Diamond-like carbon (DLC) film exhibits an extreme hardness, low friction coefficient, chemical stability, heat conductivity, high resistance, and high optical transparency. There properties lead to remarkable on industrial applications Diamond-like carbon films were deposited onto the silicon (100) and ITO glass substrates. Under the same deposition conditions, the characteristics of DLC films were evaluated by the variations of deposited parameters such as the applied voltage, deposition temperature the concentrations of electrolyte; acetic acid. The properties due to the different substrate were compared and discussed in detail. In experimental work, the properties of DLC film were conducted by various measurements. Scanning electron microscopy can make an insight into the surface morphology also to reveal the uniformity of the DLC films. For the I-t curves of DLC film growth, it can be used to study of the growth mechanism by correlation the surface morphology observed by Scanning electron microscopy (SEM). The transmission, refraction index and optical band gap of DLC film was measured by the N &K analyzer. Finally, the hydrogen content, composition and microstructure of DLC films were characterized by the FTIR and XPS analyze According to above results, DLC film using the electrolyte of acetic acid was more difficult to deposit on Silicon substrate because the very high activation energy and the high hydrogen ion existing in DI water firstly deposited on the surface of Si substrate. For FTIR measurement, The absorption wavenumber of various bonding observed were positioned at 610 cm-1,680 cm-1,1100 cm-1 and 3600 cm-1~3800 cm-1and to be cauterized as the bonding of Si-H¡BSi-O and O-H, respectively. The absorption peaks within the range from 2800 cm-1 to 3100 cm-1 were missing. Peaks observed were attributed to the bonding of Si-C¡B SP3 C-C¡B C-O¡BC-C¡BC=O and C=C and the CHn bonding was missing on the surface of substrate. The reaction mechanism of DLC deposition can be suggested from the results of measurements. As bias voltage applied, the acetic ion; CH3COO- were attracted by the Anode as the state of C¡]Anode¡^-OOCCH3, and then to give electron and form the CH3+ion»PCO2. The hydrogen ion and methyl ion were attracted by cathode. The competitive reaction was built between ions to deposited DLC films and/or to form Si-H. However, experimental results show that the last was preferred and for forming the DLC film was forbidden.

Recently, synthesis of Diamond-Like Carbon (DLC) films has received considerable interest. Owing to their similar characteristics of diamonds, such as extreme hardness, chemical stability, and high heat conductivity etc, DLC films are regarded as one of the most promising materials. But the practical applications have been limited due to their high internal stress and insufficient adhesion at the interface between DLC film and substrate. Several methods used to the deposition of Me-DLC films have been proposed. Studies have shown that the internal stress was released and the adhesion also improved by doping metallic element into DLC films. Conventionally, metal incorporation in DLC films were prepared by vapor deposition. The requirement of high vacuum equipment makes the process complicated. Besides, there are many merits in electrodeposition, such as low cost, simplicity of experimental set up, and availability for deposition on complex shapes substrate in large area. In this study, electrodepositing technique was used to synthesize the amorphous Cu-DLC films deposited on ITO substrate, in which the pH value of electrolyte varied, to study the characteristics and the composition of DLC films. According to the I-t curves of deposition, the end of current density was used for the impedance comparison of films. With the addition of Cu, the resistance of the electron transportation in Cu-DLC was reduced, and the awl-shaped surface morphology was observed by AFM measurement, which could enhance the electron field emission properties of thin films. For Raman analysis, the effect of Cu addition would promote the sp2 bonding¡F this result corresponds with the increasing ID/IG value. It indicates that film becomes graphitization due to the addition of Cu and leads the shift of G-peak position toward lower wavenumber. ESCA spectra of C1s and Cu2p indicate no obvious evidence of Cu-C formation. The sp2/(sp2+sp3) ratio increases with the pH value. In addition, we found that Cu-DLC in acidic environmental condition, or doping as [Cu(NH3)n]2+ complex is more conducive to the growth of copper metal in DLC films, and has the lowest optical band gap value deduced by n&k analyzer. Finally, we discussed the thin film growth mechanisms and the characteristic of electron field emission for the applications in the future.

This thesis is to study the microstructure and its related surface properties of CrxNy¡BDLC and TiO2 films prepared by different plasma energy processes such as physical vapor deposition (PVD) and plasma immersion ion implantation (PIII) processes. In the first chapter of this thesis, the different Cr-based coatings (Cr, Cr2N and CrN) were prepared by PVD process with lower plasma energy condition (less than 100 eV). The surface energy properties of these Cr-based coating were studied and the relationship between the surface properties and adhesion forces with respect to epoxy molding compound (EMC) were also investigated. It is found that the PVD-Cr2N coating has the largest contact angle of water and the lowest polar components and surface energy. The low surface energy is attributed to the high density of surface reactive sites and the formation of N-H bonds and O-H bonds on the film surface that act as the effective adsorption sites for carbon. The second chapter focuses on the modification of the electrical and mechanical properties of DLC films with high plasma energy (5¡ã45 keV) , PIII post-treatment. It is found that using high ion energy and a short treating time can modify the sp3 content, the nano-hardness and the Young¡¦s modulus of the DLC films. The formation mechanisms of sp3 C-C bonds in the implanted DLC film involve the reaction among compressive stress field, bonds breakage and recombination process induced by hydrogen and carbon ion. The third chapter deals with the structure and photocatalytic properties of nitrogen-doped TiO2 film prepared by PVD process with lower plasma energy condition (less than 100 eV). The TiO2-xNx films are indirect transition type and optical band gap energy is various with increasing nitrogen flow rate. The TiO2-xNx film with high crystallinity and porous surface morphology shows the best degradation rate of methylene blue solution.

The purpose of this project was to determine how Dlc-2 and Rho signaling modulate the ceramide induction of PGP synthase. This induction was studied at the transcriptional, post-transcriptional, and post-translational levels using cell culture, Real-Time RT-PCR, protein purification, phage display, and western blotting techniques. We have demonstrated that the PGP synthase gene is not controlled at the transcriptional level by ceramide and Rho, nor is the mRNA stability of PGP synthase affected. However, ceramide and Rho do seem to exhibit translational or post-translational control over the PGP synthase protein. The relationships between Dlc-2 (and Rho), ceramide, and PGP synthase (and CL) are important to understand. All three are involved in cancer and apoptotic responses. The knowledge gained by the experiments discussed in this thesis will contribute to an understanding of how these proteins and lipids interact. This knowledge may then be used in the future to develop cancer treatments.

The purpose of this project was to determine how Dlc-2 and Rho signaling modulate the ceramide induction of PGP synthase. This induction was studied at the transcriptional, post-transcriptional, and post-translational levels using cell culture, Real-Time RT-PCR, protein purification, phage display, and western blotting techniques. We have demonstrated that the PGP synthase gene is not controlled at the transcriptional level by ceramide and Rho, nor is the mRNA stability of PGP synthase affected. However, ceramide and Rho do seem to exhibit translational or post-translational control over the PGP synthase protein. The relationships between Dlc-2 (and Rho), ceramide, and PGP synthase (and CL) are important to understand. All three are involved in cancer and apoptotic responses. The knowledge gained by the experiments discussed in this thesis will contribute to an understanding of how these proteins and lipids interact. This knowledge may then be used in the future to develop cancer treatments.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-10-11T20:39:41Z No. of bitstreams: 1 LauraCamilaDinizDosSantos_TESE.pdf: 4073170 bytes, checksum: 92574134d5de6d9b67507f43f4493cef (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-10-13T23:43:45Z (GMT) No. of bitstreams: 1 LauraCamilaDinizDosSantos_TESE.pdf: 4073170 bytes, checksum: 92574134d5de6d9b67507f43f4493cef (MD5) / Made available in DSpace on 2016-10-13T23:43:45Z (GMT). No. of bitstreams: 1 LauraCamilaDinizDosSantos_TESE.pdf: 4073170 bytes, checksum: 92574134d5de6d9b67507f43f4493cef (MD5) Previous issue date: 2015-07-09 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O grande interesse no uso de filmes de carbono tipo diamante (DLC) ? justificado por suas not?veis propriedades mec?nicas e tribol?gicas, como alta dureza, elevada resist?ncia ao desgaste, in?rcia qu?mica, e baix?ssimo coeficiente de atrito. Essa combina??o de propriedades ?nicas confere ao revestimento aplica??es nas mais diversas ?reas. No entanto, o elevado n?vel de tens?es compressivas, que se originam durante o crescimento do filme, dificulta a obten??o de alta ader?ncia. Al?m disso, a elevada dureza do filme aliada ? diferen?a no coeficiente de expans?o t?rmica em rela??o ao substrato faz com que o DLC n?o acompanhe facilmente a deforma??o do substrato, o que pode provocar a delamina??o e falha total do revestimento, especialmente em a?os. O principal objetivo deste trabalho ? o estudo e desenvolvimento do processo de deposi??o de filmes finos de DLC com a inser??o de nanopart?culas de prata, atrav?s da t?cnica de gaiola cat?dica, seguido da avalia??o de sua ader?ncia ao substrato e seu comportamento tribol?gico. Neste trabalho, filmes de DLC dopados com prata (DLC-Ag) foram depositados sobre o a?o AISI 301, utilizando a t?cnica de deposi??o qu?mica na fase vapor assistida por plasma (PECVD). A parte inovadora desse trabalho refere-se a deposi??o do filme de DLC, onde a prata ? introduzida utilizando uma gaiola cat?dica de prata (GCP) durante o processo de deposi??o. Testes tribol?gicos foram realizados para se analisar a ader?ncia entre filme e substrato, o coeficiente de atrito, e a resist?ncia ao desgaste. A t?cnica de espectroscopia Raman foi utilizada para verificar o arranjo estrutural dos ?tomos de carbono e obter par?metros importantes. Os filmes foram adicionalmente caracterizados por microscopia eletr?nica de varredura (MEV). Os resultados mostraram que a utiliza??o da GCP foi bastante eficaz, pois produziu um filme com maior homogeneidade e dependendo se a gaiola foi utilizada durante todo o processo ou n?o, h? uma propor??o maior ou menor de prata no filme de DLC. / Diamond-like carbon (DLC) films have awaked growing interest due to its outstanding mechanical and tribological properties such as high hardness, high wear resistance, chemical inertness, and low friction coefficient. This is unique combination of specific properties enable film application in several fields. However, the high level of compressive stresses, which arise during film growth, hinders to obtain high adhesion. Furthermore, the high film hardness coupled to the difference in thermal expansion coefficient regarding the substrate, results in the DLC does not easily follow the deformation of the substrate. It may result in film delamination and complete failure, especially in case of substrates made of steels. The main objective of this work is to study and develop the deposition process of DLC thin films with inclusion of silver nanoparticles by cathodic cage technology. The adhesion between film and substrate as well as the tribological behavior were analyzed in detail. Doped Diamond-like carbon films with silver (DLC-Ag) were deposited on AISI 301 steel by plasma-enhanced chemical vapor deposition (PECVD). The innovative part of this PhD thesis is the DLC-Ag film deposition by using a silver cathodic cage. Tribologicals tests were performed to evaluate the adhesion between film and substrate, friction coefficient and wear resistance. Raman spectroscopy was applied to verify the structural arrangement of carbon atoms and to obtain important parameters. The films were further characterized by scanning electron microscopy (SEM). The results proved that the deposition by silver cathodic cage is an effective technique for production of more homogeneous DLC-Ag films. The amount of Ag in the film can be adjusted by dwell time of the cathodic cage process.

Die spinale Muskelatrophie (SMA) ist eine monogenetische Erkrankung, bei der es durch den Verlust des SMN Proteins zur Degeneration der α-Motoneurone im Rückenmark kommt. Abhängig vom Schweregrad zeigen die Patienten bereits innerhalb der ersten Lebensmonate ausgeprägte Lähmungen der Skelettmuskulatur und eine Zwerchfellparese einhergehend mit einer reduzierten Lebenserwartung. Mithilfe von Mausmodellen für die SMA konnte gezeigt werden, dass der Motoneuronenverlust bei Smn-defizienten Mäusen mit Störungen der Neurotransmission an der motorischen Endplatte und mit Differenzierungsstörungen der Motoneurone einhergeht. Die Differenzierungs-störungen primärer Smn-defizienter Motoneurone sind eng gekoppelt mit einer verminderten Clusterbildung spannungsabhängiger Kalziumkanäle im distalen axonalen Bereich. Dies wiederum führt zu einer verminderten Frequenz spontaner Kalziumeinströme am Axonterminus und hat eine veränderte axonale Elongation zur Folge. Es wurden folgende Aspekte in Bezug auf die Verstärkung und die Induktion spontaner Kalziumeinströme in Mausmodellen für spinale Muskelatrophien in dieser Arbeit adressiert: 1) Lassen sich spontane Kalziumeinströme in Smn-defizienten Motoneuronen durch die externe Applikation von Kalziumkanalagonisten verstärken? 2) Sind spontane Kalziumeinströme in primären Motoneuronen durch den Brain-derived-neurotrophic-factor (BDNF) induzierbar? 3) Zeigen primäre Motoneurone eines Mausmodells für spinale Muskelatrophie mit Ateminsuffizienz Typ 1 (SMARD1) ebenfalls veränderte Kalziumtransienten? Die Ergebnisse meiner Arbeit zeigen, dass durch den Kalziumkanalagonisten R-Roscovitine die Frequenz der spontanen Kalziumeinströme im distalen Axon von Smn-defizienten Motoneuronen signifikant erhöht wird. Dies hat wiederum einen regulierenden Effekt auf die Differenzierung der SMA Motoneurone zur Folge. Smn-defiziente Motoneurone zeigen somit keine Unterschiede mehr in Bezug auf Axonlängen und Wachstumskegelflächen im Vergleich zu Kontrollzellen. Für R- 10 Roscovitine ist neben der agonistischen Wirkung am Kalziumkanal auch ein inhibitorischer Effekt auf die Cyclin-abhängige Kinase 5 beschrieben. Es konnte jedoch gezeigt werden, dass die erhöhten Kalziumtransienten unter der Behandlung mit R-Roscovitine durch eine direkte Bindung an die Cav2 Kalziumkanäle verursacht werden und nicht durch eine Cdk5 Blockade. Dafür spricht die schnelle und reversible Wirkung von R-Roscovitine, sowie die Aufhebung des R-Roscovitines Effekts bei gleichzeitiger Gabe des Cav2.2 Antagonisten ω-Conotoxin MVIIC. Der zweite Aspekt dieser Arbeit behandelt den Einfluss der neurotrophen Faktoren BDNF, CNTF und GDNF auf die Kalziumtransienten am Wachstumskegel wildtypischer Motoneurone. Der Vergleich der neurotrophen Faktoren zeigt, dass nur BDNF eine induzierende Wirkung auf spontane Kalziumtransienten am Wachstumskegel hat. Der letzte Abschnitt dieser Arbeit beschäftigt sich mit den Kalziumtransienten bei Motoneuronen aus dem Nmd2J (SMARD1) Mausmodell. Die SMARD1 gilt als eigenständige Form der spinalen Muskelatrophien mit unterschiedlicher Genetik und unterschiedlichen klinischen Merkmalen. Die Motoneurone weisen in Bezug auf die Kalziumtransienten keine Unterschiede zwischen Wildtyp und Nmd2J Mutante auf. Es ergibt sich somit kein Hinweis darauf, dass die Degeneration der Motoneurone bei der SMARD1 von einer Störung der Kalziumhomöostase im distalen axonalen Bereich ausgeht. / Spinal muscular atrophy (SMA) is a monogenetic disorder which is caused by the loss of the SMN Protein and leads to the degeneration of α-motoneurons. Within the first few months of life most patients are clinically affected with severe motor deficits of skeletal muscles and a diaphragm paralysis, going along with a reduced life expectancy depening on the degree of severity. With the aid of SMA mouse models it was shown that the loss of motoneurons with Smn deficiancy lies in an impaired neurotransmission of the motoneuron endplat leading to a differentiation disorder of the motoneurons. This differentiation disorder is strongly connected to a reduced cluster formation of voltage-dependent calcium channels in the distal axonal area. The impaired cluster formation in turn leads to a reduced frequency of spontanous calcium transients at the axon terminus, followed by an altered axonal elongation. In this work the following aspects concerning the enhancement and induction of spontanous calcium transients in mouse models of spinal muscular atrophy were adressed: 1) Does the external application of calcium channel agonists increase spontanous calcium transients in Smn-deficient motoneurons? 2) Is the neurotrophic factor Brain-derived neurotrophic factor (BDNF) able to induce spontanous calcium transients in primary motoneurons? 3) Do primary motoneurons of a mouse model for spinal muscular atrophy with respiratory distress (SMARD1) show altered calcium transients as well? The results of my work show that the calcium channel agonist R-Roscovitine significantly increases the frequency of spontanous calcium transients in growth cones of Smn-deficient motoneurons which in turn has a regulatory effect on the differentiation of SMA motoneurons. Smn-deficient motoneurons treated with R-Roscovitine do not show any differences concerning axon length and growth cone size compared to control cells. Apart from the agonist effect on the calcium channels, R-Roscovitine also has an inhibitory impact on the cyclin-dependant kinase 5. The results of this work show that the positive effect on the calcium 12 transients under R-Roscovitine treatment is because R-Roscovitine binds directly to the calcium channel rather than due to an inhibition of cdk5. Arguments supporting this idea are the rapid and reversible channel kinetics of R-Roscovitine. Plus, the effect of R-Roscovitine can be repealed when the Cav2 channal antagonist ω-conotoxin is given simultaneously. In the second part of this work the influence of the neurotrophic factors BDNF, CNTF and GDNF on the calcium transients of wildtype motoneurons is investigated. Comparing these neurotrophic factors show that only BDNF has an impact on local calcium channel kinetics in growth cones of motoneurons. The last part of this work deals with the investigation of calcium transients in motoneurons from the Nmd2J (SMARD1) mouse model. SMARD1 is an independent form of spinal muscular atrophies with different genetical and clinical aspects compared to proximal SMA. The results of this work show that Nmd2J motoneurons do not show any difference in growth cone calcium influx between wildtype and mutant. Thus, there is no indication that the degeneration of SMARD1 motoneurons has any pathophysiological similarities with motoneurons from the proximal SMA mouse model. Hence, there are also no indications that the reason for motoneuron degeneration in SMARD1 lies in an impaired calcium homeostasis in the distal axonal area.

O projeto de novos materiais para aplicações tecnológicas em biomateriais e bioengenharia é altamente dependente de como as células aderem à superfície de um material. A adesão e crescimento em biomateriais depende de propriedades do substrato, tais como molhabilidade da superfície, a topografia e a composição química de superfície. O objetivo deste estudo foi investigar as interações de diversos materiais com culturas celulares de células epitelial CHO (Ovário de Hamster Chinês). Os materiais utilizados foram SU-8 2005 (elétron-resiste, Microchem), PDMS (Poli (dimetil siloxano), Down Corning), DLC (Diamond-like Carbon) e vidro foi utilizado como referência. Superfícies de vidro, SU-8, PDMS e DLC lisas (planas) e isentas de modificação ou tratamento específico foram avaliadas quanto ao cultivo de células CHO. Valores médios dos fatores de forma (Ff) de 450 células foram calculados para cada uma das culturas realizadas sobre os 4 substratos. Foram obtidos Ff próximos a 0,52 para o vidro, o SU-8 liso e o DLC, demonstrando um bom espraiamento das células nessas superfícies. A superfície de PDMS apresentou valor unitário para o fator de forma (Ff), que está relacionado a um baixo espraiamento das células. A energia de superfície (ES) obtida para o PDMS é compatível com o resultado de fator de forma (Ff), uma vez que o menor valor para ES é coerente com a baixa adesão celular, o que gerou células com elevado fator de forma (Ff). O SU-8 foi modificado por implantação iônica com uma dose de 1,2x1016 átomos/cm2 e a energia de implantação foi de 8 keV, como referência foi utilizada uma superfície lisa de SU-8 sem implantação. Os resultados mostraram que o número de células vivas por unidade de área foi superior na superfície de SU-8 com prata implantada, mostrando o bom desempenho da cultura nesse substrato. As superfícies de DLC modificadas por tratamento com plasma de oxigênio (DLC-O) e com plasma de hexafluoreto de enxofre (DLC-F) foram utilizadas para cultura celular, os resultados de três experimentos independentes de contagem de número de núcleos (marcados com DAPI) por unidade de área confirmaram os resultados obtidos através do teste de viabilidade (marcados com trypan blue). A superfície de DLC-O, apresentou um maior número de núcleos por unidade de área, quando comparado à superfície DLC-F, da mesma forma que nos resultados obtidos pelo teste de viabilidade. As energias de superfície para as amostras de DLC-F e DLC-O indicaram que a superfície DLC-O é mais hidrofílica do que a superfície DLC-F, que está coerente com o que é conhecido da literatura e com os resultados obtidos em nosso trabalho. Cultura de células CHO foram realizadas em superfícies litografadas com estruturas hexagonais periódicas com o parâmetro 2R (diâmetro do círculo inscrito) sendo 12 µm, 30 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. Estas superfícies foram caracterizadas por microscopia óptica de fluorescência com relação ao número de núcleos (marcados com o fluoróforo DAPI) por unidade de área, isto é, núcleos/mm2. Obteve-se histogramas com o número médio de núcleos por mm2 em três experimentos independentes, onde o número núcleos/mm2 foi consideravelmente maior para 80 µm. As superfícies contendo cavidades periódicas de 12 µm e 30 µm apresentaram dificuldade para as células CHO aderirem à superfície. Em uma outra etapa realizou-se culturas celulares em triplicata dos substratos com as superfícies 12 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. As células em cada uma das superfícies foram analisadas por microscopia óptica (MO) para avaliação da viabilidade celular, utilizando marcador trypan blue. Obteve-se histogramas com os valores médios para o número de células vivas/mm2 para as culturas celulares que corrobora os resultados obtidos no histograma da cultura celular que tiveram os núcleos marcados pelo fluoróforo DAPI. Assim, fica confirmado o melhor desempenho da cultura celular no substrato 80 µm que apresentou o maior número de células vivas/mm2 As micrografias obtidas através de marcação por DAPI foram analisadas através da função de correlação com intuito de se entender como as células estavam organizadas. Isso foi feito para cada uma das superfícies litografadas, 12 µm, 30 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. As superfícies dos substratos 80 µm apresentaram os menores valores de distâncias para primeiros e segundos vizinhos, ou seja, as células estão mais próximas umas das outras. As demais superfícies tendem a separar mais as células. Obteve-se também os valores de raio de aglomerado (rc), distância entre os aglomerados (dc) e o número de primeiros vizinhos (Np) através do ajuste da função de correlação. A análise de correlação mostrou com clareza o que não era evidenciado apenas visualizando-se as imagens. Ela mostra que as células, mesmo em SU-8 liso tem a forte tendência de formar aglomerados de células com raio de aproximadamente 45 µm. No caso de substratos lisos, células CHO apresentaram a melhor adesão na superfície do SU-8, seguido do DLC, enquanto que o PDMS foi a pior situação, devido à baixa molhabilidade do material. No caso de superfícies com microestrutura, SU-8 contendo microcavidades hexagonais de 12 e 30 µm mostraram ser as situações mais adversas para o crescimento de células CHO, provavelmente por causa da topografia das cavidades serem de menor tamanho quando comparadas ao tamanho das células CHO. Em vez disso, SU-8, contendo microcavidades hexagonais de 80 µm foi a superfície mais favorável para o crescimento de células CHO. / The design of new materials for technological applications in biomaterials and bioengineering is highly dependent on how the cells adhere to the material surface. The cells adhesion and growth on biomaterials depends on substrate properties such as surface wettability, topography and the chemical composition. The aim of this study was to investigate the interactions of various materials with cell cultures of epithelial cells CHO (Chinese Hamster Ovary). The materials used were SU-8 2005 (electron resists, Microchem), PDMS (poly (dimethyl siloxane), Dow Corning), DLC (Diamond-like Carbon) and glass was used as reference. Unmodified and flat surfaces of glass, SU-8, PDMS and DLC were evaluated for the culture of CHO cells. Form factor (Ff) values were calculated as average of 450 cells for each of the cultures performed on the four substrates. Ff close to 0.52 was obtained for flat surfaces of glass, SU-8 and DLC, showing a good cell spreading on these surfaces. The surface of PDMS presented a form factor (Ff) near unity, which is related to low spreading cell. The surface energy (ES) obtained for the PDMS is coherent with the Ff result, since the smallest value of ES is consistent with the low cell adhesion, which resulted in cells with a high Ff. The SU-8 was modified by ion implantation using a dose of 1.2x1016 atoms/cm2 and an implantation energy of 8 keV, unmodified flat SU-8 was used as a reference. The cell culture results showed that the number of live cells per unit area was greater in the SU-8 surface implanted with silver, showing a good performance in the culture substrate. The DLC surfaces modified by plasma treatment with oxygen (DLC-O) and sulfur hexafluoride (DLC-F) were used for cell culture. The results of three independent experiments, counting the number of nuclei (marked with DAPI) per unit area, confirmed the results obtained by the viability test (marked with trypan-blue). The surface of the DLC-O had higher number of nuclei per unit area when compared to the surface of the DLC-F, similarly to the results obtained for the viability test. The surface energies of the DLC-F and DLC-O samples indicated that the DLC-O surface is more hydrophilic than the DLC-F surface, which is consistent with results obtained with our work and with the literature. CHO cell culture were performed on surfaces with periodic hexagonal structures with the diameter of inscribed circle (2R) given by 12 µm, 30 µm, 80 µm, 280 µm, 560 µm and also on flat SU-8. These surfaces were characterized by fluorescence optical microscopy with respect to the number of nuclei (marked with fluorophore DAPI) per unit area, i.e. nuclei/mm2. Histograms were obtained for the average number of nuclei per mm2 in three independent experiments, where the substrate with periodic hexagonal structures with 2R = 80 µm presented considerably higher nuclei/mm2. Surfaces containing periodic cavities of 2R =12 µm and 30 µm were adverse for CHO cells adhesion. In another approach, cell culture were analyzed by light microscopy (LM) for evaluation of cell viability using trypan-blue marker. This was carried out in triplicate cell culture on substrates with surfaces 12 µm, 80 µm, 280 µm, 560 µm and also on flat SU-8. Histograms were generated for average number of living cells/mm2 for each substrate, which corroborates with the results obtained for the cell culture marked with fluorophore DAPI. Thus, it is confirmed the better performance of the cell culture on substrates with 2R = 80 µm, presenting the highest number of living cells/mm2. The micrographs obtained with cells marked with DAPI were analyzed through the correlation function with the aim of understanding how the cells were organized. This was performed for each of the lithographed surfaces 12 µm, 30 µm, 80 µm, 280 µm, 560 µm and also flat SU-8. The surfaces of the substrates with 2R = 80 µm had the lowest values for length between its neighbors, that is, the cells are closer to each other. The remaining surfaces tend to separate the cells. Also were obtained the cluster radius values (rc), the distance between the clusters (dc) and the number of nearest neighbors (Np) through the correlation function fitting. The correlation analysis clearly showed what was not possible to observe by viewing the images. It shows that the cells, even in flat SU-8, have a strong tendency to form clusters of cells within about 45 micrometers. In the case of flat substrates, CHO cells exhibited better adhesion to the surface of SU-8, followed by the DLC, while the PDMS was worse due to low wettability of the material. In the case of surfaces with microstructures, SU-8 containing hexagonal microstructures of 12 and 30 µm showed to be the most adverse conditions for the CHO cell growth, probably because of the topography of the cavities being smaller in size compared to the size of CHO cells. SU-8 with 80 µm hexagonal microstructures was more favorable surface for the growth of CHO cells.

Downloadable content (DLC) distribution models employ various methods to keep consumers interested. Some methods encourage consumers to purchase and engage with a DLC model and some methods discourage consumers from engaging with the model. This study seeks to determine which models keep consumers interested and which discourage consumers from purchasing DLC. We do this by reaching out to multiplayer communities of four different games that are affected by different models and have them fill out a survey regarding their purchasing behaviour. After analysing the results, we concluded that models that require players to own the same DLC in order to play together or that utilize watermarks for DLC that are not owned, are less effective at incentivizing players to purchase when compared to models that either allow players to share DLC, or acquire it through in-game means. / Distributionsmodeller för nedladdningsbart innehåll använder olika metoder för att hålla konsumenterna intresserade. Vissa metoder uppmuntrar konsumenter att köpa och engagera sig i en DLC-modell och vissa metoder avskräcker konsumenterna från att engagera sig med modellen. Denna studie syftar till att bestämma vilka modeller som håller konsumenterna intresserade och som avskräcker konsumenterna från att köpa DLC. Vi gör det genom att nå ut till multiplayer-grupper i fyra olika spel som påverkas av olika modeller och få dem att fylla i en undersökning om deras inköpssätt. Efter att ha analyserat resultaten drog vi slutsatsen att modeller som kräver att spelare äger samma DLC för att kunna spela tillsammans eller använda vattenstämplar för DLC som inte ägs, är mindre effektiva för att stimulera spelare att köpa jämfört med modeller som antingen tillåter spelare att dela DLC, eller skaffa det via in-game-medel.

O projeto de novos materiais para aplicações tecnológicas em biomateriais e bioengenharia é altamente dependente de como as células aderem à superfície de um material. A adesão e crescimento em biomateriais depende de propriedades do substrato, tais como molhabilidade da superfície, a topografia e a composição química de superfície. O objetivo deste estudo foi investigar as interações de diversos materiais com culturas celulares de células epitelial CHO (Ovário de Hamster Chinês). Os materiais utilizados foram SU-8 2005 (elétron-resiste, Microchem), PDMS (Poli (dimetil siloxano), Down Corning), DLC (Diamond-like Carbon) e vidro foi utilizado como referência. Superfícies de vidro, SU-8, PDMS e DLC lisas (planas) e isentas de modificação ou tratamento específico foram avaliadas quanto ao cultivo de células CHO. Valores médios dos fatores de forma (Ff) de 450 células foram calculados para cada uma das culturas realizadas sobre os 4 substratos. Foram obtidos Ff próximos a 0,52 para o vidro, o SU-8 liso e o DLC, demonstrando um bom espraiamento das células nessas superfícies. A superfície de PDMS apresentou valor unitário para o fator de forma (Ff), que está relacionado a um baixo espraiamento das células. A energia de superfície (ES) obtida para o PDMS é compatível com o resultado de fator de forma (Ff), uma vez que o menor valor para ES é coerente com a baixa adesão celular, o que gerou células com elevado fator de forma (Ff). O SU-8 foi modificado por implantação iônica com uma dose de 1,2x1016 átomos/cm2 e a energia de implantação foi de 8 keV, como referência foi utilizada uma superfície lisa de SU-8 sem implantação. Os resultados mostraram que o número de células vivas por unidade de área foi superior na superfície de SU-8 com prata implantada, mostrando o bom desempenho da cultura nesse substrato. As superfícies de DLC modificadas por tratamento com plasma de oxigênio (DLC-O) e com plasma de hexafluoreto de enxofre (DLC-F) foram utilizadas para cultura celular, os resultados de três experimentos independentes de contagem de número de núcleos (marcados com DAPI) por unidade de área confirmaram os resultados obtidos através do teste de viabilidade (marcados com trypan blue). A superfície de DLC-O, apresentou um maior número de núcleos por unidade de área, quando comparado à superfície DLC-F, da mesma forma que nos resultados obtidos pelo teste de viabilidade. As energias de superfície para as amostras de DLC-F e DLC-O indicaram que a superfície DLC-O é mais hidrofílica do que a superfície DLC-F, que está coerente com o que é conhecido da literatura e com os resultados obtidos em nosso trabalho. Cultura de células CHO foram realizadas em superfícies litografadas com estruturas hexagonais periódicas com o parâmetro 2R (diâmetro do círculo inscrito) sendo 12 µm, 30 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. Estas superfícies foram caracterizadas por microscopia óptica de fluorescência com relação ao número de núcleos (marcados com o fluoróforo DAPI) por unidade de área, isto é, núcleos/mm2. Obteve-se histogramas com o número médio de núcleos por mm2 em três experimentos independentes, onde o número núcleos/mm2 foi consideravelmente maior para 80 µm. As superfícies contendo cavidades periódicas de 12 µm e 30 µm apresentaram dificuldade para as células CHO aderirem à superfície. Em uma outra etapa realizou-se culturas celulares em triplicata dos substratos com as superfícies 12 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. As células em cada uma das superfícies foram analisadas por microscopia óptica (MO) para avaliação da viabilidade celular, utilizando marcador trypan blue. Obteve-se histogramas com os valores médios para o número de células vivas/mm2 para as culturas celulares que corrobora os resultados obtidos no histograma da cultura celular que tiveram os núcleos marcados pelo fluoróforo DAPI. Assim, fica confirmado o melhor desempenho da cultura celular no substrato 80 µm que apresentou o maior número de células vivas/mm2 As micrografias obtidas através de marcação por DAPI foram analisadas através da função de correlação com intuito de se entender como as células estavam organizadas. Isso foi feito para cada uma das superfícies litografadas, 12 µm, 30 µm, 80 µm, 280 µm, 560 µm e também em SU-8 liso. As superfícies dos substratos 80 µm apresentaram os menores valores de distâncias para primeiros e segundos vizinhos, ou seja, as células estão mais próximas umas das outras. As demais superfícies tendem a separar mais as células. Obteve-se também os valores de raio de aglomerado (rc), distância entre os aglomerados (dc) e o número de primeiros vizinhos (Np) através do ajuste da função de correlação. A análise de correlação mostrou com clareza o que não era evidenciado apenas visualizando-se as imagens. Ela mostra que as células, mesmo em SU-8 liso tem a forte tendência de formar aglomerados de células com raio de aproximadamente 45 µm. No caso de substratos lisos, células CHO apresentaram a melhor adesão na superfície do SU-8, seguido do DLC, enquanto que o PDMS foi a pior situação, devido à baixa molhabilidade do material. No caso de superfícies com microestrutura, SU-8 contendo microcavidades hexagonais de 12 e 30 µm mostraram ser as situações mais adversas para o crescimento de células CHO, provavelmente por causa da topografia das cavidades serem de menor tamanho quando comparadas ao tamanho das células CHO. Em vez disso, SU-8, contendo microcavidades hexagonais de 80 µm foi a superfície mais favorável para o crescimento de células CHO. / The design of new materials for technological applications in biomaterials and bioengineering is highly dependent on how the cells adhere to the material surface. The cells adhesion and growth on biomaterials depends on substrate properties such as surface wettability, topography and the chemical composition. The aim of this study was to investigate the interactions of various materials with cell cultures of epithelial cells CHO (Chinese Hamster Ovary). The materials used were SU-8 2005 (electron resists, Microchem), PDMS (poly (dimethyl siloxane), Dow Corning), DLC (Diamond-like Carbon) and glass was used as reference. Unmodified and flat surfaces of glass, SU-8, PDMS and DLC were evaluated for the culture of CHO cells. Form factor (Ff) values were calculated as average of 450 cells for each of the cultures performed on the four substrates. Ff close to 0.52 was obtained for flat surfaces of glass, SU-8 and DLC, showing a good cell spreading on these surfaces. The surface of PDMS presented a form factor (Ff) near unity, which is related to low spreading cell. The surface energy (ES) obtained for the PDMS is coherent with the Ff result, since the smallest value of ES is consistent with the low cell adhesion, which resulted in cells with a high Ff. The SU-8 was modified by ion implantation using a dose of 1.2x1016 atoms/cm2 and an implantation energy of 8 keV, unmodified flat SU-8 was used as a reference. The cell culture results showed that the number of live cells per unit area was greater in the SU-8 surface implanted with silver, showing a good performance in the culture substrate. The DLC surfaces modified by plasma treatment with oxygen (DLC-O) and sulfur hexafluoride (DLC-F) were used for cell culture. The results of three independent experiments, counting the number of nuclei (marked with DAPI) per unit area, confirmed the results obtained by the viability test (marked with trypan-blue). The surface of the DLC-O had higher number of nuclei per unit area when compared to the surface of the DLC-F, similarly to the results obtained for the viability test. The surface energies of the DLC-F and DLC-O samples indicated that the DLC-O surface is more hydrophilic than the DLC-F surface, which is consistent with results obtained with our work and with the literature. CHO cell culture were performed on surfaces with periodic hexagonal structures with the diameter of inscribed circle (2R) given by 12 µm, 30 µm, 80 µm, 280 µm, 560 µm and also on flat SU-8. These surfaces were characterized by fluorescence optical microscopy with respect to the number of nuclei (marked with fluorophore DAPI) per unit area, i.e. nuclei/mm2. Histograms were obtained for the average number of nuclei per mm2 in three independent experiments, where the substrate with periodic hexagonal structures with 2R = 80 µm presented considerably higher nuclei/mm2. Surfaces containing periodic cavities of 2R =12 µm and 30 µm were adverse for CHO cells adhesion. In another approach, cell culture were analyzed by light microscopy (LM) for evaluation of cell viability using trypan-blue marker. This was carried out in triplicate cell culture on substrates with surfaces 12 µm, 80 µm, 280 µm, 560 µm and also on flat SU-8. Histograms were generated for average number of living cells/mm2 for each substrate, which corroborates with the results obtained for the cell culture marked with fluorophore DAPI. Thus, it is confirmed the better performance of the cell culture on substrates with 2R = 80 µm, presenting the highest number of living cells/mm2. The micrographs obtained with cells marked with DAPI were analyzed through the correlation function with the aim of understanding how the cells were organized. This was performed for each of the lithographed surfaces 12 µm, 30 µm, 80 µm, 280 µm, 560 µm and also flat SU-8. The surfaces of the substrates with 2R = 80 µm had the lowest values for length between its neighbors, that is, the cells are closer to each other. The remaining surfaces tend to separate the cells. Also were obtained the cluster radius values (rc), the distance between the clusters (dc) and the number of nearest neighbors (Np) through the correlation function fitting. The correlation analysis clearly showed what was not possible to observe by viewing the images. It shows that the cells, even in flat SU-8, have a strong tendency to form clusters of cells within about 45 micrometers. In the case of flat substrates, CHO cells exhibited better adhesion to the surface of SU-8, followed by the DLC, while the PDMS was worse due to low wettability of the material. In the case of surfaces with microstructures, SU-8 containing hexagonal microstructures of 12 and 30 µm showed to be the most adverse conditions for the CHO cell growth, probably because of the topography of the cavities being smaller in size compared to the size of CHO cells. SU-8 with 80 µm hexagonal microstructures was more favorable surface for the growth of CHO cells.