A retrospective case study of 3 patients that developed epithelial ingrowth after laser in situ keratomileusis (LASIK). This study was conducted at Boston Eye Group in Brookline, Massachusetts. The Oculus Pentacam was used to study corneal densitometry for each patient. Corneal densitometry readings were obtained for each patient pre-operatively and post-operatively after ingrowth was discovered. Densitometry was recorded at the central nest of opacity and at the leading edges of the ingrowth. For all patients, the most severe stages of epithelial ingrowth observed on slit lamp photographs correlated to the highest densitometry readings, with peak densitometry ranging from 73.3 - 95.1. These values were much higher than pre-operative densitometry readings, which ranged from 21.8 - 27.2. In two cases, the Pentacam densitometry map revealed progression of ingrowth towards the visual axis that was only faintly detectable or not detectable at all on corresponding slit lamp photographs. Corneal densitometry can be used as an objective measure of the severity and progression of epithelial ingrowth.

The success of orthopaedic implants fixed in the skeletal system using bone ingrowth into porous surfaces is critically dependent on the extent and quality of the initial bone ingrowth and the subsequent long-term maintenance of the bone within the porous structure. Biologically-significant elements (Ca, Mg, Mn) were incorporated at concentrations up to 5 mol% in solid solution in yttia-stabilised tetragonal zirconia polycrystal (3YTZP), whilst controlling microstructure and phase composition, to investigate the effect of ceramic chemistry on cellular behaviour in vitro and bone ingrowth into porous structures in vivo. Cellular attachment, proliferation, and migration on the ceramics were investigated using in vitro assays using fibroblasts. Cells were able to adhere strongly and proliferate on all ceramic surfaces, exhibiting maximal proliferation and minimal migration on 3YTZP but significantly faster migration on doped-3YTZP. The TZP ceramics were therefore considered to support normal cellular processes and thus were suitable for further study in vivo. A technique based on pressure casting ceramic slurry into a polymer preform of the desire pore structure, followed by polymer burnout and then sintering, was developed for fabricating porous bioceramics containing highly-controlled three-dimensional pore geometries. The ability of a selected pore structure to support bone ingrowth was tested using hydroxyapatite by implanting samples into femoral cortical bone of adult sheep for 4 and 12 weeks. Bone was able to rapidly colonise the porous structure and remodel such that, by 12 weeks implantation time, the majority of the porosity was filled with mature lamellar bone. Porous scaffolds of pure 3YTZP and 3YTZP doped with 1 mol% Mg, 1 mol% Mn, 1 mol% Ca, or 5 mol% Ca were fabricated and tested in the sheep model. Bone ingrowth into the doped compositions was significantly greater than that into pure 3YTZP, and was similar to that into the porous hydroxyapatite, indicating that the dopants significantly promoted osteogenesis within the bioinert scaffolds. This finding has application in clinical applications in that the initial bone ingrowth and, potentially, the long-term maintenance of bone within the porous structure may be improved by the incorporation of small amounts of biologically significant elements.

Submitted by Gizele Lima (gizele.lima@inpa.gov.br) on 2017-04-06T13:49:08Z No. of bitstreams: 2 Versão_FINAL_Erick_Oblitas_Tese_CFT_30_Agosto_2016.pdf: 2324664 bytes, checksum: af0011498c382832eb3b3dcc609c3bda (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-06T13:49:08Z (GMT). No. of bitstreams: 2 Versão_FINAL_Erick_Oblitas_Tese_CFT_30_Agosto_2016.pdf: 2324664 bytes, checksum: af0011498c382832eb3b3dcc609c3bda (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-08-30 / Fundação de Amparo à Pesquisa do Estado do Amazonas - FAPEAM / Understanding the primary productivity in many ecosystems, including tropical forests is necessary due to the scarcity of information on many of its components, as well as its importance in the carbon cycle in terrestrial ecosystems. Most studies have focused on investigating the productivity of forests in their above-ground components, so that little is known yet about the dynamics of productivity and nutrient cycling made by the roots below ground, which is mainly linked to difficult to have to study this compartment in forest ecosystems. Moreover, the few studies on the productivity of existing roots in the Amazon, were made in a small number of plots and on short time, so that was given little attention to the spatial variability and temporal productivity of fine roots. In this context stocks and productivity of fine roots (Rp) were evaluated in 9 plots of 0,5 ha in upland forests located in the Forest Reserve Adolfo Ducke. The soils had their physical and chemical properties evaluated over time. Having been to Rp analyzed in relation to the role of controlling environmental factors and regional topographic position (plateaux, slope and valley bottoms). The areas of plateaux had a lower value of Rp (1,58±0,12 Mg C ha-1 yr-1) when compared with the valley bottoms (2,48±0,28 Mg C ha-1 yr-1). The slope areas behaved similarly to plateaux areas with 1,57±0.08 Mg C ha-1 yr-1. As to the residence time in the soil fine roots in the areas of valley bottoms have an average time of 5 months and the areas of plateaux 3 on average three months. The Rp was not sensitive to the availability of nutrients in the soil, but it seems to respond to the levels of water availability, but in a complex way. The forest structure has also strong influence on levels of Rp. Net primary productivity (NPP) was calculated with literature data of assistance to the same study plots in 8,34 Mg C ha-1 yr-1 to plateaux, 7,82 Mg C ha-1 yr-1 for slope area and 7,81 Mg C ha-1 yr-1 for the velley bottoms respectively. However allocation between above and below ground was different with valley bottoms average allocating the NPP 32% below ground, while plateaux + slope area allocated only 19,8%. The morphology of the fine roots of the slopes and valley bottoms showed no changes regarding the availability of water in as the plateaus in the dry season had higher mortality. The information presented here has the potential to contribute to the improvement of regional estimates of primary productivity in the Amazon, allowing modeling its dynamics from other variables more easily measurable, especially precipitation and forest biomass. / A compreensão da produtividade primária em muitos ecossistemas, incluindo florestas tropicais faz-se necessária, devido à escassez de informações em vários de seus componentes, assim como sua importância no ciclo do carbono em ecossistemas terrestres. A maior parte dos estudos tem enfocado em investigar a produtividade das florestas nos seus componentes acima do solo, de forma que pouco se conhece ainda sobre a dinâmica da produtividade e da ciclagem de nutrientes feitas pelas raízes abaixo do solo, o que está ligado principalmente à dificuldade de se tem em estudar esse compartimento em ecossistemas florestais. Além disso, os poucos estudos sobre produtividade de raízes existentes na Amazônia, foram feitos em um número pequeno de parcelas e sobre curto intervalo de tempo, de forma que foi dada pouca atenção à variabilidade espacial ou temporal da produtividade de raízes finas. Neste contexto os estoques e a produtividade de raízes finas (RP) foram avaliadas em 9 parcelas de 0,5 ha em florestas de terra firme localizadas na Reserva Florestal Adolfo Ducke. Os solos tiveram suas propriedades físicas e químicas avaliadas ao longo do tempo. Tendo sido a Rp analisadas em relação ao papel controlador dos fatores ambientais e da posição topográfica regional (platô, vertente e baixio). As áreas de platô tiveram um menor valor de RP (1,58±0,12 Mg C ha-1 ano- 1) quando comparada com os valores de baixio (2,48±0,28 Mg C ha-1 ano-1). As áreas de vertente se comportaram de forma semelhante às áreas do platô com 1,57±0,08 Mg C ha-1 ano -1. Em quanto ao tempo de residência no solo as raízes finas nas áreas de baixio têm em media um tempo de 5 meses e as áreas de platô 3 em media três meses. A RP não se mostrou sensível à disponibilidade de nutrientes no solo, mas parece responder aos níveis de disponibilidade de água, porém de forma complexa. A estrutura da floresta tem também forte influência nos níveis de Rp. A produtividade primária líquida (PPL) foi calculada, com auxilio de dados da literatura para as mesmas parcelas de estudo, em 8,34 Mg C ha-1 ano-1 para platô, 7,82 Mg C ha-1 ano-1 para a vertente e 7,81 Mg C ha-1 ano-1 para o baixio, respectivamente. Entretanto a alocação entre acima e abaixo do solo foi diferente, com baixios alocando em média 32% da PPL abaixo do solo enquanto platô+vertente alocaram apenas 19,8%. A morfologia das raízes finas das vertentes e baixios não apresentaram mudanças em relação a disponibilidade de agua, em quanto os platôs na estação seca tiveram maior mortalidade. As informações aqui apresentadas tem potencial para contribuir para o aperfeiçoamento das estimativas regionais de produtividade primária na Amazônia, permitindo modelar sua dinâmica a partir de outras variáveis mais facilmente mensuráveis, notadamente a precipitação e a biomassa florestal.

Additive manufactured porous titanium scaffolds have been extensively investigated for orthopaedic applications. The quantification of tissue response to the biomaterial implants is primarily achieved by analysing a two-dimensional (2D) stained histological section. More recently, three-dimensional X-ray micro-computed tomography (μCT) has become increasingly applied. Although histology is the gold standard, μCT allows non- destructive quantification of 3D tissue structures with minimal sample preparation and high contrast. A methodology to correlate information from both histology and μCT of a single sample might provide greater insights than either examining the results separately. However, this task is challenging because histology and μCT provide different types of information (stained tissue morphology vs. greyscale dependent on the X-ray absorption of material) and dimensionality (2D vs 3D). A semi-automated methodology was developed to directly quantify tissue formation and efficacy within an additive manufactured titanium implant using histology and μCT. This methodology was then extended to correlatively integrate nano-scale elemental information from nano- secondary ion mass spectroscopy (NanoSIMS). The correlative information was applied to investigate the impact of silver release on bone formation within a nano-silver coated additive manufactured implant. The correlative imaging methodology allowed for the quantification of the significant volumetric shrinkage (~15%) that occurs on histology slice preparation. It also demonstrated the importance of the location of the histological sectioning of the tissue and implant, revealing that up to 30% differences in bone ingrowth can be found along the entire length of the porous implant due to preferential bone ingrowth from the periphery to the centre. The quality and quantity of newly formed bone were found to be comparable between the uncoated and nano-silver coated Ti-implants, suggesting that the layer of silver nanoparticles on the Ti-implant does not negatively impact bone formation. Further, the newly formed bone at 2 weeks had a trabecula morphology with bone at the interface of Ti-implant as well as at a distant. This indicates that both contact (bone apposition on implant) and distance (bone ingrowth from host bone) osteogenesis were present in both types of implants. Finally, nanoscale elemental mapping showed silver was present primarily in the osseous tissue and was co-localised to sulphur suggesting that silver sulphide may have formed.

This thesis summarizes the results concerning the manufacture of medical implants for bone replacement using electron beam melting (EBM) which is an additive manufacturing (AM) technology, and aims to satisfy the engineering needs for the medical functionality of manufacturing technology. This thesis has focused on some microscopic properties for surfaces and bone integration. The process parameters of EBM manufacturing were studied to ascertain whether they have impacts on surface appearance, as surface properties have impacts on bone integration and implant performance. EBM manufacturing uses an electron beam to melt metal powder onto each layer in a manner akin to welding. The electron beam is controlled by process parameters that may be altered to a certain extent by the operator. There are individual process parameters for every material, and new parameters are set when developing new materials. In this thesis, process parameters in default settings were altered to ascertain whether it was possible to specify process parameters for implant manufacturing. The blood chamber model was used for thromboinflammation validation, using human whole blood. The model is used to identify early reactions of coagulation and immunoreactions. The material used in this study was Ti6Al4V-ELI, which is corrosion resistant and has the same surface oxide layers as titanium, and CoCr-F75, which has high stiffness, is wear-resistant and is commonly used in articulating joints. The study shows that among the process parameters researched, a combination of speed and current have the most impact on surface roughness and an interaction of parameters were found using design of experiment (DOE). As-built EBM surfaces show thrombogenicity, which in previous studies has been associated with bone ingrowth. Surface structure of as-build EBM manufactured surfaces are similar to implants surfaces described by Pilliar (2005), but with superior material properties than those of implants with sintered metals beads. By altering the process parameters controlling the electron beam, surface roughness of as-build parts may be affected, and the rougher EBM manufactured surfaces tend to be more thrombogen than the finer EBM manufactured surfaces. As-build EBM manufactured surfaces in general show more thrombogenicity than conventional machined implants surfaces. / Denna avhandling behandlar tillverkning av medicinska implantat för integration i ben. I fokus är den additiva tillverkningstekniken ”elektronstrålesmältning” ( Electron Beam Melting –EBM), en av flera tekniker som populärt beskrivs med termen 3D-skrivare. Avhandlingen fokuserar på mikroskopiska ytegenskaper och dess inverkan på benintegration. Processparametrarna för EBM-tillverkning studerades för att fastställa hur de påverkar ytans utseende, efter som ytegenskaper har effekt på implantatens funktion. EBM-tillverkning använder en elektronstråle som likt svetsning smälter ihop metallpulver. Elektronstrålen styrs av processparametrar som till viss mån kan justeras av maskinoperatören. Det finns individuella processparametrar för varje material och nya parametrar utvecklas till varje ny legering. I denna avhandling har ”grundinställningarnas processparametrar” studerats för att ta reda på om det är möjligt att ställa in specifika parametrar till implantattillverkning. Med hjälp av blodkammarmetoden, som använder humant blod, har thromboinflammatoriska egenskaper undersökts. Metoden identifierar tidiga koagulations- och immunologiska reaktioner. Legeringarna som undersökts i denna studie var Ti6Al4V-ELI, som är korrosionsbeständigt med samma uppsättning oxider på ytan som titan har, och CoCr-F75, en legering som har hög styvhet, är slitstarkt och är vanligt förekommande i implantat för leder. Bland de undersökta processparametrarna visar en kombination av hastighet och ström ha mest inverkan på ytjämnhet och en interaktion mellan parametrar identifierades med hjälp av försöksplanering. EBM-tillverkade ytor visade på thrombogena egenskaper som i tidigare studier kan relateras till god integration i benvävnad. Ytstrukturen hos EBM-tillverkade ytor liknar de implantatytor som Pilliar (2005) beskriver, men materialegenskaperna är bättre än de materialegenskaper som implantat, med sintrad yta, har. Genom att ändra processparametrarna som styr elektronstrålen kan ytstrukturen påverkas. Grövre EBM-tillverkade ytor tenderar att vara mer thrombogena än de finare EBM-tillverkade ytorna är. Obehandlade EBM-tillverkade ytor i allmänhet är mer thrombogena än vad konventionellt framställda implantatytor är.

Grasslands are characterized by recurring disturbances such as fire and grazing occurring against a background of topoedaphic heterogeneity and climatic variability. The result is a complex, multi-scaled disturbance regime, in which fire and grazing often have interactive roles, yet they have usually been studied independently. Relationships between climate, fire and simulated grazing (=mowing) were explored to determine the roles these disturbances play in shaping patterns and processes in southern mixed-grass prairie. A field experiment investigated the potential effects of these disturbances on above and belowground plant productivity, patch dynamics, and soil respiration over a 2-year period characterized by drought (1998) and normal (1999) rainfall. Spring burning and mowing had interactive effects on aboveground net primary production (ANPP). Consistent with published single factor studies, burning without mowing doubled ANPP, whereas mowing in the absence of burning had neutral effects. However, subsequent mowing on burned plots reduced ANPP gains to levels comparable with all unburned plots. Drought reduced ANPP by 22% relative to a normal rainfall year. In contrast to the traditional model of root response to defoliation, burning and mowing each stimulated root length recruitment measured with minirhizotrons. However, subsequent mowing on burned plots did not produce additional root recruitment. Fire and mowing appear to interact by affecting different components of root recruitment (production and mortality, respectively). Root biomass recovered from ingrowth cores were not correlated with minirhizotron results, and responded only to drought, suggesting that methodological differences have contributed to the varied root responses reported in the literature. Drought suppressed soil respiration, diminished soil moisture, and enhanced soil temperature, whereas fire and/or mowing had little effect. Results suggest that any fire or mowing effects on soil respiration in southern mixed-grass prairie may be highly constrained by moisture limitations during dry periods. In summary, patch level response to fire is a pulse of root recruitment followed by increased ANPP, unless subsequent grazing offsets these gains. Grazing alone produces a pulse of root recruitment, perhaps to replace consumed foliage. This study demonstrates the interactive nature of fire and grazing in grasslands, and the perils of single-factor studies.

Subscription articles: Theses and dissertations which contain embedded PJAs as part of the formal submission can be posted publicly by the awarding institution with DOI links back to the formal publications on ScienceDirect.doi:10.1016/j.msec.2015.10.069 / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19578号 / 医博第4085号 / 新制||医||1013(附属図書館) / 32614 / 京都大学大学院医学研究科医学専攻 / (主査)教授 安達 泰治, 教授 開 祐司, 教授 妻木 範行 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

A comprehensive understanding of the interactions between orthopaedic and dental implant surfaces with the surrounding host tissue is essential in the design of advanced biomaterials that better promote bone growth and osseointegration of implants. Dental implants with roughened surfaces and high surface energy are well known to promote osteoblast differentiation in vitro and promote increased bone-to-implant contact in vivo. In addition, increased surface roughness increases osteoblasts response to the vitamin D metabolite 1α,25(OH)2D3. However, the exact mechanisms mediating cell response to surface properties and 1α,25(OH)2D3 are still being elucidated. The central aim of the thesis is to investigate whether integrin signaling in response to rough surface microtopography enhances osteoblast differentiation and responsiveness to 1α,25(OH)2D3. The hypothesis is that the integrin α5β1 plays a role in osteoblast response to surface microtopography and that 1α,25(OH)2D3 acts through VDR-independent pathways involving caveolae to synergistically enhance osteoblast response to surface roughness and 1α,25(OH)2D3. To test this hypothesis the objectives of the studies performed in this thesis were: 1) to determine if α5β1 signaling is required for osteoblast response to surface microstructure; 2) to determine if increased responsiveness to 1α,25(OH)2D3 requires the vitamin D receptor, 3) to determine if rough titanium surfaces functionalized with the peptides targeting integrins (RGD) and transmembrane proteoglycans (KRSR) will enhance both osteoblast proliferation and differentiation, and 4) to determine whether caveolae, which are associated with integrin and 1α,25(OH)2D3 signaling, are required for enhance osteogenic response to surface microstructure and 1α,25(OH)2D3. The results demonstrate that integrins, VDR, and caveolae play important roles in mediating osteoblast response to surface properties and 1α,25(OH)2D3. Silencing of the β1 integrin in osteoblast-like MG63 cells significantly reduced osteogenic response to surface topography and 1α,25(OH)2D3. Silencing of the α5 subunit did not alter the response of MG63 cells to changing surface roughness or chemistry, although future work must confirm these results given similar cell surface α5 integrin expression observed in control and α5-silenced cells. Multifunctional RGD, KRSR, and KSSR coated surfaces show that RGD increased osteoblast proliferation and reduced differentiation, KRSR had no affect on osteoblast phenotype, and KSSR increased osteoblast differentiation. These results suggest that titanium surfaces can be modified to manipulate proliferation and differentiation and that RGD/KSSR functionalized surfaces could be further investigated for use as osteointegrative surfaces. The results using VDR deficient osteoblasts demonstrate that 1α,25(OH)2D3 acts via VDR-dependent mechanisms in cells cultured on titanium surfaces that support terminal differentiation. In caveolae deficient osteoblasts, 1α,25(OH)2D3 affected cell number, alkaline phosphatase activity, and TGF-β1 levels, although levels of osteocalcin and PGE2 were not affected. These results are consistent with the hypothesis that VDR is required for the actions of 1α,25(OH)2D3, but that caveolae-dependent membrane 1α,25(OH)2D3 signaling modulates traditional VDR signaling. The exact mechanisms for this interaction remain to be shown. Overall, these results are important in better understanding the role of β1 integrin partners in mediating osteoblast response to implant surfaces and in understanding how integrin signaling can alter osteoblast differentiation and responsiveness to 1α,25(OH)2D3 via genomic and non-genomic pathways.

The effect of partially saturated conditions on aqueous diffusion was investigated on the Upper Ordovician Queenston Formation shale from the Michigan Basin of southwest Ontario, Canada. Effective diffusion coefficients (De) were determined for iodide tracer on duplicate cm-scale samples from a core segment. Partially saturated conditions were created with a new gas-ingrowth method that takes advantage of the variability of N2 solubility with pressure. The method is designed to create partially saturated pores, quantify the level of partial gas/brine saturation within the tracer-accessible pore space, and measure De under fully porewater-saturated and partially gas-saturated conditions for the same sample. X-ray radiography is used with an iodide tracer for quantifying the degree of partial saturation and measuring De. The saturated De values range from 2.8 x 10-12 to 3.1 x 10-12 m2/s. Following generation of a gas phase in the pores (average gas saturations of 4 to 6.7 %), De values decrease by 20 to 22 % relative to the porewater-saturated condition, indicating that the tortuosity factor (ratio of constrictivity to tortuosity) is sensitive to saturation. Suggesting that a small volume of the pore fraction is responsible for majority of the transport. The gas-ingrowth method was successful for generating partial gas saturation, but the distribution of the gas phase is non-uniform, with relatively high gas saturations near boundaries and lower saturations in the interior of the samples.

Made available in DSpace on 2016-06-02T19:30:04Z (GMT). No. of bitstreams: 1 6174.pdf: 1522034 bytes, checksum: 23f91c32606478def4ff95b2a113c91d (MD5) Previous issue date: 2014-07-11 / Universidade Federal de Minas Gerais / Plant biomass and productivity are ecological properties that affect community functioning. The belowground biomass of cerrado is underestimated and, therefore, it is important that we know how it is related to biotic and abiotic variables. In the first chapter, we tested for the relationship between different diversity indices and aboveB and belowground biomass. Species diversity and functional divergence positively affected the aboveground biomass, but not the belowground biomass, both in the cerrado and in the seasonal forest. Resource use complementarity led to a better community functioning, but did not predict all the community biomass production, as it disregarded the belowground component. Inclusion of environmental variables and functional traits, in the second chapter, was important to generate models that predicted the belowground biomass. The models were significant, even tough they showed low explanatory power for the cerrado. Foraging for limiting nutrients, altitude, and functional traits related to disturbance were selected in the models predicting the belowground biomass. In the third chapter, we separated fine and coarse roots in two depths. We used structural equation modeling to test for the effects of environmental variables on the belowground biomass in each root category and each depth. We identified soil fertility causing less fine root biomass and recent fire causing less coarse root in the deep soil layer. Shallow root biomass was not caused by any of the ecological processes we studied. Also, aluminum content led to low soil fertility and recent fire caused higher soil fertility, as we expected. The carbon stock of the cerrado and the seasonal forest is large and should not be neglected when estimating the impacts caused by climate and landBuse changes. / A biomassa e a produtividade das plantas são propriedades ecológicas importantes para o funcionamento das comunidades. A biomassa hipógea do cerrado é subestimada, por isso, é importante sabermos sua dimensão e como ela se relaciona com fatores bióticos e abióticos. No primeiro capítulo, testamos a relação entre índices de diversidade e as biomassas epígea e hipógea. A diversidade de espécies e a divergência funcional estiveram relacionadas com a biomassa epígea, mas não com a biomassa hipógea, tanto no cerrado quanto na floresta estacional. A complementaridade no uso dos recursos levou a um melhor funcionamento das comunidades, mas não explicou toda a produção de biomassa vegetal. A inclusão de variáveis ambientais e traços funcionais, no segundo capítulo, gerou modelos que explicaram a alocação da biomassa e produtividade hipógeas. Os modelos foram significativos, apesar de terem baixo poder preditivo no cerrado. O forrageamento por nutrientes, a altitude e os traços funcionais relacionados aos distúrbios foram selecionados nos modelos prevendo a biomassa hipógea. No terceiro capítulo, separamos as raízes em finas e grossas e em dois estratos de profundidade. Usamos modelos de equações estruturais para testar os efeitos das variáveis ambientais na biomassa das raízes de cada estrato. Identificamos a fertilidade do solo causando menor biomassa de raízes finas e fogos recentes levando a menor biomassa de raízes grossas profundas. A biomassa das raízes superficiais não foi causada por nenhum dos processos ecológicos estudados e deve estar relacionada a interações bióticas. Há também relação entre a quantidade de alumínio e menor fertilidade do solo, e fogos recentes causaram maior fertilidade do solo. O estoque de carbono no cerrado e na floresta estacional semidecidual são grandes e não devem ser ignorados quando estimamos o impacto causado por mudanças climáticas e no uso da terra.