Indiana University-Purdue University Indianapolis (IUPUI) / Pyk2: Potential Regulator of Post-Menopausal Bone Loss H.W. LARGURA1,2*, P. ELENISTE2, S. HUANG2, S. LIU1, M. ALLEN3, A. BRUZZANITI2. 1Indiana University School of Dentistry Department Orthodontics and Oral Facial Development, 2Indiana University School of Dentistry Department of Oral Biology, 3Indiana University School of Medicine Department of Anatomy and Cell Biology, Indianapolis, Indiana, USA Osteoporosis is a pathologic condition of bone, commonly found in post-menopausal women, which occurs from an imbalance between bone formation and resorption. Following menopause, the bone resorbing activity of osteoclasts exceeds bone formation by osteoblasts, resulting in decreased trabecular and cortical bone and a subsequent decrease in bone mass. Reduced bone mass increases the risk of pathologic fracture of bones. Due to adverse effects associated with current treatment protocols for bone loss, alternative treatment modalities with reduced adverse effects are needed. Estrogen plays a role in maintaining balance in the bone remodeling cycle by controlling remodeling activation, osteoblast and osteoclast numbers, and their respective effectiveness in formation and resorption. With declining estrogen levels, this elegantly balanced interaction is altered and bone resorption exceeds bone formation, resulting in bone loss and increased bone fragility. Pyk2 is a protein tyrosine kinase that plays an important role in regulating bone resorption by osteoclasts, as well as osteoblast proliferation and differentiation. Deletion of the Pyk2 gene in mice leads to an increase in bone mass, in part due to dysfunctional osteoclast and osteoblast activity. In this study, we examined the role of Pyk2 in the effects of estrogen on bone mass. We used wild type (WT) and Pyk2 knock-out (KO) mice that had been ovariectomized (OVX) and treated with or without estrogen (E2)-releasing pellets. Control mice included sham OVX surgery receiving placebo pellet. We found that deletion of Pyk2 conferred increased bone mass in sham, OVX and OVX+E2 mice. In addition, Pyk2 KO mice supplemented with 17estradiol exhibited a marked increase in bone volume/trabecular volume, trabecular number, and trabecular thickness, but not cortical bone parameters compared to WT mice. Results of this study provide evidence for the role of Pyk2 in the effects of estrogen on bone mass. Understanding the role of Pyk2 in bone could lead to the development of new pharmaceutical targets for the treatment of bone loss associated with osteoporosis.

<p> This thesis focuses on an examination of cortical and trabecular bone density and geometry at the radius and tibia in postmenopausal women, primarily women with history of breast carcinoma, while also assessing musculoskeletal changes in postmenopausal breast cancer patients after treatment with the Aromatase Inhibitor, Anastrozole. The first sub-study is an investigation of the reproducibility of the pQCT measurement parameters at the radius and tibia in healthy pre-and postmenopausal women. Results indicated that the reproducibility was good at the radius and even better at the tibia for all parameters measured. The second study is an appraisal of the level of osteoporosis knowledge in a cohort of postmenopausal women. The participants were assessed via the Facts on Osteoporosis Quiz, a well validated questionnaire, and the data revealed significantly lower test scores among the breast cancer subjects in comparison with healthy postmenopausal women. In the remaining group of studies, pQCT technology was utilized to describe trabecular and cortical bone at the radius and tibia in postmenopausal women and women with a history of breast carcinoma whom had been prescribed Anastrozole. The following measurement sites were significantly lower in the breast cancer subjects: TOT_DEN and TOT_CNT at the 4% radius; CRT_DEN, TOT_CNT, and CRT_CNT at the 20% radius; TOT_DEN at the 4% tibia; and CRT_DEN at the 38% tibia. With respect to time on Anastrozole, TOT_CNT at the 4% radius (r=-0.36); TOT_CNT (r=-0.33), CRT_CNT (r=-0.34) and CRT_DEN (r=-0.44) at the 20% radius; and CRT_DEN (r=-0.39) and CRT_CNT (r=-0.27) at the 38% tibia were significantly negatively correlated with days on Anastrozole. Furthermore, after two years of Anastrozole treatment in a small cohort of breast cancer subjects, there was a significant decrease in CRT_DEN (p=0.025) at the 20% diaphyseal radius and also at the 38% diaphyseal tibia (p=0.051). Together, the sub-studies that comprise this thesis demonstrate that there are noteworthy deficiencies in osteoporosis knowledge among postmenopausal women, particularly those with a history of breast carcinoma, and yet, these are the same women that have an increased need to understand the preventative and treatment options regarding this disease as they demonstrate reduced bone density at all measurement sites. It also appears that time on Anastrozole primarily affects cortical bone density in these women. In summary, this thesis provides novel details regarding cortical bone in breast cancer subjects and emphasizes the need for a normative database of bone quality parameters at different skeletal sites in order to gain a better understanding of the utility of each skeletal site with regard to fracture risk prediction. </p> / Thesis / Doctor of Philosophy (PhD)

No description available.

Os bisfosfonatos são medicamentos amplamente e efetivamente utilizados para o tratamento de doenças osteolíticas. Entretanto, na cavidade oral, é de particular relevância, pois possuem como efeito adverso a osteonecrose dos maxilares induzida pelo uso de bisfosfonatos. Sua etiopatogenia ainda não é bem estabelecida, os métodos de detecção são insatisfatórios e as terapias recomendadas são por vezes, medidas paliativas e ineficazes. Pouco ainda é sabido sobre o efeito do Ácido Zoledrônico na microestrutura óssea, portanto, propusemo-nos a realizar um estudo em modelo animal que analisasse o trabeculado ósseo da mandíbula através da Micro-CT. Foram utilizados 24 ratos machos (Rattus novergicus, albinus, Wistar), com 12 semanas de vida, divididos em 2 grupos: grupo controle (cloreto de sódio 0,9%) e grupo ácido zoledrônico (ácido zoledrônico 0,6mg/kg). As substâncias foram administradas via intraperitoneal a cada 28 dias em um total de 5 doses. Após 150 dias do início do experimento, foi realizada a eutanásia dos animais e então as amostras foram preparadas e escaneadas (Skyscan 1174) para análise da microestrutura óssea através da Micro- CT. O teste t-student demonstrou diferença estatisticamente significativa (p<0,05) em todos os fatores: volume ósseo, densidade óssea, fator de padrão trabecular, índice de modelo estrutural, espessura trabecular, separação trabecular, porosidade total exceção de número de trabéculas e volume tecidual, demonstrando que há alterações significativas na estrutura trabecular pelo uso de bisfosfonatos. O grupo medicado com ácido zoledrônico comparado ao grupo controle demonstrou trabéculas mais espessas, menos separadas e com menores ligações. / Bisphosphonates are widely and effectively drugs used for the treatment of osteolytic disorders. However, in the oral cavity, this situation is of particular relevance as it can lead to bisphosphonate related osteonecrosis of the jaws. Its etiopathogenesis is still not established, detection methods are unsatisfactory and recommended therapies are sometimes palliative and often ineffective. Little is known about the effect of zoledronic acid on the quality of trabecular bone, therefore, we proposed to conduct a study in an animal model to examine the trabecular bone of the jaw through the Micro-CT. 24 male rats were used (Rattus norvegicus, Albinus, Wistar), with 12 weeks old, divided into 2 groups: control group (sodium chloride 0.9%) and group with zoledronic acid (zoledronic acid 0.6 mg / kg). The substances were administered intraperitoneally every 28 days for a total of 5 doses. After 150 days from the beginning of the experiment, the animals were sacrificed and then the samples were prepared and scanned (Skyscan 1174) for analysis of the bone microstructure through Micro-CT. The \"t-student\" test demonstrated statistically significant differences (p<0.05) in all factors: bone volume, osseous density, trabecular pattern, structure model index, trabecular thickness, trabecular separation, total porosity except trabecular number and tissue volume, demonstrating that there are significant changes in the trabecular structure of the bisphosphonates. Zoledronic Acid compared to control group shows thicker, less separate and lower connected trabeculae.

Ossos trabeculares possuem uma microestrutura porosa e podem ser modelados como um sólido elástico linear, heterogêneo e anisotrópico. A microtomografia tridimensional por raios-x (&#956CT) tem sido mundialmente utilizada para a caracterização de osso trabecular em pesquisas relacionadas à qualidade óssea e a doenças do metabolismo ósseo como a osteoporose. Na literatura existem poucas investigações sobre a influência de diferentes subvolumes amostrais na caracterização de propriedades mecânicas de osso trabecular humano através de &#956CT e ensaio mecânico computacional pelo método de Elementos Finitos. Nesta investigação doze vértebras humanas da região lombar foram utilizadas para a caracterização das propriedades mecânicas do osso trabecular através de &#956CT e modelagem computacional por elementos finitos. Uma amostra cúbica virtual com 18,5 mm de lado foi extraída de cada vértebra e quatro cubos menores centrais foram obtidos a partir dela com reduções de 20%, 40%, 60%, 80% do volume original de cada cubo. A abordagem Direct Mechanics por meio de análise de elementos finitos foi realizada através do Software FAIM (v6.0, Numerics88 Solutions Ltd). Os valores médios nas três direções principais de carregamento para os módulos de Young, Poisson e Torção obtidos foram, respectivamente, E1 = 294 MPa , E2 = 258 MPa , E3 = 153 MPa, G23 = 86 MPa, G31 = 103 MPa, G12 = 100 MPa, v21 = 0,121, v31 = 0,076, v12 = 0,137, v32 = 0,077, v13 = 0,141 e v23 = 0,140. O teste estatístico de Kruskal - Wallis ANOVA fator único foi aplicado com os procedimentos de comparação aos pares (teste de Tukey) mostrou que E1 &#8800 E3, E2 &#8800 E3 e E1 = E2. Isso indica que há a tendência para duas direções diferentes de carregamento mecânico nas amostras de osso trabecular das vértebras humanas analisadas, caracterizando uma simetria transversalmente isotrópica. Entretanto, os autovalores da matriz M (tensor anisotropia) expressaram uma tendência da microestrutura trabecular das vértebras humanas para a simetria ortotrópica, mostrando que a análise do fabric não classifica adequadamente a simetria da estrutura. A avaliação dos parâmetros microestruturais mostrou a tendência de um aumento do grau de conectividade das trabéculas a medida que ocorre uma redução dos subvolumes analisados (100% para 20% ou em milímetros de 18,5 para 3,7) acompanhado de um acréscimo dos valores da fração de volume ósseo. Esta tendência reforça a ideia de analisar-se as possíveis variações dos parâmetros morfométricos e mecânicos em domínios específicos. Em outras palavras, uma avaliação local com a escolha de volumes menores dos parâmetros microestruturais fração de volume, conectividade, espessura trabecular, separação trabecular, número trabecular e parâmetros mecânicos (Módulos de Young, Poisson e Torção) podem melhorar o prognóstico da resistência óssea, a qual prediz o risco de fratura de estruturas de osso esponjoso com precisão. Quando as propriedades mecânicas estão associadas com informações microestruturais por &#956CT são gerados mais parâmetros para se avaliar a qualidade óssea no diagnóstico de doenças do metabolismo ósseo. Portanto, a microtomografia de raios-X e análise de elementos finitos oferecem uma técnica não-invasiva com grande potencial para a avaliação da qualidade óssea. / Trabecular bone structures have a porous microstructure and can be modeled as linear elastic solid with a heterogeneous and anisotropic structure. The X-ray microtomography (&#956CT) has been used worldwide for the characterization of trabecular bone and its relationship with bone quality in metabolism diseases such as osteoporosis. In the literature there are few investigations regarding the influence of trabecular samples subvolumes in the assessment of microstructure and mechanical properties using &#956CT and finite element analysis. In this investigation twelve human vertebraes were used for the characterization of mechanical properties of trabecular bone using CT and computational modeling by finite elements. A cube sample with 18.5 mm sides was computationally extracted from each vertebrae and four smaller central cubes were obtained from it, with a 20%, 40%, 60%, 80% reduction from the original volume. The Direct Mechanics approach by finite element analysis was performed through the FAIM Software (v6.0, Numerics88 Solutions Ltd.). The mean values on three main directions of loading for Youngs Modulus, Poissons Modulus and Shears Modulus, were: E1 = 294 MPa , E2 = 258 MPa , E3 = 153 MPa G23 = 86 MPa, G31 = 103 MPa e G12 = 100 MPa, v21 = 0,121, v31 = 0,076, v12 = 0,137, v32 = 0,077, v13 = 0,141 e v23 = 0,140. The Kruskal-Wallis One Way Analysis of Variance on Ranks was applied with pairwise multiple comparison procedures (Tukey Test) showing that E1 &#8800 E3, E2 &#8800 E3 e E1 = E2. This indicates that there are two different main directions of loading on the trabecular bone samples of human vertebrae which is related to a transversely isotropic symmetry. However the eigenvalues of the matrix M (tensor anisotropy) expressed a tendency to orthotropic symmetry of the trabecular microstructure of human vertebrae, showing that the fabric assumption does not adequately classify the symmetry of the structure. The assessment of microstructural properties showed a tendency to increase the connectivity of the trabeculae with the reduction of the analyzed subvolumes (100% to 20% or 18.5 mm to 3.7 mm) as well as an increase of the bone volume fraction values. Those results highlight the idea that mechanical properties are better described in local regions. In other words, a local assessment of the microstructure parameters volume fraction, connectivity, trabecular thickness, trabecular separation, trabecular number and mechanical parameters (Youngs Modulus, Poissons Modulus and Shears Modulus) can improve the prediction of bone strength, which predicts the risk of cancellous bone fracture accurately. When the mechanical properties are associated with microstructural information more bone quality parameters are generated for the diagnosis of bone metabolism diseases and they can predict the fracture risk of cancellous bone structures with higher accuracy. Therefore, the X-ray microtomography and finite element analysis offer a non-invasive technique with great potential for the assessment of bone quality.

Os bisfosfonatos são medicamentos amplamente e efetivamente utilizados para o tratamento de doenças osteolíticas. Entretanto, na cavidade oral, é de particular relevância, pois possuem como efeito adverso a osteonecrose dos maxilares induzida pelo uso de bisfosfonatos. Sua etiopatogenia ainda não é bem estabelecida, os métodos de detecção são insatisfatórios e as terapias recomendadas são por vezes, medidas paliativas e ineficazes. Pouco ainda é sabido sobre o efeito do Ácido Zoledrônico na microestrutura óssea, portanto, propusemo-nos a realizar um estudo em modelo animal que analisasse o trabeculado ósseo da mandíbula através da Micro-CT. Foram utilizados 24 ratos machos (Rattus novergicus, albinus, Wistar), com 12 semanas de vida, divididos em 2 grupos: grupo controle (cloreto de sódio 0,9%) e grupo ácido zoledrônico (ácido zoledrônico 0,6mg/kg). As substâncias foram administradas via intraperitoneal a cada 28 dias em um total de 5 doses. Após 150 dias do início do experimento, foi realizada a eutanásia dos animais e então as amostras foram preparadas e escaneadas (Skyscan 1174) para análise da microestrutura óssea através da Micro- CT. O teste t-student demonstrou diferença estatisticamente significativa (p<0,05) em todos os fatores: volume ósseo, densidade óssea, fator de padrão trabecular, índice de modelo estrutural, espessura trabecular, separação trabecular, porosidade total exceção de número de trabéculas e volume tecidual, demonstrando que há alterações significativas na estrutura trabecular pelo uso de bisfosfonatos. O grupo medicado com ácido zoledrônico comparado ao grupo controle demonstrou trabéculas mais espessas, menos separadas e com menores ligações. / Bisphosphonates are widely and effectively drugs used for the treatment of osteolytic disorders. However, in the oral cavity, this situation is of particular relevance as it can lead to bisphosphonate related osteonecrosis of the jaws. Its etiopathogenesis is still not established, detection methods are unsatisfactory and recommended therapies are sometimes palliative and often ineffective. Little is known about the effect of zoledronic acid on the quality of trabecular bone, therefore, we proposed to conduct a study in an animal model to examine the trabecular bone of the jaw through the Micro-CT. 24 male rats were used (Rattus norvegicus, Albinus, Wistar), with 12 weeks old, divided into 2 groups: control group (sodium chloride 0.9%) and group with zoledronic acid (zoledronic acid 0.6 mg / kg). The substances were administered intraperitoneally every 28 days for a total of 5 doses. After 150 days from the beginning of the experiment, the animals were sacrificed and then the samples were prepared and scanned (Skyscan 1174) for analysis of the bone microstructure through Micro-CT. The \"t-student\" test demonstrated statistically significant differences (p<0.05) in all factors: bone volume, osseous density, trabecular pattern, structure model index, trabecular thickness, trabecular separation, total porosity except trabecular number and tissue volume, demonstrating that there are significant changes in the trabecular structure of the bisphosphonates. Zoledronic Acid compared to control group shows thicker, less separate and lower connected trabeculae.

Afin de mieux comprendre le comportement mécanique de l’os trabéculaire et d’améliorer la prédiction du risque de fracture, l’objectif de cette thèse est de développer un modèle numérique « bio-fidèle » prenant en compte l’hétérogénéité tissulaire, et de déterminer les contraintes mécaniques au sein des travées osseuses, dans le domaine de déformation élastique. À l’échelle tissulaire, une étude par nanoindentation a permis de dissocier les comportements élastiques et plastiques de l’os en fonction de sa composition (minéral/collagène). Ainsi, le comportement élastique du tissu osseux serait principalement lié à sa quantité de minéral alors que ses propriétés plastiques seraient davantage liées à la phase organique. Une loi reliant le degré de minéralisation de l’os (DMB) au module élastique a été déterminée dans l’os humain. La création d’un modèle numérique reproduisant de manière rigoureuse le comportement élastique de l’os trabéculaire, nécessite la prise en compte de l’hétérogénéité de la quantité de minéral (DMB) et donc son acquisition en 3D. Grâce à une méthode de recalage d’image 2D/3D, les acquisitions de microtomographie ont été comparées aux valeurs obtenues par microradiographie quantitative, méthode de référence de mesure du DMB. Sous certaines conditions, la microtomographie permet une évaluation correcte de l’hétérogénéité minérale. La création et l’analyse d’un modèle numérique par éléments finis de l’os trabéculaire, à partir des images de tomographie, a montré l’importance des paramètres du modèle (taille et formulation des éléments) ainsi que le rôle de l’hétérogénéité minérale sur l’évaluation des contraintes locales appliquées aux travées osseuses. / Finite element modeling has become more and more suitable to estimate the mechanical properties of trabecular bone. Such models tend to be used to evaluate bone fracture risk. The main goal of this study was to create a bio-faithful model of trabecular bone to evaluate elastic stresses fields in trabeculae. In a first part, a nanoindentation study lead to a dissociation of elastic and plastic behavior of bone tissue, depending of its composition (mineral/collagen). At osteon level, bone elastic behavior mainly depends on mineral quantity whereas its plastic behavior mainly depends on collagen maturity. The relation between degree of mineralization of bone (DMB) and elastic modulus have been determined for human bone. The purpose of second part of the study was to evaluate DMB heterogeneity inside a 3D model of trabecular bone. Using 2D/3D registration, we compare the results obtain with high resolution microtomography to those from quantitative microradiography, the goldstandard method used to measure DMB. We prove that it was possible to obtain a good evaluation of mineral heterogeneity in trabecular bone by tomography. The last part of this study, is dedicated to the creation of a finite element model of trabecular bone. After analyzing the influence of finite element modeling parameter on the assessment of mechanical response (size and element formulation), we showed that the integration of mineral heterogeneity at the tissue level lead to strong modifications of stress fields in bone trabeculae. The results of this study prove that bone mineral heterogeneity is an important parameter and should be taken into account when evaluating trabecular bone mechanical properties.

L'analyse de la microstructure osseuse joue un rôle important pour étudier des maladies de l'os comme l'ostéoporose. Des nouveaux scanners périphériques haute résolution (HR-pQCT) permettent de faire des acquisitions de la micro-architecture osseuse in-vivo sur l'homme. Toutefois la résolution spatiale de ces appareils reste comparable à la taille des travées osseuses, ce qui limite leur analyse quantitative. L'objectif de cette thèse est de proposer de nouvelles approches jointes super-résolution/ segmentation pour une analyse quantitative plus fine des images HR-pQCT in-vivo de la structure osseuse trabéculaire. Dans une première étape nous nous sommes concentrés sur des méthodes 2D de super-résolution avec régularisation par variation totale (TV) puis par variation totale d'ordre plus élevé (Higher Degree TV), avec minimisation par un algorithme ADMM (Alternating Direction Method of Multipliers). Ensuite, nous avons proposé une méthode itérative combinant le principe de Morozov et la méthode de Newton pour estimer le paramètre de régularisation TV. Comparé à la méthode UPRE (Unbiased Predictive Risk Estimator), la méthode proposée est plus rapide et ne requiert pas un balayage exhaustif des valeurs des paramètres. Nous avons développé dans une deuxième étape une méthode de super-résolution/segmentation conjointe avec un a priori basé sur la Variation Totale et une relaxation convexe (Tvbox), qui permet d'améliorer les paramètres quantitatifs de l'os et de la connectivité 3D. La méthode a été validée sur des images expérimentales micro-CT déteriorées artificiellement. Finalement, en vue de l'application à des images réelles HR-pQCT, nous nous sommes intéressés à une approche conjointe semi-aveugle super-résolution/segmentation qui vise à estimer à la fois l'image binaire super-résolue et le noyau de convolution. Des résultats sur des images micro-CT et HR-pQCT sont présentés. En conclusion, notre travail montre que les méthodes d'optimisation basées sur la régularisation TV sont prometteurs pour améliorer la quantification de la micro-architecture osseuse sur des images HR-pQCT. / The investigation of trabecular bone micro-architecture provides relevant information to determine the bone strength, an important parameter in osteoporosis investigation. While the spatial resolution of clinical CT is not sufficient to resolve the trabecular structure, the High Resolution peripheral Quantitative CT (HR-pQCT) has been developed to investigate bone micro-architecture in-vivo at peripheral sites (tibia and radius). Despite this considerable progress, the quantification of 3D trabecular bone micro-architecture in-vivo remains limited due to a lack of spatial resolution compared to the trabeculae size. The objective of this thesis is to propose new joint super-resolution/segmentation approaches for improving the quantitative analysis of in-vivo HR-pQCT images of the trabecular bone structure. To begin with, we have investigated 2D super-resolution methods based on Total Variation (TV) and Higher Degree Total Variation (HDTV) and Alternating Direction Method of Multipliers (ADMM) minimization. Afterwards, an iterative method combining the Morozov principle and the Newton method was proposed in order to estimate the TV regularization parameter. The proposed method provides a very good regularization parameter only in few iterations compared with the UPRE method that requires an extensive scanning of parameter values. Furthermore, we have developed a 3D joint super-resolution/segmentation method based on a TV a prior with a convex relaxation (TVbox). The validation of the proposed methods was made on experimental micro-CT bone images artificially deteriorated. The results showed an improvement of the bone parameters and 3D connectivity with the TVbox method. Moreover, we have investigated a semi-blind joint super-resolution/ segmentation approach aiming to estimate both the binary super-resolved image and the assumed Gaussian blurring kernel that is not known for the real HR-pQCT images. Results on micro-CT and HR-pQCT experimental bone images were presented. In conclusion, our work has shown that TV based regularization methods promise to improve the quantification of bone micro-architecture from HR-pQCT images.

Ossos trabeculares possuem uma microestrutura porosa e podem ser modelados como um sólido elástico linear, heterogêneo e anisotrópico. A microtomografia tridimensional por raios-x (&#956CT) tem sido mundialmente utilizada para a caracterização de osso trabecular em pesquisas relacionadas à qualidade óssea e a doenças do metabolismo ósseo como a osteoporose. Na literatura existem poucas investigações sobre a influência de diferentes subvolumes amostrais na caracterização de propriedades mecânicas de osso trabecular humano através de &#956CT e ensaio mecânico computacional pelo método de Elementos Finitos. Nesta investigação doze vértebras humanas da região lombar foram utilizadas para a caracterização das propriedades mecânicas do osso trabecular através de &#956CT e modelagem computacional por elementos finitos. Uma amostra cúbica virtual com 18,5 mm de lado foi extraída de cada vértebra e quatro cubos menores centrais foram obtidos a partir dela com reduções de 20%, 40%, 60%, 80% do volume original de cada cubo. A abordagem Direct Mechanics por meio de análise de elementos finitos foi realizada através do Software FAIM (v6.0, Numerics88 Solutions Ltd). Os valores médios nas três direções principais de carregamento para os módulos de Young, Poisson e Torção obtidos foram, respectivamente, E1 = 294 MPa , E2 = 258 MPa , E3 = 153 MPa, G23 = 86 MPa, G31 = 103 MPa, G12 = 100 MPa, v21 = 0,121, v31 = 0,076, v12 = 0,137, v32 = 0,077, v13 = 0,141 e v23 = 0,140. O teste estatístico de Kruskal - Wallis ANOVA fator único foi aplicado com os procedimentos de comparação aos pares (teste de Tukey) mostrou que E1 &#8800 E3, E2 &#8800 E3 e E1 = E2. Isso indica que há a tendência para duas direções diferentes de carregamento mecânico nas amostras de osso trabecular das vértebras humanas analisadas, caracterizando uma simetria transversalmente isotrópica. Entretanto, os autovalores da matriz M (tensor anisotropia) expressaram uma tendência da microestrutura trabecular das vértebras humanas para a simetria ortotrópica, mostrando que a análise do fabric não classifica adequadamente a simetria da estrutura. A avaliação dos parâmetros microestruturais mostrou a tendência de um aumento do grau de conectividade das trabéculas a medida que ocorre uma redução dos subvolumes analisados (100% para 20% ou em milímetros de 18,5 para 3,7) acompanhado de um acréscimo dos valores da fração de volume ósseo. Esta tendência reforça a ideia de analisar-se as possíveis variações dos parâmetros morfométricos e mecânicos em domínios específicos. Em outras palavras, uma avaliação local com a escolha de volumes menores dos parâmetros microestruturais fração de volume, conectividade, espessura trabecular, separação trabecular, número trabecular e parâmetros mecânicos (Módulos de Young, Poisson e Torção) podem melhorar o prognóstico da resistência óssea, a qual prediz o risco de fratura de estruturas de osso esponjoso com precisão. Quando as propriedades mecânicas estão associadas com informações microestruturais por &#956CT são gerados mais parâmetros para se avaliar a qualidade óssea no diagnóstico de doenças do metabolismo ósseo. Portanto, a microtomografia de raios-X e análise de elementos finitos oferecem uma técnica não-invasiva com grande potencial para a avaliação da qualidade óssea. / Trabecular bone structures have a porous microstructure and can be modeled as linear elastic solid with a heterogeneous and anisotropic structure. The X-ray microtomography (&#956CT) has been used worldwide for the characterization of trabecular bone and its relationship with bone quality in metabolism diseases such as osteoporosis. In the literature there are few investigations regarding the influence of trabecular samples subvolumes in the assessment of microstructure and mechanical properties using &#956CT and finite element analysis. In this investigation twelve human vertebraes were used for the characterization of mechanical properties of trabecular bone using CT and computational modeling by finite elements. A cube sample with 18.5 mm sides was computationally extracted from each vertebrae and four smaller central cubes were obtained from it, with a 20%, 40%, 60%, 80% reduction from the original volume. The Direct Mechanics approach by finite element analysis was performed through the FAIM Software (v6.0, Numerics88 Solutions Ltd.). The mean values on three main directions of loading for Youngs Modulus, Poissons Modulus and Shears Modulus, were: E1 = 294 MPa , E2 = 258 MPa , E3 = 153 MPa G23 = 86 MPa, G31 = 103 MPa e G12 = 100 MPa, v21 = 0,121, v31 = 0,076, v12 = 0,137, v32 = 0,077, v13 = 0,141 e v23 = 0,140. The Kruskal-Wallis One Way Analysis of Variance on Ranks was applied with pairwise multiple comparison procedures (Tukey Test) showing that E1 &#8800 E3, E2 &#8800 E3 e E1 = E2. This indicates that there are two different main directions of loading on the trabecular bone samples of human vertebrae which is related to a transversely isotropic symmetry. However the eigenvalues of the matrix M (tensor anisotropy) expressed a tendency to orthotropic symmetry of the trabecular microstructure of human vertebrae, showing that the fabric assumption does not adequately classify the symmetry of the structure. The assessment of microstructural properties showed a tendency to increase the connectivity of the trabeculae with the reduction of the analyzed subvolumes (100% to 20% or 18.5 mm to 3.7 mm) as well as an increase of the bone volume fraction values. Those results highlight the idea that mechanical properties are better described in local regions. In other words, a local assessment of the microstructure parameters volume fraction, connectivity, trabecular thickness, trabecular separation, trabecular number and mechanical parameters (Youngs Modulus, Poissons Modulus and Shears Modulus) can improve the prediction of bone strength, which predicts the risk of cancellous bone fracture accurately. When the mechanical properties are associated with microstructural information more bone quality parameters are generated for the diagnosis of bone metabolism diseases and they can predict the fracture risk of cancellous bone structures with higher accuracy. Therefore, the X-ray microtomography and finite element analysis offer a non-invasive technique with great potential for the assessment of bone quality.

Cette thèse porte sur le développement d’une nouvelle technique de modélisation des problèmes IFS utilisant les méthodes particulaires. Ce travail s’inscrit dans la continuité des travaux de recherche de l’équipe biomécanique du LAMIH, concernant la compréhension du comportement de l’os humain dans son environnement de moelle osseuse. La méthode SPH a été utilisée pour la modélisation des travées osseuses, supposées dans une première approche comme des solides élastiques. La méthode LB a été développée pour la modélisation des écoulements de moelle considérée comme un fluide visqueux incompressible. L’efficacité et la performance de ces deux méthodes ont été démontrées grâce aux benchmarks académiques évalués et les résultats comparés à ceux de la littérature ou ceux obtenus par des logiciels commerciaux. A l’issue d’une revue de l’état de l’art des techniques de couplage fluide-structure, une approche partitionnée en temps a été choisie, permettant d’utiliser deux codes distincts basés sur des algorithmes de résolution de type dynamique explicite. La discrétisation spatiale est faite par une technique spécifique basée sur les domaines fictifs, cette technique est très efficace car elle ne nécessite pas de rediscrétisation des domaines. L’approche de couplage développée a été appliquée à des benchmarks académiques ainsi qu’à une application en biomécanique, ayant permis d’aboutir à des résultats numériques satisfaisants. Plusieurs pistes d’amélioration sont maintenant nécessaires afin d’aller vers des modélisations plus biofidèles telles que la prise en compte du contact et de l’endommagement. / The objective of this thesis is the development of a new technique for the FSI problems modelling using particulars methods. This work is in the continuity of the LAMIH biomechanics team research works, regarding the comprehension of behavior of bone in its environment of marrow. The SPH method was used for the trabeculae modelling, supposed in a first attempt as an elastic solid. The LB method was developed for the marrow flow modelling considered as a viscous incompressible liquid. The efficacy and performance of these two methods were demonstrated using academics benchmarks which were evaluated and the results were compared of those of literature and of those obtained from commercials softwares. Following a bibliographic review of FSI coupling techniques, a partitioned approach in time was chosen, allowing the use of two separates codes, both based on a dynamic explicit algorithm resolution scheme. The special discretization was done based on a specific technique of fictional domain, this technique is very efficient because it doesn’t require an additional domain discretization. The coupling approach developed was applied on academic benchmarks and on a biomechanical application, leading to satisfactory numerical results. Many Improvement track are now necessary to go towards more biofidelic modeling as taking into account the contact and the damage.