Canalicular Fluid Research Articles

Computer model of non-Newtonian canalicular fluid flow in lacunar-canalicular system of bone tissue.

Brittle bone diseases are a global healthcare problem for orthopaedic clinicians, that reduces bone strength and promotes bone fracture risk. In vivo studies reported that loading-induced fluid flow through the lacunar-canalicular channel (LCS) of bone tissue inhibit such bone loss and encourages osteogenesis i.e. new bone formation. Canalicular fluid flow converts mechanical signals into biological signals and regulates bone reconstruction by releasing signalling molecules responsible for mechanotransduction. In-silico model mostly considers canalicular fluid is Newtonian, however, physiological canalicular fluid may be non-Newtonian in nature as it contains nutrients and supplements. Accordingly, this study attempts to develop a two-dimensional in-silico model to compute loading-induced non-Newtonian canalicular fluid flow in a complex LCS of bone tissue. Moreover, canalicular fluid is considered as a Jeffery fluid, that can easily be reduced to Newtonian fluid as a special case. The results show that physiological loading modulates the canalicular fluid flow, wall shear stress (WSS) and streamline in bone LCS. Fluid velocity and WSS increases with increase in non-dimensional frequency and non-Newtonian parameter (Jeffery fluid parameters) and reduce with change in permeability. The outcomes of this study may provide new insights in the role of mechanical loading-induced non-Newtonian canalicular fluid flow dynamics in bone LCS. The key findings of this study can be used to improve the understanding of osteocyte mechanobiology involved inside the bone tissue.

A computational study on electric field-induced canalicular fluid flow in bone tissue

A computational study on electric field-induced canalicular fluid flow in bone tissue

Perlecan, A Multi-Functional, Cell-Instructive, Matrix-Stabilizing Proteoglycan With Roles in Tissue Development Has Relevance to Connective Tissue Repair and Regeneration.

This review highlights the multifunctional properties of perlecan (HSPG2) and its potential roles in repair biology. Perlecan is ubiquitous, occurring in vascular, cartilaginous, adipose, lymphoreticular, bone and bone marrow stroma and in neural tissues. Perlecan has roles in angiogenesis, tissue development and extracellular matrix stabilization in mature weight bearing and tensional tissues. Perlecan contributes to mechanosensory properties in cartilage through pericellular interactions with fibrillin-1, type IV, V, VI and XI collagen and elastin. Perlecan domain I - FGF, PDGF, VEGF and BMP interactions promote embryonic cellular proliferation, differentiation, and tissue development. Perlecan domain II, an LDLR-like domain interacts with lipids, Wnt and Hedgehog morphogens. Perlecan domain III binds FGF-7 and 18 and has roles in the secretion of perlecan. Perlecan domain IV, an immunoglobulin repeat domain, has cell attachment and matrix stabilizing properties. Perlecan domain V promotes tissue repair through interactions with VEGF, VEGF-R2 and α2β1 integrin. Perlecan domain-V LG1-LG2 and LG3 fragments antagonize these interactions. Perlecan domain V promotes reconstitution of the blood brain barrier damaged by ischemic stroke and is neurogenic and neuroprotective. Perlecan-VEGF-VEGFR2, perlecan-FGF-2 and perlecan-PDGF interactions promote angiogenesis and wound healing. Perlecan domain I, III and V interactions with platelet factor-4 and megakaryocyte and platelet inhibitory receptor promote adhesion of cells to implants and scaffolds in vascular repair. Perlecan localizes acetylcholinesterase in the neuromuscular junction and is of functional significance in neuromuscular control. Perlecan mutation leads to Schwartz-Jampel Syndrome, functional impairment of the biomechanical properties of the intervertebral disc, variable levels of chondroplasia and myotonia. A greater understanding of the functional working of the neuromuscular junction may be insightful in therapeutic approaches in the treatment of neuromuscular disorders. Tissue engineering of salivary glands has been undertaken using bioactive peptides (TWSKV) derived from perlecan domain IV. Perlecan TWSKV peptide induces differentiation of salivary gland cells into self-assembling acini-like structures that express salivary gland biomarkers and secrete α-amylase. Perlecan also promotes chondroprogenitor stem cell maturation and development of pluripotent migratory stem cell lineages, which participate in diarthrodial joint formation, and early cartilage development. Recent studies have also shown that perlecan is prominently expressed during repair of adult human articular cartilage. Perlecan also has roles in endochondral ossification and bone development. Perlecan domain I hydrogels been used in tissue engineering to establish heparin binding growth factor gradients that promote cell migration and cartilage repair. Perlecan domain I collagen I fibril scaffolds have also been used as an FGF-2 delivery system for tissue repair. With the availability of recombinant perlecan domains, the development of other tissue repair strategies should emerge in the near future. Perlecan co-localization with vascular elastin in the intima, acts as a blood shear-flow endothelial sensor that regulates blood volume and pressure and has a similar role to perlecan in canalicular fluid, regulating bone development and remodeling. This complements perlecan’s roles in growth plate cartilage and in endochondral ossification to form the appendicular and axial skeleton. Perlecan is thus a ubiquitous, multifunctional, and pleomorphic molecule of considerable biological importance. A greater understanding of its diverse biological roles and functional repertoires during tissue development, growth and disease will yield valuable insights into how this impressive proteoglycan could be utilized successfully in repair biology.

Electromagnetic field induced alterations in fluid flow through lacuno-canalicular system of bone

Osteoporosis, bone-muscle disuse, and long-term exposure to space microgravity cause bone loss and promote bone fractures. Therapeutic devices and interventions for the prevention of bone loss and regulation of fracture healing are serious challenges for orthopaedic clinicians and researchers. Electromagnetic field exposure in combination with mechanical loading has the potential to prevent disuse bone loss as it promotes osteogenesis i.e., new bone formation. Loading-induced fluid motion in lacunar space acts as a stimulus for osteo-activities. Nevertheless, how the electromagnetic field regulates biomechanical stimulus such as fluid motion required for osteogenesis within bone tissue is not well understood. Accordingly, this paper attempts to establish a mathematical model to investigate the role of electromagnetic fields to alter the cantilever bending-induced lacunar fluid flow behaviour. A more realistic curvy canalicular channel subjected to electrohydrodynamics (EHD) mechanism is modelled to study poromechanical and canalicular fluid flow parameters such as pore-pressure gradient, fluid velocity, wall shear, and streamline patterns. Computational results indicate that electromagnetic fields modulate the fluid motion required for osteogenic activities even in bone tissues with higher porosity and permeability. The present study explains how electromagnetic fields possibly modulate mechanobiological stimulus in bone tissue. Based on the outcomes, an optimal loading regimen along with the electromagnetic field can be selected to enhance osteogenic response to prevent and mitigate irreversible bone loss that occurred due to microgravity and disuse.

Physiological Loading-Induced Interstitial Fluid Dynamics in Osteon of Osteogenesis Imperfecta Bone.

Osteogenesis imperfecta (OI), also known as "brittle bone disease," is a genetic bone disorder. OI bones experience frequent fractures. Surgical procedures are usually followed by clinicians in the management of OI. It has been observed physical activity is equally beneficial in reducing OI bone fractures in both children and adults as mechanical stimulation improves bone mass and strength. Loading-induced mechanical strain and interstitial fluid flow stimulate bone remodeling activities. Several studies have characterized strain environment in OI bones, whereas very few studies attempted to characterize the interstitial fluid flow. OI significantly affects bone micro-architecture. Thus, this study anticipates that canalicular fluid flow reduces in OI bone in comparison to the healthy bone in response to physiological loading due to altered poromechanical properties. This work attempts to understand the canalicular fluid distribution in single osteon models of OI and healthy bone. A poromechanical model of osteon is developed to compute pore-pressure and interstitial fluid flow as a function of gait loading pattern reported for OI and healthy subjects. Fluid distribution patterns are compared at different time-points of the stance phase of the gait cycle. It is observed that fluid flow significantly reduces in OI bone. Additionally, flow is more static than dynamic in OI osteon in comparison to healthy subjects. This work attempts to identify the plausible explanation behind the diminished mechanotransduction capability of OI bone. This work may further be extended for designing better biomechanical therapies to enhance the fluid flow in order to improve osteogenic activities in OI bone.

Perlecan in Pericellular Mechanosensory Cell-Matrix Communication, Extracellular Matrix Stabilisation and Mechanoregulation of Load-Bearing Connective Tissues.

In this study, we review mechanoregulatory roles for perlecan in load-bearing connective tissues. Perlecan facilitates the co-acervation of tropoelastin and assembly of elastic microfibrils in translamellar cross-bridges which, together with fibrillin and elastin stabilise the extracellular matrix of the intervertebral disc annulus fibrosus. Pericellular perlecan interacts with collagen VI and XI to define and stabilize this matrix compartment which has a strategic position facilitating two-way cell-matrix communication between the cell and its wider extracellular matrix. Cues from the extracellular matrix are fed through this pericellular matrix back to the chondrocyte, allowing it to perceive and respond to subtle microenvironmental changes to regulate tissue homeostasis. Thus perlecan plays a key regulatory role in chondrocyte metabolism, and in chondrocyte differentiation. Perlecan acts as a transport proteoglycan carrying poorly soluble, lipid-modified proteins such as the Wnt or Hedgehog families facilitating the establishment of morphogen gradients that drive tissue morphogenesis. Cell surface perlecan on endothelial cells or osteocytes acts as a flow sensor in blood and the lacunar canalicular fluid providing feedback cues to smooth muscle cells regulating vascular tone and blood pressure, and the regulation of bone metabolism by osteocytes highlighting perlecan’s multifaceted roles in load-bearing connective tissues.

Anatomical variations in cortical bone surface permeability: Tibia versus femur

Cortical bone surfaces (periosteal and endosteal) exhibit differential (re)modelling response to mechanical loading. This poses a serious challenge in establishing an in silico model to predict site-specific new bone formation as a function of mechanical stimulus. In this regard, mechanical loading-induced fluid motion in lacunar-canalicular system (LCS) is assumed osteogenic. Micro-architectural properties, especially permeability regulate canalicular fluid motion within the bone. The knowledge of these properties is required to compute flow distribution. Along the same line, it is possible that cortical surfaces may experience differential fluid distribution due to anatomical variations in microarchitectural properties which may induce distinct new bone response at cortical surfaces. Nevertheless, these properties are not well reported for cortical surfaces in the literature. Accordingly, the present study aims to measure microarchitectural properties especially permeability at different anatomical locations (medial, lateral, anterior, and posterior) of periosteal and endosteal surfaces using nanoindentation. A standard poroelastic optimization technique was used to estimate permeability, shear modulus, and Poisson's ratio. The properties are also compared for two weight-bearing bones i.e. tibia and femur. Endosteal surface was found more permeable as compared to the periosteal surface. Tibial endosteal surface had shown greater permeability values at most of the anatomical locations as compared to femoral endosteal surface. The outcomes may be used to precisely predict site-specific osteogenesis in cortical bone as a function of canalicular flow distribution. This work may ultimately be beneficial in designing the loading parameters to stimulate desired new bone response for the prevention and the cure of bone loss.

Canal of Hering loss is an initiating step for primary biliary cholangitis (PBC): A hypothesis

The origin and initiating features of PBC remain obscure despite decades of study. However, recent papers have demonstrated loss of canals of Hering (CoH) to be the earliest histologic change in liver biopsy specimens from patients with primary biliary cholangitis (PBC). We posit that CoH loss prior to significant inflammation or evidence of bile duct injury might be a very early, perhaps even an initiating lesion of PBC. As a potential target of inflammatory or toxic injury, CoH loss may initiate rather than follow the cascade of events leading to duct injury and loss and their sequelae. Toxins may be exogenous in origin, such as environmental toxins or drug exposures, or endogenous, resulting from genetic or epigenetic alterations in canalicular bile transporters upstream from the CoH. In turn, this hypothesis suggests that loss of CoH would lead to altered bile flow and composition injurious to downstream bile ducts, because bile composition has not been modulated by normal CoH physiologic functions or because, in the absence of CoH, canalicular fluid flow into the biliary tree is disrupted interfering with soluble trophic factors important for bile duct integrity. Regardless of the pathogenic mechanism causing CoH loss, only following such loss would the characteristic diagnostic findings of PBC become evident: damage to downstream interlobular and sub-lobular bile ducts. To the extent that the causal mechanisms for CoH loss can be identified, clinical identification (as through early identification of CoH loss) and intervention (depending on the inciting cause) may offer promise for treatment of this enigmatic disease.

Estimation of Loading-Driven Fluid-Flow Pattern in Lacunar-Canalicular Space Using Fluid Structure Interaction

Bone cells namely osteoblasts and osteocytes are assumed responsible for loading-induced osteogenesis. Osteocytes lie within the bone and connect to each other via cell process passing through canalicular spaces which forms a canalicular network. Fluid motion across this network acts as a medium of communication with neighbouring cells. Mechanical loading-induced pressure gradients in lacunar canalicular space (LCS) causes canalicular fluid to flow. However, it remains unclear how canalicular fluid motion and solid structure interact with each other under loading derived strain environment. In the present study, a two-way fluid structure interaction model is developed to estimate canalicular fluid flow behaviour. Flow streamlines and wall shear stress are computed. Results indicates that wall shear due to fluid flow on the pores wall is also in the same pattern as the velocity streams and velocity is maximum at those regions where the wall shear is also maximum. The outcomes provide a better understanding for developing strategies to enhance the fluid flow in bone, which may ultimately be useful in the development of effective countermeasure for the reversal of bone loss.

The role of fluid flow on bone mechanobiology: mathematical modeling and simulation

The effect of fluid flow on tissue adaptation was the focus of many research works during the last years. Moreover, the use of poroelasticity models to simulate and understand the interstitial flow movement has taken interest due to the possibility to include the fluid effect on mechanical simulations. In particular, shear stresses induced by bone canalicular fluid flow are suggested to be one of the mechanical stimulus controlling bone remodeling processes. Due to the high difficulty to measure canalicular fluid flow and shear stresses, computational poroelastic models can be used in order to estimate these parameters. In this work, a finite element dual porosity model based on Russian doll poroelasticity is developed. Two experiments with a turkey ulna and a human femur are simulated. Bone lacuno-canalicular fluid flow is computed and compared with the experimental results, focusing on the zones of bone remodeling and showing a relation between this flow and the bone formation process.

Canalicular Junctions in the Osteocyte Lacuno-Canalicular Network of Cortical Bone.

The osteocyte lacuno-canalicular network (LCN) is essential for bone remodeling because osteocytes regulate cell recruitment. This has been proposed to occur through liquid-flow-induced shear forces in the canaliculi. Models of the LCN have thus far assumed that it contains canaliculi connecting the osteocyte lacunae. However, here, we reveal that enlarged spaces occur at places where several canaliculi cross; we name these spaces canalicular junctions. We characterize them in detail within mice cortical bone using synchrotron nanotomography at two length scales, with 50 and 130 nm voxel size, and show that canalicular junctions occur at a density similar to that of osteocyte lacunae and that canalicular junctions tend to cluster. Through confocal laser scanning microscopy, we show that canalicular junctions are widespread as we have observed them in cortical bone from several species, even though the number density of the canalicular junctions was not universal. Fluid flow simulations of a simple model system with and without a canalicular junction clearly show that liquid mass transport and flow velocities are altered by the presence of canalicular junctions. We suggest that these canalicular junctions may play an important role in osteocyte communication and possibly also in canalicular fluid flow. Therefore, we believe that they constitute an important component in the bone osteocyte network.

Canalicular fluid flow induced by loading waveforms: A comparative analysis

Dynamic loading on the bone is beneficial in prevention and cure of bone loss as it encourages osteogenesis (i.e., new bone formation). Loading parameters such as strain magnitude, frequency, cycles, and strain rate (depending on loading waveform) affect the new bone formation. In-vivo studies suggested an optimal and osteogenic range of strain magnitude, frequency, and cycles to elicit the maximum new bone response. Still, there is no consensus on the selection of loading waveform. Animal studies on bone adaptation considered sinusoidal, and non-sinusoidal (e.g., trapezoidal, sawtooth, and triangular) loading waveforms according to physiological loadings (e.g., walking, running, and jumping etc.) without considering the relative effect of these waveforms on the loading-induced mechanical environment. The present study attempts to bridge this gap. Accordingly, this work hypothesizes that bone being a biphasic material (solid and fluid phases) experiences the same strain distribution for the different loading waves of the same amplitude, however, other components of the mechanical environment such as pore-pressure and interstitial fluid motion regulating the bone adaptation may differ. An in-vivo cantilever bending study is selected to substantiate the hypothesis. A poroelastic model is used to estimate the pore pressure and fluid motion developed in mouse tibia subjected to the: (i) trapezoidal, (ii) sawtooth, and (iii) triangular bending waves. Furthermore, poroelastic response of pore-pressure and fluid motion induced by these loading waveforms are compared and analyzed. This work also investigates how bone loss associated alterations in the microstructural environment of cortical bone affect the canalicular fluid motion induced by these waveforms. Overall results may be useful in designing optimal biomechanical interventions such as physical exercises to improve the bone health.

In silico modeling of bone adaptation to rest-inserted loading: Strain energy density versus fluid flow as stimulus

In vivo studies suggest that cyclic and low-magnitude loading can be useful over pharmaceutical drugs in normalizing bone loss as it encourages osteogenesis (i.e. new bone formation) at the sites of elevated strain magnitude. In silico models assumed normal strain or strain energy density (SED) as the stimulus to predict loading-induced osteogenesis, however, these models may have limited success in fitting the in vivo new bone formation at several instances. For example, rest-inserted cyclic loading amplifies the new bone formation as compared to continuous-cyclic loading even though similar strain magnitude were induced in both the cases. It is also believed that loading-induced interstitial fluid flow can also be a potential stimulus of osteogenesis. The present study hypothesizes that fluid motion as osteogenic stimulus may explain the afore-mentioned anomalies. Accordingly, this work studies osteogenesis as functions of SED and canalicular fluid motion using an in silico model. Therefore, the new bone formation is considered roughly proportional to stimuli above their osteogenic thresholds. This model attempts to simulate in vivo new bone formation noticed in rest-inserted cantilever loading studies. The model's prediction of site-specific new bone formation improves when fluid flow is considered as the stimulus. It is also noticed that fluid motion as the stimulus closely fits the new bone formation for another in vivo study where the effects of aging on osteogenesis were examined. These attempts to establish fluid flow as a potential osteogenic stimulus can be useful in the prediction of site-specific new bone formation. The findings will ultimately be useful in designing biomechanical interventions such as prophylactic exercises to cure bone loss.

Mathematically modeling fluid flow and fluid shear stress in the canaliculi of a loaded osteon.

BackgroundMechanical load-induced intraosseous pressure gradients may result in some fluid stimuli effects, such as fluid flow and fluid shear stress (FSS), which may enable bone cells to detect external mechanical signals. Interstitial bone fluid flow is known to occur in lacunar–canalicular porosity (PLC).MethodsIn order to characterize lacunar–canalicular fluid flow behavior, a hierarchical osteon system is developed. The osteon is modeled as a poroelastic annular cylinder with two types of impermeable boundary cases considered on its outer wall: one is elastic restrained (Case I), whereas the other is displacement confined (Case II). Analytical solutions such as canalicular fluid velocity, pressure, fluid flow rate (FFR), and shear stress are obtained.ResultsResults show that the amplitudes of FFR and FSS are proportional to strain amplitude and frequency. However, the key loading factor governing canalicular fluid flow behavior is the strain rate. The larger canalicular radius is, the larger amplitudes of FFR and FSS generalized, especially, the FSS amplitude is proportional to canalicular radius. In addition, both FFR and FSS amplitudes produced in case II are larger than those of case I.ConclusionStrain rate can be acted as a representative loading parameter governing the canalicular fluid flow behavior under a physiological state. This model can facilitate better understanding the load induced the fluid permeation in the PLC. The approach can also be used to analyze the structure of the proteoglycan matrix in the fluid space surrounding the osteocytic process in the canaliculus.

Computer modelling of bone's adaptation: the role of normal strain, shear strain and fluid flow.

Bone loss is a serious health problem. In vivo studies have found that mechanical stimulation may inhibit bone loss as elevated strain in bone induces osteogenesis, i.e. new bone formation. However, the exact relationship between mechanical environment and osteogenesis is less clear. Normal strain is considered as a prime stimulus of osteogenic activity; however, there are some instances in the literature where osteogenesis is observed in the vicinity of minimal normal strain, specifically near the neutral axis of bending in long bones. It suggests that osteogenesis may also be induced by other or secondary components of mechanical environment such as shear strain or canalicular fluid flow. As it is evident from the literature, shear strain and fluid flow can be potent stimuli of osteogenesis. This study presents a computational model to investigate the roles of these stimuli in bone adaptation. The model assumes that bone formation rate is roughly proportional to the normal, shear and fluid shear strain energy density above their osteogenic thresholds. In vivo osteogenesis due to cyclic cantilever bending of a murine tibia has been simulated. The model predicts results close to experimental findings when normal strain, and shear strain or fluid shear were combined. This study also gives a new perspective on the relation between osteogenic potential of micro-level fluid shear and that of macro-level bending shear. Attempts to establish such relations among the components of mechanical environment and corresponding osteogenesis may ultimately aid in the development of effective approaches to mitigating bone loss.

Hierarchical model for strain generalized streaming potential induced by the canalicular fluid flow of an osteon

A hierarchical model is developed to predict the streaming potential (SP) in the canaliculi of a loaded osteon. Canaliculi are assumed to run straight across the osteon annular cylinder wall, while disregarding the effect of lacuna. SP is generalized by the canalicular fluid flow. Analytical solutions are obtained for the canalicular fluid velocity, pressure, and SP. Results demonstrate that SP amplitude (SPA) is proportional to the pressure difference, strain amplitude, frequency, and strain rate amplitude. However, the key loading factor governing SP is the strain rate, which is a representative loading parameter under the specific physiological state. Moreover, SPA is independent of canalicular length. This model links external loads to the canalicular fluid pressure, velocity, and SP, which can facilitate further understanding of the mechanotransduction and electromechanotransduction mechanisms of bones.

Interstitial fluid flow within bone canaliculi and electro-chemo-mechanical features of the canalicular milieu

Canalicular fluid flow is acknowledged to play a major role in bone functioning, allowing bone cells' metabolism and activity and providing an efficient way for cell-to-cell communication. Bone canaliculi are small canals running through the bone solid matrix, hosting osteocyte's dendrites, and saturated by an interstitial fluid rich in ions. Because of the small size of these canals (few hundred nanometers in diameter), fluid flow is coupled with electrochemical phenomena. In our previous works, we developed a multi-scale model accounting for coupled hydraulic and chemical transport in the canalicular network. Unfortunately, most of the physical and geometrical information required by the model is hardly accessible by nowadays experimental techniques. The goal of this study was to numerically assess the influence of the physical and material parameters involved in the canalicular fluid flow. The focus was set on the electro-chemo-mechanical features of the canalicular milieu, hopefully covering any in vivo scenario. Two main results were obtained. First, the most relevant parameters affecting the canalicular fluid flow were identified and their effects quantified. Second, these findings were given a larger scope to cover also scenarios not considered in this study. Therefore, this study gives insight into the potential interactions between electrochemistry and mechanics in bone and provides the rational for further theoretical and experimental investigations.

Loss of the NHE2 Na+/H+ Exchanger in Mice Results in Dilation of Folliculo‐Stellate Cell Canaliculi

Genetic ablation of the NHE2 Na+/H+ exchanger causes gastric achlorhydria, absorptive defects in kidney and colon, and low fertility. Here we show that NHE2 is expressed in the pituitary, with the highest mRNA expression in pars distalis and lower expression in pars intermedia. In pars distalis of NHE2-null mice, prominent cyst-like dilatations of folliculo-stellate (FS) cell canaliculi developed with age, and there were increased FS cell area, accumulation of lipid in FS cell cytoplasm, redundancies in FS cell basement membrane, and other changes. The expansion of the canaliculi indicates that NHE2 is a major absorptive Na+/H+ exchanger in the luminal membranes lining the extensive network of channels formed by FS cells, which may provide a means of intrapituitary communication. The results suggest that NHE2 contributes to homeostatic regulation of the volume and composition of the canalicular fluid and may counter the secretory activity of the CFTR Cl− channel, which is known to be expressed in pituitary.

Decrease in the osteocyte lacunar density accompanied by hypermineralized lacunar occlusion reveals failure and delay of remodeling in aged human bone

Aging decreases the human femur's fatigue resistance, impact energy absorption, and the ability to withstand load. Changes in the osteocyte distribution and in their elemental composition might be involved in age-related bone impairment. To address this question, we carried out a histomorphometric assessment of the osteocyte lacunar distribution in the periosteal and endosteal human femoral cortexes of 16 female and 16 male donors with regard to age- and sex-related bone remodeling. Measurements of the bone mineral density distribution by quantitative backscattered electron imaging and energy dispersive X-ray analysis were taken to evaluate the osteocyte lacunar mineral composition and characteristics. Age-dependent decreases in the total osteocyte lacunar number were measured in all of the cases. This change signifies a risk for the bone's safety. Cortical subdivision into periosteal and endosteal regions of interest emphasized that, in both sexes, primarily the endosteal cortex is affected by age-dependent reduction in number of osteocyte lacunae, whereas the periosteal compartment showed a less pronounced osteocyte lacunar deficiency. In aged bone, osteocyte lacunae showed an increased amount of hypermineralized calcium phosphate occlusions in comparison with younger cases. With respect to Frost's early delineation of micropetrosis, our microanalyses revealed that the osteocyte lacunae are subject to hypermineralization. Intralacunar hypermineralization accompanied by a decrease in total osteocyte lacunar density may contribute to failure or delayed bone repair in aging bone. A decreased osteocyte lacunar density may cause deteriorations in the canalicular fluid flow and reduce the detection of microdamage, which counteracts the bone's structural integrity, while hypermineralized osteocyte lacunae may increase bone brittleness and render the bone fragile.

Real-time measurement of solute transport within the lacunar-canalicular system of mechanically loaded bone: Direct evidence for load-induced fluid flow

Since proposed by Piekarski and Munro in 1977, load-induced fluid flow through the bone lacunar-canalicular system (LCS) has been accepted as critical for bone metabolism, mechanotransduction, and adaptation. However, direct unequivocal observation and quantification of load-induced fluid and solute convection through the LCS have been lacking due to technical difficulties. Using a novel experimental approach based on fluorescence recovery after photobleaching (FRAP) and synchronized mechanical loading and imaging, we successfully quantified the diffusive and convective transport of a small fluorescent tracer (sodium fluorescein, 376 Da) in the bone LCS of adult male C57BL/6J mice. We demonstrated that cyclic end-compression of the mouse tibia with a moderate loading magnitude (–3 N peak load or 400 µɛ surface strain at 0.5 Hz) and a 4-second rest/imaging window inserted between adjacent load cycles significantly enhanced (+31%) the transport of sodium fluorescein through the LCS compared with diffusion alone. Using an anatomically based three-compartment transport model, the peak canalicular fluid velocity in the loaded bone was predicted (60 µm/s), and the resulting peak shear stress at the osteocyte process membrane was estimated (∼5 Pa). This study convincingly demonstrated the presence of load-induced convection in mechanically loaded bone. The combined experimental and mathematical approach presented herein represents an important advance in quantifying the microfluidic environment experienced by osteocytes in situ and provides a foundation for further studying the mechanisms by which mechanical stimulation modulates osteocytic cellular responses, which will inform basic bone biology, clinical understanding of osteoporosis and bone loss, and the rational engineering of their treatments. © 2011 American Society for Bone and Mineral Research.