O objetivo do presente estudo foi sintetizar e caracterizar um material reparador para uso endodôntico com propriedades anti-inflamatória, antimicrobiana e remineralizante. A pasta experimental tem como propósito ser um sistema carreador de fármacos para regiões de difícil acesso em Odontologia. A apresentação do material é em forma de pó:líquido. No pó se encontra α-fosfato tricálcico, tungstato de cálcio e microesferas de amoxicilina (AMX-MS), já no líquido estão contidas nanocápsulas de indometacina (IndOHNC). A pasta experimental foi testada em relação a suas características físicoquímicas e biológicas. As AMX-MS obtiveram tamanho de 1,604 μm ± 0,08, forma esférica confirmada por MEV e teor da droga foi 1,63 mg g-1. As IndOHNC obtiveram tamanho de 162 ± 7,5 nm e forma esférica confirmada por MET. O teor do fármaco foi de 1 mg mL-1 ± 0,02. O escoamento da pasta foi de 18.56 ± 0.29, a espessura de película obtida foi 33 μm e radiopacidade de 1,81 mmAl. A pasta experimental demonstrou atividade antibacteriana contra o Enterococcus faecalis. A maior concentração de pasta experimental apresentou o maior valor em relação à viabilidade celular, com 187,03% no teste SRB. A atividade da enzima fosfatase alcalina e a formação de nódulos mineralizados obtiveram um gradual aumento em função do tempo. A migração celular demonstrou fechamento da ferida, e a pasta experimental foi capaz de acelerar o processo (p<0.05). Em conclusão, a pasta experimental demonstrou propriedades físico químicas e biológicas confiáveis, podendo ser um material promissor para o reparo da região periapical. / The aim of this study was to synthesize and characterize a new reparative material with anti-inflammatory, antimicrobial and remineralizing properties. The reparative material was developed to be a drug delivery system for regions with difficult access in Dentistry. The formulation is presented in powder/liquid. The powder is composed of α-tricalcium phosphate, calcium tungstate and amoxicillin microspheres (AMX-MS). The liquid is composed of nanocapsules containing indomethacin (IndOH-NC). The physicochemical and biological properties of the experimental endodontic paste were evaluated. The AMX-MS obtained a mean size of 1.604 μm ± 0.08, spherical shape and the encapsulation capacity was 1.63 mg g-1. IndOH-NCs obtained a mean size of 162 ± 7.5 nm and spherical shape confirm by MET. The content of the encapsulated drug was 1 mg mL-1 ± 0.02. The experimental paste flow was 18.56 ± 0.29 mm, mean film thickness was 33 μm and radiopacity equivalent to 1.81 mmAl. The experimental paste showed antibacterial activity against Enterococcus faecalis. The highest concentration of experimental paste presented the highest value in cell viability (187.03% in SRB test). The activity of the phosphatase alkaline enzyme and the formation of mineralized nodules showed a gradual increase as a function of time. Cell proliferation showed continuous wound closure, and the experimental paste was able to accelerate the process (p<0.05). In conclusion, the experimental paste demonstrated reliable physicochemical and biological properties, and it could be a promising material for periapical region repair.

L’objet de ce travail est l’étude de la biocompatibilité in vitro des matériaux à base de COC, afin d’évaluer leur potentiel de biomatériaux implantables.Cette évaluation est réalisée par le suivi de plusieurs paramètres : viabilité cellulaire/cytotoxicité, évaluation des phénomènes de stress oxydant, inflammatoires et hémocompatibilité. Une relation entre ces réponses biologiques et les propriétés physico chimiques des matériaux étudiés a été appréhendée.Les résultats montrent une bonne biocompatibilité des films testés avec un impact significatif de la présence des additifs (anti-oxydant et lubrifiant) sur les paramètres biologiques et physico-chimiques évalués.L’effet simulé du vieillissement biologique de ces matériaux sur leur biocompatibilité et leurs caractères physico-chimiques a également été étudié. Des conditions de pH et d’oxydation extrêmes, ainsi que le contact avec des macrophages pendant 1 mois, ont un effet sur la surface et sur l’interaction des films de COC avec l’environnement biologique sans compromettre leur biocompatbilité. La présence d’additifs a également eu un impact sur ces modifications.En suivant une logique de management du risque, la systématisation de l’ensemble des méthodes développées a permis d’obtenir une approche simplifiée et validée au sein du laboratoire, applicable à l’ensemble des matériaux naturels ou synthétiques susceptibles d’être utilisés dans la fabrication des DM implantables / Abstract : The purpose of this work is the study of the in vitro biocompatibility of COC-based materials in order to evaluate their potential as implantable biomaterials.This evaluation is carried out by monitoring several parameters: cell viability / cytotoxicity, evaluation of oxidative stress, inflammatory reactions and hemocompatibility. A relationship between these biological responses and physicochemical properties of the studied materials has been apprehended.Results show a good biocompatibility of the tested films with a significant impact of the presence of additives (anti-oxidant and lubricant) on the evaluated biological and physicochemical parameters.The simulated effect of biological aging of these materials on their biocompatibility and physico-chemical characteristics has also been studied. Extreme pH and oxidation conditions, as well as contact with macrophages during 1 month, affect the surface and interaction of COC films with the biological environment without compromising their biocompatibility. The presence of additives also had an impact on these changes.Following a risk management logic, the systematization of the developed methods within the laboratory made it possible to obtain a simplified and validated approach, applicable to all natural or synthetic materials that could be used for manufacturing implantable medical devices.

Tato práce je zaměřena na studium vlastností organických polovodivých materiálů se zaměřením na jejich vodivost a pohyblivost nosičů náboje. Hlavním cílem této práce je objasnit vztah mezi chemickou strukturou organických polovodičů a jejich vlastnostmi. Teoretická část práce je zaměřena na základy organických polovodičů, transport náboje a přehled vlastností organických polovodivých materiálů, které vedly k jejich aplikaci v polních a elektrochemických tranzistorech. Experimentální část představuje přehled použitých materiálů, způsoby jejich přípravy a charakterizační metody. V rámci výsledkové části bylo vyvinuto nebo optimalizováno několik metod pro přípravu tenkých vrstev a následně byl studován jejich vliv na výkon organických polem řízených tranzistorů. Byly charakterizovány a diskutovány relevantní vlastnosti nových organických polovodivých materiálů se zaměřením na pohyblivost nosičů náboje. Byla charakterizována a diskutována biokompatibilita několika organických polovodičů. Elektrické vlastnosti, stabilita a biokompatibilita elektroaktivních polymerních inkoustů na bázi PEDOT:PSS byla charakterizována a diskutována s ohledem na jejich možné použití v bioelektronice. Nakonec byl zkonstruován organický bioelektronický senzor pro detekci fyziologické odpovědi kardiomyocytů na základě studovaných materiálů.

Die Anwendung von körperfremden Materialien zur Behandlung verschiedenster Krankheitsbilder, wie zum Beispiel als Zahnersatz oder Knochenstabilisierung, ist seit Jahrtausenden fester Bestandteil in der Medizin. Während damals hauptsächlich stabile Materialien genutzt wurden, die möglichst wenig mit dem menschlichen Körper interagieren, wird heutzutage ein anderer Ansatz verfolgt. Intelligente Materialien können nicht nur passiv die Heilung unterstützen, sondern aktiv zu ihr beitragen. Ein berühmtes Beispiel hierfür ist das Formgedächtnismaterial Nitinol, das in Stents zur Behandlung verengter Arterien eingesetzt wird. Diese Arbeit beschäftigt sich mit Eisen-Palladium, einem neuen Formgedächtnismaterial, bei dem der Effekt nicht wie bei Nitinol über eine Temperaturänderung sondern durch ein äußeres Magnetfeld induziert wird. Da man somit körpertemperaturbedingte Restriktionen in biomedizinischen Anwendungen umgehen kann, birgt Eisen-Palladium ein hohes Potential für Drug-Delivery Systeme oder mikromechanische Pumpen. Da eine optimale Verträglichkeit des Materials mit seiner biologischen Umgebung absolut unabdingbar ist, untersucht diese Arbeit verschiedene Möglichkeiten, die Oberfläche zu modifizieren und somit die Adhäsion biologischer Zellen zu unterstützen. Zu diesem Zweck wurden das Peptid RGD als spezifische Zelladhäsionssequenz, ein Plasmapolymer auf L-Lysin Basis als unspezifische Beschichtung und die Nanostrukturierung der Eisen-Palladium Oberfläche durch Glanzwinkeldeposition untersucht. Die verwendeten Methoden beinhalten Immunofluoreszenztests zur Quantifizierung der fokalen Kontakte zwischen Zellen und Material, theoretische Dichtefunktionaltheorie Rechnungen, sowie Kontraktilitätsmessungen mittels eines selbst entwickelten Biegebalkenaufbaus. Somit gelingt es in dieser Arbeit, die gegenseitigen Beziehungen des Materials mit der jeweiligen Oberflächenmodifikation mit den lebenden Zellen aus verschiedenen Blickwinkeln zu analysieren. Durch eine Kombination aus experimentellen und theoretischen Methoden werden die Stärken und Schwächen der einzelnen Funktionalisierungsmethoden beleuchtet und die Bildung fokaler Kontakte für eine verbesserte Zelladhäsion wird maßgeblich verbessert.

Titanium has been used in bio-medical implants for decades due to its superior biocompatibility. To improve the osseointegration of dental and orthopaedic implants, various surface modification techniques have been used including laser surface texturing. In particular, short-pulsed lasers, such as femtosecond and picosecond lasers, are widely used for surface modification. In this thesis, commercially pure Ti surfaces are modified by a femtosecond laser to explore the relationship between surface topography, surface chemistry, surface wettability, and biocompatibility with the goal of improving the osseointegration of implants. The laser textured surfaces consist of 1μm wide grooves spaced 10 μm, 4.8 μm, 2.4 μm and 1.2 μm apart. Gradient configurations where the groove spacing varies are also investigated. Surface morphology was characterized using Optical Microscopy (OM), Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). A custom-build contact angle measurement apparatus is used to investigate the wettability of the laser textured surfaces using the sessile drop method. Freshly laser-treated commercially pure Ti surfaces are found to be super-hydrophilic and become hydrophobic over time when exposed to air. The presence of grooves can accelerate the evolution of the contact angle over time, and introduces anisotropy in the wetting behavior (along vs. across the grooves). The hydrophilicity of laser treated surfaces can be retained by storing samples in ethanol. X-ray Photoelectron Spectroscopy (XPS) shows that the relative carbon content increases over time when Ti samples are exposed to air, which results in the subsequent evolution of the contact angle and cell response to laser textured Ti surfaces. Besides, laser treatment promotes the oxidation of pure Ti, and the product, TiO2, is responsible for the better biocompatibility. In vitro experiments using MG 63s osteoblast-like cells are implemented on laser-treated Ti surfaces and polished surfaces (control) with 1 day, 3 days and 7 days of cell culture. The best cell outcome was obtained by storing samples in air for 1 week, where storing for shorter or longer times resulted in the worst outcome, especially in the early stages of cell adhesion. There does not appear to be a direct link between wettability and the fate of cells on Ti surfaces. Indeed, while samples stored in air and ethanol have drastically different contact angle measurements (the former being hydrophobic and the latter hydrophilic), the cell behavior was unaffected. In addition, while wettability and laser treatment can affect the early stages of cell adhesion, they do not have a strong effect on the number of cells at longer incubation times (3 and 7 days). Laser machining does however affect the cell morphology and alignment, where cells preferentially align themselves parallel to the direction of the laser machined grooves with an elongated morphology.

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M. Tech. (Department of Metallurgical Engineering, Faculty of Engineering and Technology), Vaal University of Technology. / Elemental titanium (Ti), niobium (Nb), zirconium (Zr), and iron (Fe) powders were used to fabricate four near-β alloys with non-toxic of composition Ti-34Nb-25Zr, Ti-34Nb-25Zr-0.4Fe, Ti-34Nb-25Zr-1.2Fe, and Ti-34Nb-25Zr-2Fe (wt. %) (TNZ and TNZF) using spark plasma sintering (SPS) of nano-crystalline powders attained by high energy ball milling. The fabricated alloys were compared to Ti-34Nb-25Zr (used as a benchmark alloy in this study) and comparison was made with the commercially used Ti base alloys produced either by conventional methods or powder metallurgy. The powder mixtures were milled for 5 hours using a Simoloyer high energy ball mill with a ball to powder ratio of 10:1 and a rotational speed of 1000 rpm. This was followed by sintering the mechanically alloyed powders at 1100 ºC for 10 minutes with a pressure of 50 MPa and a heating rate of 100 ºC/min using an H-HP D25 spark plasma sintering furnace (FCT System, Germany). The powders were characterised for particle size and crystal structure using SEM and XRD. The consolidated components were characterised with regards to density, microstructure, mechanical properties. The electrochemical behaviour of the alloys was investigated using a Digi Ivy DY2300 series potentiostat. Three corrosion medium, Sodium chloride (NaCl), phosphate buffered saline solution (PBS) and Dulbecco’s modified eagle’s medium that mimic the conditions in the human body were used. Mouse myoblast cell line (C2C12) was used to investigate the biocompatibility of the sintered alloys in 1010x5 mm specimens using standard colorimetric assay MTT. Both electrochemical and biocompatibility test were conducted in triplicates and the results compared with that of the benchmark. Results of mechanical alloying of powder mixtures demonstrated an inhomogeneous structure. Milling for 5 hours resulted in agglomeration of small Fe and Zr particles. Milling for 3 hours resulted in a better distribution of elements compared to longer milling times. Therefore, sintering powders milled for 3 hours would have yielded better results. The densification results were acceptable and ranged between 97-99% of theoretical densities. Although some porosity was observed, especially on the un-etched microstructure. An insignificant decrease in density was observed when 1.2 (wt. %) Fe was added. The sintered samples had microstructures which were not homogenous. However, the addition of Fe yielded a more homogeneous microstructure compared to the one with less Fe. Therefore, TNZF with 2 (wt. %) Fe had a more homogenous microstructure. Sintering at 1100 ºC resulted in undissolved niobium and titanium which were observed in the microstructure as dark and white areas. The hardness of the TNZF alloys were comparable and lied between 373 and 432 Hv. These hardness values are higher than other similar titanium-based alloys fabricated using conventional methods. The addition of Fe to TNZ showed an insignificant decrease in hardness. The addition of Fe was found to decrease the Young’s Modulus of TNZ from 119.1 to 80 GPa with an addition of 2 wt.% Fe. However, an unacceptable reduction (230.91 to 158.2 MPa) in strength was also noticed. Pseudo passivation was observed when the alloys were immersed in 0.9 % Sodium Chloride (NaCl) which could be attributed to the inhomogeneity in the microstructure. The possibility of pitting corrosion was also observed. The alloy containing 2 Fe (wt.%) was found to be more corrosion resistant than the other alloys. The TNZF alloys exhibited better corrosion resistance in 0. 9% NaCl compared to phosphate buffered solution (PBS) and DMEM. The corrosion behaviour in PBS and DMEM cannot clearly be explained from the graphs. The morphology of the corroded samples was almost the same for all the alloys in different corrosion media. The microstructures showed pits which could have been from the pores that acted as initiation sites for pitting. In cell culture for 1 and 7 days, the cell viability for TNZF alloys was greater than that of the control group (TNZ). A significant decrease in cell viability for TNZF was observed in cell culture for 4 days. The addition of Fe on TNZ do not cause toxic effects and show good cell adhesion, indicating in-vitro cytocompatibility. The greatest cell viability of 102±3.0 % for Ti-34Nb-25Zr-2Fe. The analysis of cell morphology indicated good cell-substrate interaction. The TNZF alloys developed in this study can be suitable candidates for orthopaedic implant application due to their low Young’s modulus, corrosion resistance and superior biocompatibility. However, the strength needs significant improvement. The advantage of this biomaterial, when compared to commercial alloys, is the absence of cytotoxicity elements such as Al and V.

Proteomics is the large-scale study of proteins, particularly their structures and functions. Proteomics has been utilized in medicine for investigation of disease mechanisms and biomarker discovery. Instrumental methods cover sample preparation, protein and peptide separation and mass spectrometry. At present, there is no proteomic method that can be used as universal for every sample. Analytical methods need to be adapted and optimized for certain samples. The aim of this work was to create methodic procedures and to interpret results of experimental and clinical research. The first part of the thesis includes experiments utilizing proteomics to study changes in the plasma proteome clinically relevant porcine model of sepsis-induced peritonitis. Proteomic analyzes were also starting methodological strategies in experiments aimed at kidney physiology and pathophysiology of acute kidney injury during sepsis. Renal biopsies were analyzed in order to study the time course of proteome changes caused by sepsis and surgery. The second part of the thesis contains experiments studying biocompatibility. A method for elution of proteins interacting with adsorbents used in extracorporeal liver support system and with hemodialyzer capillaries was prepared. Analysis of proteins adsorbed to polysulfone...

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