Ossos trabeculares possuem uma microestrutura porosa e podem ser modelados como um sólido elástico linear, heterogêneo e anisotrópico. A microtomografia tridimensional por raios-x (uCT) tem sido mundialmente utilizada para a caracterização de osso trabecular em pesquisas relacionadas à qualidade óssea e à doenças do metabolismo ósseo como a osteoporose. Na literatura há poucas investigações comparando as análises morfométricas 2D e 3D de osso trabecular. Nesta investigação amostras de osso trabeculares cilíndricas extraídas da cabeça femoral bovina foram utilizadas para a comparação dessas análises por \'mü\'CT. O software CT-Analyser foi utilizado para medidas em três direções (crânio-caudal, lateral-medial e anterior-posterior) de diversos parâmetros da microestrutura trabecular tais como espessura trabecular, separação trabecular, número trabecular e os autovalores do tensor de anisotropia (M). A comparação entre os valores dos parâmetros medidos por análises morfométricas 2D e 3D foi realizada pelo teste-t pareado com nível de significância p < 0,05 e por correlação linear de Pearson. Os autovalores da matriz M mostram que a microestrutura trabecular bovina tem uma tendência para a simetria transversalmente isotrópica. Os resultados obtidos mostram que a microtomografia tridimensional por raios-X é uma técnica de grande potencial para caracterização da qualidade óssea gerando bons parâmetros para o diagnóstico de doenças do metabolismo ósseo. / Trebecular bones have a porous microstructure and can be modeled as a linear elastic solid, heterogenous and anisotropic. The x-ray tridimensional microtomography (\'mü\'CT) has been worldwide used for the assessment of trabecular bone in investigations related to bone quality and aimed to the diagnostic of bone metabolic diseases such as osteoporosis. In the literature few investigations have compared the 2D and 3D morphometric analysis of trabecular bone. In this investigation cylindrical trabecular bone samples were harvested from bovine head femur to carry out that comparison by \'mü\'CT. The CT-Analyser software was used to measure in three directions (superior-inferior, lateral-medial and anterior-posterior) several microstructural trabecular parameters such as trabecular thickness, trabecular separation, trabecular number and the eigenvalues of the fabric tensor (M). The comparison between the parameters values measured by 2D and 3D morphometric analysis was performed by the paired-sample t test with a level of significance p < 0,05 and the Pearson\'s linear correlation. The eigenvalues show that the bovine trabecular microstructure has a tendence to transverse isotropy symmetry. The results show that x-ray tridimensional microtomography (\'mü\'CT) is a technique of great potential for characterization of generating good bone quality parameters for the diagnosis of diseases of bone metabolism.

<p>The trabecular meshwork (TM) is comparable to a bioactive filter that plays a major role in regulating outflow of aqueous humor of the eye and setting intraocular pressure (IOP). TM dysfunction may lead to ocular hypertension which is the major risk factor in glaucoma. Although the outflow properties of the TM have been assessed over the last sixty years, very little work has been done assessing its mechanical properties. Therefore, the major goals of these studies were two-fold: (1) to determine the relationship between mechanical properties of TM, specifically the bulk Young's modulus, and outflow function in normal and glaucomatous eyes, and (2) to establish a method and possible animal model for future testing of this relationship.</p><p>Outflow function was assessed by constant pressure perfusion in enucleated eyes at four pressure levels (10, 20, 30, and 40 mmHg) to determine outflow facilities and variability in outflow resistance with pressure elevation. A micro-strain analyzer (MSA) was used to determine the circumferential bulk Young's modulus of the TM post-perfusion. Based on their relative ease of availability, pigs and rats were explored as possible animal models. Due to the small size of rat eyes, atomic force microscopy (AFM) was used to assess the Young's modulus of TM rather than with a MSA.</p><p>We found that there was a relationship with better outflow function and a stiffer TM in normal eyes. Additionally, glaucomatous TM was found to be much softer and more variable than normal TM. Unfortunately, porcine TM did not serve as a good model for the bulk Young's modulus of human TM, presumably due to anatomical difference in its outflow pathway. Lastly, we were able to establish a new method for measuring the Young's modulus of rat TM for future work to determine potential mechanism for evaluating stiffness changes that may be associated with glaucoma.</p> / Dissertation

L'ostéoporose est un problème majeur de santé publique (150000 fractures par an en France), qui se traduit par une diminution de la masse et de la « qualité » osseuse entraînant une augmentation du risque fracturaire. Basés uniquement sur une mesure de densité minérale osseuse, les moyens cliniques actuels de diagnostic de cette pathologie apparaissent insuffisants pour quantifier précisément la résistance mécanique de l'os. Dans cette optique, il est primordial de pouvoir évaluer non seulement l'évolution de masse osseuse au cours du temps, mais également ses propriétés mécaniques tant macro que micro. Cette thèse est consacrée au développement d'un modèle numérique de remodelage osseux intégrant des paramètres mécano-biologiques permettant de simuler à la fois la dégradation d'une microstructure d'os trabéculaire au cours du temps ainsi que son adaptation mécanique. Une modification de ces paramètres permet de simuler des cas pathologiques avec ou sans traitements anti-ostéoporotiques. Le remodelage est ensuite couplé au processus de mécanotransduction, en intégrant l'influence du chargement mécanique sur la réponse cellulaire. Différents cas de chargement ont été ainsi modélisés, en particulier faible chargement tel que l'alitement, ou surchargement tel que la pratique d'activités sportives. L'hyperminéralisation ainsi que l'hétérogénéité du tissu osseux ont été intégrés au modèle et leur influence sur la localisation des sites de remodelage a été également analysée. Les résultats des simulations obtenus à partir de microarchitectures d'os humain sur plusieurs années sont encourageants car ils concordent avec les observations cliniques. / Osteoporosis is characterized by a low bone mass density but also an alteration of mechanical properties. The clinical diagnostic is made from the measure of the bone mineral density (BMD) but this examen seems insufficient to quantify bone resistance. In this work, a numerical model of cancellous bone degradation, aging and mechanical adaptation is proposed. Based on hypermineralization, this model simulates the cancellous bone remodeling process over many years. This model allows to predict the behavior of cancellous adaptation in a mechanical low loading case for instance. Results are similar to clinicial tendancy.

<p>Osteoporosis and fractures are debilitating skeletal problems. Accumulating the highest peak bone mass in both cortical and trabecular bone (Tb) as well as developing strong Tb microarchitecture play an integral role in preventing bone loss and osteoporotic fractures later in life. Because Tb is modulated by genetics (G) and environment (e.g. diet, D), my dissertation research focuses on the influence of dietary calcium (Ca) intake, genetics as well as GxD interaction controlling Tb phenotypes in two clinically relevant skeletal sites, i.e. the femur and the L5 vertebra. Male mice from 11 in bred lines and 51 BXD recombinant inbred (RI) lines were fed either adequate (Basal, 0.5%) or low (0.25%) Ca diets from 4-12 weeks of age. We used micro-computed tomography to measure Tb mass and microarchitecture phenotypes. We systematically proved that there are site-specific effects of diet, genetic, and GxD interactions influencing Tb phenotypes. This indicates that there are unique genetic effects modulating Tb at each bone site. Therefore, we conducted a genetic mapping experiment using the 51 BXD RI lines separately for each bone site. We coupled genetic mapping analysis with bioinformatics analysis to identify novel genetic variation and candidate genes accounting for the variation in each phenotypes. The findings from this work serve as a foundation for future research to identify novel pathways and genes underlying the development of Tb as well as an adaptation to Ca insufficiency.</p> <br> <p> </p>

Les buts de cette thèse étaient de définir les déterminants de la santé osseuse chez les jeunes femmes en surpoids et obèses et d'explorer les effets de deux programmes longitudinaux d'entraînement physique (force vs. endurance) sur les paramètres osseux chez les jeunes femmes en surpoids et obèses. Deux études préliminaires et six études principales ont été menées. La première étude préliminaire a montré que la VO2 max (L/mn) est un déterminant positif de la masse osseuse chez les enfants en surpoids. La deuxième étude préliminaire a démontré que l'IMC est un déterminant négatif du CSI, du BSI et de l'ISI chez les jeunes femmes. La première étude principale a montré que l'obésité est associée à de faibles valeurs de CSI, de BSI et d'ISI chez les jeunes femmes. La deuxième étude principale a montré que la VO2 max (ml/mn/kg) est positivement corrélée au CSI et à l'ISI chez les jeunes femmes. La troisième et la quatrième étude ont montré que la surcharge pondérale est associée à une augmentation du CMO et de la DMO mais pas du TBS et que le TBS est corrélé positivement à la VO2 max (L/mn). La cinquième étude principale chez 68 jeunes femmes en surpoids et obèses a montré que la détente verticale, la VO2 max (L/mn), la puissance maximale (watts) et la force maximale en demi-squat sont positivement corrélées aux paramètres osseux chez les jeunes femmes en surpoids et obèses. La sixième étude principale a montré que les deux types d'entrainement physique (force et endurance) sont efficaces pour augmenter le CMO CE et la DMO lombaire et diminuer le poids et la masse grasse. L'entrainement de force était aussi efficace pour l'augmentation du TBS et les indices de résistance osseuse du col fémoral (CSI, BSI et ISI). / The aims of this thesis were to define the determinants of bone health in young overweight and obese men and to explore the effects of two physical training programs (strength vs. endurance) on bone parameters in young overweight and obese women. Two preliminary studies and six main studies have been conducted. The first preliminary study has shown that VO2 max (L/mn) is a positive determinant of bone mass in overweight children. The second preliminary study has shown that BMI is a negative determinant of CSI, BSI and ISI in young women. The first main study has shown that obesity is associated with lower CSI, BSI and ISI values in young women. The second main study has shown that VO2 max (L/mn) is positively correlated to CSI and ISI in young women. The third and the fourth study have shown that being overweight is associated with higher BMC and BMD values but not with TBS values and that TBS is positively associated with VO2 max (L/mn). The fifth main study conducted on 68 young overweight and obese women has shown that vertical jump, VO2 max (L/min), maximal power (watts) and 1-RM half-squat are positively correlated with bone parameters in overweight and obese adult women. The sixth main study has shown that the two training programs (strength and endurance) are effective to increase whole body BMC and lumbar spine BMD, and to decrease body weight and fat mass. Strength training was also effective to increase TBS and composite indices of femoral neck strength (CSI, BSI and ISI)

Trabecular bone has a complex and porous microstructure. This study develops approaches to determine the mechanical behaviour of this material at the macroscopic level through the use of homogenisation-based multiscale methods using micro-finite element simulations. In homogenisation-based finite element methods, a simulation involving a representative volume element of the microstructure of the considered material is performed with a specific set of boundary conditions. The macroscopic stresses and strains are retrieved as averaged quantities defined over this domain. Most of the homogenisation-based work on trabecular bone has been performed to study its macroscopic elastic regime, and therefore define its constant macroscopic stiffness tensor. The rod and plate-shaped microstructure of trabecular bone can be precisely identified with advanced scanning tools, such as micro-computed tomography devices. Taking into account the size requirements to achieve a certain independence of boundary conditions for trabecular bone in a homogenisation-based multiscale setting, the resulting stack of images can have around ten million solid voxels after binarisation. Although a completely linear finite element simulation with such a large system may be feasible with commercial packages (with the proper time and memory requirements), it is not possible to perform a nonlinear simulation for such a mesh in a reasonable time frame, and the amount of required memory may not be available. A highly scalable parallel driver program which solves finite strain elastoplastic systems was developed within the framework of the existing parallel code ParaFEM. This code was used throughout this study to evaluate the yield and post-yield properties of trabecular bone. It was run on cutting edge high performance computing platforms (BlueGene/Q at the Hartree Centre, Science and Technology Facilities Council; and ARCHER, UK National Supercomputing Service, at Edinburgh Parallel Computing Centre). Micro-finite element simulations require definition of properties at the microscopic scale and it is unclear how these properties affect the macroscopic response. This study examines the effect of compressive hydrostatic yield at the microscopic scale on the macroscopic behaviour. Two different microscopic yield criteria, one permitting yielding at compressive hydrostatic stresses and the other not, were considered. A large number of load cases were examined. It was found that these two microscopic yield criteria only influence macroscopic yield behaviour in load scenarios which are compression-dominated; for other load cases, macroscopic response is insensitive to the choice of the microscopic yield criterion, provided it has an appropriate strength asymmetry. Also, in compression-dominated load cases, high density bone is much more sensitive as it is more like a continuum, resulting in the microscopic properties being more directly upscaled. Only a few previous studies have employed homogenisation to evaluate the macroscopic yield criterion of trabecular bone. However, they either used a simplified microscopic yield surface or examined only a small number of load cases. A thorough multiaxial evaluation of the macroscopic yield surface was performed by applying a wide range of loading scenarios (160 load cases) on trabecular bone samples. Closed-form yield surfaces with different symmetries (isotropy, orthotropy and full anisotropy) were fitted to the numerically obtained macroscopic yield points in strain space, and the fitting errors were evaluated in detail for different subsets of load cases. Although orthotropy and full anisotropy showed the smallest fitting errors, they were not significantly superior to the isotropic fit. Thus, isotropy in strain space presents itself as the most suitable option due to the simplicity of its implementation. The study showed that fitting errors do depend on the chosen set of load cases and that shear load cases are extremely important as it was found that even for these highly aligned samples, trabecular bone presents some degree of shear asymmetry, i.e. different strength in clockwise and counter-clockwise shear directions. There have been no previous attempts to evaluate the post-yield behaviour of trabecular bone through homogenisation-based studies on detailed micro-finite element trabecular bone meshes. A damage and plasticity constitutive law for the microscale based on existing data in the literature was considered. A homogenisation-based multiscale approach was used to evaluate the hardening and stiffness reduction at the macroscale when uniaxial load scenarios are applied to trabecular bone samples, for a small range of plastic strain Euclidean norms. Results show that damage progression at the macroscale for trabecular bone is not isotropic, which is contrary to what has been assumed previously, and that both the evolution of the yield surface and damage are different for tension, compression and shear. Nonetheless, they can be correlated with plastic strain Euclidean norms by using linear relationships. It was also observed that macroscopic damage in a specific load case affects differently the on-axis orthotropic stiffness and the off-axis orthotropic stiffness components. The findings of this study will permit the use of a more rigorous definition of the post-elastic macroscopic behaviour of trabecular bone in finite element settings.

Bone quality influences bone strength with important consequences for osteoporosis, fracture risk and dental implant success. Whilst imaging that is capable of capturing bone structure in 3D is becoming more common, quantitative clinical measures of bone quality rely on bone quantity, not structure. If bone quality could be more accurately measured, and the influence of bone architecture better understood, strength may be better predicted. This thesis presents methods for making structural comparisons between successive micro-CT images of loaded bone and explores the limitations of these. I present a novel method to detect where damage occurs in loaded rat vertebrae based on multiscale rigid registration and difference measures. Together these methods represent a quantitative approach to image guided failure analysis. Time-lapsed micro-CT images of 14 successively loaded rat vertebrae were acquired and damaged regions found using these. Using a random forest classifier I tested whether the damaged regions could be predicted by several commonly used structural measures (bone area and volume), three-dimensional texture measures (co-occurrence matrices and fractal dimension) and a more novel type of architectural measure (based on the structure tensor). A combination of parameters was able to predict damage regions with specificities in the range 70-90% and sensitivities of 60-70%.Using ovariectomised rats as a model of osteoporosis I have performed a pilot experiment to investigate how changes in bone quality might effect our results. The wider applicability of my methods are demonstrated by applying them to dental cone beam images of healthy and osteoporotic patients.

Nous avons évalué l'apport de l'analyse de la texture de radiographies de l'os trabéculaire sur le diagnostic de l'ostéoporose et sur le pronostic de l'arthrose. Tout d'abord, la combinaison de la densité minérale osseuse et de l'analyse de texture de la microarchitecture osseuse a été étudiée dans la perspective d'améliorer la détection des fractures ostéoporotiques. La texture a été analysée avec le paramètre fractal H et le TBS (Trabecular Bone Score) calculés sur des régions d’intérêt de radiographies du calcanéum et de densitométries osseuses aux vertèbres, respectivement. L'information supplémentaire apportée par l'analyse de texture a permis de discerner des femmes avec des fractures de fragilité dans le groupe des patientes non ostéoporotiques. Dans un second temps, des techniques de prétraitement ou de filtrage des images ont été exploitées afin d'améliorer l'analyse de la texture osseuse. L'une des dernières méthodes de décomposition variationnelle a été comparée avec des techniques classiques de filtrage. Les prétraitements améliorent certaines caractéristiques de texture, mais parmi celles-ci, le paramètre fractal H était plus robuste puisqu'il restait au même niveau de performance sans nécessiter de prétraitement. Enfin, les capacités prédictives de l'analyse de texture dans la progression de l'arthrose ont été étudiées à partir de radiographies numérisées du genou. La progression de l'arthrose a été évaluée par la variation du score radiographique de Kellgren et Lawrence à 4 ans d'intervalle. La signature fractale et l'entropie mesurées sur le compartiment tibio-fémoral latéral étaient significativement associées à la progression de la gonarthrose. / We have evaluated the added value of trabecular bone radiography's texture analysis to improve osteoporosis diagnosis and osteoarthritis prognosis. Firstly, fracture discrimination by combined Bone Mineral Density (BMD) and microarchitectural texture analysis has been studied. The Spine Trabecular Bone Score (TBS) and calcaneus mean Hurst parameter (H) has been assessed. The texture parameters have helped to classify a subset of non-osteoporotic women into a group with a higher risk of fractures. Secondly, preprocessing or filtering methods have been used to improve bone texture analysis. One of the latest variational decomposition model has been compared to classical filtering methods. Preprocessing improve some texture features but among them the fractal parameter H is more reliable since it doesn't need preprocessing to obtain the same performance. At last, the predictive ability of texture analysis with respect to knee osteoarthritis progression has been studied from digitalized radiography. The progression of knee osteoarthritis has been evaluated by the change in Kellgren and Lawrence scale at 4 years. The fractal signature and entropy assessed on lateral tibiofemoral compartment have been significantly associated with knee osteoarthritis progression.

In the Kuehn lab, it has been shown that inducible pluripotent stems cells that have been induced to be trabecular meshwork cell-like (iPSC-TM) have a unique ability to regenerate dysfunctional trabecular meshwork (TM) cells by sharing specific unknown factors. In this thesis will discuss the novel means by which I isolate primary human Trabecular Meshwork (pTMs) and efficiently prepare cell cultures for experimentation, such as a sequencing experiment in which I studied expression changes that arose when the TM cell culture’s cell cycle control is manipulated. Previous research has shown that pTM grow atypical when 100% confluent compared to other epithelial cells creating an interesting time frame by which to observe their unique cell cycle control. Using newly isolated TM cell cultures I investigated expression of mRNA and miRNA to understand their roles in cell cycle control of these atypical cultures. With regards to the isolation of TM cell cultures were able to show that the “Crawling Out” methodology is an effective way to establish a pure TM cell line with both a low contamination rate and less passages/time. With these cultures we were able to establish 50 mRNAs and 19 miRNAs that were differential expressed in the TM cell cultures that were atypically grown. When reviewing the literature many of these expression changes were linked to carcinogenics, and the progression/prognosis of various cancer types.

Although there are now many theories describing empirical relationships between strength properties of bone and various explanatory variables, the need for improved non-invasive diagnostic techniques to assess bone fragility is of core importance in clinical problems such as osteoporosis. The aim of this thesis was to develop non-invasive radiological methods to assess trabecular bone architecture. Measures of structural anisotropy and bone structure from X-ray or radiological projections have been developed. The first measure, the projected mean intercept length (PMIL), allows extraction of the total bone surface (BS/TV) and the mean intercept length (MIL) from projections of trabecular structure. The second measure, the line projection deviation (LPD), is a technique that quantifies the preferential alignment of trabecular bone from projections of the trabecular structure. Hence, in combination, the PMIL and LPD allow non-invasive extraction of BS/TV and more detailed preferential alignment from projections of the trabecular structure. In this thesis the PMIL and LPD are introduced and their properties explored. The PMIL and LPD are used to examine the anisotropy and architectural properties of a number of human vertebral body trabecular bone samples. When used in combination with clinical densitometry, these measures improve explanation of the variance in strength, elastic modulus and toughness of vertebral body trabecular bone samples by up to 40% when compared to densitometric values alone. While µCT can provide the information needed to access trabecular architecture, it cannot be used in clinical settings since its high radiation dose makes it only applicable to small objects ex-vivo. At present, clinically available CT does not provide sufficient resolution to resolve trabecular structures. Thus, the methods described in this thesis will allow estimates of structural parameters from plain X-rays, providing for the first time, the possibility of clinical use of such estimates. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1311218 / Thesis (Ph.D.) -- School of Medical Sciences, 2008