Challenges in treating injuries, together with an increased need for repair of damaged tissues and organs, have made regenerative medicine a major research area today. Biomaterials such as silk fibroin (SF) have proven to be excellent tissue scaffolds possessing properties essential in tissue engineering such as biocompatibility, biodegradability and exceptional mechanical properties. SF nanofibres are especially attractive due to their large surface-to-volume ratio and high porosity which is beneficial in regenerative medicine. However, to design biomaterial scaffolds, chemical and physical properties of SF have to be sufficiently known. The thesis aims to contribute to knowledge by characterizing silk fibroin from the African wild silkworm species Gonometa rufobrunnae, Gonometa postica, Argema mimosae, Epiphora bahuniae and Anaphe panda. Moreover, the feasibility of producing nanofibrous biomaterial scaffolds from these fibroins is explored. The chemical composition of degummed fibres was investigated using Capillary electrophoresis whilst Infrared (IR) and Raman spectroscopic techniques were utilized to determine structural characteristics of the fibroin. In addition, thermal behaviour and mechanical properties of the fibroins were also investigated. Nanofibres were fabricated via electrospinning. The effects of solution concentration, voltage, polymer flow rate and tip to collector distance were studied to give optimum electrospinning conditions. IR spectroscopy was also utilized to observe the conformational structure of the degummed and electrospun fibres whilst scanning electron microscopy (SEM) provided information on the size and morphology of the fibres. The use of the nanofibres as biomaterials was evaluated using cytotoxicity tests. Results showed that glycine, alanine and serine constituted over 70% of the amino acid composition of all the fibroins. Gonometa fibroin had more glycine than alanine whilst the opposite was true for Argema mimosae, Epiphora bahuniae and Anaphe panda fibroin. The abundance of basic amino acids in Gonometa rufobrunnae, Gonometa postica, Argema mimosae and Epiphora bahuniae fibroin makes them prime candidates for cell and tissue culture. The amino acid composition of the fibroins influenced secondary structure as the β-sheet structure. Anaphe panda, Argema mimosae and Epiphora bahuniae silks was made up of mostly alanine-alanine (Ala-Ala)n polypeptides whilst Gonometa fibroin had an interesting mixture of both glycine-alanine (Gly-Ala)n and (Ala-Ala)n units. The unique structures impacted the mechanical and thermal properties of the fibroins. Production of Gonometa nanofibres was mainly dependent on fibroin solution concentration. A minimum of 27 % w/v was needed to produce defect free nanofibres. Diameters of the electrospun fibres produced ranged from 300 to 2500 nm. IR spectroscopy data highlighted that the β-sheet conformation of degummed fibroin was degraded during the formation of the nanofibres rendering them water soluble. It was however possible to regenerate the β-sheet structure in the nanofibres by exposing them to various solvents. Cytotoxicity tests using Sulforhodamine B (SRB) assay demonstrated that the nanofibres were not toxic to cells, a major prerequisite for use as a biomaterial. This thesis successfully provides useful data in an area that has been minimally explored. Results suggest that SF from African silkworm species offers diversity in properties and are therefore attractive for use as biomaterials, especially in cell and tissue engineering. As far as we could determine, we are the first to extend the use of fibroin from African silk species by producing Gonometa SF nanofibres that are of potential use as biomaterial scaffolds. / Chemistry / D. Phil. (Chemisty)

published_or_final_version / abstract / Mechanical Engineering / Doctoral / Doctor of Philosophy

Silk fibroin has created a surge of interest for use as organic material due to its optical transparency, biocompatibility, biodegradability, and excellent physical properties. However, the implementation of silk films and structures into biomedical and sensing devices has been relatively low due to a lack of understanding of the mechanisms involved in such implementation. Increasing need for multifunctional high-performance organic materials has caused an emphasis on the ability of researchers to spatiotemporally pattern and control the structure and consequently functional properties of materials. Silk fibroin displays high potential for use as a controllable biomaterial that can be formed into a myriad of different structures for various applications. By implementation of an aqueous silk solution approach combining various fabrication techniques, several different pristine-silk and silk-composite membranes have been developed to investigate the importance of internal structuring. Different methods of investigation include: 1) incorporation of reinforcing nanoparticles within the silk matrix; 2) neutron reflectivity measurements of ultrathin silk films; 3) film patterning with nanoscale features followed by boundary organized surface mineralization of inorganic nanoparticles. The ultimate goal will be to provide fundamental data assisting in an increased knowledge of silk fibroin-based membranes and the effect of secondary structures on properties of interest.

Challenges in treating injuries, together with an increased need for repair of damaged tissues and organs, have made regenerative medicine a major research area today. Biomaterials such as silk fibroin (SF) have proven to be excellent tissue scaffolds possessing properties essential in tissue engineering such as biocompatibility, biodegradability and exceptional mechanical properties. SF nanofibres are especially attractive due to their large surface-to-volume ratio and high porosity which is beneficial in regenerative medicine. However, to design biomaterial scaffolds, chemical and physical properties of SF have to be sufficiently known. The thesis aims to contribute to knowledge by characterizing silk fibroin from the African wild silkworm species Gonometa rufobrunnae, Gonometa postica, Argema mimosae, Epiphora bahuniae and Anaphe panda. Moreover, the feasibility of producing nanofibrous biomaterial scaffolds from these fibroins is explored. The chemical composition of degummed fibres was investigated using Capillary electrophoresis whilst Infrared (IR) and Raman spectroscopic techniques were utilized to determine structural characteristics of the fibroin. In addition, thermal behaviour and mechanical properties of the fibroins were also investigated. Nanofibres were fabricated via electrospinning. The effects of solution concentration, voltage, polymer flow rate and tip to collector distance were studied to give optimum electrospinning conditions. IR spectroscopy was also utilized to observe the conformational structure of the degummed and electrospun fibres whilst scanning electron microscopy (SEM) provided information on the size and morphology of the fibres. The use of the nanofibres as biomaterials was evaluated using cytotoxicity tests. Results showed that glycine, alanine and serine constituted over 70% of the amino acid composition of all the fibroins. Gonometa fibroin had more glycine than alanine whilst the opposite was true for Argema mimosae, Epiphora bahuniae and Anaphe panda fibroin. The abundance of basic amino acids in Gonometa rufobrunnae, Gonometa postica, Argema mimosae and Epiphora bahuniae fibroin makes them prime candidates for cell and tissue culture. The amino acid composition of the fibroins influenced secondary structure as the β-sheet structure. Anaphe panda, Argema mimosae and Epiphora bahuniae silks was made up of mostly alanine-alanine (Ala-Ala)n polypeptides whilst Gonometa fibroin had an interesting mixture of both glycine-alanine (Gly-Ala)n and (Ala-Ala)n units. The unique structures impacted the mechanical and thermal properties of the fibroins. Production of Gonometa nanofibres was mainly dependent on fibroin solution concentration. A minimum of 27 % w/v was needed to produce defect free nanofibres. Diameters of the electrospun fibres produced ranged from 300 to 2500 nm. IR spectroscopy data highlighted that the β-sheet conformation of degummed fibroin was degraded during the formation of the nanofibres rendering them water soluble. It was however possible to regenerate the β-sheet structure in the nanofibres by exposing them to various solvents. Cytotoxicity tests using Sulforhodamine B (SRB) assay demonstrated that the nanofibres were not toxic to cells, a major prerequisite for use as a biomaterial. This thesis successfully provides useful data in an area that has been minimally explored. Results suggest that SF from African silkworm species offers diversity in properties and are therefore attractive for use as biomaterials, especially in cell and tissue engineering. As far as we could determine, we are the first to extend the use of fibroin from African silk species by producing Gonometa SF nanofibres that are of potential use as biomaterial scaffolds. / Chemistry / D. Phil. (Chemisty)

by Andreas Kyriacou. / Thesis (Ph.D.)--Florida Atlantic University, 2012. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2012. Mode of access: World Wide Web. / The effect of Fe substitution on the crystal structure of hydroxyapatite (HAp) is studied by applying simultaneous Rietveld refinements of powder x-ray and neutron diffraction patterns. Fe is one of the trace elements replacing Ca in HAp, which is the major mineral phase in bones and teeth. The morphology and magnetic properties of the Fe-HAp system are also studied by transmission electron microscopy and magnetization measurements. Samples of Ca(5-x)Fex(PO4)3OH with 0< x < 0.3 were prepared. Single phase HAp was identified in x-ray diffraction patterns (XRD) of samples with x < 0.1 inferring that the solubility limits are less than 0.1. Hematite ((Sa(B-Fe2O3) is identified as a secondary phase for higher Fe content. The refined parameters show that Fe is incorporated in the HAp structure by replacing Ca in the two crystallographic sites with a preference at the Ca2 site. This preference explains the small effect of the Fe substitution on the lattice constants of HAp. The overall decrease of the lattice constants is explained by the ionic vi size difference of Ca and Fe. The increasing trend of the a-lattice constant with x in the Fe substituted samples is attributed to a lattice relaxation caused by the substitution of the 4- and 6-fold Fe at the 7- and 9-fold Ca1 and Ca2 sites. This Ca local geometry reduction is indicated by a slight increase of the Ca1-O3 and Ca2-O1 bond lengths. Above the solubility limit x = 0.05, the Fe is partitioned in and out of the HAp structure with increasing nominal Fe content x. The excess Fe is oxidized to hematite. The TEM analysis and magnetic measurements support the results of the simultaneous Rietveld refinements. The TEM images show no significant effect on the morphology and size of the HAp particles upon Fe incorporation. The particles are either spheres or short rods of dimensions 20-60 nm. Hematite particles are imaged in the samples with x exceeding the solubility limit. These particles

Orientador: Rondinelli Donizetti Herculano / Coorientador: Eduardo Maffud Cilli / Coorientador: Ricardo José de Mendonça / Banca: Reinaldo Marchetto / Banca: Renê de Oliveira Beleboni / Resumo: O látex extraído da seringueira Hevea brasiliensis têm mostrado promissores resultados em aplicações biomédicas, onde m embranas produzidas dest e material têm sido utilizadas como próteses e enxertos médicos devido às suas características de biocompatibilidade e estímulo natural à angiogênese. O objetivo desse trabalho consiste na incorporação de peptídeos em membranas de látex natural e no estudo de suas liberações. Esperamos com os resultados, viabilizar o uso do látex natural como sistema de liberação sustentada de peptídeos e/ou proteínas. A taxa de liberação dos peptídeos foi monitorada e quantificada por Cromatografia Líquida de Alta Eficiênc ia (CLAE) em modo analítico. A incorporação do peptídeo às membranas de látex acompanhou diferentes caracterizações, dentre elas: Espectroscopia de Infravermelho por Transformada de Fourier (FTIR) que identificou a estabilidade dos peptídeos após incorpora ção ao látex natural pela preservação de bandas características para os peptídeos, dentre elas, as bandas 1643 - 1638 cm - 1 deformação NH de aminas e C=O de amidas; 1524 - 1514 cm - 1 deformação C=C de aromáticos; e 542 - 498 cm - 1 deformação S - S, Capacidade d e absorção de fluídos, Perda de massa em soluções aquosas, CLAE e Microscopia Eletrônica de Varredura (MEV). Durante os ensaios de liberação dos peptídeos pelas membra nas de látex natural, observou - se a instabi lidade dos peptídeos em solução, com degradaçã o total em períodos que variaram de 48 a 120 horas nos valores de pH 7,4; 12,0 ; e água destilada (pH 5,6 - 6,4) . Entretanto, para a solução de tampão acetato pH 4,0 não houve instabilidade dos peptídeos em solução. Desta forma, estes resultados sugerem a p ossível interferência causada por proteases alcal inas presentes no látex natural . Adicionalmente, com o propósito de eliminar das membranas o agente c... / Abstract: The latex extracted from Hevea brasiliensis rubber tree have shown promising results in biomedical applications, where membranes produced from this material have been used as medical implants and grafts, due to its biocompatibility and natural stimulation of angiogenesis. Th e aim of this work was t o incorporate peptides in to natural latex membranes and the study of its release. We look forward to the results, enabling the use of natural latex as Controlled Release System of peptides and/or proteins. The rate of peptide releas e were monitored and quantified by High Performance Liquid Chromatography (HPLC) in an analytical way. The peptides incorporation into latex membranes followed different characterizations, such as: Fourier Transform Infrared Spectroscopy (FTIR) which ident ified the stability of peptides after incorporation into the natural latex by the preservation of characteristic bands for peptides, among them, 1643 - 1638 cm - 1 NH strain of amines and amides C=O; 1524 - 1514 cm - 1 strain C=C aromatic; and 542 - 498 cm - 1 s train S - S, fluid absorption capacity, the loss in weight in aqueous solutions, HPLC and Scanning Electron Microscopy (SEM). During the release tests of peptides by the natural latex membranes, it was observed instability of the peptides in solution, with t otal extinction in periods that ranged between 48 and 120 hours in pH 7.4; 12.0; and distilled water (pH 5.6 - 6.4). However, for acetate buffer solution (pH 4.0) was not observed instability of peptides. Therefore, these results suggest that there are pos sible interferences caused by alkaline proteases present in the natural latex. Additionally, in order to remove the causative agent of degradation, were prepared new membranes subjected to washing treatment, and the peptides posteriorly adsorbed on the mem branes. During the release tests, i t was possible to observe the release and stability of peptides wi... / Mestre

by Tang Wing Chi = 碳-碳化鈦生物陶器之合成與表徵 / 鄧詠芝. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references. / Text in English; abstracts in English and Chinese. / by Tang Wing Chi = Tan-tan hua tai sheng wu tao qi zhi he cheng yu biao zheng / Deng Yongzhi. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Table of contents --- p.vi / List of figures --- p.xi / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Biomorphic materials --- p.1-1 / Chapter 1.2 --- Synthesis of biomorphic ceramics --- p.1-1 / Chapter 1.2.1 --- Pyrolysis --- p.1-2 / Chapter 1.2.1.1 --- Pyrolysis conditions --- p.1-3 / Chapter 1.2.2 --- Infiltration --- p.1-4 / Chapter 1.2.2.1 --- Vapor or gas infiltration --- p.1-4 / Chapter 1.2.2.2 --- Liquid infiltration --- p.1-4 / Chapter 1.2.2.3 --- Chemical vapor infiltration (CVI) --- p.1-5 / Chapter 1.2.2.4 --- Sol-gel processing --- p.1-5 / Chapter 1.2.3 --- Sintering --- p.1-6 / Chapter 1.3 --- Potential applications of biomorphic ceramics --- p.1-7 / Chapter 1.3.1 --- Homogenous porosity structures --- p.1-7 / Chapter 1.3.2 --- Heterogeneous porosity structures --- p.1-7 / Chapter 1.4 --- Methods and raw materials --- p.1-8 / Chapter 1.4.1 --- Infiltration --- p.1-8 / Chapter 1.4.2 --- Starting materials --- p.1-8 / Chapter 1.4.2.1 --- Ti source --- p.1-8 / Chapter 1.4.2.2 --- Biotemplates --- p.1-9 / Chapter 1.5 --- Objectives and approaches --- p.1-9 / Chapter 1.6 --- Thesis layout --- p.1-10 / References --- p.1-11 / Figures --- p.1-13 / Chapter Chapter 2 --- Methodology and Instrumentation / Chapter 2.1 --- Introduction --- p.2-1 / Chapter 2.2 --- Synthesis --- p.2-1 / Chapter 2.2.1 --- Biomorphic C-TiC ceramics from dragon tree --- p.2-1 / Chapter 2.2.2 --- Biomorphic C-TiC ceramics from wool sponge --- p.2-2 / Chapter 2.3 --- Characterization methods --- p.2-3 / Chapter 2.3.1 --- Differential thermal analyzer (DTA) --- p.2-3 / Chapter 2.3.2 --- Scanning electron microscopy (SEM) --- p.2-4 / Chapter 2.3.3 --- X-Ray powder diffractometry (XRD) --- p.2-4 / Chapter 2.3.4 --- Transmission electron microscopy (TEM) --- p.2-4 / Chapter 2.3.5 --- X-ray photoelectron spectroscopy (XPS) --- p.2-6 / Chapter 2.3.6 --- Compression tests --- p.2-6 / References --- p.2-9 / Figures --- p.2-8 / Chapter Chapter 3 --- The starting materials / Chapter 3.1 --- Introduction --- p.3-1 / Chapter 3.2 --- Fresh dragon tree --- p.3-1 / Chapter 3.2.1 --- SEM results --- p.3-1 / Chapter 3.2.2 --- Discussions --- p.3-2 / Chapter 3.3 --- Fresh sea wool sponge --- p.3-2 / Chapter 3.3.1 --- SEM results --- p.3-3 / Chapter 3.3.2 --- Discussions --- p.3-3 / Chapter 3.4 --- Tyzor-LA solution --- p.3-3 / Chapter 3.4.1 --- Physical properties --- p.3-4 / Chapter 3.4.2 --- DTA results --- p.3-4 / Chapter 3.4.3 --- XRD results --- p.3-5 / Chapter 3.4.4 --- Discussions --- p.3-5 / Chapter 3.5 --- Conclusions --- p.3-6 / References --- p.3-7 / Figures --- p.3-8 / Chapter Chapter 4 --- Results and discussions of biomorphic products fabricated from dragon tree / Chapter 4.1 --- Introduction --- p.4-1 / Chapter 4.2 --- Fabrication of biomorphic C-TiC ceramics --- p.4-1 / Chapter 4.2.1 --- Biotemplates from dragon tree --- p.4-1 / Chapter 4.2.1.1 --- XRD results --- p.4-1 / Chapter 4.2.1.2 --- SEM results --- p.4-2 / Chapter 4.2.1.3 --- Discussions --- p.4-2 / Chapter 4.2.2 --- Biomorphic C-TiC ceramics --- p.4-3 / Chapter 4.2.2.1 --- XRD results --- p.4-3 / Chapter 4.2.2.1.1 --- Effects of sintering temperature --- p.4-3 / Chapter 4.2.2.1.2 --- Effects of time duration --- p.4-3 / Chapter 4.2.2.2 --- SEM results --- p.4-4 / Chapter 4.2.2.3 --- TEM results --- p.4-4 / Chapter 4.2.2.3.1 --- Sample sintered at 1400。C for 6 hours --- p.4-4 / Chapter 4.2.2.3.2 --- Sample sintered at 1200°C for 6 hours --- p.4-5 / Chapter 4.2.2.3.3 --- Sample sintered at 1100。C for 6 hours --- p.4-5 / Chapter 4.2.2.3.4 --- Sample sintered at 900°C for 32 hours --- p.4-6 / Chapter 4.2.2.4 --- XPS results --- p.4-6 / Chapter 4.2.2.5 --- Results of compression tests --- p.4-7 / Chapter 4.2.2.6 --- Discussions --- p.4-7 / Chapter 4.3 --- Biomorphic C-TiC ceramics by repeated infiltration --- p.4-10 / Chapter 4.3.1 --- XRD results --- p.4-10 / Chapter 4.3.2 --- Discussions --- p.4-10 / Chapter 4.4 --- Conclusions --- p.4-11 / References --- p.4-12 / Figures --- p.4-13 / Chapter Chapter 5 --- Results and discussions of biomorphic products fabricated from sea wool sponges / Chapter 5.1 --- Introduction --- p.5-1 / Chapter 5.2 --- Fabrication of C-TiC biomorphic ceramics with sea wool sponges --- p.5-1 / Chapter 5.2.1 --- XRD results --- p.5-1 / Chapter 5.2.2 --- SEM results --- p.5-2 / Chapter 5.2.3 --- TEM results --- p.5-3 / Chapter 5.2.4 --- Discussions --- p.5-3 / Chapter 5.3 --- Dilution of Tyzor-LA solution --- p.5-5 / Chapter 5.3.1 --- XRD results --- p.5-5 / Chapter 5.3.2 --- SEM results --- p.4-5 / Chapter 5.3.3 --- Discussions --- p.5-6 / Chapter 5.4 --- Further annealing of biomorphic C-TiC ceramics in air --- p.5-6 / Chapter 5.4.1 --- XRD results --- p.5-7 / Chapter 5.4.2 --- SEM results --- p.5-7 / Chapter 5.4.3 --- TEM results --- p.5-8 / Chapter 5.4.4 --- Discussions --- p.5-9 / Chapter 5.5 --- Conclusions --- p.5-11 / References --- p.5-12 / Figures --- p.5-13 / Chapter Chapter 6 --- Conclusions and future works / Chapter 6.1 --- Conclusions --- p.6-1 / Chapter 6.2 --- Future works --- p.6-2 / References --- p.6-4

Orientador: Roberta Okamoto / Coorientador: Tetuo Okamoto / Banca: Idelmo Rangel Garcia Júnior / Banca: Thallita Pereira Queiroz / Resumo: (1) Objetivos: Avaliar por meio da análise histométrica e imunoistoquímica, o potencial osteocondutor de grânulos de hidroxiapatita e beta- tricálcio fosfato (Bone Ceramic - Straumann®) no processo de reparo de defeitos ósseos críticos em calvária de ratos. (2) Métodos: Foram utilizados ratos divididos em três grupos (n=8), submetidos à eutanásia aos 14 dias e 28 dias pós-cirúrgicos. Em cada animal foi realizado um defeito ósseo de 5mm na calvária, sendo o Grupo Coágulo (GC) com o defeito preenchido somente com coágulo; Grupo Osso Autógeno (GA), preenchido com osso autógeno e Grupo Bone Ceramic (GBC), com osso aloplástico (Straumann®). Foi avaliada a área de tecido ósseo presente na região central dos defeitos ósseos. Para a comparação entre os valores obtidos, foi realizado o teste ANOVA e como pós-teste, Tukey. Foi adotado como nível de significância p<0,05. Foi realizada análise imunoistoquímica, com os anticorpos primários contra OC, RUNX 2, TRAP e VEGF, para a avaliação dos diferentes estágios do processo de reparo ósseo. Como análise adicional, foram obtidads imagens 3D das cálvarias em microtomografia. (3) Resultados: O local do defeito ósseo originalmente tratado com BC foi totalmente preenchido por um conglomerado de biomateriais e osso neoformado, o qual foi estatisticamente superior ao GC, em ambos os períodos analisados (p<0,05). Remanescentes de BC eram visíveis e estavam em íntimo contato com o tecido ósseo. Além disso, no grupo GA as observações foram semelhantes, porém foi notado fechamento total do defeito aos 28 dias e houve maior área de neoformação óssea em todos os períodos analisados, comparando-se aos demais biomateriais (p<0,05). Defeitos que foram preenchidos com CO apresentaram apenas um tecido conjuntivo delgado fechando o defeito... / Abstract: (1)Objectives: To evaluate by histometric and immunohistochemical analysis, the osteoconductive potential of hydroxyapatite granules and beta-tricalcium phosphate (Bone Ceramic - Straumann ®) in the repair of critical bone defects in rat calvaria process. (2)Methods: The rats were divided into three groups (n = 8) and submitted to euthanasia at 14 days and 28 days post-surgery. In each animal a 5 mm calvaria bone defect was performed. In Blood Clot Group (CG) the defect was filled only with clot; in the Autogenous Bone Group (AG) the defect was filled with autogenous bone and in Bone Ceramic Group (BC), with alloplastic bone (Straumann ®).The bone tissue area was evaluated in the central region of bone defects. The results were submitted to ANOVA and Tukey's test at 5% level of significance. For the evaluation of the different stages of the bone healing process, immunohistochemical analysis was performed with primary antibodies against OC, RUNX 2, TRAP and VEGF. As an additional analysis, calvaria's 3D images were obtained in microtomography. (3)Results: The bone defect treated with BC was filled with a conglomerate of biomaterials and bone formation, which was statistically higher than CG in both periods analyzed. Remnants of BC were visible and were in intimate contact with bone tissue. Furthermore, the AG group observations were similar, but it was observed a complete defect closure after 28 days and there was a greater area of bone formation in all periods analyzed, comparing to other biomaterials (p <0.05). The defects filled with CO showed only a slender conjunctive tissue closing the bone defect. The TRAP protein was lightly immunomarked, while immunostaining of VEGF was moderate. The osteocalcin immunostaining was present in all analyzed periods (14 and 28 days). The AG group's 3D image showed full compatibility bone... / Mestre

Orientador: Ana Paula Rosifini Alves Claro / Coorientador: Rodrigo França / Coorientadora: Ana Lúcia do Amaral Escada / Banca: Roberto Zenhei Nakazato / Banca: Gilbert Silva / Banca: Paulo Cézar Soares Júnior / Banca: Cecília Amelia de Carvalho Zavaglia / Resumo: O presente trabalho tem como objetivo a obtenção de revestimentos poliméricos bactericidas formados por ácido hialurônico/quitosana/ZnO na superfície da liga Ti15Zr15Mo após crescimento de nanotubos de TiO2 empregando oxidação anódica. A liga Ti15Zr15Mo foi processada em forno a arco voltaico com atmosfera controlada e tratados termicamente. Os nanotubos de TiO2 foram obtidos por meio de oxidação anódica. Para obtenção dos revestimentos, as amostras foram recobertas com camadas de ácido hialurônico/quitosana/ZnO. As superfícies foram avaliadas empregando microscopia eletrônica de varredura (MEV), analise por difração de raios-X (DRX), espectroscopia de fotoelétrons excitados por raios-X (XPS) e ângulo de contato. A condição ideal para obtenção dos nanotubos ordenados de TiO2 na superfície da liga foi 30V, eletrólito formado por glicerol, água deionizada (1:1 em volume) e fluoreto de amônio (2,7 g/L) e tempo de 24 horas. Para esses parâmetros, os nanotubos formados apresentaram diâmetro médio de 130 nm e espessura de 1290 nm. O revestimento polimérico escolhido foi de 2 bicamadas de ácido hialurônico/quitosana contendo partículas de ZnO. Os estudos in vitro realizados para avaliar adesão bacteriana de Staphylococcus aureus mostraram que não houve diferença significativa entre as amostras com e sem recobrimento / Abstract: Nowadays the development of new materials for biomedical application has become of extreme importance due to the need of new materials with excellent mechanical properties, physicochemical and biocompatibility for use in the manufacture of biomedical devices. In addition to the material presenting the necessary bulk properties, it is of utmost importance that they present a response from the host to the implant. For this, the surface of the material must have characteristics that can be obtained with superficial treatments. This work aims at the analysis of the surface of nanotubes grown on the Ti15zr15Mo alloy functionalized with thin film of hyaluronic acid, chitosan and ZnO nanoparticles. The surface was analyzed by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and contact angle. The nanotubes formed had a mean diameter of 130 nm and a thickness of 1290 nm. The coating with the polymeric bilayers, with a number of more than 4 bilayers, closed the nanotubes and increased the contact angle, being used 2 biacamadas for the incorporation of ZnO. The concentration of the ZnO solution was very low for incorporation and bactericidal effect / Doutor

Physical factors in the environment of a cell regulate cell function and behavior and are involved in the formation and maintenance of tissue. There is strong evidence that substrate rigidity plays a key role in determining cell response in culture. Previous studies have demonstrated the importance of rigidity in numerous cellular processes including migration and adhesion and stem cell differentiation. Immune cells have been shown to respond differently to surfaces having different rigidities. Atypical response to rigidity is also a characteristic of cancerous cells. Understanding the mechanisms that support cellular rigidity sensing can lead to new tissue engineering strategies and potential new therapies based on rigidity modulation. A new technique was developed for the creation of biomimetic surfaces comprising regions of heterogeneous rigidity on the micro- and nanoscale. The surfaces are formed by exposing an elastomeric film of polydimethylsiloxane (PDMS) to a focused electron beam to form patterned regions of micro- and nanoscale spots. This thesis involves the formation of theses surfaces, characterization of their physical and chemical properties as a consequence of the electron beam exposure and investigation of how cells behave when plated on these surfaces. Cellular response to different patterns of heterogeneous rigidity is performed for several cell types. Human mesenchymal stem cells plated upon electron beam-exposed PDMS in a pattern of spots with diameters ranging from 2 µm to 100 nm display differential focal adhesion co-localization to the exposed features, depending on both rigidity and feature size. This behavior persists as the area of the exposed regions is reduced below ~1 µm. On spots with diameters of ~ 250 nm and smaller, focal adhesion co-localization is lost. This supports the notion that there is a length scale for cellular rigidity sensing, with the critical length in the range of a few hundred nanometers. When the heterogeneous rigidity surfaces are applied to CD4+ T cells, accumulations of proteins including TCR and pCasL on the exposed features are observed as a function of feature size. The pCasL appeared to significantly accumulate on 2 µm spots; For spots ~ 1 µm and below, cells appeared unable to identify the rigid regions. Further, Ca2+ release, a functional indicator of immunoresponse, is significantly enhanced on mixed-rigidity patterned PDMS relative to both soft and hard PDMS. Possible signaling pathways of TCR activation have been verified on e-beam exposed PDMS substrates with heterogeneous rigidity. These results are suggestive of possible new approaches to adoptive immunotherapy based on rigidity modulation. Studies on breast cancer cells indicate that on patterned substrates, sub-cellular processes are also significantly modulated. Integrin recruitment is enhanced on the rigid regions. Understanding the role of geometry in cellular rigidity response will point the way toward revealing its functional response and will shed light on the mechanistic underpinnings of this process.