published_or_final_version / Mechanical Engineering / Doctoral / Doctor of Philosophy

Orientador: Antonio Carlos Guastaldi / Banca: César Antunes de Freitas / Banca: Luiz Augusto Sousa Marques de Rocha / Resumo: O Ti e suas ligas são os materiais mais utilizados como biomateriais nas áreas biomédicas. Sua biocompatibilidade é resultado da estabilidade do filme de TiO2 formado sobre a superfície do material, protegendo-o contra o processo contínuo de corrosão, agravado por íons agressivos. Neste trabalho, foram utilizadas ligas de Ti fundidas contendo 6, 10 e 15% (m/m) de Mo, Ti cp e Mo metálico. As concentrações de Mo eram específicas às diferentes fases do titânio (para 6%, a fase  era a mais estável; em 10%, a fase  + ; e em 15%, a fase ). Todos os corpos-de-prova foram polidos, e, por meio de técnicas eletroquímicas (potencial de circuito aberto E com o tempo t; voltametria cíclica; espectroscopia de impedância eletroquímica), o comportamento eletroquímico deles foi analisado em soluções simulando o fluido biológico, uma vez que é comum na sociedade moderna o uso de dentifrícios contendo 1450 ppm de flúor e o uso de águas fluoretadas contendo entre 0,6 e 0,8 ppm de flúor para prevenção de cáries. Para a caracterização morfológica e análise do filme formado sobre a superfície do Ti, foram utilizadas, respectivamente, as técnicas de microscopia eletrônica de varredura e espectroscopia de fotoelétrons excitados por raios X. A liga de Ti-15Mo apresentou maior resistência à corrosão, com valores de potenciais menos negativos, seguida das ligas de Ti-10Mo e Ti-6Mo. A maior estabilidade do filme foi observada em solução de Na2SO4 0,15 mol L-1, seguida das soluções de Ringer 0,15 mol L-1, Ringer 0,15 mol L-1 mais NaF 0,15 mol L-1 e NaF 0,036 mol L-1 (voltamogramas). A resistência do filme Rp aumentou com o tempo de imersão para todas as ligas, porém, a resistência da solução Rs aumentou na presença de Cl- e F-, principalmente em F- (diagramas de Bode e Nyquist), o que diminuiu a resistência das... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ti and its alloys are the materials most used as biomaterials in the biomedical fields. Its biocompatibility is due to the stability of TiO2 film formed on this material surface, protecting it against the continuous corrosion process, injured by aggressive ions. In this work were used merged Ti alloys containing 6, 10 and 15% (w / w) of Mo, Ti cp and Mo metal. The Mo concentrations was specific for different phases of titanium (for 6%, the phase  is more stable; for 10%, it is the phase  + ; and for 15%, it is the phase ). All the specimens were polished, and by electrochemical techniques (open circuit potential E with time t; cyclic voltammetry; electrochemical impedance spectroscopy), the electrochemical behavior of them was analyzed in solutions simulating the biological fluid, as it is common in modern society the use of dentifrices containing 1450 ppm of fluoride and the use of fluoridated water containing between 0.6 and 0.8 ppm of fluoride for caries prevention. For the morphological characterization and analysis of the film formed on the Ti surface were used, respectively, techniques of scanning electron microscopy and X-ray photoelectron spectroscopy. The Ti-15Mo alloy showed a higher corrosion resistance, with less negative potential values (variation of E vs. t), followed by the Ti-10Mo and Ti-6Mo alloy. The higher film stability was observed in 0.15 mol L-1 Na2SO4 solution, followed by solutions of 0.15 mol L-1 Ringer, 0.15 mol L-1 Ringer plus 0.036 mol L -1 NaF and 0.036 mol L-1 NaF (voltammograms). The film resistance Rp increased with the immersion time for all alloys, however, the solution resistance Rs increased in the presence of Cl- and F-, mainly F- ions (Bode plots and Nyquist), reducing the alloys resistance against corrosion. There was no pitting corrosion of alloys (microscopy). The film formed on their... (Complete abstract click electronic access below) / Mestre

A series of interesting core/shell silver/phenol formaldehyde resin (PFR) nano/microstructures were also synthesized through an efficient microwave process by self-assembly growth. Various morphologies, including monodispersed nanospheres, nanocables, and microcages were obtained by changing the fundamental experimental parameters, such as the reaction time and the surfactants (Pluronic P123 or CTAB). The results indicated that the presence of triblock copolymer Pluronic P123 would result in hollow silver/PFR microcages, while CTAB would prefer the formation of ultralong silver/PFR coaxial nanocables. In the absence of surfactants, monodispersed core/shell silver/PFR nanospheres could be obtained. The size of the nanospheres can be controlled in the range of 110 to 450 nm by changing the molar ratio of reagents (phenol:hexamine). The morphology and composition of the as-prepared products were characterized. The formation mechanism of the products was discussed based on the obtained results. / Bifunctional mesoporous core/shell Ag FeNi3 nanospheres were synthesized by reducing iron(III) chloride, nickel(II) chloride and silver nitrate with hydrazine in ethylene glycol under microwave irradiation. The efficient microwave-hydrothermal process significantly shortened the synthesis time to one minute. The toxicity of Ag FeNi3 nanospheres were tested by exposing to zebrafish, they were less toxic than silver nanoparticles. In vitro MRI confirmed the effectiveness of the Ag FeNi3 nanospheres as sensitive MRI probes. The interaction of Rhodamine Band nanospheres showed greatly enhanced fluorescence over the FeNi3 nanoparticles. / Finally, a series of ZnO microarchitectures including monodispersed spindles, branches, flowers, paddies, and sphere-like clusters were prepared by an efficient microwave-hydrothermal process. The ZnO mophologies could be effectively controlled by changing the reaction conditions such as the reaction temperature, the reactant concentrations and the solvent system. Simple microspindles, interesting flowers and paddies could be obtained in the presence of hexamine, and the more attractive sphere-like clusters could be synthesized by introducing phenol. The formation mechanisms of different morphologies are discussed in detail. These interesting ZnO structures may have potential applications in electronic and optoelectronic devices. / Inorganic nanostrucured materials have attracted much attention owing to their unique features and important applications in biomedicine. This thesis describes the development of rapid and efficient approaches to synthesize inorganic nanostructures, and introduces some potential applications. / Magnetic nanostructures, such as necklace-like FeNi3 magnetic nanochains and magnetite nanoclusters, were synthesized by an efficient microwave-hydrothermal process. They were used as magnetic resonance imaging (MRI) contrast agents. Magnetic FeNi3 nanochains were synthesized by reducing iron(III) acetylacetonate and nickel(II) acetylacetonate with hydrazine in ethylene glycol solution without any template under microwave irradiation. This was a rapid and economical route based on an efficient microwave-hydrothermal process which significantly shortened the synthesis time to mins. The morphologies and size of the materials could be effectively controlled by adjusting the reaction conditions, such as, the reaction time, temperature and concentrations of reactants. The morphology and composition of the as-prepared products were characterized by field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The size of the aligned nanospheres in the magnetic FeNi 3 chains could be adjusted from 150nm to 550nm by increasing the amounts of the precursors. Magnetic measurements revealed that the FeNi3 nanochains showed enhanced coercivity and saturation magnetization. Toxicity tests by exposure of FeNi3 nanochains to the zebrafish larvae showed that the as-prepared nanochains were biocompatible. In vitro magnetic resonance imaging (MRI) confirms the effectiveness of the FeNi 3 nanochains as sensitive MRI probes. Magnetite nanoclusters were synthesized by reducing iron(III) acetylacetonate with hydrazine in ethylene glycol under microwave irradiation. The nanoclusters showed enhanced T2 relaxivity. In vitro and in vivo MRI confirmed the effectiveness of the magnetite nanoclusters as sensitive MRI probes. We also investigated the biodistribution of the nanoclusters in rat liver and spleen. / Jia, Juncai. / Adviser: Jimmy C. Yu. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Hydroxyapatite (HA) is a highly biocompatible calcium phosphate which closely resembles the mineral component of bone. Porous structures composed of biocompatible materials are believed to enhance fixation (and integration) as they encourage the ingrowth of bone into the implant. Consequently there is great interest in the potential of porous HA as an alternative to bone homo grafts and auto grafts. However, despite the interest in the field, there has been no thorough investigation into the physical and structural properties of porous HA and their effects on bone ingrowth. The material used in this study was a carbonated apatite (containing < 0.9 % levels of trace element impurities) with a trabecular macrostructure, that had been converted from bovine cancellous bone. The apparent density of the material ranged from 0.35 -1.45 g.cm-3 and the macrostructural morphology varied from an open equiaxed foam to a columnar honeycomb-like structure. The ultimate compressive stress was strongly related (r = 0.9) to the square of the apparent density, while compressive modulus was influenced by both apparent density and macrostructural morphology. Transmission electron microscopy of human osteoblast-like cells cultured on the material demonstrated that cells were closely associated with the surface. Specimens with densities of 0.6.0.9 and 1.2 g.cm-3 were then implanted in a lapine cancellous site for periods of 10 days, 3, 5, 13 and 26 weeks. After implantation all specimens elicited a highly biocompatible response, with active areas of bone deposition, remodelling and revascularization and no fibrous encapsulation. The amount of bone ingrowth within the implant (25-10%) after.5 weeks was found to vary with apparent density (0.6-1.2 g.cm-3) indicating that osseointegration was a function of macrostructural morphology. Pushout testing of retrieved spedmens indicated that all implants were securely fixed by 5 weeks (2-3 MPa). Compression testing demonstrated that after 5 weeks low density implants were sufficiently reinforced by bone ingrowth to equal the compressive strength of the host tissue (6 MPa) which increased to approximately 20 MPa at 3 and 6 months.

Lung disease is the third leading cause of death worldwide. The shortage of transplantable donor organs has profound consequences, especially for patients with end-stage lung disease, where transplantation remains the only definitive treatment. Donor organ demand far exceeds supply; and currently four out of five donor lungs are deemed unacceptable for transplantation at the time of donation, making lung the least utilized solid organ. To address the donor organ shortage, the recovery of lungs unacceptable for transplantation and the development of lung substitutes or biologic constructs capable of long-term gas exchange are under investigation. However, significant challenges remain due to a limited understanding of lung development, stem cell biology, and the mechanisms of lung injury and repair. Accordingly, novel biomaterials capable of elucidating underlying mechanisms of lung regeneration as well as facilitating strategies in lung tissue engineering, regenerative medicine, and therapeutic cell delivery would have great utility. Furthermore, robust biosystems capable of prolonged whole organ support would enable extended organ recovery times, advanced therapeutic interventions, and recipient-specific organ manipulation. The integration of new mechanistic insights into lung regeneration, novel biomaterial therapeutics, and a platform for prolonged extracorporeal organ support will enable the functional recovery and use of donor lungs initially unacceptable for transplantation. Herein is described the development and validation of (i) tissue-specific extracellular matrix biomaterials as biomedical research tools, (ii) ex vivo lung perfusion systems in small and large animal models, and (iii) a clinical-scale organ support and recovery system with novel strategies for the delivery of pulmonary therapeutics.

Orientador: Luana Marotta Reis de Vasconcellos / Banca: Luciane Dias de Oliveira / Banca: Gabriela de Fátima Santana-Melo / Resumo: O objetivo do trabalho foi analisar in vitro o comportamento de osteoblastos MG-63 em contato com membranas reabsorvíveis porosas de poli (ácido lático) (PLA) e policaprolactona (PCL), incorporadas com fibras cerâmicas de silicato de cálcio (CaSiO3), visando aplicação na regeneração óssea guiada. Foram utilizados seis grupos experimentais, a partir da concentração do teor de CaSiO3, PLA P; PLA + 5% CaSiO3; PLA + 10% CaSiO3; PCL P; PCL + 5% CaSiO3; PCL + 10% CaSiO3, e o grupo controle. Foram avaliadas viabilidade celular, genotoxicidade; produção de proteína total, atividade de fosfatase alcalina e formação matriz mineralizada. Bem como, a influência das membranas poliméricas na redução de biofilmes monotípicos de Enterococcus faecalis quando associadas ou não a solução de gluconato de clorexidina a 0,12%. Os dados foram analisados por ANOVA e Tukey (p<0.05%). Os resultados mostraram que nenhum grupo experimental foi citotóxico, mas os grupos PCL P, PLA 5%, PLA P e PCL 10% apresentaram maior viabilidade celular com diferença estatística dos grupos PCL 5% e PLA 10% (p<0.05). No teste de genotoxicidade os grupos experimentais não foram genotóxicos, já que apresentaram números de micronúcleos semelhantes ou menores ao grupo controle (p>0.05). Todos os grupos experimentais apresentaram proteína total e atividade de fosfatase alcalina semelhante estatisticamente ao controle (p>0.05) e permitiram a formação de matriz mineralizada. Com relação à redução de biofilme, os grupos experim... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this study was to analyze the behavior of MG-63 osteoblasts in contact with poly (lactic acid) (PLA) and polycaprolactone (PCL) porous resorbable membranes, incorporated with calcium silicate ceramic fibers (CaSiO3), aiming application in bone regeneration guided in periapical lesions. Six experimental groups were used, from the concentration of CaSiO3, PLA P; PLA + 5% CaSiO3; PLA + 10% CaSiO3; PCL P; PCL + 5% CaSiO3; PCL + 10% CaSiO3, and the control group. Cell viability, genotoxicity; total protein production, alkaline phosphatase activity and mineralized matrix formation. As well as the influence of polymer membranes on the reduction of monotypic biofilms of Enterococcus faecalis when associated with 0.1% chlorhexidine gluconate solution. Data were analyzed by ANOVA and Tukey (p <0.05%). The results showed that no experimental group was cytotoxic, but the PCL P, PLA 5%, PLA P and PCL 10% groups presented higher cell viability with a statistical difference between PCL 5% and PLA 10% (p <0.05). In the genotoxicity test, the experimental groups were not genotoxic, since they had similar or smaller numbers of micronuclei to the control group (p> 0.05). All experimental groups presented total protein and alkaline phosphatase activity statistically similar to the control (p> 0.05) and allowed the formation of mineralized matrix. Regarding the biofilm reduction, the PCL P and PCL 10% experimental groups showed a higher biofilm reduction for Enterococcus faecalis... (Complete abstract click electronic access below) / Mestre

Doctor of Philosophy(PhD) / Biomedical materials are being extensively researched, and many different types such as metals, metal alloys, and polymers are being used. Currently used biomedical materials are not perfect in terms of corrosion resistance, biocompatibility, and surface properties. It is not easy to fabricate from scratch new materials that can fulfill all requirements and an alternative approach is to modify the surface properties of current materials to cater to the requirements. Plasma immersion ion implantation (PIII) is an effective and economical surface treatment technique and that can be used to enhance the surface properties of biomaterials. The unique advantage of plasma modification is that the surface properties and functionalities can be enhanced selectively while the favorable bulk attributes of the materials such as strength remain unchanged. In addition, the non-line of sight feature of PIII is appropriate for biomedical devices with complex geometries such as orthopedic implants. However, care must be exercised during the plasma treatment because low-temperature treatment is necessary for heat-sensitive materials such as polymers which typically have a low melting point and glass transition temperature. Two kinds of biomedical materials will be discussed in this thesis. One is nickel titanium (NiTi) alloy which is a promising orthopedic implant material due to its unique shape memory and superelastic properties. However, harmful ions may diffuse from the surface causing safety hazards. In this study, we investigate the properties and performance of NiTi after nitrogen and oxygen PIII in terms of the chemical composition, corrosion resistance, and biocompatibility. The XPS results show that barrier layers mainly containing TiN and TiOx are produced after nitrogen and oxygen PIII, respectively. Based on the simulated in vitro and electrochemical corrosion tests, greatly reduced ion leaching and improved corrosion resistance are accomplished by PIII. Porous NiTi is also studied because the porous structure possesses better bone ingrowth capability and compatible elastic modulus with human bones. These advantages promote better recovery in patients. However, higher risks of Ni leaching are expected due to the increased exposed surface area and rougher topography than dense and smooth finished NiTi. We successfully apply PIII to porous NiTi and in vitro tests confirm good cytocompatibility of the materials. The other type of biomedical materials studied here is ultra-high molecular weight polyethylene (UHMWPE) which is a potential material for use in immunoassay plates and biosensors. In these applications, active antibodies or enzymes attached to a surface to detect molecules of interests by means of specific interactions are required. Moreover, the retention of enzyme activity is crucial in these applications. Therefore, the aim of this study is to investigate the use of PIII to prepare UHMWPE surfaces for binding of active proteins in terms of the binding density and ‘shelf life’ of the treated surfaces. Argon and nitrogen PIII treatments are attempted to modify the surface of UHMWPE. Horseradish peroxidase (HRP) is selected to conduct the protein binding test since it is a convenient protein to assay. Experimental results show that both PIII treated surfaces significantly improve the density of active HRP bound to the surface after incubation in buffer containing HRP. Furthermore, the PIII treated surfaces are found to perform better than a commercially available protein binding surface and the shelf life of the PIII treated surfaces under ambient conditions is at least six months. In conclusion, a biocompatible barrier layer on NiTi and a protein binding surface on UHMWPE is synthesized by PIII. The surface properties such as corrosion resistance and functionality on these two different types of substrates are improved by PIII.

Patients who suffer from severe joint destruction caused by arthritis often undergo total joint arthroplasty (TJA). A major limitation of this treatment and common long-term complication is the development of aseptic loosening of the prosthesis in as many as 20% of patients. The current paradigm to explain aseptic loosening proposes that wear debris generated from the prosthesis initiates a macrophage-mediated inflammatory response by resident macrophages, leading to osteoclast activation and bone resorption at the implant interface. This can then lead to the development of a peri-prosthetic fracture. The principal aim of fracture healing is to restore the bone to its original form and strength. However, this ultimate goal can be altered if the healing is impaired. This impairment may be due to bone disease (osteoporosis) or even the introduction of a foreign material such as PE wear debris that could have migrated from the articulating surface to the fracture site. A standard closed unilateral fracture of the right femur was performed in both normal and oestrogen deficient rats following fixation with a k-wire. Ceridust (PE wear debris) was combined with hyaluronic acid and saline and injected directly into the fracture site. Femurs were assessed using radiographs, histology and immunohistochemistry. Histological analysis revealed that complete remodelling was achieved in all control groups by 6 weeks post-fracture with mechanical strength returning to normal values. The mechanical properties of the fractures were not influenced by the presence of PE wear debris in the dose and timing examined. Histology and immunohistochemistry however, did reveal a local effect of the presence of PE wear debris. The histology adjacent to the PE particles was inferior to the controls but did not manifest itself in a reduction in the mechanical properties except in the oestrogen deficient bone at 6 weeks post-fracture. The levels of MMP-1 and TNF-?? correlated to the presence of PE particles. In this thesis, I have shown the mechanism by which bone remodelling in fracture healing could be retarded due to the presence of PE wear debris, by increased matrix degradation in both normal and oestrogen deficient animals.

The aims of this study were to prepare poly (ethylcyanoacrylate) (PECA) nanoparticles on the basis of different types of microemulsions, to investigate the entrapment within and release of a bioactive from these particles and to establish a set of delivery systems with varying entrapment and release characteristics, thereby giving the formulator the opportunity of a more tailor-made approach in the development of a delivery system. Furthermore the scale up of particle preparation and the possible enhancement of the immunogenic properties of PECA particles by incorporation of the adjuvant Quil A was investigated. Methods: Four phase triangles were established and microemulsion samples, used as a template to prepare nanoparticles, were characterised by viscosity and conductivity measurements, polarising light microscopy, freeze fracture transmission electron microscopy (TEM), cryo field emission electron microscopy (cryo FESEM) and self-diffusion NMR to determine their microemulsion type (droplet, bicontinuous, solution type). PECA nanoparticles were prepared from different types of microemulsions by interfacial polymerisation. Particle size, polydispersity index (PI) and [zeta]-potential were measured by photon correlation spectroscopy and electrophoretic mobility respectively. Normal scanning electron microscopy (SEM) and cryo FESEM were used to visualise particles. Fluorescently labelled ovalbumin (FITC-OVA) was used as a model protein/antigen and entrapment within and release from nanoparticles was investigated. To scale up nanoparticle preparation an instrumental set-up with reactor, peristaltic pump and stirrer was used. A 2⁷ fractional factorial study was designed to observe possible factors or their interactions that could influence particle formation under scale up conditions. For an immunological study freeze dried formulations of PECA nanoparticles, having FITC-OVA and Quil A entrapped, were prepared, and activation and uptake of formulations by murine bone marrow derived dendritic cells (DCs) and T cells in vitro were monitored. Results: Results obtained from the measurements described above, for formulations from the four different phase triangles, indicated that microemulsions of w/o droplet, bicontinuous or solution type could be formed. It was possible to prepare PECA nanoparticles from all of the different types of microemulsions. Particles had an average size of 265 nm � 24, with an average PI of 0.18 � 0.05 and an average negative [zeta]-potential of -17 mV � -5. Particle size, PI and [zeta]-potential were not influenced by the type of microemulsion that was used as a polymerisation template. Entrapment and release were however influenced by the type of microemulsion and although entrapment of FITC-OVA was generally high for PECA particles, it was highest for particles prepared from a droplet type microemulsion. Entrapment could also be increased by increasing amounts of monomer. The rate of release was dependent on the amount of monomer used for polymerisation and the type of microemulsion used for particle preparation, with nanoparticles prepared from a w/o droplet type microemulsion showing the slowest release. Furthermore it was shown that particle preparation could be scaled-up with the instrumental set-up used in this study, but conditions need to be refined as the average particle size and polydispersity index were considerably larger (441 nm � 101, 0.68 � 0.14) when compared to particles prepared by the beaker-pipette method (see above). The adjuvant Quil A could efficiently be entrapped into PECA nanoparticles together with FITC-OVA. Incubation of DCs and T cells with the various formulations did, however, not result in increased uptake or activation. Conclusions: PECA nanoparticles with high entrapment efficiency of antigen and adjuvant can be prepared from different types of microemulsions. Particles show different rates of entrapment and release depending on the type of microemulsion used as a polymerisation template, possibly because two different types of nanoparticles form. Nanocapsules are believed to form on the basis of droplet type microemulsions and nanospheres form on the basis of bicontinuous and solution type microemulsions. Freeze dried formulations of PECA nanoparticles, containing Quil A and FITC-OVA, were not able to induce an immune response, which might be due to charge repulsion effects between the negatively charged PECA nanoparticles and the negatively charged surface of dendritic cells. Moreover, no adjuvant effect of Quil A was apparent, perhaps caused by total encapsulation of the compound into the particle matrix with no active groups extending out displaying adjuvanticity.

University of Macau / Faculty of Science and Technology / Department of Electromechanical Engineering