Tissue strain amplification at the osteocyte lacuna: a microstructural finite element analysis

A parametric finite element model of an osteocyte lacuna was developed to predict the microstructural response of the lacuna to imposed macroscopic strains. The model is composed of an osteocyte lacuna, a region of perilacunar tissue, canaliculi, and the surrounding bone tissue. A total of 45 different simulations were modeled with varying canalicular diameters, perilacunar tissue material moduli, and perilacunar tissue thicknesses. Maximum strain increased with a decrease in perilacunar tissue modulus and decreased with an increase in perilacunar tissue modulus, regardless of the thickness of the perilacunar region. An increase in the predicted maximum strain was observed with an increase in canalicular diameter from 0.362 to 0.421 microm. In response to the macroscopic application of strain, canalicular diameters increased 0.8% to over 1.0% depending on the perilacunar tissue modulus. Strain magnification factors of over 3 were predicted. However, varying the size of the perilacunar tissue region had no effect on the predicted perilacunar tissue strain. These results indicate that the application of average macroscopic strains similar to strain levels measured in vivo can result in significantly greater perilacunar tissue strains and canaliculi deformations. A decrease in the perilacunar tissue modulus amplifies the perilacunar tissue strain and canaliculi deformation while an increase in the local perilacunar tissue modulus attenuates this effect.     

Strain Concentrations Surrounding an Ellipsoid Model of Lacunae and Osteocytes

Direct cell sensing of tissue matrix strains is one possible signaling mechanism for mechanically mediated bone adaptation. We utilized homogenization theory lo estimate bone tissue matrix strains surrounding osteocytes using two sets of models. The first set of models estimated the strain levels surrounding the lacunae and canaliculi, taking into account variations in lamellar properties. The second set estimated strain levels in the osteocyte and the surrounding matrix for different cellular mechanical properties. The results showed that the strain levels found in and surrounding osteocytes, 1700 to 2700 microstrain (denoted as μe; 1 =.0001% strain), were significantly greater than the trabecular tissue level strains of [1325 μe, 287 μe, 87 μe] used for model input. Variation in lamellar properties did not affect strain levels, except at lamellar boundaries. Strain in and surrounding the osteocyte was not significantly affected by cellular stiffness ranging between 28 and 28,000 Pascals (Pa). Strain levels surrounding lacunae and canaliculi were approximately equivalent.     

Osteocyte lacunae tissue strain in cortical bone

Current theories suggest that bone modeling and remodeling are controlled at the cellular level through signals mediated by osteocytes. However, the specific signals to which bone cells respond are still unknown. Two primary theories are: (1) osteocytes are stimulated via the mechanical deformation of the perilacunar bone matrix and (2) osteocytes are stimulated via fluid flow generated shear stresses acting on osteocyte cell processes within canaliculi. Recently, much focus has been placed on fluid flow theories since in vitro experiments have shown that bone cells are more responsive to analytically estimated levels of fluid shear stress than to direct mechanical stretching using macroscopic strain levels measured on bone in vivo. However, due to the complex microstructural organization of bone, local perilacunar bone tissue strains potentially acting on osteocytes cannot be reliably estimated from macroscopic bone strain measurements. Thus, the objective of this study was to quantify local perilacunar bone matrix strains due to macroscopically applied bone strains similar in magnitude to those that occur in vivo. Using a digital image correlation strain measurement technique, experimentally measured bone matrix strains around osteocyte lacunae resulting from macroscopic strains of approximately 2000 microstrain are significantly greater than macroscopic strain on average and can reach peak levels of over 30,000 microstrain locally. Average strain concentration factors ranged from 1.1 to 3.8, which is consistent with analytical and numerical estimates. This information should lead to a better understanding of how bone cells are affected by whole bone functional loading.     

Strain Concentrations Surrounding an Ellipsoid Model of Lacunae and Osteocytes

Direct cell sensing of tissue matrix strains is one possible signaling mechanism for mechanically mediated bone adaptation. We utilized homogenization theory to estimate bone tissue matrix strains surrounding osteocytes using two sets of models. The first set of models estimated the strain levels surrounding the lacunae and canaliculi, taking into account variations in lamellar properties. The second set estimated strain levels in the osteocyte and the surrounding matrix for different cellular mechanical properties. The results showed that the strain levels found in and surrounding osteocytes, 1700 to 2700 microstrain (denoted as μe; 1 μe =.0001% strain), were significantly greater than the trabecular tissue level strains of {1325 μe, 287 μe, 87 μe} used for model input. Variation in lamellar properties did not affect strain levels, except at lamellar boundaries. Strain in and surrounding the osteocyte was not significantly affected by cellular stiffness ranging between 28 and 28, 000 Pascals (Pa). Strain levels surrounding lacunae and canaliculi were approximately equivalent.     

Modeling microdamage behavior of cortical bone

Bone is a complex material which exhibits several hierarchical levels of structural organization. At the submicron-scale, the local tissue porosity gives rise to discontinuities in the bone matrix which have been shown to influence damage behavior. Computational tools to model the damage behavior of bone at different length scales are mostly based on finite element (FE) analysis, with a range of algorithms developed for this purpose. Although the local mechanical behavior of bone tissue is influenced by microstructural features such as bone canals and osteocyte lacunae, they are often not considered in FE damage models due to the high computational cost required to simulate across several length scales, i.e., from the loads applied at the organ level down to the stresses and strains around bone canals and osteocyte lacunae. Hence, the aim of the current study was twofold: First, a multilevel FE framework was developed to compute, starting from the loads applied at the whole bone scale, the local mechanical forces acting at the micrometer and submicrometer level. Second, three simple microdamage simulation procedures based on element removal were developed and applied to bone samples at the submicrometer-scale, where cortical microporosity is included. The present microdamage algorithm produced a qualitatively analogous behavior to previous experimental tests based on stepwise mechanical compression combined with in situ synchrotron radiation computed tomography. Our results demonstrate the feasibility of simulating microdamage at a physiologically relevant scale using an image-based meshing technique and multilevel FE analysis; this allows relating microdamage behavior to intracortical bone microstructure.     

Homogenization theory and digital imaging: A basis for studying the mechanics and design principles of bone tissue

Bone tissue is a complex multilevel composite which has the ability to sense ad respond to its mechanical environment. It is believed that bone cells called osteocytes within the bone matrix sense the mechanical environment and determine whether structural alterations are needed. At present it is not known, however, how loads are transferred from the whole bone level to cells. A computational procedure combining representative volume element (RVE) based homogenization theory with digital imaging is proposed to estimate strains at various levels of bone structure. Bone tissue structural organization and RVE based analysis are briefly reviewed. The digital image based computational procedure was applied to estimate strains in individual trabeculae (first-level microstructure). Homogenization analysis of an idealized model was used to estimate strains at one level of bone structure around osteocyte lacunae (second-level trabecular microstructure). The results showed that strain at one level of bone structure is amplified to a broad range at the next microstructural level. In one case, a zeor-level tensile principal strain of 495 muE engendered strains ranging between -1000 and 7000 muE in individual trabeculae (first-level microstructure). Subsequently, a first-level tensile principal strains of 1325 muE within an inidividual trabecula engendered strains ranging between 782 and 2530 muE around osteocyte lacunae. Lacunar orientation was found to influence strains around osteocyte lacunae much more than lacunar ellipticity. In conclusion, the computational procedure combining homogenization theory with digital imaging can proveide estimates of cell level strains within whole bones. Such results may be used to bridge experimental studies of bone adaptation at the whole bone and cell culture level. (c) 1994 John Wiley & Sons, Inc.     

Measurement of microstructural strain in cortical bone

It is well known that mechanical factors affect bone remodeling such that increased mechanical demand results in net bone formation, whereas decreased demand results in net bone resorption. Current theories suggest that bone modeling and remodeling is controlled at the cellular level through signals mediated by osteocytes. The objective of this study was to investigate how macroscopically applied bone strains similar in magnitude to those that occur in vivo are manifest at the microscopic level in the bone matrix. Using a digital image correlation strain measurement technique, experimentally determined bone matrix strains around osteocyte lacuna resulting from macroscopic strains of approximately 2,000 microstrain (0.2%) reach levels of over 30,000 microstrain (3%) over fifteen times greater than the applied macroscopic strain. Strain patterns were highly heterogeneous and in some locations similar to observed microdamage around osteocyte lacuna indicating the resulting strains may represent the precursors to microdamage. This information may lead to a better understanding of how bone cells are affected by whole bone functional loading.     

Osteocyte lacuna size and shape in women with and without osteoporotic fracture

Osteocytes have been hypothesized to control the amount and location of bone tissue which is resorbed or formed, based on the strain magnitude they perceive, and therefore may play a role in the bone loss of osteoporosis. The shape of osteocyte lacunae influences the mechanical strain applied to the osteocyte; thus, it is important to quantify their shape to further understand the mechanical environment of this cell. Previous studies of the size and shape of lacunae have been contradictory and limited to two-dimensional measurements on iliac crest biopsies. This investigation measured the size and shape of osteocyte lacunae in trabecular bone near a typical fracture site from three-dimensional image sets obtained by confocal microscopy. Bone tissue specimens were obtained from individuals undergoing hip replacement subsequent to fracture, and matched cadaveric specimens without fracture. After extensive image processing to differentiate the lacunae from the matrix, the volume and anisotropy of the lacuna were determined. No significant difference was found in the size (volume) or shape (anisotropy) of the lacunae between women with and without osteoporotic fracture, although there was a large range of sizes and shapes in both groups. These results suggest that the size or shape of the lacunae, which influences the strain in osteocytes, does not play a role in osteoporotic fracture. In addition, this study provides geometric measures of lacunae that are important in computational modeling of the mechanical environment of osteocytes.     

Mechanosensation and fluid transport in living bone

The mechanosensory mechanisms in bone include (i) the cell system that is stimulated by external mechanical loading applied to the bone; (ii) the system that transduces that mechanical loading to a communicable signal; and (iii) the systems that transmit that signal to the effector cells for the maintenance of bone homeostasis and for strain adaptation of the bone structure. The effector cells are the osteoblasts and the osteoclasts. These systems and the mechanisms that they employ have not yet been unambiguously identified. The candidate systems will be reviewed. It will be argued that the current theoretical and experimental evidence suggests that osteocytes are the principal mechanosensory cells of bone, that they are activated by shear stress from fluid flowing through the osteocyte canaliculi, and that the electrically coupled three-dimensional network of osteocytes and lining cells is a communications system for the control of bone homeostasis and structural strain adaptation. The movement of bone fluid from the region of the bone vasculature through the canaliculi and the lacunae of the surrounding mineralized tissue accomplishes three important tasks. First, it transports nutrients to the osteocytes in the lacunae buried in the mineralized matrix. Second, it carries away the cell waste. Third, the bone fluid exerts a force on the cell process, a force that is large enough for the cell to sense. This is probably the basic mechanotransduction mechanism in bone, the way in which bone senses the mechanical load to which it is subjected. The mechanisms of bone fluid flow are described with particular emphasis on mechanotransduction. Also described is the cell to cell communication by which higher frequency signals might be transferred, a potential mechanism in bone by which the small whole tissue strain is amplified so the bone cells can respond to it. One of the conclusions is that higher frequency low amplitude strains can maintain bone as effectively as low frequency high amplitude strains. This conclusion leads to a paradigm shift in how to treat osteoporosis and how to cope with microgravity.     

High-resolution image-based simulation reveals membrane strain concentration on osteocyte processes caused by tethering elements

Osteocytes are vital for regulating bone remodeling by sensing the flow-induced mechanical stimuli applied to their cell processes. In this mechanosensing mechanism, tethering elements (TEs) connecting the osteocyte process with the canalicular wall potentially amplify the strain on the osteocyte processes. The ultrastructure of the osteocyte processes and canaliculi can be visualized at a nanometer scale using high-resolution imaging via ultra-high voltage electron microscopy (UHVEM). Moreover, the irregular shapes of the osteocyte processes and the canaliculi, including the TEs in the canalicular space, should considerably influence the mechanical stimuli applied to the osteocytes. This study aims to characterize the roles of the ultrastructure of osteocyte processes and canaliculi in the mechanism of osteocyte mechanosensing. Thus, we constructed a high-resolution image-based model of an osteocyte process and a canaliculus using UHVEM tomography and investigated the distribution and magnitude of flow-induced local strain on the osteocyte process by performing fluid–structure interaction simulation. The analysis results reveal that local strain concentration in the osteocyte process was induced by a small number of TEs with high tension, which were inclined depending on the irregular shapes of osteocyte processes and canaliculi. Therefore, this study could provide meaningful insights into the effect of ultrastructure of osteocyte processes and canaliculi on the osteocyte mechanosensing mechanism.

     

Metabolic pathways of the fossil dinosaur bones. Part V. Morphological differentiation of osteocyte lacunae and bone canaliculi and their significance in the system of extracellular communication

The study was carried out on dinosaur bones nearly 80 million years old. Samples for examination were prepared with specially elaborated methods. The light and transmission electron microscopic images permitted two kinds of bone lacunae and two types of paralacunar canalicular endings to be distinguished. The lacunae of the first kind were characterized by their elongated shape, their length exceeding their width several times, their dimensions being 31.2/9.4 microns. The lacunae of the other kind were not so long, their mean measurements amounting to 21.32/9.7 microns. Among the paralacunar canalicular endings those of small diameter were more numerous. The canaliculi of wider, funnel-shaped endings amounted to two or three, they were usually localized in the polar part of the lacuna, and were defined as the axial canaliculi. These were canaliculi of a large diameter. The canalicular wall was constructed of collagen fibres. The same fibres were found in the lacunar wall. Also a relationship between the structure of the lacunar wall and the localization of an osteocyte in the lacuna was analysed in the light and electron microscopes. In regard to the structure of the bone lacuna and the localization of an osteocyte in it, zones A and B were distinguished. Zone A had a characteristic loose and disorderly system of collagen fibres building the lacunar wall. The fibres in this area were by nature open to view. Besides, this region of the lacunar wall revealed specific terraced hollows. Zone B was distinguished by a compact system of parallelly arranged collagen fibres, which formed characteristic ridges in the lacunar wall. The localization of the osteocyte in the lacuna was irregular, the pericellular space around it being of variable width. This space was shown to contain mucopolysaccharides. The images obtained from dinosaur bone were compared with those already known for modern bone. These comparisons permitted it to be ascertained that zone A corresponds to a spot in the lacuna in which the osteocyte exhibits a decreased activity. Zone B is the area of the actual direction of the osteocyte's activity aiming at the shaping of the wall of its lacuna. It can be supposed that the widened endings of the paralacunar canaliculi perform more important functions in conveyance, this being evident from comparisons of analogous areas in modern bone.     

Comparison of strain measurement in the mouse forearm using subject-specific finite element models,strain gaging,and digital image correlation

Mechanical loading in bone leads to the activation of bone-forming pathways that are most likely associated with a minimum strain threshold being experienced by the osteocyte. To investigate the correlation between cellular response and mechanical stimuli, researchers must develop accurate ways to measure/compute strain both externally on the bone surface and internally at the osteocyte level. This study investigates the use of finite element (FE) models to compute bone surface strains on the mouse forearm. Strains from three FE models were compared to data collected experimentally through strain gaging and digital image correlation (DIC). Each FE model was assigned subject-specific bone properties and consisted of one-dimensional springs representing the interosseous membrane. After three-point bending was performed on the ulnae and radii, moment of inertia was determined from microCT analysis of the bone region between the supports and then used along with standard beam analyses to calculate the Young’s modulus. Non-contact strain measurements from DIC were determined to be more suitable for validating numerical results than experimental data obtained through conventional strain gaging. When comparing strain responses in the three ulnae, we observed a 3–14% difference between numerical and DIC strains while the strain gage values were 37–56% lower than numerical values. This study demonstrates a computational approach for capturing bone surface strains in the mouse forearm. Ultimately, strains from these macroscale models can be used as inputs for microscale and nanoscale FE models designed to analyze strains directly in the osteocyte lacunae.     

Microstructural strain near osteocyte lacuna in cortical bone in vitro

Mechanical factors affect bone remodeling such that increased mechanical demand results in net bone formation, whereas decreased demand results in net bone resorption. Two proposed mechanical signals are stress-generated fluid flow forces acting on cells and bone matrix deformation itself. A prominent current theory is that bone cells are more responsive to fluid flow than to mechanical strain. Recent experiments support this conclusion: bone cells increase their production of osteopontin (OPN) mRNA, prostaglandin (PGE(2)), and nitric oxide (NO) in response to fluid flow in contrast to cells stimulated by mechanical strain levels similar to those measured in vivo. However, when cells are subjected to substrate strains levels many times greater than those measured in vivo, increased biological activity again results. We assert that it is neither fluid flow nor matrix deformation per se, but rather the resulting cell deformation that causes cell biological response. Machined specimens of undamaged bovine cortical bone were subjected to increasing levels of macroscopic strain while observed under an optical microscope at 220X. Continuum level strain was measured using a standard foil strain gauge attached to the back of the specimen and ranged from 500 to 6,000 microstrain. Images of the specimen surface at each strain level were captured. To determine the level of osteocyte deformation that results from fluid flow in vitro, MLO-Y4 cells were cultured on collagen coated 190 cm2 plastic sheets and subjected to steady fluid flow at 16 dynes/cm(2). Images representing the initial undisturbed cell configuration and the configuration of the cells after ten minutes of fluid flow were acquired from a videotape of the flow experiment. The captured unloaded vs. loaded image pairs were analyzed to determine the local deformation and strain fields using a digital stereoimaging system. When subjected to a nominal continuum strain level approximately equal to that measured in humans in vivo during rigorous activity (2,000 microstrain), the local, osteocyte level strains can be as high as 12,000 to 15,000 microstrain (1.2% to 1.5%). Average osteocyte strains due to fluid flow in vitro increase from 7,972 microstrains after 16 seconds of flow to 22,856 microstrains after 64 seconds of flow. In contrast, maximum strains measured in vivo are approximately 1,800 microstrain in humans and up to 3,000 microstrain in other species. These data may help to explain why bone cells are more sensitive to fluid flow than substrate strain; fluid forces result in cell deformations much higher than those considered to be "physiological".     

Orientation of collagen at the osteocyte lacunae in human secondary osteons

This work characterizes an aspect of human bone micro-structure, pertinent to fracture initiation and arrest. It addresses how the orientation of elementary components proximate to osteocyte lacunae influences secondary osteon micro-biomechanics. New data at the perilacunar region concerning orientation of collagen-apatite, and prior data on collagen orientation outside the perilacunar region, are incorporated in a novel simulation of osteons to investigate how orientation relates to strains and stresses during mechanical testing. The perilacunar region was observed by confocal microscopy within single lamellar specimens, isolated from osteons. The specimens were separated by extinct or bright appearance in transverse section under circularly polarizing light. This is because synchrotron diffraction and confocal microscopy had established that each type, away from the perilacunar region, corresponds to specific dominant collagen orientation (extinct lamellae's dominant collagen forming small angles with the original osteon axis, while the bright lamellae's forms larger angles). Morphometry of serial confocal images of each perilacunar region showed collagen orientation generally following the orientation of canaliculi, circumambiently-perpendicular to the lacuna. The lacunae tilted relative to the lamellar walls were more numerous in extinct than in bright lamella. Their apices were less likely in extinct than bright lamella to show collagen following the canalicular orientation. The simulation of osteocyte lacunae in osteons, under tension or compression loading, supports the hypothesis that collagen orientation affects strains and stresses at the equatorial perilacunar region in conjunction with the presence of the lacuna. We further conjecture that collagen orientation diverts propagation of micro-cracks initiating from apices.     

Three‐dimensional characterization of osteocyte volumes at multiple scales,and its relationship with bone biology and genome evolution in ray‐finned fishes

Osteocytes, cells embedded within the bone mineral matrix, inform on key aspects of vertebrate biology. In particular, a relationship between volumes of the osteocytes and bone growth and/or genome size has been proposed for several tetrapod lineages. However, the variation in osteocyte volume across different scales is poorly characterized and mostly relies on incomplete, two‐dimensional information. In this study, we characterize the variation of osteocyte volumes in ray‐finned fishes (Actinopterygii), a clade including more than half of modern vertebrate species in which osteocyte biology is poorly known. We use X‐ray synchrotron micro‐computed tomography (SRµCT) to achieve a three‐dimensional visualization of osteocyte lacunae and direct measurement of their size (volumes). Our specimen sample is designed to characterize variation in osteocyte lacuna morphology at three scales: within a bone, among the bones of one individual and among species. At the intra‐bone scale, we find that osteocyte lacunae vary noticeably in size between zones of organized and woven bone (being up to six times larger in woven bone), and across cyclical bone deposition. This is probably explained by differences in bone deposition rate, with larger osteocyte lacunae contained in bone that deposits faster. Osteocyte lacuna volumes vary 3.5‐fold among the bones of an individual, and this cannot readily be explained by variation in bone growth rate or other currently observable factors. Finally, we find that genome size provides the best explanation of variation in osteocyte lacuna volume among species: actinopterygian taxa with larger genomes (polyploid taxa in particular) have larger osteocyte lacunae (with a ninefold variation in median osteocyte volume being measured). Our findings corroborate previous two‐dimensional studies in tetrapods that also observed similar patterns of intra‐individual variation and found a correlation with genome size. This opens new perspectives for further studies on bone evolution, physiology and palaeogenomics in actinopterygians, and vertebrates as a whole.     

Machine vision photogrammetry: a technique for measurement of microstructural strain in cortical bone.

Understanding local microstructural deformations and strains in cortical bone may lead to a better understanding of cortical bone damage development, fracture, and remodeling. Traditional experimental techniques for measuring deformation and strain do not allow characterization of these quantities at the microstructural level in cortical bone. This study describes a technique based on digital stereoimaging used to measure the microstructural strain fields in cortical bone. The technique allows the measurement of material surface displacements and strains by comparing images acquired from a specimen at two distinct stress states. The accuracy of the system is investigated by analyzing an undeformed image set; the test image is identical to the reference image but translated by a known pixel amount. An increase in the correlation sub-image train parameter results in an increase in displacement measurement accuracy from 0.049 to 0.012 pixels. Errors in strain calculated from the measured displacement field were between 39 and 564 microstrain depending upon the sub-image train size and applied image displacement. The presence of a microcrack in cortical bone results in local strain at the crack tip reaching 0.030 (30,000 microstrain) and 0.010 (10,000 microstrain) near osteocyte lacunae. It is expected that the use of this technique will allow a greater understanding of bone strength and fracture as well as bone mechanotransduction.     

Function of osteocytes in bone

Although the structural design of cellular bone (i.e., bone containing osteocytes that are regularly spaced throughout the bone matrix) dates back to the first occurrence of bone as a tissue in evolution, and although osteocytes represent the most abundant cell type of bone, we know as yet little about the role of the osteocyte in bone metabolism. Osteocytes descend from osteoblasts. They are formed by the incorporation of osteoblasts into the bone matrix. Osteocytes remain in contact with each other and with cells on the bone surface via gap junction–coupled cell processes passing through the matrix via small channels, the canaliculi, that connect the cell body–containing lacunae with each other and with the outside world. During differentiation from osteoblast to mature osteocyte the cells lose a large part of their cell organelles. Their cell processes are packed with microfilaments. In this review we discuss the various theories on osteocyte function that have taken in consideration these special features of osteocytes. These are (1) osteocytes are actively involved in bone turnover; (2) the osteocyte network is through its large cell-matrix contact surface involved in ion exchange; and (3) osteocytes are the mechanosensory cells of bone and play a pivotal role in functional adaptation of bone. In our opinion, especially the last theory offers an exciting concept for which some biomechanical, biochemical, and cell biological evidence is already available and which fully warrants further investigations. © 1994 Wiley-Liss, Inc.     

Quantification of a rat tail vertebra model for trabecular bone adaptation studies

A feedback controlled loading apparatus for the rat tail vertebra was developed to deliver precise mechanical loads to the eighth caudal vertebra (C8) via pins inserted into adjacent vertebrae. Cortical bone strains were recorded using strain gages while subjecting the C8 in four cadaveric rats to mechanical loads ranging from 25 to 100 N at 1 Hz with a sinusoidal waveform. Finite element (FE) models, based on micro computed tomography, were constructed for all four C8 for calculations of cortical and trabecular bone tissue strains. The cortical bone strains predicted by FE models agreed with strain gage measurements, thus validating the FE models. The average measured cortical bone strain during 25-100 N loading was between 298 +/- 105 and 1210 +/- 297 microstrain (muepsilon). The models predicted average trabecular bone tissue strains ranging between 135 +/- 35 and 538 +/- 138 mu epsilon in the proximal region, 77 +/- 23-307 +/- 91 muepsilon in the central region, and 155 +/- 36-621 +/- 143 muepsilon in the distal region for 25-100 N loading range. Although these average strains were compressive, it is also interesting that the trabecular bone tissue strain can range from compressive to tensile strains (-1994 to 380 mu epsilon for a 100 N load). With this novel approach that combines an animal model with computational techniques, it could be possible to establish a quantitative relationship between the microscopic stress/strain environment in trabecular bone tissue, and the biosynthetic response and gene expression of bone cells, thereby study bone adaptation.     

Estimation of bone permeability using accurate microstructural measurements

While interstitial fluid flow is necessary for the viability of osteocytes, it is also believed to play a role in bone's mechanosensory system by shearing bone cell membranes or causing cytoskeleton deformation and thus activating biochemical responses that lead to the process of bone adaptation. However, the fluid flow properties that regulate bone's adaptive response are poorly understood. In this paper, we present an analytical approach to determine the degree of anisotropy of the permeability of the lacunar-canalicular porosity in bone. First, we estimate the total number of canaliculi emanating from each osteocyte lacuna based on published measurements from parallel-fibered shaft bones of several species (chick, rabbit, bovine, horse, dog, and human). Next, we determine the local three-dimensional permeability of the lacunar-canalicular porosity for these species using recent microstructural measurements and adapting a previously developed model. Results demonstrated that the number of canaliculi per osteocyte lacuna ranged from 41 for human to 115 for horse. Permeability coefficients were found to be different in three local principal directions, indicating local orthotropic symmetry of bone permeability in parallel-fibered cortical bone for all species examined. For the range of parameters investigated, the local lacunar-canalicular permeability varied more than three orders of magnitude, with the osteocyte lacunar shape and size along with the 3-D canalicular distribution determining the degree of anisotropy of the local permeability. This two-step theoretical approach to determine the degree of anisotropy of the permeability of the lacunar-canalicular porosity will be useful for accurate quantification of interstitial fluid movement in bone.     

Uptake of horseradish peroxidase by bone cells during endochondral bone development

Summary To investigate the mechanisms whereby bone cells absorb organic bone-matrix components during endochondral bone development, rat humeri were examined, employing horseradish peroxidase as a soluble protein tracer.Intravenously-injected peroxidase filled the osteoid layer and penetrated into the osteocyte lacunae and canaliculi, but did not enter the mineralized bone matrix. Whereas osteocytes rarely took up exogenous peroxidase, osteoblasts and osteoclasts actively endocytosed peroxidase in pinocytotic coated vesicles, tubular structures, and vacuoles. They also formed endocytotic vacuoles containing peroxidase in the Golgi area. The Golgi apparatus and dense bodies of these bone cells were, however, free of reaction products. Osteoclast ruffled borders were responsible for peroxidase absorption. In the osteoblast, osteocyte and osteoclast, endogenous peroxidatic reaction was detected only in mitochondria and not in other membrane-bounded vesicles and bodies. These results strongly suggest that both osteoblasts and osteoclasts participate in the resorption of bone-matrix organic components during bone remodelling.