissant les techniques de production adaptées et les paramètres d'élaborations optimisés, puis, la mise en œuvre d'un processus de dissolution assisté par un courant et la maîtrise des paramètres de dissolution. Le magnésium et le calcium étant des métaux très réactifs au contact de l'atmosphère, l'élaboration d'alliages Mg-Ca se fait sous atmosphère neutre. Il est possible d'obtenir une microstructure stable avec des teneurs en calcium comprises entre 0% et 33% atomique, lorsque le calcium et le magnésium forment l'intermétallique Mg2Ca. Lors du refroidissement du mélange métallique à fusion, il se forme une microstructure d'équilibre constituée de dendrites (de Mg si 0 at.% < Ca < 10 at.% et de Mg2Ca si 10 at.% < Ca < 33 at.%) et d'un agrégat eutectique lamellaire composé de Mg et de Mg2Ca. La dissolution anodique d'alliages Mg-Ca s'effectue par l'application d'une différence de potentiel entre une électrode constituée de l'alliage à dissoudre et une contre électrode constituée d'un métal plus noble. Il a été mis en évidence que les relargages d'ions Mg2+ et Ca2+ en solution peuvent être augmentés en diminuant la chute ohmique entre les électrodes (distance minimale, tension appliquée maximale) et en augmentant la réactivité du processus (alliages riches en calcium, conductivité et minéralité de la solution, durée de dissolution et surfaces exposées importantes). Cependant, les forts relargages d'ions Mg2+ et Ca2+, qui alcalinisent fortement la solution, favorisent la formation de précipités limitant la dissolution tels que le tartre, les oxydes et hydroxydes de magnésium et de calcium. Plusieurs solutions pour réguler le pH de la solution et optimiser les relargages de cations Mg2+ et Ca2+ ont été envisagées. / The Mg-Ca alloys are biocompatible and biodegradable alloys widely used for biomedical applications such as bioresorbable implants because of their corrosion rate and degradation behaviour into human body. These solutions have inspired the SEB Group to develop a new process for the mineralization of water by dissolving Mg-Ca alloys. The objective of this thesis is to design new Mg-Ca alloys choosing the appropriate production techniques and optimizing elaborations settings and then, to work on the dissolution process assisted by a current controling of dissolution parameters. Magnesium and calcium being very reactive metal in contact with the atmosphere, the development Mg-Ca alloys was done in a neutral atmosphere. It is possible to obtain a stable microstructure with calcium contents between 0% and 33 atomic% through the intermetallic form Mg2Ca. It's possible to obtain equilibrium microstructures consisting of dendrites (Mg if 0 at.% <Ca <10 at.% And Mg2Ca if 10 at.% <Ca <33 at.%) and a lamellar eutectic aggregate made from Mg and Mg2Ca. Anodic dissolution of Mg-Ca alloys is made by applying a potential between one electrode made from the Mg-Ca alloy and a counter electrode made from a more noble metal. It has been demonstrated that the releases of Mg2+ and Ca2+ in solution may be controlled throught the decrease of the resistance drop between the electrodes (minimum distance, maximum applied voltage) and the increase of the process reactivity (rich alloys calcium, mineral and conductivity of the solution, dissolving time and significant exposed surfaces). However, strong releases of Mg2+ and Ca2+, which strongly alkalize the solution, promote the formation of precipitates limiting dissolution rate such as scale, oxides and hydroxides formation. Several solutions to regulate the pH of the solution and optimize the releases of cation Mg2+ and Ca2+ were considered.

Background: Collagens of marine origin are applied increasingly as alternatives to mammalian collagens in tissue engineering. The aim of the present study was to develop a biphasic scaffold from exclusively marine collagens supporting both osteogenic and chondrogenic differentiation and to find a suitable setup for in vitro chondrogenic and osteogenic differentiation of human mesenchymal stroma cells (hMSC). Methods: Biphasic scaffolds from biomimetically mineralized salmon collagen and fibrillized jellyfish collagen were fabricated by joint freeze-drying and crosslinking. Different experiments were performed to analyze the influence of cell density and TGF-β on osteogenic differentiation of the cells in the scaffolds. Gene expression analysis and analysis of cartilage extracellular matrix components were performed and activity of alkaline phosphatase was determined. Furthermore, histological sections of differentiated cells in the biphasic scaffolds were analyzed. Results: Stable biphasic scaffolds from two different marine collagens were prepared. An in vitro setup for osteochondral differentiation was developed involving (1) different seeding densities in the phases; (2) additional application of alginate hydrogel in the chondral part; (3) pre-differentiation and sequential seeding of the scaffolds and (4) osteochondral medium. Spatially separated osteogenic and chondrogenic differentiation of hMSC was achieved in this setup, while osteochondral medium in combination with the biphasic scaffolds alone was not sufficient to reach this ambition. Conclusions: Biphasic, but monolithic scaffolds from exclusively marine collagens are suitable for the development of osteochondral constructs.

Beim Tissue Engineering ist die Vaskularisierung von größeren Zell-Matrix-Konstrukten nach Implantation bis heute ein großes Problem. Durch das initiale Fehlen eines mikrovaskulären Netzwerkes kommt es zu einem raschen Zellsterben im Scaffold. Aufgrund dessen war das Ziel dieser Arbeit, im Sinne des in situ-Tissue Engineering ein Scaffold auf Basis von mineralisiertem Kollagen zu entwickeln, welches mit dem angiogenen Wachstumsfaktor VEGF funktionalisiert wird, um den Prozess der Vaskularisierung – die Einsprossung von Blutgefäßen – zu fördern und gleichzeitig durch Chemoattraktion in vivo Zellen aus dem umliegenden Knochengewebe in das Innere des Scaffolds migrieren zu lassen, so dass eine beschleunigte Defektheilung erzielt wird. Poröse Scaffolds aus mineralisiertem Kollagen wurden durch zwei unterschiedliche Strategien funktionalisiert und durch in vitro-Testungen charakterisiert. Die erste Strategie umfasste die Heparin-Modifizierung der gesamten Scaffolds, während die zweite Strategie die Injizierung eines zentralen VEGF-haltiges Depots in das Scaffoldinnere darstellte. Neben der Charakterisierung der Scaffolds wurde die Freisetzungskinetik des Modellwachstumsfaktors VEGF aus den modifizierten Scaffolds untersucht und die biologische Aktivität des freigesetzten Faktors auf Endothelzellen getestet. Zusätzlich wurde bei der 2. Strategie, der Injizierung eines Wirkstoffdepots, die Ausbildung eines Wirkstoffgradienten und die zielgerichtete Migration von Endothelzellen in Richtung des Wirkstoffdepots analysiert.

Beim Tissue Engineering ist die Vaskularisierung von größeren Zell-Matrix-Konstrukten nach Implantation bis heute ein großes Problem. Durch das initiale Fehlen eines mikrovaskulären Netzwerkes kommt es zu einem raschen Zellsterben im Scaffold. Aufgrund dessen war das Ziel dieser Arbeit, im Sinne des in situ-Tissue Engineering ein Scaffold auf Basis von mineralisiertem Kollagen zu entwickeln, welches mit dem angiogenen Wachstumsfaktor VEGF funktionalisiert wird, um den Prozess der Vaskularisierung – die Einsprossung von Blutgefäßen – zu fördern und gleichzeitig durch Chemoattraktion in vivo Zellen aus dem umliegenden Knochengewebe in das Innere des Scaffolds migrieren zu lassen, so dass eine beschleunigte Defektheilung erzielt wird. Poröse Scaffolds aus mineralisiertem Kollagen wurden durch zwei unterschiedliche Strategien funktionalisiert und durch in vitro-Testungen charakterisiert. Die erste Strategie umfasste die Heparin-Modifizierung der gesamten Scaffolds, während die zweite Strategie die Injizierung eines zentralen VEGF-haltiges Depots in das Scaffoldinnere darstellte. Neben der Charakterisierung der Scaffolds wurde die Freisetzungskinetik des Modellwachstumsfaktors VEGF aus den modifizierten Scaffolds untersucht und die biologische Aktivität des freigesetzten Faktors auf Endothelzellen getestet. Zusätzlich wurde bei der 2. Strategie, der Injizierung eines Wirkstoffdepots, die Ausbildung eines Wirkstoffgradienten und die zielgerichtete Migration von Endothelzellen in Richtung des Wirkstoffdepots analysiert.