RESUMO
Mucopolysaccharidosis-I (MPS-I) is a lysosomal storage disease (LSD) caused by inactivating mutations of IDUA, encoding the glycosaminoglycan-degrading enzyme α-l-iduronidase. Although MPS-I is associated with skeletal abnormalities, the impact of IDUA deficiency on bone remodeling is poorly defined. Here we report that Idua-deficient mice progressively develop a high bone mass phenotype with pathological lysosomal storage in cells of the osteoblast lineage. Histomorphometric quantification identified shortening of bone-forming units and reduced osteoclast numbers per bone surface. This phenotype was not transferable into wild-type mice by bone marrow transplantation (BMT). In contrast, the high bone mass phenotype of Idua-deficient mice was prevented by BMT from wild-type donors. At the cellular level, BMT did not only normalize defects of Idua-deficient osteoblasts and osteocytes but additionally caused increased osteoclastogenesis. Based on clinical observations in an individual with MPS-I, previously subjected to BMT and enzyme replacement therapy (ERT), we treated Idua-deficient mice accordingly and found that combining both treatments normalized all histomorphometric parameters of bone remodeling. Our results demonstrate that BMT and ERT profoundly affect skeletal remodeling of Idua-deficient mice, thereby suggesting that individuals with MPS-I should be monitored for their bone remodeling status, before and after treatment, to avoid long-term skeletal complications.
Assuntos
Remodelação Óssea , Iduronidase/uso terapêutico , Mucopolissacaridose I/fisiopatologia , Mucopolissacaridose I/terapia , Animais , Transplante de Medula Óssea , Proliferação de Células , Células Cultivadas , Criança , Terapia Combinada , Modelos Animais de Doenças , Terapia de Reposição de Enzimas , Feminino , Humanos , Iduronidase/deficiência , Iduronidase/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucopolissacaridose I/patologia , Osteoclastos/enzimologiaRESUMO
BACKGROUND/AIM: Le Fort lines have traditionally been considered as zones of weakness in the mid-facial skeleton although the structural basis of increased bone fragility at these sites has not yet been investigated. Considering recent findings of occlusal loading-related regional heterogeneity in the mid-facial bone micro-architecture, the aim of this study was to explore whether such heterogeneity in cortical and cancellous bone micro-architecture may contribute to increased fragility at Le Fort fracture sites. MATERIALS AND METHODS: Twenty-five cortical and thirteen cancellous bone specimens were harvested from a dry skull and analyzed by micro-CT. Specimens were classified into Le Fort or Non-Le Fort groups based on their location in the mid-facial skeleton. RESULTS: Cortical bone along Le Fort lines showed tendencies toward lower thickness (1.5±0.63 vs 1.75±0.79; P=.39) and greater porosity (11.48±5.67 vs 10.28±5.28; P=.59). A significant difference was detected in the trabecular degree of anisotropy which was higher in cancellous bone from Le Fort fracture sites (2.14±0.69 vs 1.58±0.34; P=.02). CONCLUSIONS: Regional heterogeneity in cortical bone micro-architecture could not fully explain increased fragility of the mid-facial skeleton at the Le Fort lines. However, regional differences in trabecular bone anisotropy may contribute to increased bone fragility.
Assuntos
Ossos Faciais/ultraestrutura , Pontos de Referência Anatômicos , Densidade Óssea/fisiologia , Humanos , Técnicas In Vitro , Masculino , Microtomografia por Raio-XRESUMO
The structure of human cortical bone evolves over multiple length scales from its basic constituents of collagen and hydroxyapatite at the nanoscale to osteonal structures at near-millimeter dimensions, which all provide the basis for its mechanical properties. To resist fracture, bone's toughness is derived intrinsically through plasticity (e.g., fibrillar sliding) at structural scales typically below a micrometer and extrinsically (i.e., during crack growth) through mechanisms (e.g., crack deflection/bridging) generated at larger structural scales. Biological factors such as aging lead to a markedly increased fracture risk, which is often associated with an age-related loss in bone mass (bone quantity). However, we find that age-related structural changes can significantly degrade the fracture resistance (bone quality) over multiple length scales. Using in situ small-angle X-ray scattering and wide-angle X-ray diffraction to characterize submicrometer structural changes and synchrotron X-ray computed tomography and in situ fracture-toughness measurements in the scanning electron microscope to characterize effects at micrometer scales, we show how these age-related structural changes at differing size scales degrade both the intrinsic and extrinsic toughness of bone. Specifically, we attribute the loss in toughness to increased nonenzymatic collagen cross-linking, which suppresses plasticity at nanoscale dimensions, and to an increased osteonal density, which limits the potency of crack-bridging mechanisms at micrometer scales. The link between these processes is that the increased stiffness of the cross-linked collagen requires energy to be absorbed by "plastic" deformation at higher structural levels, which occurs by the process of microcracking.
Assuntos
Envelhecimento/fisiologia , Osso e Ossos/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Fenômenos Biomecânicos , Produtos Finais de Glicação Avançada/análise , Humanos , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios XRESUMO
Notch signaling is a key pathway controlling various cell fate decisions during embryogenesis and adult life. It is activated by binding of specific ligands to four different Notch receptors that are subsequently cleaved by presenilins to release an intracellular domain that enters the nucleus and activates specific transcription factors. While the skeletal analysis of various mouse models with activated or inactivated Notch signaling has demonstrated a general impact of this pathway on bone remodeling, the more recent identification of NOTCH2 mutations in individuals with Hajdu-Cheney syndrome (HCS) has highlighted its human relevance. Since HCS is primarily characterized by skeletal defects, these latter findings led us to analyze the specific role of Notch2 in skeletal remodeling. After observing Notch2 expression in osteoblasts and osteoclasts, we utilized Runx2-Cre and Lyz2-Cre mice to inactivate Notch2 in cells of the osteoblast or osteoclast lineage, respectively. Whereas Notch2(fl/fl)/Lyz2-Cre mice did not display significant alterations of skeletal growth, bone mass or remodeling, Notch2(fl/fl)/Runx2-Cre mice progressively developed skeletal abnormalities in long bones. More specifically, these mice displayed a striking increase of trabecular bone mass in the proximal femur and the distal tibia at 6 and 12months of age. Whereas undecalcified sectioning of the respective regions did not reveal impaired osteocyte differentiation as a potential trigger for the observed phenotype, ex vivo experiments with bone marrow cells identified an increased osteogenic capacity of Notch2(fl/fl)/Runx2-Cre cultures. Collectively, our findings demonstrate that Notch2 physiologically regulates bone remodeling by inhibiting trabecular bone formation in the appendicular skeleton. Understanding the underlying mechanisms may help to improve diagnosis and therapy of HCS.
Assuntos
Osso Esponjoso/metabolismo , Osso Esponjoso/patologia , Osteoblastos/metabolismo , Receptor Notch2/metabolismo , Animais , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Fêmur/patologia , Perfilação da Expressão Gênica , Integrases/metabolismo , Camundongos , Tamanho do Órgão , Especificidade de Órgãos , Osteoclastos/metabolismo , Osteogênese , Fenótipo , Tíbia/patologiaRESUMO
Although the concept of the occlusal load transfer through the facial skeleton along the buttresses has been extensively studied, there has been no study to link microarchitecture of the mid-facial bones to the occlusal load distribution. The aim of this study was to analyze micro-structural properties of the mid-facial bones in relation to occlusal stress. The study was performed by combining the three-dimensional finite element analysis (3D FEA) and micro-computed tomography analysis (micro-CT). Clenching was simulated on the computer model of the adult male human skull which was also used as a source of bone specimens. After the FEA was run, stress was measured at the specific sites in cortical shell and trabecular bone of the model along and between the buttresses. From the corresponding sites on the skull, twenty-five cortical and thirteen cancellous bone specimens were harvested. The specimens were classified into high stress or low stress group based on the stress levels measured via the FEA. Micro-architecture of each specimen was assessed by micro-CT. In the high stress group, cortical bone showed a tendency toward greater thickness and density, lower porosity, and greater pore separation. Stress-related differences in microstructure between the groups were more pronounced in trabecular bone, which showed significantly greater bone volume fraction (BV/TV) and trabecular thickness (Tb.Th) in the high stress group. Our results suggest that the mid-facial bones in the adult dentate male skull exhibit regional variations in cortical and trabecular bone micro-architecture that could be a consequence of different occlusal stress.
Assuntos
Ossos Faciais/anatomia & histologia , Ossos Faciais/fisiologia , Estresse Mecânico , Densidade Óssea/fisiologia , Humanos , Masculino , Suporte de CargaRESUMO
To unravel the origins of decreased bone strength in the superolateral femoral neck, we assessed bone structural features across multiple length scales at this cortical fracture initiating region in postmenopausal women with hip fracture and in aged-matched controls. Our combined methodological approach encompassed atomic force microscopy (AFM) characterization of cortical bone nano-structure, assessment of mineral content/distribution via quantitative backscattered electron imaging (qBEI), measurement of bone material properties by reference point indentation, as well as evaluation of cortical micro-architecture and osteocyte lacunar density. Our findings revealed a wide range of differences between the fracture group and the controls, suggesting a number of detrimental changes at various levels of cortical bone hierarchical organization that may render bone fragile. Namely, mineral crystals at external cortical bone surfaces of the fracture group were larger (65.22nm±41.21nm vs. 36.75nm±18.49nm, p<0.001), and a shift to a higher mineral content and more homogenous mineralization profile as revealed via qBEI were found in the bone matrix of the fracture group. Fracture cases showed nearly 35% higher cortical porosity and showed significantly reduced osteocyte lacunar density compared to controls (226±27 vs. 247±32#/mm(2), p=0.05). Along with increased crystal size, a shift towards higher mineralization and a tendency to increased cortical porosity and reduced osteocyte lacunar number delineate that cortical bone of the superolateral femoral neck bears distinct signs of fragility at various levels of its structural organization. These results contribute to the understanding of hierarchical bone structure changes in age-related fragility.
Assuntos
Colo do Fêmur/ultraestrutura , Fraturas do Quadril/patologia , Fraturas por Osteoporose/patologia , Idoso , Idoso de 80 Anos ou mais , Fenômenos Biomecânicos , Densidade Óssea/fisiologia , Cálcio/análise , Estudos de Casos e Controles , Cristalização , Feminino , Colo do Fêmur/química , Colo do Fêmur/fisiopatologia , Fraturas do Quadril/metabolismo , Fraturas do Quadril/fisiopatologia , Humanos , Microscopia de Força Atômica/métodos , Osteócitos/patologia , Osteoporose Pós-Menopausa/patologia , Osteoporose Pós-Menopausa/fisiopatologia , Fraturas por Osteoporose/metabolismo , Fraturas por Osteoporose/fisiopatologia , Microtomografia por Raio-X/métodosRESUMO
Wnt signalling is a key pathway controlling bone formation in mice and humans. One of the regulators of this pathway is Dkk1, which antagonizes Wnt signalling through the formation of a ternary complex with the transmembrane receptors Krm1/2 and Lrp5/6, thereby blocking the induction of Wnt signalling by the latter ones. Here we show that Kremen-2 (Krm2) is predominantly expressed in bone, and that its osteoblast-specific over-expression in transgenic mice (Col1a1-Krm2) results in severe osteoporosis. Histomorphometric analysis revealed that osteoblast maturation and bone formation are disturbed in Col1a1-Krm2 mice, whereas bone resorption is increased. In line with these findings, primary osteoblasts derived from Col1a1-Krm2 mice display a cell-autonomous differentiation defect, impaired canonical Wnt signalling and decreased production of the osteoclast inhibitory factor Opg. To determine whether the observed effects of Krm2 on bone remodeling are physiologically relevant, we analyzed the skeletal phenotype of 24 weeks old Krm2-deficient mice and observed high bone mass caused by a more than three-fold increase in bone formation. Taken together, these data identify Krm2 as a regulator of bone remodeling and raise the possibility that antagonizing KRM2 might prove beneficial in patients with bone loss disorders.