The osteocytes are the most abundant and longest living bone cells, embedded in the bone matrix. They are interconnected with each other through dendrites, located in slender canals called canaliculi. The osteocyte lacunae, cavities in which the cells are located, together with the canaliculi form a communication network throughout the bone matrix, permitting transport of nutrients, waste and signals. These cells were firstly considered passive, but lately it has become increasingly clear their role as mechanosensory cells and orchestrators of bone remodeling. Despite recent advances in imaging techniques, none of the available methods can provide an adequate 3D assessment of the lacuno-canalicular network (LCN). The aims of this thesis were to achieve 3D imaging of the LCN with synchrotron radiation X-ray computed tomography (SR-CT) and to develop tools for 3D detection and segmentation of this cell network, leading towards automatic quantification of this structure. We demonstrate the feasibility of parallel beam SR-CT to image in 3D the LCN (voxel~300 nm). This technique can provide data on both the morphology of the cell network and the composition of the bone matrix. Compared to the other 3D imaging methods, this enables imaging of tissue covering a number of cell lacunae three orders of magnitude greater, in a simpler and faster way. This makes possible the study of sets of specimens in order to reach biomedical conclusions. Furthermore, we propose the use of divergent holotomography, to image the ultrastructure of bone tissue (voxel~60 nm). The image reconstruction provides phase maps, obtained after the application of a suitable phase retrieval algorithm. This technique permits assessment of the cell network with higher accuracy and it enables the 3D organization of collagen fibres organization in the bone matrix, to be visualized for the first time. In order to obtain quantitative parameters on the geometry of the cell network, this has to be segmented. Due to the limitations in spatial resolution, canaliculi appear as 3D tube-like structures measuring only 1-3 voxels in diameter. This, combined with the noise, the low contrast and the large size of each image (8 GB), makes the segmentation a difficult task. We propose an image enhancement method, based on a 3D line filter combined with bilateral filtering. This enables improvement in canaliculi detection, reduction of the background noise and cell lacunae preservation. For the image segmentation we developed a method based on variational region growing. We propose two expressions for energy functionals to minimize in order to detect the desired structure, based on the 3D line filter map and the original image. Preliminary quantitative results on human femoral samples are obtained based on connected components analysis and a few observations related to the bone cell network and its relation with the bone matrix are presented.

Le groupe des KDIGO a défini un nouveau syndrome regroupant les troubles minéraux et osseux et les calcifications vasculaires au cours de la maladie rénale chronique {TMO-MRC). Les TMO-MRC sont associés à une augmentation du risque de fracture, de calcification vasculaire et à une surmortalité. La quête d'un biomarqueur fiable de la maladie osseuse et des calcifications vasculaires reste le défi du néphrologue. Au cours des dernières années, de nombreuses protéines de l'os ont été associées aux calcifications vasculaires chez les patients urémiques, tels que les phosphatases alcalines osseuses et, plus récemment, la sclérostine. Cette petite protéine est secrétée par l'ostéocyte et inhibe l'ostéoformation en bloquant la voie de signalisation Wnt dans l'ostéoblaste. Il a récemment été suggéré que la sclérostine aurait une activité catabolique sur l'os et serait impliquée dans la déminéralisation du squelette. L'objectif de ce travail était d'étudier la sclérostine au cours de la MRC. Nous avons tout d'abord montré que les concentrations sériques de sclérostine augmentaient avec la baisse du débit de filtration glomérulaire mesurée par la clairance de l'inuline et ce dès le stade 3 de la MRC, indépendamment de l'âge. De plus, cette étude nous a permis de montrer pour la première fois que la phosphorémie était indépendamment et positivement associée à la sclérostine sérique. Ensuite, nous avons retrouvé une association positive et forte entre la concentration sérique de sclérostine et les calcifications vasculaires chez des patients en hémodialyse chronique. Enfin, nous avons montré que les calcifications artérielles étaient significativement associées à une qualité osseuse corticale altérée étudiée en scanner quantitatif de haute résolution. Ces résultats suggèrent que la sclérostine pourrait constituer un messager important dans la relation entre l'os et la paroi vasculaire calcifiée des patients atteints d'une insuffisance rénale chronique terminale / The KDlGO group identified a new syndrome involving mineral and bone disorders and vascular calcification in chronic kidney disease (CKD-MBD}. CKD-MBD are associated with an increased risk of fracture, vascular calcification and increased mortality. The search for a reliable biomarker of bone disease and vascular calcification remains the challenge of the nephrologist. ln recent years, many bone proteins have been associated with vascular calcification in uremic patients, such as bone alkaline phosphatase and more recently sclerostin. This small protein is secreted by the osteocyte and is an inhibitor of bone formation by blocking the Wnt signaling in osteoblasts. lt has recently been suggested that sclerostin has a catabolic activity on bone and is involved in the demineralization of the skeleton. The aim of this research was to study the sclerostin during the course of CKD. We first showed that serum levels of sclerostin increased with the decrease of glomerular filtration rate measured by inulin clearance already from stage 3 chronic kidney disease, regardless of age. Furthermore, in this study, we showed for the first time that serum phosphorus was independently and positively associated with serum sclerostin. Subsequently we found a positive and strong association between serum sclerostin and vascular calcification in maintenance hemodialysis patients. Finally, we have shown that arterial calcification were significantly associated with an altered cortical bone quality studied by high resolution peripheral quantitative computed tomography. These results suggest that sclerostin could be an important messenger in the cross-talk between the bone and the calcified vascular wall in end stage renal disease

L’ostéoporose ménopausique est généralement traitée par une stratégie médicamenteuse à visée anti-résorptive, couplée à des exercices de musculation/proprioception relativement efficaces pour maintenir la masse musculaire et prévenir le risque de chute. Ces dernières années, bien que de nombreuses études animales et cliniques aient suggéré un effet ostéogénique des vibrations corps entier (VCE), les résultats pas toujours concluants et les protocoles très hétérogènes rendent l’interprétation difficile. Dans ce travail, nous avons étudié l’importance de la fréquence de la vibration dans les effets osseux induits par les VCE, sur des rats adultes. Nous avons montré que la stimulation osseuse était dose dépendante de la fréquence, avec un effet ostéogénique d’autant plus important que la fréquence est élevée, et un effet délétère des basses fréquences. Le régime à haute fréquence (90Hz) améliore la micro-macroarchitecture de l’os cortical (épaississement cortical, diminution de la porosité) et de l’os trabéculaire (augmentation du volume osseux, nombre de travées), et stimule la formation osseuse (taux de formation osseuse augmenté), alors que le régime basse fréquence (8Hz) découple les activités de formation et minéralisation, responsable d’une diminution des DMO corticales et trabéculaires, caractéristiques d’une ostéomalacie. Dans cette étude, la réponse ostéogénique plus marquée aux vertèbres vs les os longs (Tibia et fémur) suggère un rôle de la moelle grasse dans la réponse osseuse aux VCE. L’étude clinique réalisée par la suite chez des femmes ménopausées, confirme la capacité des VCE à stimuler la formation osseuse corticale et trabéculaire (épaississement cortical, augmentation de l’aire corticale, maintien de la porosité, augmentation du volume osseux trabéculaire) avec un effet systémique des VCE (os porteurs et non porteurs). L’étude de la propagation du signal vibratoire chez l’animal et l’humain montre une amplification du signal dans les basses fréquences, caractéristique d’un effet de résonance, et une transmission plus importante des vibrations au-delà de 40Hz, expliquant en partie les effets des VCE en fonction de la fréquence. Nos résultats suggèrent l’utilisation des VCE comme moyen non pharmacologique de prévention voir traitement de la fragilité osseuse / The postmenopausal osteoporosis is generally handled by a medicinal strategy with anti-résorptive aim, coupled with relatively effective exercises of body-building / proprioception to maintain the muscular mass and prevent the risk of fall. These last years, although numerous animal and clinical studies suggested an osteogenic effect of the whole-body vibrations (WBV), the not always decisive results and the very heterogeneous protocols make difficult the interpretation. In this work, we studied the importance of the frequency of vibration in the bone effects induced by WBV on adult rats. We have shown that bone stimulation was dependent on the dose frequency, with a particularly important that the frequency is high osteogenic effect and a detrimental effect of low frequency. The high frequency system (90Hz) improves the control of the microarchitecture cortical bone (cortical thickening, reduced porosity) and trabecular bone (bone augmentation, number of spans), and stimulate bone formation (bone formation rate increased), while the low frequency regime (8Hz) decouples training and mineralization, causing a decrease in cortical and trabecular BMD, characteristic of osteomalacia. In this study, the more pronounced the vertebrae vs long bones (femur and tibia) osteogenic response suggests a role of the fat in the bone marrow response to WBV. The clinical study later postmenopausal women, confirms the ability of WBV to stimulate cortical and trabecular bone formation (cortical thickening, increased cortical area, maintaining porosity, trabecular bone volume increase) with a systemic effect of WBV (bearing and non-bearing bones). The study of the propagation of the vibration signal in animals and humans shows an amplification of the signal in the low frequency characteristic of a resonance effect, and a greater transmission of vibrations beyond 40Hz, explaining part of the effects according to the WBV frequency. Our results suggest the use of WBV as non-pharmacological means of prevention, or even treatment, of bone fragility

Les effets des agents anti-ostéoporotiques sur le tissu osseux sont évalués au niveau de la crête iliaque (CI) mais les réponses aux traitements peuvent varier selon le site osseux. Le but de cette étude était de comprarer les effets de l’acide zolérodronique (ZOL) et du tériparatide (TPTD) au niveau de la crête iliaque et de la vertèbre lombaire L1 (VL1) chez la brebis âgée. Le ZOL a induit une forte diminution du remodelage osseux et une augmentation des microendommagements au niveau des deux sites et une modification des crosslinks du collagène surtout au niveau de l’os cortical de la CI. Trois mois de TPTD ont augmenté le remodelage osseux uniquement au niveau de la VL1. En conclusion, les délais et les amplitudes de réponses au ZOL ou au TPTD diffèrent entre la CI et la VL1 chez la brebis. Ces résultats montrent l’importance de prendre en compte le site osseux pour évaluer les effets des agents anti-ostéoporotiques / The effects of anti-osteoporotic agents on bone tissue are evaluated on iliac crest (IC) but the answers to treatments may vary according to the skeletal site. The purpose of this study was to compare the effect of zoledronic acid (ZOL) and teriparatide (TPTD) on IC and lumbar vertebrae (LV1) in ewes. ZOL has induced a high decrease of bone remodeling, an increase in microdamages in both sites and a modification of collagen crosslinks mainly in cortical bone of IC. Three months of TPTD has increased the bone remodeling only in LV1. In conclusion, the delays and the magnitudes of responses to ZOL or to TPTD differ between IC and LV1 in ewes. These results show that the distinction of bone sites to study the early effects of antiosteoporotic therapies appears meaningful

La microarchitecture osseuse est un des déterminants de la qualité osseuse qui peut maintenant être évaluée in vivo au radius et au tibia distaux avec une résolution isotropique de 82μm par un nouveau scanner à haute résolution (XtremeCT, SCANCO Medical AG). Par ailleurs, l’utilisation d’analyse en éléments finis sur les volumes 3D obtenus permet d’évaluer les propriétés biomécaniques de l’os comme la résistance osseuse. Nous avons montré qu’il s’agissait d’une technique prometteuse pour évaluer la densité, la microarchitecture et les propriétés biomécaniques osseuses au niveau des sites périphériques, notamment parce que ces mesures étaient associées chez la femme avec des fractures ostéoporotiques de toutes sortes. Nous avons également montré que les mêmes mesures étaient tout aussi pertinentes chez l’homme, alors qu’il est moins sujet à l’ostéoporose. Les résultats étaient associés aux fractures ostéoporotiques de toutes sortes, notamment les fractures vertébrales. L’analyse en éléments finis permet donc la mesure in vivo de la résistance osseuse, ce qui pourrait fournir des informations sur la fragilité osseuse et le risque de fracture non accessible par les seules mesures de densité ou de microarchitecture osseuse. / Bone microarchitecture is one of the determinants of bone quality that can now be evaluated in vivo at the distal radius and tibia with an isotropic resolution of 82μm with a new high-resolution peripheral scanner (XtremeCT, SCANCO Medical AG). Moreover, the use of finite element analysis on the 3D bone volume acquired allows the assessment of bone biomechanical properties such as bone strength. Our studies show that this technique is promising to assess bone density, microarchitecture and strength at peripheral skeletal sites. Indeed those measures were associated with osteoporotic fractures of all kinds in women. We also demonstrated that those same measures were associated with osteoporotic fractures of all kinds, including vertebral fractures, in men, who are less prone to be affected by osteoporosis. Finite element analysis allows in vivo measurement of bone strength, which might provide additional information about bone fragility and fracture risk that are not assessed by measures of density or microarchitecture.

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.

La consommation d’alcool a des effets sur le tissu osseux. L’alcoolisme est une des causes d’ostéoporose secondaire chez l’homme. Dans ce travail nous avons mené différentes expérimentations chez le rat afin d’étudier les effets d’une consommation chronique d’alcool combinée ou non à un entraînement aérobie sur le tissu osseux. Nous avons montré qu’une faible dose d’alcool administrée pendant une période courte peut avoir un effet positif sur la densité minérale osseuse et l’épaisseur trabéculaire. En revanche, la combinaison activité physique et consommation modérée d’alcool n’a pas d’effet additif sur la potentialisation du tissu osseux. Nous avons également démontré un effet dose de l’alcool indiquant des effets délétères majorés sur la densité minérale osseuse (DMO), la microarchitecture corticale et la résistance osseuse avec des apports croissants (25%, 30% et 35% v/v). La modification de DMO s’accompagne d’un changement de composition corporelle et d’une diminution de la leptine systémique. Cependant, le nombre d’adipocytes augmente dans la moelle osseuse. Nous avons mis en évidence dans ce modèle d’ostéoporose secondaire due à l’alcool une augmentation de l’apoptose des ostéocytes, corrélée à la diminution de la DMO et à l’augmentation de l’adiposité médullaire. Nous avons de plus mis en évidence une incorporation de lipides dans les ostéocytes, incorporation fortement corrélée à l’apoptose de ces cellules. Enfin, nos résultats montrent qu’un exercice physique régulier combiné à une consommation chronique et excessive d’alcool permet de prévenir les effets délétères de l’alcool sur les paramètres osseux (porosité corticale, épaisseur corticale) et limite la diminution de la DMO. Cette diminution est associée à une régulation de l’apoptose des ostéocytes. / Heavy chronic alcohol consumption has deleterious effects on bone tissue. It is one of the major causes of secondary osteoporosis in men. In this work, we draw several experimentations to assess the effects of chronic alcohol consumption on bone, combined or not to an aerobic training in the rat. We showed that light to moderate chronic alcohol consumption during a short time lead to an increase of bone mineral density (BMD) and trabecular thickness, whith no additive effects of physical exercise on bone tissue. When the alcohol doses were increased, we showed deleterious effects on BMD, microarchitecture, bone resistance with a dose effect with increasing alcohol doses (25%, 30% and 35% v/v): the more alcohol was concentrated and the more the bone parameters were decreased. The BMD decrease was associated with a change in body composition, and with a decrease in serum leptin. However, the number of lipid droplets in the bone marrow was increased dramatically. We demonstrated that there was a huge increase in osteocyte apoptosis with alcohol (35% v/v) in this alcohol-induced osteoporosis model, which was correlated with BMD and bone marrow adiposity. We have also shown that there was lipid incorporation in bone micro vessels and in osteocytes, which was correlated with osteocyte apoptosis. Lastly, we showed that when regular exercise was associated with heavy chronic alcohol consumption, the bone parameters were normal (trabecular, cortical thickness, femur length) and the BMD was less decreased compared to alcohol-fed and sedentary rats. These effects were associated with a regulation of osteocyte apoptosis.

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.

L’opération morphologique de squelettisation transforme chaque objet d’une image en une forme linéique qui préserve la topologie de ce dernier (propriété d’homotopie). Elle est largement utilisée en biométrie mais aussi dans la reconnaissance des caractères ainsi que pour l’extraction de la microarchitecture osseuse. L’objectif de cette thèse est de développer une méthode de squelettisation appliquée directement sur les niveaux de gris de l’image, ce qui a pour large avantage de s’affranchir de prétraitement comme la binarisation. Une revue des méthodes de squelettisation en niveaux de gris permet de constater que l’amincissement est l’une des approches les plus usitées de par sa propriété d’homotopie. Cependant, cette approche est sensible au bruit de l’image et produit des squelettes sur-connectés. Un premier paramétrage de l’amincissement a été proposé dans la littérature afin d’abaisser des configurations de pixels liées au bruit. La première contribution de ce travail est de proposer un ajustement de ce paramètre basé sur une décision statistique. Il s’agit d’identifier les tests d’hypothèses correspondants aux différentes configurations d’abaissement en vue de fixer ce paramètre de façon locale. Ceci conduit à la mise en place d’une squelettisation appelée Self Contrast Controlled Thinning (SCCT) puisque robuste au bruit tout en s’adaptant automatiquement au contraste de l’image. La squelettisation SCCT est rendue accessible aux domaines d’application grâce à son implantation optimisée basée sur les files d’attente hiérarchiques. Ayant noté le peu d’efforts consacrés à l’évaluation de la squelettisation en niveaux de gris, la deuxième contribution de ce travail est de proposer un protocole visant à évaluer l’opération de squelettisation sur la base des propriétés requises à savoir la préservation de la topologie et de la géométrie. Ce protocole est déroulé sur une base d’images synthétiques et nous permet de comparer notre approche à celles de la littérature. La troisième contribution est de proposer une structuration du squelette en graphe donnant accès aux descripteurs structurels et morphométriques des objets étudiés en vue d’une exploitation du squelette par les experts des domaines d’applications. Dans le cadre du projet Voxelo coordonné par le laboratoire B2OA de l’Université Paris Diderot, cette structuration est exploitée pour extraire les descripteurs de la qualité de la microarchitecture osseuse à partir d’images RX haute résolution. / Skeletonization is an image transformation that aims to represent objects by their medial axis while preserving their topological characteristics (homotopy). It is widely used in biometrics, character recognition and also in the extraction of bone microarchitecture. The objective of this thesis is to develop a skeletonization method applied directly on image gray levels. This has the large advantage of freeing the operation from preprocessing techniques such as binarization. A review of grayscale skeletonization methods shows that the morphological thinning is one of the most used approaches for its topology preservation property. However, this approach is sensitive to image noise and produces inexploitable skeletons. A first parameterization of the thinning process has been proposed in the literature to reduce noise-related information. The first contribution of this work is to propose an adjustment of this parameter based on a statistical decision. To this end, a hypothesis test is identified for each lowering criterion in order to set the thinning parameter locally. This leads us to propose the Self Contrast Controlled Thinning method SCCT that is robust to noise and is automatically adjusted to image contrast. The SCCT is made available to application domains through its optimized implementation based on hierarchical queues. Noticing the lack of efforts to assess grayscale skeletonization, the second contribution of this work is to propose a quantitative evaluation protocol assessing skeletonization with regard to its fundamental properties that are namely the preservation of topology and geometry. This protocol is conducted over a synthetic images database and allows us to compare SCCT to approaches from the literature. The third contribution consists in structuring the skeleton into a graph that gives access to objects structural and morphometric descriptors and enables the exploitation of the skeleton by experts from various fields of application. This structuring is applied in the context of Voxelo project which is coordinated by B2OA laboratory of the University Paris Diderot. In this context, descriptors of bone microarchitecture quality are extracted from X-ray high resolution images.

<p>The risk of fracture is higher in adults with type 2 diabetes compared to controls without type 2 diabetes, despite normal or higher than normal bone mineral density (BMD). In addition to BMD, bone strength depends on other factors such as structural and material <em>bone qualities</em>, which are not accounted for in BMD measurements. The objective was to determine whether structural and material <em>bone qualities</em> are different in adults with type 2 diabetes compared to controls without type 2 diabetes. First, a cross-sectional study was undertaken using MRI to investigate distal radius trabecular bone microarchitecture, a structural <em>bone quality.</em> In women with type 2 diabetes, trabecular bone holes were larger compared to controls, which is important because greater trabecular bone hole size is related to reduced bone strength. Next, a two year prospective study was conducted with the participants involved in the cross-sectional study to determine whether changes in trabecular bone microarchitecture are different in women with type 2 diabetes compared to controls. There was a greater increase in the number of trabecular bone holes in women with type 2 diabetes compared to controls, which provides early evidence of trabecularization of cortical bone in women with type 2 diabetes. In the third study, quantitative backscattered electron imaging (qBEI) was used to derive bone mineralization density distribution (BMDD) outcomes for bone samples from adults with and without type 2 diabetes to compare material <em>bone quality. </em>There was evidence of elevated bone calcium concentration and reduced mineralization heterogeneity in bone samples from adults with type 2 diabetes compared to controls, which may contribute to bone brittleness. In summary, differences in structural and material <em>bone qualities </em>identified in this body of work provide explanations for elevated fracture risk in adults with type 2 diabetes.</p> / Doctor of Science (PhD)