Bone remodeling theory applied to the study of n unit-elements model.

The aim of this paper is to illustrate the application of mathematical tools for the analysis of non-linear dynamical systems to the study of global stability of one kind of bone remodeling scheme applied to n unit-elements model. The particular aspects analyzed here are the stationary states related to this theory and a condition of their stability. The non-linear equations governing the remodeling process are solved by finite-difference method and the well-known results on the heterogeneous spatial organizations have been retrieved and confirm the analytical study. This kind of remodeling theory is useful for investigating the effects of physiological parameters on the development, maintenance, and adaptation of bone under mechanical loading.     

The application of topology optimization on the quantitative description of the external shape of bone structure

The aim of this paper was to introduce the idea of topology optimization in engineering to the simulation of bone morphology. The external shape of bone structure was predicted with the quantitative bone functional adaptation theory. The high-order nonlinear equation of bone remodeling proposed by Zhu et al. (J. Biomech. 35(7)(2002)951) combining with the finite element method was adopted to a rectangular design domain, which occupies a larger space than the external shape of bone structure. It was at this point we imported the idea of topology optimization in engineering. The proximal femur was used here as an example, whose external shape and internal density distribution were simultaneously simulated quantitatively to validate that the external shape of bone structure could be successfully predicted in this way. Then the growth of vertebral body from young to old was simulated numerically in its coronal section to discuss the significance of the prediction of external shape. The study in this paper provides computational basis for further studies on osteophyte formation, osteoporosis, osteoarthritis, bone growth and even bone evolution, etc.     

An Analysis and Comparison of Convergence and Uniqueness of Time-Independent Bone Adaptation Models

Several stimuli are proposed in the bone remodeling theory. It is not clear, if a unique solution exists and if the result is convergent using a certain stimulus. In this study, the strain stimulus, strain energy stimulus and the von Mises stress stimulus for bone remodeling are compared and applied to a square plate model using the finite element method. In the plane stress state, the remodeling equilibrium equations are transformed into functions of only the principal strains and the graphs of these functions are drawn in a diagram using the principal strains as the variables of two coordinate axes. The equation of the sum of principal strain squared equal to a constant is a circle in the diagram. The remodeling equilibrium equation of the strain stimulus is a quadrangle fitting into the circle, the remodeling equilibrium equation of the strain energy stimulus is an ellipse and the remodeling equilibrium equation of the von Mises stress stimulus is also an ellipse close to the principal strains circle when we take the same constants in the above equations. Using the finite element method, two models are performed with the uniform initial elastic properties and with the semi-random initial distribution of the elastic properties. The principal strains as the final finite element results converge within 2% of the objective constant for all the different stimuli. The obtained Young's moduli of two models as the adaptation object are different but in equilibrium, i.e. the equilibrium solution of adaptation model is not unique. The principal strains can not be used to examine the uniqueness of solution, since two different solutions can have the same results of principal strains. Using a certain stimulus, certain initial properties and a certain iterative equation, the solution is unique in equilibrium. The results using the model in this study show also that the same results can be obtained using any of the three stimuli when a proper constant in each remodeling equilibrium equation is chosen.     

Analysing the mechanical performance and growth adaptation of Norway spruce using a non-linear finite-element model and experimental data

Thirteen Norway spruce [Picea abies (L.) Karst.] trees of different size, age, and social status, and grown under varying conditions, were investigated to see how they react to complex natural static loading under summer and winter conditions, and how they have adapted their growth to such combinations of load and tree state. For this purpose a non-linear finite-element model and an extensive experimental data set were used, as well as a new formulation describing the degree to which the exploitation of the bending stress capacity is uniform. The three main findings were: material and geometric non-linearities play important roles when analysing tree deflections and critical loads; the strengths of the stem and the anchorage mutually adapt to the local wind acting on the tree crown in the forest canopy; and the radial stem growth follows a mechanically high-performance path because it adapts to prevailing as well as acute seasonal combinations of the tree state (e.g. frozen or unfrozen stem and anchorage) and load (e.g. wind and vertical and lateral snow pressure). Young trees appeared to adapt to such combinations in a more differentiated way than older trees. In conclusion, the mechanical performance of the Norway spruce studied was mostly very high, indicating that their overall growth had been clearly influenced by the external site- and tree-specific mechanical stress.     

Comparative Mechanical Properties and Histology of Bone

Different bone tissues differ in their amounts of porosity,mineralization,reconstruction, and preferred orientation. Allthese have important effects on mechanical properties. Veryporous, cancellous bone is always weaker and morecompliant thancompact bone on a weight for weight basis, yet it occurs inplaceswhere its energyabsorbing ability, or its low density,is advantageous. Bonevaries considerably in its mineralization,and such variations have quite disproportionate effects on mechanicalproperties. These variations can be shown to be adaptive. Inparticular, there must always be a compromise between stiffnessandresistanceto fracture; these two properties run contrary to each other.The reason for secondary remodeling is an unresolved problem,though in a few places the role of such remodeling in changingthe grain of the bone is clearly mechanically adaptive. Themechanical properties of non-mammalian bone are obscure, andas the histology of such bone is often quite different fromthat of mammalian bone, we are no doubt in for some surpriseswhen the mechanical properties ofnonmammalian bone are discovered.     

Interstitial fluid flow in canaliculi as a mechanical stimulus for cancellous bone remodeling: in silico validation

Cancellous bone has a dynamic 3-dimensional architecture of trabeculae, the arrangement of which is continually reorganized via bone remodeling to adapt to the mechanical environment. Osteocytes are currently believed to be the major mechanosensory cells and to regulate osteoclastic bone resorption and osteoblastic bone formation in response to mechanical stimuli. We previously developed a mathematical model of trabecular bone remodeling incorporating the possible mechanisms of cellular mechanosensing and intercellular communication in which we assumed that interstitial fluid flow activates the osteocytes to regulate bone remodeling. While the proposed model has been validated by the simulation of remodeling of a single trabecula, it remains unclear whether it can successfully represent in silico the functional adaptation of cancellous bone with its multiple trabeculae. In the present study, we demonstrated the response of cancellous bone morphology to uniaxial or bending loads using a combination of our remodeling model with the voxel finite element method. In this simulation, cancellous bone with randomly arranged trabeculae remodeled to form a well-organized architecture oriented parallel to the direction of loading, in agreement with the previous simulation results and experimental findings. These results suggested that our mathematical model for trabecular bone remodeling enables us to predict the reorganization of cancellous bone architecture from cellular activities. Furthermore, our remodeling model can represent the phenomenological law of bone transformation toward a locally uniform state of stress or strain at the trabecular level.     

Lactoferrin and bone; structure-activity relationships.

The maintenance of the mechanical integrity of the skeleton depends on bone remodeling, the well-coordinated balance between bone formation by osteoblasts and bone resorption by osteoclasts. The coupled action of osteoblasts and osteoclasts is regulated by the action of many local and circulating hormones and factors as well as central regulation by a neurological mechanism. We have previously shown that lactoferrin can promote bone growth. At physiological concentrations, lactoferrin potently stimulates the proliferation and differentiation of primary osteoblasts and acts as a survival factor. Lactoferrin also affects osteoclasts, potently inhibiting their formation. In vivo, local injection of lactoferrin results in substantial increases in bone formation and bone area. In a critical bone-defect model in vivo, lactoferrin was also seen to promote bone growth. The mitogenic effect of lactoferrin in osteoblast-like cells is mediated mainly through low-density lipoprotein-receptor protein-1 (LRP1), a member of the low-density lipoprotein-receptor-related proteins that are primarily known as endocytic receptors; however, LRP1 is not necessary for the anti-apoptotic actions of lactoferrin. Lactoferrin also induces the activation of p42/44 mitogen-activated protein kinase (MAPK) signalling and the PI3-kinase-dependent phosphorylation of Akt in osteoblasts. In this study, we examined other properties of lactoferrin and the way they affect osteogenic activity. The degree of glycosylation, iron-binding, and the structure-activity relationships indicate that lactoferrin maintains osteogenic activity in deglycosylated, holo, and apo forms, and in with various small fragments of the molecule. These data suggest that lactoferrin signals through more than 1 membrane-bound receptor to produce its anabolic skeletal effects, and that it signals through diverse pathways. We conclude that lactoferrin might have a physiological role in bone growth and healing and a potential therapeutic role as an anabolic factor in osteoporosis.     

Skeletal adaptations during mammalian reproduction

Remarkable changes occur in the mammalian skeleton prior to, during and after the reproductive cycle. Skeletal changes occur with ovarian maturation and initiation of menses and estrus in adolescence, which may result in a greater accumulation of skeletal mineral in the female vs the male skeleton. There is also some evidence to suggest an excess skeletal mass in young female experimental animals. In early pregnancy, growth, modeling and perhaps suppressed remodeling promote the accumulation of calcium. Some changes may also occur with the transition from pituitary to placental control of the pregnancy. In later pregnancy, an increase in bone turnover appears to coincide with fetal skeletal mineralization. Rapid and important changes occur in the skeleton and mineral metabolism in the transition from pregnancy to lactation as the mammary gland rather than the uterus draws on the maternal calcium stores. Lactational demands are met at least partially by a temporary demineralization of the skeleton, which is associated with increased bone modeling and remodeling. Endochondral growth almost ceases during lactation, but envelope-specific bone modeling and remodeling are greatly increased. This is generally associated with a loss of skeletal mass and density, more apparent at sites with less of a mechanical role (e.g. central metaphysis regions and the endocortical envelope). The post-lactational period is profoundly anabolic with substantial increases in bone formation, but blunted resorption at almost all skeletal envelopes. Skeletal mass is increased during this period and it is associated with improved skeletal mechanical properties. There are several important observations. 1) The nulliparous animal appears to have an excess skeletal mass to perhaps compensate for maternal metabolic inefficiency of the first reproductive cycle. 2) Changes in growth, modeling and remodeling occur at different times and at different skeletal envelopes during the reproductive cycle. These site-specific, temporal changes appear to be adaptations that facilitate the use of skeletal mineral while preserving mechanical competence. 3) After the first reproductive cycle, modeling and remodeling optimize the existing skeletal mass into a structure that better accommodates the prevailing mechanical environment. 4) The post-lactational period is profoundly anabolic and may provide new strategies for preservation of skeletal mass when reproductive capacity ceases.     

Differential Actions of the Endocytic Collagen Receptor uPARAP/Endo180 and the Collagenase MMP-2 in Bone Homeostasis

A well-coordinated remodeling of uncalcified collagen matrices is a pre-requisite for bone development and homeostasis. Collagen turnover proceeds through different pathways, either involving extracellular reactions exclusively, or being dependent on endocytic processes. Extracellular collagen degradation requires the action of secreted or membrane attached collagenolytic proteases, whereas the alternative collagen degradation pathway proceeds intracellularly after receptor-mediated uptake and delivery to the lysosomes. In this study we have examined the functional interplay between the extracellular collagenase, MMP-2, and the endocytic collagen receptor, uPARAP, by generating mice with combined deficiency of both components. In both uPARAP-deficient and MMP-2-deficient adult mice the length of the tibia and femur was decreased, along with a reduced bone mineral density and trabecular bone quality. An additional decrease in bone length was observed when combining the two deficiencies, pointing to both components being important for the remodeling processes in long bone growth. In agreement with results found by others, a different effect of MMP-2 deficiency was observed in the distinct bone structures of the calvaria. These membranous bones were found to be thickened in MMP-2-deficient mice, an effect likely to be related to an accompanying defect in the canalicular system. Surprisingly, both of the latter defects in MMP-2-deficient mice were counteracted by concurrent uPARAP deficiency, demonstrating that the collagen receptor does not support the same matrix remodeling processes as the MMP in the growth of the skull. We conclude that both uPARAP and MMP-2 take part in matrix turnover processes important for bone growth. However, in some physiological situations, these two components do not support the same step in the growth process.     

Craniofacial sutures: Signaling centres integrating mechanosensation,cell signaling,and cell differentiation

Cranial sutures are dynamic structures in which stem cell biology, bone formation, and mechanical forces interface, influencing the shape of the skull throughout development and beyond. Over the past decade, there has been significant progress in understanding mesenchymal stromal cell (MSC) differentiation in the context of suture development and genetic control of suture pathologies, such as craniosynostosis. More recently, the mechanosensory function of sutures and the influence of mechanical signals on craniofacial development have come to the forefront. There is currently a gap in understanding of how mechanical signals integrate with MSC differentiation and ossification to ensure appropriate bone development and mediate postnatal growth surrounding sutures. In this review, we discuss the role of mechanosensation in the context of cranial sutures, and how mechanical stimuli are converted to biochemical signals influencing bone growth, suture patency, and fusion through mediation of cell differentiation. We integrate key knowledge from other paradigms where mechanosensation forms a critical component, such as bone remodeling and orthodontic tooth movement. The current state of the field regarding genetic, cellular, and physiological mechanisms of mechanotransduction will be contextualized within suture biology.     

Mechanical and electrical interactions in bone remodeling

The natural remodeling and adaptation of skeletal tissues in response to mechanical loading is a classic example of physical regulation in biology. It is largely because it involves forces that do not seem to fit into the familiar schemes of biochemical controls that bone adaptation mechanisms have intrigued us for at least a century. The effect of electromagnetic fields on organisms is another example of this, and the two have become linked in an attempt to explain bone remodeling (“Yasuda's hypothesis”). This paper re-examines the roles of endogenous and exogenous electromagnetic fields in the response of bone to mechanical forces. A series of experiments is reviewed in which mechanical and electrical stimuli were applied to implants in the medullary canal of rabbit long bones. The results suggest that endogenously generated electrical currents are not required to initiate mechanically stimulated bone formation, but that direct mechanical effects on bone cells is the more likely scenario. Based on this and other evidence from the literature, it is suggested that when exogenous electromagnetic stimuli are applied, bone cells respond by modulating the activity of more primary activators such as hormones, growth factors, cytokines, and mechanical forces. Bioelectromagnetics 18:193–202, 1997. © 1997 Wiley-Liss, Inc.     

The turnover of mineralized growth plate cartilage into bone may be regulated by osteocytes

During endochondral ossification, growth plate cartilage is replaced with bone. Mineralized cartilage matrix is resorbed by osteoclasts, and new bone tissue is formed by osteoblasts. As mineralized cartilage does not contain any cells, it is unclear how this process is regulated. We hypothesize that, in analogy with bone remodeling, osteoclast and osteoblast activity are regulated by osteocytes, in response to mechanical loading. Since the cartilage does not contain osteocytes, this means that cartilage turnover during endochondral ossification would be regulated by the adjacent bone tissue. We investigated this hypothesis with an established computational bone adaptation model. In this model, osteocytes stimulate osteoblastic bone formation in response to the mechanical bone tissue loading. Osteoclasts resorb bone near randomly occurring microcracks that are assumed to block osteocyte signals. We used finite element modeling to evaluate our hypothesis in a 2D-domain representing part of the growth plate and adjacent bone. Cartilage was added at a constant physiological rate to simulate growth. Simulations showed that osteocyte signals from neighboring bone were sufficient for successful cartilage turnover, since equilibrium between cartilage remodeling and growth was obtained. Furthermore, there was good agreement between simulated bone structures and rat tibia histology, and the development of the trabecular architecture resembled that of infant long bones. Additionally, prohibiting osteoclast invasion resulted in thickened mineralized cartilage, similar to observations in a knock-out mouse model. We therefore conclude that it is well possible that osteocytes regulate the turnover of mineralized growth plate cartilage.     

A homogenized constrained mixture (and mechanical analog) model for growth and remodeling of soft tissue

Most mathematical models of the growth and remodeling of load-bearing soft tissues are based on one of two major approaches: a kinematic theory that specifies an evolution equation for the stress-free configuration of the tissue as a whole or a constrained mixture theory that specifies rates of mass production and removal of individual constituents within stressed configurations. The former is popular because of its conceptual simplicity, but relies largely on heuristic definitions of growth; the latter is based on biologically motivated micromechanical models, but suffers from higher computational costs due to the need to track all past configurations. In this paper, we present a temporally homogenized constrained mixture model that combines advantages of both classical approaches, namely a biologically motivated micromechanical foundation, a simple computational implementation, and low computational cost. As illustrative examples, we show that this approach describes well both cell-mediated remodeling of tissue equivalents in vitro and the growth and remodeling of aneurysms in vivo. We also show that this homogenized constrained mixture model suggests an intimate relationship between models of growth and remodeling and viscoelasticity. That is, important aspects of tissue adaptation can be understood in terms of a simple mechanical analog model, a Maxwell fluid (i.e., spring and dashpot in series) in parallel with a “motor element” that represents cell-mediated mechanoregulation of extracellular matrix. This analogy allows a simple implementation of homogenized constrained mixture models within commercially available simulation codes by exploiting available models of viscoelasticity.     

Osteopenic mice: animal models of the aging skeleton

While our understanding of the developmental biology of the skeleton, like that of virtually every other subject in biology, has been transformed by recent advances in human and mouse genetics, we still know very little, in molecular and genetic terms, about skeletal physiology. Thus, among the many questions that are largely unexplained are the following: why is osteoporosis mainly a women's disease? How is bone mass maintained nearly constant between the end of puberty and the arrest of gonadal functions? Molecular genetics has emerged as a powerful tool to study previously unexplored aspects of the physiology of the skeleton. Among mammals, mice are the most promising animals for this experimental work. This has been previously demonstrated e.g. through the tremendous impact of the different osteopetrotic models on our molecular understanding of osteoclastic bone resorption. Until recently the only way of studying bone loss situations and osteoporosis in mice was by using ovariectomy with all its limitations. Today, however, we have access to more sophisticated osteoporotic mouse-models from four different origins: Transgenic mice (HSV-TK), knock-out mice (OPG), inbred-strains (SAMP6), and through physiological modulation (icv application). These new models have already taught us several important lessons. The first is, that bone remodeling is more than just an autocrine/paracrine process. Multiple experimental evidence has demonstrated that the latter regulation exists, but genetics prove that there is no functional cross-control between resorption and formation. The second lesson is, that remodeling is, at least in part, subject to central regulation. Thus, osteoporosis is partly a central or hypothalamic disease. However, the most dramatic change and the most important advantage we feel is, that today we have models to test a new hypothesis regarding the etiology of osteoporosis before it turns to dogma. Taken together, mouse-studies may lead to a shift in our physiological understanding of skeleton biology and to the emergence of novel paradigms. These, in turn, should help us to devise new treatments for degenerative diseases of the skeleton such as osteoporosis and its associated clinical problems.     

Mechanical effects on skeletal growth

The growth (i.e. increase of external dimensions) of long bones and vertebrae occurs longitudinally by endochondral ossification at the growth plates, and radially by apposition of bone at the periosteum. It is thought that mechanical loading influences the rate of longitudinal growth. The 'Hueter-Volkmann Law' proposes that growth is retarded by increased mechanical compression, and accelerated by reduced loading in comparison with normal values. The present understanding of this mechanism of bone growth modulation comes from a combination of clinical observation (where altered loading and growth is implicated in some skeletal deformities) and animal experiments in which growth plates of growing animals have been loaded. The gross effect of growth modulation has been demonstrated qualitatively and semi-quantitatively. Sustained compression of physiological magnitude inhibits growth by 40% or more. Distraction increases growth rate by a much smaller amount. Experimental studies are underway to determine how data from animal studies can be scaled to other growth plates. Variables include: differing sizes of growth plate, different anatomical locations, different species and variable growth rate at different stages of skeletal maturity. The two major determinants of longitudinal growth are the rate of chondrocytic proliferation and the amount of chondrocytic enlargement (hypertrophy) in the growth direction. It is largely unknown what are the relative changes in these key variables in mechanically modulated growth, and what are the signaling pathways that produce these changes.     

Can deterministic mechanical size effects contribute to fracture and microdamage accumulation in trabecular bone?

Failure of bone under monotonic and cyclic loading is related to the bone mineral density, the quality of the bone matrix, and the evolution of microcracks. The theory of linear elastic fracture mechanics has commonly been applied to describe fracture in bone. Evidence is presented that bone failure can be described through a non-linear theory of fracture. Thereby, deterministic size effects are introduced. Concepts of a non-linear theory are applied to discern how the interaction among bone matrix constituents (collagen and mineral), microcrack characteristics, and trabecular architecture can create distinctively differences in the fracture resistance at the bone tissue level. The non-linear model is applied to interpret pre-clinical data concerning the effects of anti-osteoporotic agents on bone properties. The results show that bisphosphonate (BP) treatments that suppress bone remodeling will change trabecular bone in ways such that the size of the failure process zone relative to the trabecular thickness is reduced. Selective estrogen receptor modulators (SERMs) that suppress bone remodeling will change trabecular bone in ways such that the size of the failure process zone relative to the trabecular thickness is increased. The consequences of these changes are reflected in bone mechanical response and predictions are consistent with experimental observations in the animal model which show that BP treatment is associated with more brittle fracture and microcracks without altering the average length of the cracks, whereas SERM treatments lead to a more ductile fracture and mainly increase crack length with a smaller increase in microcrack density. The model suggests that BPs may be more effective in cases in which bone mass is very low, whereas SERMS may be more effective when milder osteoporotic symptoms are present.     

On foundations of discrete element analysis of contact in diarthrodial joints

Information about the stress distribution on contact surfaces of adjacent bones is indispensable for analysis of arthritis, bone fracture and remodeling. Numerical solution of the contact problem based on the classical approaches of solid mechanics is sophisticated and time-consuming. However, the solution can be essentially simplified on the following physical grounds. The bone contact surfaces are covered with a layer of articular cartilage, which is a soft tissue as compared to the hard bone. The latter allows ignoring the bone compliance in analysis of the contact problem, i.e. rigid bones are considered to interact through a compliant cartilage. Moreover, cartilage shear stresses and strains can be ignored because of the negligible friction between contacting cartilage layers. Thus, the cartilage can be approximated by a set of unilateral compressive springs normal to the bone surface. The forces in the springs can be computed from the equilibrium equations iteratively accounting for the changing contact area. This is the essence of the discrete element analysis (DEA). Despite the success in applications of DEA to various bone contact problems, its classical formulation required experimental validation because the springs approximating the cartilage were assumed linear while the real articular cartilage exhibited non-linear mechanical response in reported tests. Recent experimental results of Ateshian and his co-workers allow for revisiting the classical DEA formulation and establishing the limits of its applicability. In the present work, it is shown that the linear spring model is remarkably valid within a wide range of large deformations of the cartilage. It is also shown how to extend the classical DEA to the case of strong nonlinearity if necessary.     

Computational model for the cell-mechanical response of the osteocyte cytoskeleton based on self-stabilizing tensegrity structures

The mechanism by which mechanical stimulation on osteocytes results in biochemical signals that initiate the remodeling process inside living bone tissue is largely unknown. Even the type of stimulation acting on these cells is not yet clearly identified. However, the cytoskeleton of osteocytes is suggested to play a major role in the mechanosensory process due to the direct connection to the nucleus. In this paper, a computational approach to model and simulate the cell structure of osteocytes based on self-stabilizing tensegrity structures is suggested. The computational model of the cell consists of the major components with respect to mechanical aspects: the integrins that connect the cell with the extracellular bone matrix, and different types of protein fibers (microtubules and intermediate filaments) that form the cytoskeleton, the membrane-cytoskeleton (microfilaments), the nucleus and the centrosome. The proposed geometrical cell models represent the cell in its physiological environment which is necessary in order to give a statement on the cell behavior in vivo. Studies on the mechanical response of osteocytes after physiological loading and in particular the mechanical response of the nucleus show that the load acting on the nucleus is rising with increasing deformation applied to the integrins.     

Pattern formation in prey-taxis systems

In this paper, we consider spatial predator–prey models with diffusion and prey-taxis. We investigate necessary conditions for pattern formation using a variety of non-linear functional responses, linear and non-linear predator death terms, linear and non-linear prey-taxis sensitivities, and logistic growth or growth with an Allee effect for the prey. We identify combinations of the above non-linearities that lead to spatial pattern formation and we give numerical examples. It turns out that prey-taxis stabilizes the system and for large prey-taxis sensitivity we do not observe pattern formation. We also study and find necessary conditions for global stability for a type I functional response, logistic growth for the prey, non-linear predator death terms, and non-linear prey-taxis sensitivity.     

Analysis of microstructural and mechanical alterations of trabecular bone in a simulated three-dimensional remodeling process

Bone remodeling is a complex dynamic process, which modulates both bone mass and bone microstructure. In addition to bone mass, bone microstructure is an important contributor to bone quality in osteoporosis and fragility fractures. However, the quantitative knowledge of evolution of three-dimensional (3D) trabecular microstructure in adaptation to the external forces is currently limited. In this study, a new 3D simulation method of remodeling of human trabecular bone was developed to quantitatively study the dynamic evolution of bone mass and trabecular microstructure in response to different external loading conditions. The morphological features of trabecular plate and rod, such as thickness and number density in different orientations were monitored during the remodeling process using a novel imaging analysis technique, namely Individual Trabecula Segmentation (ITS). We showed that the volume fraction and microstructures of trabecular bone including, trabecular type and orientation, were determined by the applied mechanical load. Particularly, the morphological parameters of trabecular plates were more sensitive to the applied load, indicating that they played the major role in the mechanical properties of the trabecular bone. Reducing the applied load caused severe microstructural deteriorations of trabecular bone, such as trabecular plate perforation, rod breakage, and a conversion from plates to rods.