Dissertations / Theses on the topic 'Titanium – Surfaces'

El conocimiento de las interacciones entre célula/proteína/biomaterial es fundamental para la ingeniería de superficies debido a las numerosas aplicaciones biomédicas y biotecnológicas que se están desarrollando así como al éxito clínico que han alcanzado muchos implantes. La respuesta biológica final inducida por los implantes está fuertemente influenciada por las interacciones superficiales entre los componentes biológicos y el material sintético. Las propiedades físicas y químicas de la superficie de un biomaterial, en lugar de las propiedades en su masa, influyen directamente en la capa de proteínas que se adsorben sobre el biomaterial y, como consecuencia de ello, en la respuesta celular a la misma, tanto in vitro como in vivo.

El objetivo de esta tesis doctoral es profundizar en el conocimiento de las interacciones material-biosistema, con el énfasis en el descubrimiento de relaciones entre las propiedades superficiales de las superficies de titanio y su respuesta biológica in vitro.

El titanio comercialmente puro (Ti c.p.) está siendo ampliamente utilizado con éxito durante muchos años como biomaterial para implantes en cirugía ósea. Su excelente biocompatibilidad se basa en sus adecuadas propiedades mecánicas y, con mayor importancia, en su excelente resistencia a la corrosión. Esta última se debe principalmente a la formación espontanea de una fina película de óxido de titanio que le confiere protección natural contra los ataques degradativos. La modificación de la topografía de la superficie del titanio ha sido objeto de investigación en el pasado con el fin de mejorar la osteointegración. El granallado de partículas es una de las tecnologías más utilizadas para conferir rugosidad a las superficies del titanio. La rugosidad óptima y el tipo de partículas abrasivas del granallado para una respuesta óptima in vitro e in vivo fue previamente determinada en nuestro laboratorio. Sin embargo, todavía están por determinar cuáles son las causas últimas que llevan al biomaterial a su exitosa respuesta biológica.

En este trabajo se han estudiado superficies pulidas y rugosas de Ti c.p. obtenidas mediante el granallado con partículas abrasivas de diferente composición química(Al2O3 y SiC) y diferentes tamaños (212-300μm; 425-600μm; 1000-1400μm). La completa caracterización de las propiedades física y química de la superficie, incluyendo la rugosidad, la composición química, la mojabilidad/energía libre y la carga eléctrica de las superficies ensayadas ha llevado a una serie de relevantes conclusiones. Entre ellas, cabe destacar que a) la composición química de las partículas de granallado, así como el método de esterilización fueron los principales factores que influyeron en la mojabilidad y la energía libre superficial de las superficies de titanio estudiadas, b) el método de esterilización cambió en la energía superficial el carácter de donante de electrones de las superficies mediante el cambio de la cantidad y la naturaleza de las sustancias adsorbidas, y c) la composición química de las partículas de granallado no influyó en la carga eléctrica a pH fisiológico ni en el punto isoeléctrico de las superficies.

Un segundo paso consistió en el uso de una microbalanza de cristal de cuarzo con monitorización de la energía de disipación, para el estudio de la cinética de adsorción (cantidad y conformación) y de los procesos de adsorción competitiva de tres proteínas de especial interés en los procesos de curación del hueso - la albúmina de suero bovino (BSA), el fibrinógeno (Fbg), y la fibronectina (Fn)- en sensores lisos recubiertos de TiO2. Se determinaron diferentes modelos de procesos de adsorción con una, dos o múltiples pasos distinguibles en función de las proteínas en solución. La capa adsorbida de BSA mostró los cambios más significativos en sus propiedades mecánicas, de conformación y de incorporación de agua hasta que se alcanzaron las condiciones estables de adsorción de proteínas. La BSA, la más pequeña de las proteínas ensayadas, desplazó la Fn y el Fbg cuando se ensayó en condiciones de la competencia por la adsorción, indicando su mayor afinidad por las superficies de TiO2. También se emplearon técnicas de marcaje fluorescente para el estudio de la adsorción proteica en superficies rugosas granalladas. En este estudio, por un parte, se pudo determinar que la cantidad de Fn y BSA adsorbidas en las superficies granalladas está directamente correlacionada con su energía superficial. Por otra parte, se visualizó la adsorción de fibronectina en solución sobre muestras granalladas rugosas de Ti. La Fn formó un patrón irregular de adsorción con una mayor cantidad de proteína adsorbida en los picos que en los valles de la topografía.

También se evaluó la organización espacial de la matriz extracelular de los osteoblastos, ECM, sobre superficies de Ti lisas y rugosas por medio de la visualización de las fibrillas de Fn teñidas con marcador fluorescente. Las células osteoblásticas depositaron las fibrillas de Fn con un determinado patrón organizado dentro de la matriz total secretada. Aparecen como una película que cubre la parte superior de las diferentes superficies rugosas de titanio. Un resultado relevante es que el espesor de esta capa aumentó con la rugosidad de la topografía subyacente. Sin embargo no más de la mitad de la máxima distancia pico-valle se cubrió con la proteína secretada y/o reorganizada.

Por último, teniendo en cuenta las diferencias en la organización de la ECM y la
adsorción de Fn en las superficies ensayadas de Ti, se realizó un estudio de qRT-PCR para determinar la influencia de las propiedades superficiales del titanio, con y sin preadsorción de Fn, en la respuesta osteoblástica. La expresión génica de la subunidad 5 de la integrina celular, como marcador de la adhesión celular, se incrementó en las superficies granalladas con SiC en comparación con las granalladas con alúmina. Este resultado fue correlacionado con la mayor cantidad de Fn adsorbida debido a la mayor energía superficial de las superficies granalladas con SiC. El aumento de la rugosidad, así como la presencia de partículas de alúmina en las superficies rugosas incrementó la actividad de ALP y la expresión génica de ALP mRNA por los osteoblastos, y por lo tanto su diferenciación.
The understanding of cell/protein/biomaterial interactions is critical to the engineering of substrates for numerous biomedical and biotechnological applications and to the clinical success of implants. The final biological response induced by implants is strongly influenced by the biological-components/synthetic-material surface interactions. It is well accepted that the physical and chemical surface properties of a biomaterial rather than its bulk properties will influence the protein adlayer and then the cell response to it, both in vitro and in vivo.

The aim of this PhD thesis is to gain an increased understanding of the materialbiosystem interactions, with an emphasis on establishing correlations between surface properties of titanium surfaces and its in vitro biological response.

Commercially pure titanium (c.p. Ti) is being widely and successfully used implant biomaterial in bone surgery over many years. Its excellent biocompatibility is based in its appropriate mechanical properties and, more importantly, in its excellent corrosion resistance, which is mainly due to the presence of a naturally-occurring thin protective titanium oxide film. Modification of titanium surface topography has been a subject of research in the past with the purpose of improving its osseointegration. Grit blasting is one of the most used technologies to roughen titanium surfaces for this purpose. The optimal roughness and type of abrasive blasting-particles for a better in vitro and in vivo response was previously determined in our lab. However, which and how different relevant surface properties of the blasted titanium surfaces induce that optimal biological behavior is still poorly understood.

Smooth/polished and rough c.p. Ti surfaces obtained by blasting with abrasive
particles of different chemical composition (Al2O3 and SiC) and different sizes (212-300μm; 425-600μm; 1000-1400μm) were studied. The comprehensive characterization of physical and chemical surface properties, including roughness, chemical composition, wettability/free energy and electrical charge of the tested surfaces led to a series of relevant conclusions. Among them, it is worth noting that a) the chemical composition of the grit-blasting particles as well as the method of sterilization were found the main factors influencing wettability and surface free energy of the titanium surfaces; b) the sterilization method changed the electron donor character of the surfaces by changing the amount/nature of physisorbed substances on the surfaces, and c) the chemical composition of the blasting particles did not influence on the electrical charge at physiological pH and the isoelectric point of the surfaces.

A second step consisted in the use of a quartz crystal microbalance with monitoring of the energy dissipation to study the adsorption kinetics (amount and conformation) and adsorption competition processes of three proteins of special interest in the healing processes of bone -bovine serum albumin (BSA), fibrinogen (Fbg), and fibronectin (Fn)-on smooth TiO2-coated sensors. Different patterns of adsorption with processes in one, two or multiple distinguishable steps were determined depending of the protein in solution. The BSA adlayers showed the most significant changes in their mechanical properties/conformation/incorporation of water until steady protein-adsorption conditions were reached. BSA, the smallest of the tested proteins, displaced Fn and Fbg when in competition for adsorption, which is an indication of its higher affinity for TiO2 surfaces. Fluorescent labelling techniques where used to study protein adsorption on blasted rough surfaces. Most significantly, the amount of Fn and BSA adsorbed on blasted surfaces was positively correlated with their surface energy. The adsorption of fibronectin from solution on shot-blasted rough titanium surfaces resulted in an irregular pattern of adsorption with a higher amount of protein adsorbed on peaks than on valleys of the topography.

Further, the spatial organization of the osteoblast extracellular matrix, ECM, on smooth and rough Ti surfaces was evaluated by visualizing fluorescently-stained Fn-fibrils. Osteoblast-like cells deposited Fn- fibrils in a specific facet-like pattern that was organized within the secreted total matrix. It appeared as a film overlying the top of the different rough titanium surfaces. Interestingly, the thickness of this layer increased with the roughness of the underlying topography, but no more than half of the total maximum peak-to-alley distance was covered.

Finally, taking into consideration the differences in ECM organization and Fn adsorption on the tested Ti surfaces a qRT-PCR study was carried out to elucidate the influence of titanium surface properties with and without Fn-precoatings on the osteoblast response. The expression of 5 integrin subunit gene, as a marker for cell adhesion, was increased in SiC-blasted surfaces compared to alumina-blasted surfaces. This was related to the higher amount of adhesive-protein Fn adsorbed caused by the higher surface energy of SiC-blasted surfaces. The increase of roughness as well as the presence of alumina particles on blasted surfaces increased ALP activity and ALP mRNA gene expression by osteoblasts, and so their differentiation.

This research work contribute to increase our knowledge on the interactions taking place at the bio/non-bio interface between different biological components -water, proteins, cells- and materials of clinical relevance, such as rough titanium. The
intertwined effects of the different properties of the synthetic surfaces appear as a challenge to unravel the ultimate causes that determine the fate of cells on synthetic biomaterials.

Prosthetic-associated infections are one of the most challenging postoperative complications for orthopedic implants. The consequences that infections may lead to include patient pain, high cost, prolonged hospitalization time, and usually the revision of the implant. Current prophylaxis and therapy utilizing antibiotics are facing an emergency of increasing bacterial resistance; the design of a novel anti-infectious implant surface is therefore required. Among the potential antimicrobial alternatives are the antimicrobial peptides (AMP). AMPs are a family of natural defense peptides that has not received enough recognition until recently. The complex killing mechanisms of these cationic peptides make them very unlikely to encounter resistant mutants, and their broad-spectrum activity offers them great opportunity in possible clinical applications. In this study, a novel short AMP Tet213 with prominent bactericidal activity was chosen as the antimicrobial candidate and was covalently attached to titanium surfaces through a short bifunctional linker. This designed routine was confirmed with single cysteine before being applied to the 9-mer AMP candidate. The surface density of the immobilized AMP was determined by detecting its arginine residues after a reaction with 9,10-phenanthrequenon (PHQ). The reaction between arginine and PHQ generates a fluorescent product, by the emission of which the quantity of the arginine-containing peptide can be calculated. The density of the surface-attached Tet213 was measured to be 1.30±0.55 μg/cm². A relatively large proportion of physically adsorbed Tet213 was also observed, with the net adsorbed quantity to be 0.74±0.20 μg/cm². The affinity of the cationic AMP to the bare titanium surface is believed to be a result of electrostatic interactions. Both the covalently immobilized and the physically adsorbed Tet213 showed bactericidal activities of generally > 50% against a Pseudomonas aeruginosa (P. aeruginosa) strain which constitutively expresses luminescence when alive. The inhibition rate was calculated by the luminescence reduction and confirmed by the colony counts of the surviving bacteria. Several parameters were found to be influential to the overall inhibition rate, including the selection of the AMP candidate, the dilution of the bacterial culture and the bacterial incubation time.

There exists a clinical requirement for dental implants which will enhance the speed of achievement of osseointegration, its maintenance, and biological and physical properties. Whilst commercially pure titanium remains the material of choice for implant fabrication, a promising approach to enhancing its performance is the surface incorporation of metallic ions, or alkali modification of titanium. Osteoblast behaviour adjacent to the implant is a key factor in osseointegration and it is known that the response of these cells can be modified by the surface implantation of Ca ions. This process may modify cellular behaviour via a number of physicochemical parameters, three of which were examined in this study using commercially pure titanium, into which Ca ions had been implanted: -topography, calcium ion release, and molecular adsorption. Surface topography can mediate cellular responses and may be modified by ion implantation. Laser profilometry and white light interferometry were used to measure the roughness of cp Ti surfaces implanted with either biologically active Ca or chemically inert Ar, together with the effects of nitric acid treatment, which is routinely used in implant manufacture. Ca-ion implantation may also influence cellular responses via accelerated precipitation of calcium phosphate, providing a surface with a chemical composition more similar to that of bone. This may be at least partially due to ion release from the implanted surface. Ion release into water was therefore investigated using ion chromatography and X-ray photoelectron spectroscopy (XPS). The adsorption of organic molecules (e.g. proteins and peptides) is also important in mediating cellular responses. The effects of Ca-implantation on these processes were investigated using XPS to study the surface adsorption of small model biomolecules (amino acids) from an aqueous solution. Ion implantation had little effect on surface topography, however, the implanted Ca ions were readily released into an aqueous solution and the surfaces became more receptive to the absorption of certain amino acids. It is concluded that Ca ion implantation is a potentially valuable technique for the surface enhancement of titanium dental implants.

Objective: Topography, chemistry, and energy of titanium (Ti) implants alter cell response through variations in protein adsorption, integrin expression, and downstream cell signaling. However, the contribution of surface energy on cell response is difficult to isolate because altered hydrophilicity can result from changes in surface chemistry or microstructure. Our aim was to examine a unique system of wettability gradients created on microstructured Ti on osteoblast maturation and phenotype. Method: A surface energy gradient was created on sand-blasted/acid-etched (SLA) Ti surfaces. Surfaces were treated with oxygen plasma for 2 minutes, and then allowed to age for 1, 12, 80, or 116 hours to generate a wettability gradient. Surfaces were characterized by contact angle and SEM. MG63 cells were cultured on SLA or experimental SLA surfaces to confluence on TCPS. Osteoblast differentiation (IBSP, RUNX2, ALP, OCN, OPG) and integrin subunits (ITG2, ITGA5, ITGAV, ITGB1) measured by real-time PCR (n=6 surfaces per variable analyzed by ANOVA/Bonferroni’s modified Student’s t-test). Result: After plasma treatment, SLA surface topography was retained. A gradient of wettability was obtained, with contact angles of 32.0° (SLA116), 23.3° (SLA80), 12.5° (SLA12), 7.9° (SLA1). All surfaces were significantly more hydrophilic than the original SLA surface (126.8°). Integrin expression was affected by wettability. ITGA2 was higher on wettable surfaces than on SLA, but was highest on SLA1. ITGAV and ITGB1 were decreased on hydrophilic surfaces, but ITGA5 was not affected. IBSP, RUNX2, and ALP increased and OPG decreased with increasing wettability. OCN decreased with increasing wettability, but levels on the most wettable surface were similar to SLA. Conclusion: Here we elucidated the role of surface energy on cell response using surfaces with the same topography and chemistry. The results show that osteoblastic maturation was regulated in a wettability-dependent manner and suggest that the effects are mediated by integrins.

In this thesis two studies of reactive metal adsorption upon a low index single crystal silicon dioxide surface are presented in addition to a study of sulphur adsorption upon a low index single crystal nickel surface. Chromium growth upon the a-quartz Si02(0001) (J84xJ84) Rll 0 surface is studied at three coverages, 0.25±O.08 ML, 0.5±O.16 ML and 1.0±0.33 ML, using surface extended x-ray absorption fine structure (SEXAFS). SEXAFS measurements, from the chromium K-edge, recorded at both grazing and normal incidence show that chromium growth proceeds via the formation of mesoscopic particles with a body centred cubic (b.c.c.) like structure having an average nearest neighbour Cr-Cr distance of 2.36±O.03 A. This represents a contraction of 5.6 % from the bulk b.c.c. lattice spacing of 2.49 A. There is no evidence of a surface reaction between chromium and the surface oxygen. SEXAFS was used to study titanium reactional growth on a-quartz (0001) (J84xJ84) Rll 0 and (lx1). Three nominal coverages were studied, 0.25±O.08 ML, 0.5±O.16 ML and 1.0±O.33 ML. Both normal and grazing incidence SEXAFS data were recorded and show the formation of a spatially extensive region in which an interfacial reaction has occurred between surface oxygen and adsorbate titanium atoms. Coupled with this is the formation of subnanometre titanium clusters. The metal oxide has nearest neighbour Ti-O distances close to those of both the anatase and rutile forms of titania with the metallic titanium clusters having a Ti-Ti distance within experimental error that of bulk hexagonal close packed (h.c.p.) titanium, 2.89 A. A re-examination of the surface geometry of Ni(1l0)c(2x2)S using SEXAFS has been performed. Data out to an electron wavevector of 9 A-I are analysed with a new code to assess the influence of multiple scattering. The first shell S-Ni distance is determined to be 2.20±O.02 A with the next nearest neighbour distance being 2.29±O.02 A, giving a top-layer Ni expansion of 14±3% relative to the bulk. The influence of multiple scattering does not significantly alter these values from earlier studies.

Biomaterial surface energy, chemical composition, charge, wettability and roughness all play an important role in determining the degree of the direct bone-to-implant interface, termed osseointegration. Surface chemistry, which is influenced by surface energy, wettability, and composition, is another factor that determines osteoblast phenotype and regulates osteoblast maturation. Increased surface energy is desirable for bone implants due to enhanced interaction between the implant surface and the biological environment. The extent of bone formation in vivo is also increased with increasing water wettability of implants. The physiological role of implant surface chemistry is important in determining the success of implant osseointegration because of molecular rearrangements, surface reactions, contamination, and release of toxic or biologically active ions that are determined by the starting chemistry. However, the role of surface chemistry on osteoblast response is not fully studied. Therefore, the overall goal of this dissertation is to understand how the surface chemistry, including wettability, chemical composition, and charge density, of titanium biomaterials impacts osteoblast maturation (in vitro). This study focuses on the general hypothesis that modifications of surface chemistry of titanium surfaces with sterilization or polyelectrolyte coating on titanium surfaces regulate osteoblast response.

In the current work, two different coatings, nitrocarburised (CN) and titanium carbonitride (TiCN) on M2 grade high speed tool steel, were prepared by commercial diffusion and physical vapour deposition (PVD) techniques, respectively. Properties of the coating were characterised using a variety of techniques such as Glow-Discharge Optical Emission Spectrometry (GD-OES) and Scanning Electron Microscopy (SEM). Three non-commercial, oil-based lubricants with simplified formulations were used for this study. A tribological test was developed in which two nominally geometrically-identical crossed cylinders slide over each other under selected test conditions. This test was used to evaluate the effectiveness of a pre-applied lubricant film and a surface coating for various conditions of sliding wear. Engineered surface coatings can significantly improve wear resistance of the tool surface but their sliding wear performances strongly depend on the type of coating and lubricant combination used. These coating-lubricant interactions can also have a very strong effect on the useful life of the lubricant in a tribological system. Better performance of lubricants during the sliding wear testing was achieved hen used with the nitrocarburised (CN) coating. To understand the nature of the interactions and their possible effects on the coating-lubricant system, several surface analysis techniques were used. The molecular level investigation of Fourier Transform Infrared Spectroscopy (FTIR) revealed that oxidative degradation occurred in all used oil-based lubricants during the sliding wear test but the degradation behaviour of oil-based lubricants varied with the coating-lubricant system and the wear conditions. The main differences in the carbonyl oxidation region of the FTIR spectra (1900-1600 cm-1) between different coating-lubricant systems may relate to the effective lifetime of the lubricant during the sliding wear test. Secondary Ion Mass Spectrometry (SIMS) depth profiling shows that the CN coating has the highest lubricant absorbability among the tested tool surfaces. Diffusion of chlorine (C1), hydrogen (H) and oxygen (O) into the surface of subsurface of the tool suggested that strong interactions occurred between lubricant and tool surface during the sliding wear test. The possible effects of the interactions on the performance of whole tribological system are also discussed. The study of Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) indicated that the envelope of hydrocarbons (CmHn) of oil lubricant in the positive TOF-SIMS spectra shifted to lower mass fragment after the sliding wear testing due to the breakage of long-chain hydrocarbons to short-chain ones during the degradation of lubricant. The shift of the mass fragment range of the hydrocarbon (CmHn) envelope caries with the type of both tool surface and lubricant, again confirming that variation in the performance of the tool-lubricant system relates to the changes in surface chemistry due to tribochemical interactions at the tool-lubricant interface under sliding wear conditions. The sliding wear conditions resulted in changes not only in topography of the tool surface due to mechanical interactions, as outlined in Chapter 5, but also in surface chemistry due to tribochemical interactions, as discussed in Chapters 6 and 7.

Thesis (M.S.)--University of North Carolina at Chapel Hill, 2007.
Title from electronic title page (viewed Dec. 18, 2007). "... in partial fulfillment of the requirements for the degree of Master in Science in the Department of Prosthodontics, School of Dentistry." Discipline: Prosthodontics; Department/School: Dentistry.

With increasingly higher performance requirements and health care costs associated with dental and orthopaedic bone-anchored implants there is a need to improve the osseointegration of Ti implants. Small alterations in nanotopography feature dimensions and arrangement has recently been shown able to differentiate human mesenchymal stem cells into an osteoblastic lineage, thus showing potential for using nanotopography to improve osseointegration of Ti implants.

Esta tesis queda enmarcada en el ámbito de los biomateriales metálicos, concretamente en superficies de titanio desarrolladas para la regeneración ósea. Las aplicaciones más habituales del titanio como biomaterial son los implantes dentales y las prótesis de cadera y rodilla. Estos componentes requieren, en servicio, buena estabilidad y fijación al hueso a largo plazo. El titanio es un material idóneo para el cumplimiento de estos requisitos gracias a su alta resistencia mecánica, tenacidad, resistencia a la corrosión y, sobre todo, por su alta capacidad de osteointegración. En general, el titanio es un biomaterial bioinerte donde, una vez implantado, el tejido vivo genera una fina capa de tejido fibroso alrededor del implante la cual separa el hueso del implante. Un espesor excesivo de esta capa de tejido fibroso puede comprometer la estabilidad e integración del implante y conllevar el fracaso del tratamiento. El objetivo principal de esta tesis es el desarrollo de una nueva superficie de titanio que sea capaz de controlar e inhibir la generación de tejido fibroso en la superficie del implante. De esta manera, tratamos de mejorar la osteointegración de implantes y prótesis mediante la mejora de la respuesta celular sobre la superficie del implante. Para el control del crecimiento de tejido fibroso en la superficie se han desarrollado nuevas superficies de titanio donde se han inmovilizado dos tipos de péptidos cortos capaces de inhibir la interacción de la citoquina TGF-β, la cual incrementa la producción de este tipo de tejido por parte de las células fibroblásticas. Estos péptidos, llamados P17 y P144 han sido desarrollados por el equipo de nuestro colaborador el Dr. Francisco Borrás-Cuesta, en el Centro de Investigación Médica aplicada de la Universidad de Navarra. Esta tesis está dividida en 6 capítulos donde se describe el desarrollo y caracterización de las superficies de titanio funcionalizadas con péptidos inhibidores del TGF-β: • Capítulo 1: Introducción a los ámbitos y conceptos importantes de la tesis. • Capítulo 2: Diseño y desarrollo de un método de inmovilización covalente de péptidos cortos sobre superficies de titanio. • Capítulo 3: Estudio de los factores que intervienen en la inmovilización de péptidos cortos sobre las superficies de titanio. • Capítulo 4: Caracterización físico-química de las superficies de titanio funcionalizadas con el péptido P17. • Capítulo 5: Caracterización físico-química de las superficies de titanio funcionalizadas con el péptido P144. • Capítulo 6: Respuesta biológica in vitro de las superficies de titanio funcionalizadas con P17 y P144. Los resultados más relevantes en el desarrollo de esta tesis han sido: • El desarrollo de un nuevo método de inmovilización covalente de péptidos sobre superficies de titanio obteniendo una alta densidad de péptido en superficie con una buena estabilidad mecánica y termoquímica. • La consecución de superficies de titanio capaces de inhibir la acción del TGF-β. • Las nuevas superficies desarrolladas son capaces de incrementar la diferenciación osteoblástica y así, potencialmente mejorando la capacidad de osteointegración de implantes y prótesis de titanio. Este trabajo de investigación contribuye a aumentar el conocimiento sobre la inmovilización covalente y no covalente de péptidos cortos en superficies de titanio. También contribuye en aumentar el conocimiento de la acción e inhibición del TGF-β en células fibroblasticas y osteoblásticas, estas últimas sembradas sobre superficies de titanio. El material desarrollado es un excelente candidato para su aplicación en implantología y traumatología ósea.
This thesis is framed in the field of metallic biomaterials, specifically on titanium surfaces developed for bone regeneration. The most common applications of titanium as a biomaterial are dental implants and hip and knee prostheses. These components clinically require good stability and fixation to the bone in the long term. Titanium is an ideal material for these applications as it has high mechanical strength, toughness, corrosion resistance and, above all, a high capacity for osseointegration. In general, titanium is a bioinert material where, once implanted, the living tissue generates a thin layer of fibrous tissue around the implant which separates the bone to the implant. An excessive thickness of this layer of fibrous tissue can compromise the stability and integration of the implant leading to the failure of the biomedical treatment. The main objective of this thesis is the development of a new titanium surface with control and inhibition of the generation of fibrous tissue on the surface of the implant. We aim improving the osseointegration of implants and prostheses by benefiting cellular responses on the surface of the implant. For the control of the formation of fibrous tissue on the surface we have developed new biofunctional titanium surfaces by covalently immobilizing two different short peptides on the metallic substrate. These two peptides are inhibitors of the effect of the cytokine TGF-β1, which increases the production of fibrous tissue by the activity of fibroblastic cells. These peptides, P17 and P144, have been developed by the team of our collaborators at the Dr. Francisco Borrás-Cuesta’s lab, in the Centro de Investigación Médica Aplicada of the Universidad de Navarra This thesis is divided into 6 chapters describing the development and characterization of titanium surfaces functionalized with TGF-β inhibitor peptides: * Chapter 1: Introduction to the areas and important concepts of the thesis. • Chapter 2: Design and development of a method of covalent immobilization of short peptides on titanium surfaces. • Chapter 3: Study of the factors involved in the immobilization of short peptides on the titanium surfaces. • Chapter 4: Physical-chemical characterization of titanium surfaces functionalized with the P17 peptide. • Chapter 5: Physical-chemical characterization of titanium surfaces functionalized with the P144 peptide. • Chapter 6: In vitro biological response of titanium surfaces functionalized with P17 and P144. The most relevant results in the development of this thesis are: • The development of a new method of covalent immobilization of peptides on titanium surfaces with a high density of peptide on the surface and with a good mechanical and thermal-chemical stability. • The development of titanium surfaces with inhibitory action of TGF-β activity. • The developed new surfaces are able to increase osteoblast differentiation, thereby potentially enhancing osseointegration of the biofunctionalized titanium implants and prostheses. This research work contributes to increase the knowledge on covalent and noncovalent immobilization of short peptides on titanium surfaces. It also helps in increasing the knowledge of the action and inhibition of TGF-β on fibroblastic and osteoblastic cells; the later seeded on titanium surfaces. The developed material is an excellent candidate for its application in implantology and orthopedics.

Avec le développement de nouveaux dispositifs apparait le besoin d’être capable de contrôler la chimique de surfaces de substrats multi-échelle et multi-matériaux. Plusieurs techniques font appel à de la chimie localisée via différentes technologies. Une approche consiste à exploiter les différences de réactivités chimiques entre les différents matériaux du substrat et différents groupements chimiques de manière à fonctionnaliser sélectivement chaque matériaux pour former des couches minces organiques de type « Self Assembled Monolayer »: Ce principe proposée par Pr. G. M. Whitesides est appelé chimie orthogonale. Dans le cadre de cette thèse, le but ultime était de réaliser la fonctionnalisation chimique orthogonale de substrats dont la surface était composée de SiO2/Au/TiW La première étape de ce travail a été de déterminer pour la première fois la fonction chimique la plus adaptée pour la fonctionnalisation de TiW. Pour se faire nous avons comparé la chimie des silanes, des acides phosphoniques et des catéchols. Après caractérisations (XPS, ToF-SIMS, IR) des différentes couches, la voie des acides phosphoniques semblait être celle donnant lieu à la couche la plus stable. Ensuite nous avons étudié l’orthogonalité sur de substrats bi-matériaux (SiO2/TiW ou Au/TiW), et enfin sur substrat dont la surface était constituée de Au/SiO2/TiW
The development of nanotechnologies makes it possible to manufacture the micro or nanometric-sized patterns with various materials (dielectrics, metals, semiconductors). These heterogeneous surfaces are commonly used in the electronics industry for the production of nanoelectronic structures and components: transistors, memories or sensors. The concept of orthogonal chemical functionalization was first proposed by George M. Whitesides to modify the surfaces composed of different materials at the macroscopic scale. In this context, this PhD work aimed at exploring the orthogonal chemical functionalization approach on a predefined patterned titanium tungsten (TiW) surface by lithography producing. Pattern materials (Au, SiO2) are chosen to have different chemical properties, which can be functionalized with completely independent reactions. To achieve this aim, we have studied three different chemical groups for the formation of organolayers (silane, catechol, phosphonic acid) on TiW for the first time. The three layers were characterized (XPS, IR, ToF-SIMS) and the stability of the formed organolayers was also addressed. Then we developed and ascertained the orthogonal chemical functionalization of patterned Au/TiW and Au/SiO2/TiW surfaces. It proposes a novel strategy for the orthogonal functionalization on a triple-material patterned surface. In addition, the capture of nanoparticles by electrostatic interaction at specific location on Au/TiW patterned substrate was successfully implemented to prove the interest of such method for colloids trapping

This thesis presents surface science studies, investigating several aspects of titanium dioxide at the atomic scale. The greater part of this work is devoted to the preparation by chemical vapor deposition (CVD) of titanium(IV) tetraisopropoxide (TTIP) of ultrathin TiO2 or TiOx films on Au(111). Four ordered structures were growth and characterized. It was also demonstrated how the morphology of the film (wetting film vs island) can be tailored. The acquired knowledge about the CVD process was exploited to load nano porous gold with titania, enhancing its catalytic activity. The reactivity towards water adsorption of the titania structures on Au(111) was also investigated. Finally, part of this work concerned the studying of the behavior of water on the stoichiometric rutile TiO2(110) surface, combining the experiments with density-functional theory (DFT) calculations and (kinetic) Monte Carlo simulations. The main experimental techniques used in this work are low-energy electron diffraction (LEED), scanning tunneling microscopy (STM) and photoelectron spectroscopy (PES).

An investigation of surface structures formed on polycrystalline and single crystal TiO2 (titania) samples having under gone various heat treatments in a controlled hydrogen bearing atmosphere was conducted. The study included the recreation and examination of the process discovered by Sehoon Yoo at Ohio State University to form nanofibers on the surface of polycrystalline TiO2 disks. Fibers were formed by heating samples to 700??in a 5%H2 95%N2 gas stream. The nanofibers formed during this processes are approximately 5-20 nanometers in diameter and can be 100??f nanometers long. The fibers do not actually grow on the surface, but are what remain of the surface as the material around them is removed by the gas stream V i.e., nanocarving. The mechanism of fiber formation and the effect of varying experimental parameters remained unknown and were explored within this study. This included changing gas composition, flow rate, and changes in sample preparation. The effect of isovalent doping and impurities within the starting powder were examined. Sintering temperature and time was investigated to determine the effect of grain size and surface morphologies prior to nanocarving. The effect of elevated temperature and 5%H2 95%N gas on the surface of TiO2 single-crystal wafers was also investigated. Test methods include Thermogravimetric Analysis (TGA), Mass Spectrometry (MS), Scanning Electron Microscopy (SEM), and X-ray diffraction (XRD) analysis.

The aims of this study were to, 1) investigate the surface characteristics of polished titanium plates/discs treated with acid-etching and ultraviolet irradiation (UV); and 2) investigate the change in surface characteristics of polished titanium plates/discs treated with acid-etching and UV irradiation after aging in air, saline and citric acid for six weeks. A total of one hundred and ninety-eight commercially pure grade two titanium plates and 50 titanium discs were prepared. Titanium samples were divided into four groups: 1) polished group, 2) polished and UV group, 3) acid-etched group and 4) acid-etched and UV group. Polishing was performed by abrasive silicon carbide paper grinding. UV treatment was performed by 15W germicidal UVC, 254 nm, for 48 h. Acid-etching was performed with 67% H2SO4 at 120 ºC for 75 s. The four groups were then subjected to an aging process for six weeks in sealed containers with three different media: air, physiologic saline and citric acid. They were analyzed immediately after treatment and after aging for surface characterization: topography, roughness, wettability, crystallinity, and chemistry. The polished surface showed relatively smooth surface with typical grooves from the grinding process. Acid-etching produced micro-roughened surface with sharp pits and ridges. The average surface roughness of polished, polished-UV was lower than that of the acid-etched and acid-etched-UV surfaces (p < 0.05). Storage of titanium in saline and citric acid did not provoke any morphological or roughness changes at micron scale level when compared to the samples stored in air. Immediately after preparation and treatment, the polished and acid-etched titanium surfaces appeared to be hydrophilic with similar contact angle values (p > 0.05). After UV treatment, there was a significant reduction in contact angles (superhydrophilic) in both surfaces (p < 0.05). After storage in air for six weeks, the contact angles of the four groups significantly increased (p < 0.05) and the surfaces converted to a hydrophobic state. In contrast to samples stored in air, samples stored in saline and citric acid revealed superhydrophilic surfaces regardless of the surface type. Titanium hydride crystals were present in the acid-etched and acid-etched-UV surfaces but not in the polished and polished-UV surfaces. Titanium tetrachloride crystals were present in saline-stored surfaces. The polished surface acquired significantly higher titanium and oxygen concentrations and lower carbon contaminants compared to acid-etched surface. UV treatment substantially decreased carbon contamination and increased the titanium and oxygen concentrations in the acid-etched groups (p < 0.05). After storage for six weeks in air, the four studied surfaces had no significant changes in the surface chemistry. Storage of the titanium samples in saline and citric acid relatively increased the carbon contamination and decreased titanium and oxygen concentrations. UV treatment may be an effective way to produce clean titanium surfaces with less carbon especially after roughening the titanium surface by acid-etching. Storage of the freshly prepared titanium surface in media such as saline or citric acid could preserve the hydrophilic property of these surfaces, however, it may also negatively influence the surface chemistry due to increased carbon contaminants.
published_or_final_version
Dentistry
Master
Master of Philosophy

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.

Background: As the utilization of dental implants to replace missing teeth becomes more common, so does the incidence of peri-implant diseases, peri-implant mucositis and implantitis, caused by bacterial biofilm. The aim of this study was to evaluate the efficacy of removing bacteria from smooth-surface titanium using an air-polishing device with glycine powder abrasive.

Methods: Smooth-surface titanium discs were inoculated individually with Aggregatibacter actinomycetemcomitans, Streptococcus oralis, Streptococcus mutans, and Tannerella forsythia. Discs were treated with an air-polishing device with glycine powder and water, glycine powder alone, water alone, or tetracycline and water. Bacteria remaining on the surface of the titanium disks were removed and quantified.

Results: Compared with untreated control discs, all treatment types saw a statistically significant reduction in bacteria (p <0.01) regardless of bacterial species. There were no statistically significant differences in bacterial reduction between treatment types. Although not statistically significant, air polishing with glycine powder alone tended to be least effective in reducing total residual bacteria, while Tannerella forsythia showed the least amount of bacterial reduction.

Conclusions: For smooth surface titanium, air polishing with glycine powder and water, glycine powder alone, water alone, or tetracycline and water significantly reduces the amount of Streptococcus oralis, Streptococcus mutans, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia.

Postoperative infections are one of the most feared complications following orthopedic implant procedures. Bacterial infections occur in approximately 1-2% of the patients who undergo orthopedic implant surgery. Treatment of these infections is typically done by administering antibiotics either locally or systemically. Systemic release of antibiotics from bone cement has been reasonably successful. However, it would also be desirable to develop a method of antibiotic release from porous coated implants designed for osseointegration. The principal objective of this research was to explore the feasibility of using anodizing (electrochemical oxidation) as a surface modification technique to facilitate the attachment of antibiotics to commercially pure titanium (CP Ti) and Ti-6Al-4V orthopedic implant materials. In particular the effect of anodizing conditions on the characteristics of the oxide coating such as thickness, composition and porosity has been investigated. Using microbiological methods, the efficacy of in-vitro attachment of antibiotics to anodized surfaces was determined.

Biocompatible and corrosion resistant regarding biological fluids, titanium is still an inert material: it does not participate actively at the tissue-integration around the implant. Surface modification of titanium at nanoscale can promote cell adhesion and differentiation by modulate genes expression due to a phenomenon of mechano-transduction. In this thesis, we have focused on the development, the characterization and the direct application of our nanostructured surface of medical devices. First of all, our study focused on the preparation and physicochemical characterization. After obtaining reproducible surfaces on small samples, our research has focused on the biological characterization of the surface. In-vivo studies conducted in rabbits have allowed us to show similar biomechanical attachment and good osseointegration of the nanostructured surface compared to commonly used surfaces on the market. The application of this new surface on more complex titanium implant such as tracheal prosthesis, has allowed us to observe delamination phenomenon of the nanostructure layer. Therefore, our research was oriented towards the problem of mechanical strength of the surface with the realization of nano scratch test and tribology. A topic in the zeitgeist, as a new EU regulation for medical devices which incorporate nanomaterials will take effect in 2017. To conclude, this work enable to propose a nanosurface with promising results of tissues integration required for medical device
Biocompatible et résistant à la corrosion des fluides biologiques, le titane reste cependant un matériau inerte : il ne favorise pas de manière active l'intégration osseuse autour de l'implant. La modification de surface du titane à l'échelle nanométrique permet de moduler l'expression des gènes favorisant l'adhésion et la différentiation cellulaire par un mécanisme de mécanotransduction. Dans ces travaux de thèse, nous nous sommes donc attachés développer, caractériser et appliquer une surface nanostructurée directement sur des dispositifs médicaux. Dans un premier temps, notre étude s'est concentrée sur la préparation et la caractérisation physicochimique. Après l'obtention de surface reproductible sur petits échantillons, nos recherches se sont axées sur la caractérisation biologique de la surface. Des études invivo réalisées chez le lapin ont permis de montrer une accroche osseuse renforcée et bonne ostéointégration de la surface nanostructurée en comparaison avec des surfaces couramment utilisées sur le marché. L'application de cette nouvelle surface sur pièce plus complexe comme les prothèses de trachée, nous a permis de rendre compte d'un phénomène de délamination de la couche de nanostructure. Nos recherches se sont donc orientées vers la problématique de tenue mécanique de la surface avec la réalisation de nano scratch-test et tribologie. Un sujet dans l'air du temps, puisqu'une nouvelle règlementation européenne concernant l'incorporation de nanomatériaux dans les dispositifs médicaux rentrera en vigueur en 2017. En conclusion, ces travaux nous permettent de proposer une nouvelle surface améliorant l'intégration tissulaire intéressante pour une application médicale

This thesis addresses two separate problems - an investigation of the interaction of probe molecules with crystalline rutile and an investigation of the environment of group IA and IIA elements in organometallic compounds. Ab-initio Hartree-Fock calculations have been performed, aimed at investigating the interactions between the ionic surface of a crystal and an adsorbate molecule. Titanium dioxide, a material important for catalysis, electronic components and pigments, was chosen as the substrate, with carbon monoxide as the probe molecule. The calculations were carried out using the Crystal92 program, for the (110) surface of the Rutile polymorph of TiO(_2), employing a slab with a thickness of 5 atomic layers. The calculations investigated two orientations of the CO molecule with the molecular axis perpendicular to the surface. Results are reported showing contour diagrams for slices through the energy hypersurface parallel and perpendicular to the surface of the substrate. In order to facilitate the work described above, a program 'Builder2' was developed. This provides a convenient means for generating models of slabs of material from crystal structure data. Part of the development of Builder2 was to devise computer code to decompose standard Space Group symbols into the underlying symmetry matrices. The code for Builder2 is proprietary to Oxford Materials Ltd. and forms part of a commercial product. The environment of group IA and IIA elements in crystalline materials has not been the subject of any reported investigation. These elements, and organic ligands associated with them, play a significant role in biological systems. Around 16,000 atomic environments were extracted from the Cambridge Crystallographic Database to provide an up-to-date analysis of actual environments. The results are presented as histograms and tables, and suggestions are made for future extension of the analyses.

Thesis (M.S.)--Biomedical Engineering, Georgia Institute of Technology, 2009.
Committee Chair: Boyan, Barbara; Committee Member: Eskin, Suzanne; Committee Member: Lobachev, Kirill; Committee Member: Schwartz, Zvi. Part of the SMARTech Electronic Thesis and Dissertation Collection.

Titanium (Ti) dental implants are a successful treatment modality to replace missing teeth. Success is traditionally defined as the retention of the Ti dental implant but fails to account for peri-implant inflammatory diseases such as peri-implant mucositis and peri-implantitis. Peri-implant diseases are caused by the formation of pathogenic bacteria biofilms on the implant surface and disease progression can lead to dysfunctional and unaesthetic outcomes. There is no universally accepted treatment or management protocol for peri-implant disease. The objectives were to develop methods to prevent bacterial adhesion to Ti implant surfaces or treat existing biofilms. The relationship between bacterial adhesion of common early coloniser bacteria and topological features on dental implant surfaces was studied. Reproducible model systems were identified to be used in studies of biofilm formation and disruption. Early bacterial adhesion was investigated on engineered Ti surfaces created using Scanning-Laser-Melting or on Ti nanotubule surfaces. Photoactivation of Ti oxide films was investigated on thermally or anodically oxidised Ti and demonstrated the potential to pre-treat implant surfaces to reduce bacterial attachment. Finally chemical disinfection of Ti surfaces with a novel Eucalyptus Oil (EO) based formulation was demonstrated to increase the permeation of bactericidal agents into immature biofilms formed on Ti surfaces.

Theoretical methods can be used to describe surface chemical reactions in detail and with sufficient accuracy. Advances, especially in density functional theory (DFT) method, enable to compare computational results with experiments. Quantum chemical calculations employing ONIOM DFT/B3LYP/6-31G**-MM/UFF cluster method provided in Gaussian 03 are conducted to investigate water adsorption on rutile (110), and water and ammonia adsorption on anatase (001) surfaces of titanium dioxide. Water and ammonia adsorption on anatase (001) surface is studied by also performing PW:DFT-GGA-PW91 periodic DFT method by using VASP code and the results are compared with the results of ONIOM method. The results obtained by means of ONIOM method indicate that dissociative water adsorption on rutile (110) surface is not favorable due to high activation barrier, whereas on anatase (001) surface, it is favorable since molecular and dissociative water adsorption energies are calculated to be -23.9 kcal/mol and -58.12 kcal/mol. Moreover, on anatase (001) surface, dissociative ammonia adsorption is found energetically more favorable than molecular one (-37.17 kcal/mol vs. -23.28 kcal/mol). Thermodynamic functions at specific experimental temperatures for water and ammonia adsorption reactions on anatase (001) surface are also evaluated. The results obtained using periodic DFT method concerning water adsorption on anatase (001) surface indicate that dissociative adsorption is more favorable than molecular one (-32.28 kcal/mol vs. -14.62 kcal/mol) as in ONIOM method. On the same surface molecular ammonia adsorption energy is computed as -25.44 kcal/mol. The vibration frequencies are also computed for optimized geometries of adsorbed molecules. Finally, computed adsorption energy and vibration frequency values are found comparable with the values reported in literature.

Protein phosphorylation is a very common posttranslational modification (PTM), which lately has been found to hold the keyrole in the development of many severe diseases, including cancer. Thereby, phosphoprotein analysis tools, generally based on specific enrichment of the phosphoryl group, have been a hot topic during the last decade. In this thesis, two new TiO2-based on-target enrichment methods are developed and presented together with enlightening fundamental results. Evaluation of the developed methods was performed by the analysis of: custom peptides, β-casein, drinking milk, and the viral protein pIIIa. The results show that: i) by optimizing the enrichment protocol (first method), new phosphorylated peptides can be found and ii) by the addition of a separation step after the enrichment (second method), more multi-phosphorylated peptides, which usually are hard to find, could be detected. The fundamental part, on the other hand, shows that the phosphopeptide adsorption is caused by electrostatic interactions, in general follows the Langmuir model, and the affinity increases with the phosphorylation degree. Here, however, the complexity of the system was also discovered, as the adsorption mechanism was found to be affected by the amino acid sequence of the phosphopeptide.

Selective Serotonin Reuptake Inhibitors (SSRIs) are the most frequently prescribed class of drugs worldwide and are implemented in the treatment of depression and other psychiatric disorders. SSRIs relieve depressive symptoms by modulating levels of the neurotransmitter serotonin in the brain. SSRIs block the function of the serotonin transporter, thereby increasing concentrations of extracellular serotonin. However, serotonin levels in the neurons of the brain only account for 5% while the remaining 95% is present outside the brain. Serotonin receptors and transporter are located on bone resident cells (mesenchymal stem cells (MSCs)), osteoblasts and osteoclasts, and serotonergic activity is believed to affect bone homeostasis. Consequently, alterations in serotonin levels by SSRI treatment have the potential to alter bone formation and remodeling. Clinical reports correlate increase risk of bone fractures and delayed bone healing with SSRI use. Metallic implants are commonly used as orthopedic and dental implants to fix bony defects. Surface modifications have been used to increase the level of bone to implant contact by controlling the differentiation of MSCs into an osteoblastic linage and facilitate bone production. However, it is not known if SSRIs can affect MSCs osteoblastic differentiation and bone remodeling signaling in response to microstructured biomaterials. The aims of this study were: 1) Investigate the effects of SSRIs on MSCs differentiation on microstructured titanium (Ti), 2) Determine the effects of SSRIs on bone remodeling signaling and osteoclast activation, and 3) Elucidate the effects of SSRIs on serotonin receptors and their effect on bone remodeling. To investigate this, human MSCs were grown on tissue culture polystyrene (TCPS), smooth Ti (PT) or microstructured Ti (SLA) surfaces under exposure to therapeutic concentrations of commonly prescribed antidepressants (SSRIs (fluoxetine, sertraline, paroxetine), Selective Norepinephrine Reuptake Inhibitor (SNRI) (duloxetine) and other regularly prescribed antidepressants (bupropion)) during differentiation toward osteoblasts. Osteoblastic differentiation was assessed in MSCs after treatment with the drugs (0.1μM, 1μM, 10μM) by alkaline phosphatase activity and osteocalcin levels. Antidepressant treatment decreased levels of MSC differentiation markers on microstructured Ti surfaces. Furthermore, treatment dose-dependently decreased protein levels secreted by MSCs which are important for bone formation (BMP2, VEGF, Osteoprotegerin), and increased those involved in bone resorption (RANKL). To determine the effect of SSRIs on bone remodeling signaling and osteoclast activation, human osteoclasts were either directly exposed to antidepressants or conditioned media obtained from MSCs treated with antidepressants on Ti surfaces, after which, enzymatic tartrate-resistant acid phosphatase (TRAP) activity was assessed. Antidepressants increased TRAP activity both directly and through treated MSCs, with the highest levels evident after treatment with conditioned media from MSCs on microstructured Ti surfaces. To elucidate the effects of serotonin receptors and their effect on bone remodeling, receptors were pharmacologically inhibited. Surface roughness decreased gene expression of HTR2A, HTR1B, and HTR2B, and antidepressant treatment increased their expression. Inhibition of HTR2A decreased RANKL protein levels, while inhibition of other serotonin receptors had no effect on RANKL or OPG levels. These studies suggest that antidepressants inhibit MSCs differentiation on microstructured Ti surfaces and increase levels of proteins associated with bone resorption. Additionally, our results showed that RANKL is regulated by serotonin receptor HTR2A. Taken together, our results suggest that antidepressants have a negative effect on osteoblastic differentiation, compromising bone formation and enhancing bone resorption, which can be detrimental to patients under orthopedic and dental treatment.

In this thesis, we focused on understanding the synthesis of titanium dioxide (TiO2) films and nitrogen doped TiO2 films using an atmospheric pressure Dielectric Barrier Discharge (DBD). The first part of the work was dedicated to the deposition of TiO2 films by cold plasma DBD with titanium tetraisopropoxide as precursor in a single-step process at room temperature. The deposition rate was about 70 nm·min-1. The photocatalytic degradation rate for the degradation of methylene blue (MB) under ultra violet (UV) irradiation of the TiO2 film after annealing was close to a reference anatase TiO2 spin coated film. Moreover, the TiO2 films showed a good photocatalytic stability. The second part of the study focused on the optimization and the understanding of the effect of the plasma parameters (gas flow rate and power) on the morphology of the TiO2 films and on the investigation of the deposition mechanisms. The morphology of the film changed from granular to compact film by either increasing the total flow rate or decreasing the plasma power. In other words, adapting the energy density in the plasma allowed the control of the morphology of the TiO2 films. To our knowledge, it was the first time that the energy density parameters of the plasma were used to control the morphology of TiO2 films. The photocatalytic degradation rate for the degradation of MB under UV irradiation of the annealed TiO2 film turned out to be about 2 and 15 times higher than the one of the commercial TiO2 film and the as-deposited TiO2 films, respectively. In order to extend the light utilization to the visible light range, TiO2 films were doped with nitrogen using a room temperature argon/ammonia plasma discharge. XPS and SIMS results confirmed that the nitrogen has been incorporated in the TiO2 lattice mostly in Ti-N state. This was further confirmed by Raman spectroscopy and XRD. The plasma properties and the doping mechanism were studied by Optical Emission Spectroscopy. It is suggested that the NH radicals played a key role in the doping of TiO2. The concentration of nitrogen in the N-TiO2 coatings could be tuned by adapting the ratio of NH3 in the plasma or the plasma power. The band gap of our N-TiO2 coatings is lower than the one of undoped TiO2 coating. The photocatalytic degradation rate for N-TiO2 coating was more than 4 times higher than the one of the undoped TiO2 coating.
Doctorat en Sciences
info:eu-repo/semantics/nonPublished

The objective of this project was the modification of TiO₂ electrodes with various silanes in order to evaluate the stability of modified layers when they are used on photoelectrodes in SEMICONDUCTOR LIQUID-JUNCTION SOLAR CELLS (SLJSC). To determine the nature of the reactivity of surface hydroxyl groups towards different silanes, a surface IR study was carried out on TiO₂ powders. The powder (TiO₂) was pressed into a pellet and subjected to reactions with various silanes under different reaction conditions. All of these reactions were carried out in a vacuum line under very anhydrous conditions in order to prevent polymerization of the silanes. This study provided an understanding of the reactivity of different silanes towards surface hydroxyl groups of TiO₂ and the best reaction conditions for this purpose. With this information in hand we studied the TiO₂ (rutile) single crystal electrodes. These electrodes were subjected to reaction with silanes under the same conditions as the powders. Then the modified surface was studied using ESCA. These electrodes were subsequently subjected to photoelectrochemical conditions (photocurrent generation) and were reexamined using ESCA in order to evaluate the stability of the modified layer. A reaction scheme which was devised to induce crosslinking in the modified layer was shown to enhance the stability of the surface bound silane during the photocurrent generation. In order to form more homogeneous modified surfaces electrochemically derived polymer coatings were synthesized from divinylbenzene, 4-vinylpyridine, N-methyl-4-vinylpyridinium salts, and phenol. Except for polymers formed from N-methyl-4-vinyl pyridinium salts, other coatings were shown to be neutral. An anomalous pre-wave, and potential-induced polymer swelling and shrinking phenomena were observed in these coatings. The photocorrosion of small bandgap n-type semiconductor electrodes is a serious impediment to the development of efficient and durable conversion (photoelectrochemical) devices. Our objective in this investigation was to develop newer modified surfaces that are useful for the inhibition of this photocorrosion, and enhance the performance of n-type small bandgap semiconductors in the photoelectrochemical systems.
Ph. D.
incomplete_metadata

Dans ce travail de thèse, la texturation de surface d‟alliages de titane a été étudiée en utilisant un procédé d'écriture directe par laser femtoseconde dans le but d'améliorer la mouillabilité d‟implants dentaires et orthopédiques par les fluides biologiques et la minéralisation de la matrice (formation osseuse) tout en réduisant l'adhésion bactérienne et la formation de biofilmes. Des surfaces de titane (Ti-6Al-4Vet cp Ti) ont été micro-, nano-texturées par laser femtoseconde et une biofonctionnalisation de ces surfaces a été ajoutée ou non par greffage de peptides d'adhésion cellulaire (peptides RGD) en surface de ces différents matériaux. Les textures de surface peuvent être classées comme suit: (a) structures périodiques de surface induites par laser (LIPSS); (b) étalage de nanopiliers (NP); (c) étalage de micro colonnes recouvertes de LIPSS (MC-LIPSS) formant une distribution bimodale de rugosité. Nous avons montré que la texturation de surface par laser améliore la mouillabilité des surfaces avec de l'eau ainsi qu‟une solution saline tamponnée Hank's (HBSS) et amène une anisotropie de mouillage. Une minéralisation cellulaire est observée pour toutes les surfaces des deux alliages de titane lorsque des Cellules Souches Mésenchymateuses humaines (hMSC) sont cultivées dans un milieu ostéogénique. La minéralisation de la matrice et la formation de nodules osseux sont considérablement améliorées sur les surfaces texturées LIPSS et NP. Parallèlement,l'adhésion de Staphylococcus aureus et la formation de biofilmes sont considérablement réduites pour les surfaces texturées LIPSS et NP. La biofonctionnalisation des différentes surfaces texturées (cp Ti) par laser a été réalisée et caractérisée par spectroscopie de photoélectrons (XPS) et par microscopie à fluorescence en utilisant des peptides fluorescents. L‟ensemble des résultats obtenus suggèrent que la texturation de surface d'alliages de titane (Ti-6Al-4V et cp Ti) en utilisant une technique d‟écriture directe par laser femtoseconde est un procédé prometteur pour l'amélioration de la mouillabilité de la surface d'implants dentaires et orthopédiques par les fluides biologiques et leur ostéointégration (différenciation ostéoblastique et minéralisation de la matrice), tout en réduisant l‟adhésion de Staphylococcus aureus et la formation de biofilmes. Enfin, la combinaison de la texturation par laser et du greffage covalent d‟un principe actif (ici un peptide d‟adhésion cellulaire comme le peptide RGD) amènera indéniablement une bioactivité utile pour favoriser l'adhésion des hMSC et faciliter laformation osseuse
In the present thesis the surface texturing of Ti alloys using femtosecond laser direct writing method is explored as a potential technique to enhance the wettability of dental and orthopaedic implants by biological fluids and matrix mineralisation (bone formation), while reducing bacteria adhesion and biofilmformation. The surface texture was combined with biofunctionalisation by covalent grafting of a RGD peptide sequence as well. The surface textures can be classified as follows: (a) Laser-Induced Periodic Surface Structures-LIPSS; (b) nanopillars arrays(NP); (c) arrays of microcolumns covered with LIPSS (MC-LIPSS), forming a bimodal roughness distribution. Laser texturing enhances surface wettability by water andHank‟s balanced salt solution (HBSS) and introduces wetting anisotropy, crucial incontrolling the wetting behaviour. Matrix mineralisation is observed for all surfaces of both Ti alloys when human mesenchymal stem cells (hMSCs) are cultured in osteogenic medium. Matrix mineralisation and formation of bone-like nodules are significantly enhanced on LIPSS and NP textured surfaces. On the contrary, Staphylococcus aureusadhesion and biofilm formation are significantly reduced for LIPSS and NP textured surfaces. The biofunctionalisation of the laser textured surfaces of cp Ti is sucessfully achieved. In general, these results suggest that surface texturing of Ti alloys using femtosecond laser direct writing is a promising method for enhancing surface wettability of dental and orthopaedic implants by biological fluids and their osseointegration (osteoblastic differentiation and matrix mineralisation), while reducing Staphylococcus aureus adhesion and biofilm formation. Finally, the combination of laser texturing and covalent grafting of a RGD peptide sequence may be potentially useful for increasing cell adhesion and facilitating bone formation

Thesis (Ph. D.)--Chemistry and Biochemistry, Georgia Institute of Technology, 2009.
Committee Chair: Collard, David M.; Committee Co-Chair: Garcia, Andres J.; Committee Member: France, Stefan; Committee Member: Ragauskas, Arthur; Committee Member: Temenoff, Johnna. Part of the SMARTech Electronic Thesis and Dissertation Collection.

The Tin dioxide (SnO2) and Titanium dioxide (TiO2) are very promising materials in Material science. The SnO2 is commonly used as a gas sensor while the TiO2 is used as a catalyst in many reactions. Despite of the usefulness of these two substances, their surface structures lack detail investigations in the previous years. The Low Energy Electron Diffraction (LEED) technique is commonly used to characterize surfaces in the past 40 years, it is a mature system that many researches rely on its result. However, structural analysis in LEED requires comparison with computational results based on pre-defined structure models, which is a time-consuming method and the results are not guaranteed to be found. The direct determinations of structure by Patterson function inversion methods introduced by Huasheng Wu and S. Y. Tong could provide a different path to search for surface structure. In the Patterson function, each maximum in the function corresponds to a relative position vector of atomic pairs. Multiple-angle-incident LEED has to be performed to obtain an artifact-free Patterson function. Serveal SnO2 and TiO2 surfaces have been characterized by LEED and Patterson function inversion. SnO2 (110), (100), (101) , Rutile TiO2 (110), Anatase TiO2 (110) have been prepared by argon ion sputtering and annealing cycles and the cleanness has been checked by Auger Electron Spectroscopy and LEED. Reconstruction is observed based on the study of the LEED patterns. SnO2 (110) surface shows a 4 x 1 reconstruction in UHV environment while it gives 1 x 1 under annealing in oxygen and C(2 x 2) at higher annealing temperature afterward. SnO2 (100) , (101) and Rutile TiO2 (110) surfaces show 1 x 1 reconstruction in UHV environment and the reconstruction persists for further annealing. The Anatase TiO2 (110) surface shows a 3 x 4 reconstruction in UHV environment. The 3 x 4 reconstruction of Anatase TiO2 (110) surface would raise research interests as it is quite a special reconstruction. Multiple-angle-incident LEED has been performed on the SnO2 (100), (101) and Rutile TiO2 (110) surfaces. Patterson function inversion is performed on the surfaces SnO2 (100) and Rutile TiO2 (110) . Only LEED is performed on SnO2 (110) , (101) and Anatase TiO2 (110) surfaces. From Patterson functions analysis, the surface atoms positions are determined for the surface SnO2 (100) and Rutile TiO2 (110). The results show that their reconstructions are negligible, but they have obvious relaxations.
published_or_final_version
Physics
Master
Master of Philosophy