Lacunar-canalicular System Research Articles

Progress in the Effect of Mass Transfer in the Lacunar-Canalicular System on Aging Osteoporosis

Progress in the Effect of Mass Transfer in the Lacunar-Canalicular System on Aging Osteoporosis

Aging decreases osteocyte peri-lacunar-canalicular system turnover in female C57BL/6JN mice

Osteocytes engage in bone resorption and mineralization surrounding their expansive lacunar-canalicular system (LCS) through peri-LCS turnover. However, fundamental questions persist about where, when, and how often osteocytes engage in peri-LCS turnover and how these processes change with aging. Furthermore, whether peri-LCS turnover is associated with natural variation in cortical tissue strain remains unexplored. To address these questions, we utilized confocal scanning microscopy, immunohistochemistry, and scanning electron microscopy to characterize osteocyte peri-LCS turnover in the cortical (mid-diaphysis) and cancellous (metaphysis) regions of femurs from young adult (5 mo) and early-old-age (22 mo) female C57BL/6JN mice. LCS bone mineralization was measured by the presence of perilacunar fluorochrome labels. LCS bone resorption was measured by immunohistochemical marker of bone resorption. The dynamics of peri-LCS turnover were estimated from serial fluorochrome labeling, where each mouse was administered two labels between 2 and 16 days before euthanasia. Osteocyte participation in mineralizing their surroundings is highly abundant in both cortical and cancellous bone of young adult mice but significantly decreases with aging. LCS bone resorption also decreases with aging. Aging has a greater impact on peri-LCS turnover dynamics in cancellous bone than in cortical bone. Lacunae with recent peri-LCS turnover are larger in both age groups. While peri-LCS turnover is associated with variation in tissue strain between cortical quadrants and intracortical location for 22 mo mice, these associations were not seen for 5 mo mice. The impact of aging on decreasing peri-LCS turnover may have significant implications for bone quality and mechanosensation.

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.

Impact of gravity on fluid flow and solute transport in the bone lacunar-canalicular system: a multiscale numerical simulation study

Different gravity fields have important effects on the structural morphology of bone. The fluid flow caused by loadings in the bone lacunar-canalicular system (LCS), converts mechanical signals into biological signals and regulates bone reconstruction by affecting effector cells, which ensures the efficient transport of signaling molecules, nutrients, and waste products. In this study, the fluid flow and mass transfer effects of bone lacunar-canalicular system at multi-scale were firstly investigated, and a three-dimensional axisymmetric fluid-solid coupled finite element model of the LCS within three continuous osteocytes was established. The changes in fluid pressure field, flow velocity field, and fluid shear force variation on the surface of osteocytes within the LCS were studied comparatively under different gravitational fields (0 G, 1 G, 5 G), frequencies (1 Hz, 1.5 Hz, 2 Hz) and forms of cyclic compressive loading. The results showed that different frequencies represented different exercise intensities, suggesting that high-intensity exercise may accelerate the fluid flow rate within the LCS and enhance osteocytes activity. Hypergravity enhanced the transport of solute molecules, nutrients, and signaling molecules within the LCS. Conversely, the mass transfer in the LCS may be inhibited under microgravity, which may cause bone loss and eventually lead to the onset of osteoporosis. This investigation provides theoretical guidance for rehabilitative training against osteoporosis.

Numerical simulations of fluid flow in trabecular–lacunar cavities under cyclic loading

BackgroundUnder external loading, the fluid shear stress (FSS) in the porous structures of bones, such as trabecular or lacunar–canalicular cavity, can influence the biological response of bone cells. However, few studies have considered both cavities. The present study investigated the characteristics of fluid flow at different scales in cancellous bone in rat femurs, as well as the effects of osteoporosis and loading frequency. MethodsSprague Dawley rats (3 months old) were divided into normal and osteoporotic groups. A multiscale 3D fluid–solid coupling finite element model considering trabecular system and lacunar–canalicular system was established. Cyclic displacement loadings with frequencies of 1, 2, and 4 Hz were applied. FindingsResults showed that the wall FSS around the adhesion complexes of osteocyte on the canaliculi was higher than that on the osteocyte body. Under the same loading conditions, the wall FSS of the osteoporotic group was smaller than that of the normal group. The fluid velocity and FSS in trabecular pores exhibited a linear relationship with loading frequency. Similarly, the FSS around osteocytes also showed the loading frequency-dependent phenomenon. InterpretationThe high cadence in movement can effectively increase the FSS level on osteocytes for osteoporotic bone, i.e., expand the space within the bone with physiological load. This study might help in understanding the process of bone remodeling under cyclic loading and provide the fundamental data for the development of strategies for osteoporosis treatment.

Regulation of the Osteocyte Secretome with Aging and Disease.

As the most numerous and long-lived of all bone cells, osteocytes have essential functions in regulating skeletal health. Through the lacunar-canalicular system, secreted proteins from osteocytes can reach cells throughout the bone. Furthermore, the intimate connectivity between the lacunar-canalicular system and the bone vasculature allows for the transport of osteocyte-secreted factors into the circulation to reach the entire body. Local and endocrine osteocyte signaling regulates physiological processes such as bone remodeling, bone mechanoadaptation, and mineral homeostasis. However, these processes are disrupted by impaired osteocyte function induced by aging and disease. Dysfunctional osteocyte signaling is now associated with the pathogenesis of many disorders, including chronic kidney disease, cancer, diabetes mellitus, and periodontitis. In this review, we focus on the targeting of bone and extraskeletal tissues by the osteocyte secretome. In particular, we highlight the secreted osteocyte proteins, which are known to be dysregulated during aging and disease, and their roles during disease progression. We also discuss how therapeutic or genetic targeting of osteocyte-secreted proteins can improve both skeletal and systemic health.

Numerical simulation on mass transfer in the bone lacunar-canalicular system under different gravity fields

The bone lacunar-canalicular system (LCS) is a unique complex 3D microscopic tubular network structure within the osteon that contains interstitial fluid flow to ensure the efficient transport of signaling molecules, nutrients, and wastes to guarantee the normal physiological activities of bone tissue. The mass transfer laws in the LCS under microgravity and hypergravity are still unclear. In this paper, a multi-scale 3D osteon model was established to mimic the cortical osteon, and a finite element method was used to numerically analyze the mass transfer in the LCS under hypergravity, normal gravity and microgravity and combined with high-intensity exercise conditions. It was shown that hypergravity promoted mass transfer in the LCS to the deep lacunae, and the number of particles in lacunae increased more significantly from normal gravity to hypergravity the further away from the Haversian canal. The microgravity environment inhibited particles transport in the LCS to deep lacunae. Under normal gravity and microgravity, the number of particles in lacunae increased greatly when doing high-intensity exercise compared to stationary standing. This paper presents the first simulation of mass transfer within the LCS with different gravity fields combined with high-intensity exercise using the finite element method. The research suggested that hypergravity can greatly promote mass transfer in the LCS to deep lacunae, and microgravity strongly inhibited this mass transfer; high-intensity exercise increased the mass transfer rate in the LCS. This study provided a new strategy to combat and treat microgravity-induced osteoporosis.

The effect of different gravity fields on mass transfer in the rat bone lacunar-canalicular system

The bone lacunar-canalicular system (LCS) is an important microscopic infrastructure for signaling and solute transport in bone tissue, which guarantees normal physiological processes of the tissue, but the mass transfer laws in the LCS under different gravity fields have not yet been clarified. SD rats were injected intraperitoneally with different concentrations of sodium fluorescein tracer and subjected to mass transfer experiments in the LCS under normal gravity and hypergravity. The fluorescence distribution in the osteon was observed using laser scanning confocal microscopy. The hypergravity environment was provided by a self-designed high-G loading centrifuge. The fluorescence intensity of the Haversian canal in the osteon was the highest. The fluorescence intensity of lacunae farther away from the Haversian canal was lower, and the fitted curve was parabolic. With the increasing distance from the Haversian canal, the curve first rapidly decreased, and then the decreasing trend gradually became slower. Hypergravity promoted mass transfer in the LCS, and the 10 ​G hypergravity showed varying degrees of fluorescence intensity in each layer of the osteon relative to normal gravity, with intensity enhancements in the range of 132.7–249.0%. The fluorescence intensity was also significantly increased when the tracer concentration was halved and the gravity field magnitude was increased to 10G. In conclusion, hypergravity promoted the transport of solute molecules, nutrients, and signaling molecules within the LCS. The effect of hypergravity on mass transfer in the LCS was greater than that of tracer concentration. This may help to understand bone diseases from a mass transfer perspective.

Osteocyte Remodeling of the Lacunar-Canalicular System: What's in a Name?

Osteocytes directly modify the bone surrounding the expansive lacunar-canalicular system (LCS) through both resorption and deposition. The existence of this phenomenon is now widely accepted, but is referred to as "osteocyte osteolysis," "LCS remodeling," and "perilacunar remodeling," among other names. The uncertainty in naming this physiological process reflects the many persistent questions about why and how osteocytes interact with local bone matrix. The goal of this review is to examine the purpose and nature of LCS remodeling and its impacts on multiscale bone quality. While LCS remodeling is clearly important for systemic calcium mobilization, this process may have additional potential drivers and may impact the ability of bone to resist fracture. There is abundant evidence that the osteocyte can resorb and replace bone mineral and does so outside of extreme challenges to mineral homeostasis. The impacts of the osteocyte on organic matrix are less certain, especially regarding whether osteocytes produce osteoid. Though multiple lines of evidence point towards osteocyte production of organic matrix, definitive work is needed. Recent high-resolution imaging studies demonstrate that LCS remodeling influences local material properties. The role of LCS remodeling in the maintenance and deterioration of bone matrix quality in aging and disease are active areas of research. In this review, we highlight current progress in understanding why and how the osteocyte removes and replaces bone tissue and the consequences of these activities to bone quality. We posit that answering these questions is essential for evaluating whether, how, when, and why LCS remodeling may be manipulated for therapeutic benefit in managing bone fragility.

Manipulating load-induced fluid flow in vivo to promote bone adaptation.

Mechanical stimulation is critical to maintaining bone mass and strength. Strain has been commonly thought of as the mechanical stimulus driving bone adaptation. However, numerous studies have hypothesized that fluid flow in the lacunar-canalicular system plays a role in mechanoadaptation. The role of fluid flow compared to strain magnitude on bone remodeling has yet to be characterized. This study aimed to determine the contribution of fluid flow velocity compared to strain on bone adaptation. We used finite element modeling to design in vivo experiments, manipulating strain and fluid flow contributions. Using a uniaxial compression tibia model in mice, we demonstrated that high fluid flow velocity results in significant bone adaptation even under low strain magnitude. In contrast, high strain magnitude paired with low fluid velocity does not trigger a bone response. These findings support previous hypotheses stating that fluid flow is the principal mechanical stimulus driving bone adaptation. Moreover, they give new insights regarding bone adaptative response and provide new pathways toward treatment against age-related mechanosensitivity loss in bone.

Biomechanical analysis of a lacunar-canalicular system under different cyclic displacement loading

The objective of this study is to use the finite element (FE) method to predict the mechanical signals (interstitial fluid velocity, strain, pore pressure, and pore fluid velocity) produced by osteocyte during physiological activities. The model predicts that the amplitude and distribution of the mechanical signals are mainly affected by the loading rate. The magnitude of mechanical signals in the lacunar-canalicular system increases as the amplitude, frequency and amount of direction of load increase. Collagen hillocks can effectively amplify strain signals at the process. The established model can be used for studying the mechanism of bone mechanotransduction at the micro-level.

Study on mass transfer in the bone lacunar-canalicular system under different gravity fields.

The bone lacunar-canalicular system (LCS) is an important microstructural basis for signaling and material transport in bone tissue, guaranteeing normal physiological processes in tissues. Spaceflight astronauts and elderly osteoporosis are related to its function, so it is necessary to reveal the mass transfer laws in bone microstructure under different gravity fields to provide insight for effective clinical treatment. Using the natural LCS structure of bovine tibial cortical bone as the object, the mass transfer experiments on cortical bone were conducted by using sodium fluorescein tracer through different frequency pulsating pressure provided by dynamic perfusion loading device and different high G environments provided by high-speed centrifuge to analyze the mass transfer laws under different gravity fields and different pulsating pressures. The fluorescence intensity of lacunae within the osteon was lower the farther away from the Haversian canal. As the gravity field magnitude increased, the fluorescence intensity within each lacuna enhanced, and the more distant the lacunae from the Haversian canal, the greater the fluorescence intensity enhancement. High-frequency pulsating pressure simulated high-intensity exercise in humans can improve mass transfer efficiency in the LCS. High-intensity exercise may greatly increase solute molecules, nutrients, and signaling molecules in osteocytes and improve the activity of osteocytes. Hypergravity can enhance the transport of solute molecules, nutrients, and signaling molecules in the LCS, especially promoting mass transfer to deep layer lacunae. Conversely, mass transfer to deep layer lacunae may be inhibited under microgravity, causing bone loss and ultimately leading to osteoporosis.

Expansion of the osteocytic lacunar-canalicular system involved in pharmacological action of PTH revealed by AI-driven fluorescence morphometry in female rabbits

Osteoporosis is an age-related disorder that is characterized by reduced bone mass. Its prevention and treatment are important healthcare issues for maintaining social activity in aged societies. Although bone fractures mostly occur at sites of weakened cortical bone, pathophysiological and pharmacological evaluations of bone mass have tended to be predominantly assessed in trabecular bone. To statistically characterize cortical bone remodeling, we originally established multimode fluorescence imaging and artificial intelligence (AI)-driven morphometric analyses in six-month-old female rabbits with well-defined cortical remodeling, similar to that in humans. We evaluated three distinct administration frequencies of teriparatide [TPTD; human parathyroid hormone, hPTH (1–34)]: once (1/w), twice (2/w), and seven times (7/w) a week, with the same total dose (140 μg/kg/week). Our analyses revealed significant expansions of the osteocytic lacunar-canalicular system and Haversian canals accompanied by the development of cortical porosity and endosteal naïve bone formation induced by a frequent administration regimen (7/w) of TPTD; however, once-weekly (1/w) and twice-weekly (2/w) administration of TPTD showed little effect. These findings demonstrate a clear contrast between the effects of frequent and infrequent administration of TPTD on cortical bone metabolism and suggest that osteocytic bone remodeling is involved in the pharmacological action of PTH.

A fluid flow model in the lacunar-canalicular system under the pressure gradient and electrical field driven loads

A fluid flow model in the lacunar-canalicular system under the pressure gradient and electrical field driven loads

Interactive effects of various loading parameters on the fluid dynamics within the lacunar-canalicular system for a single osteocyte.

The osteocyte lacunar-canalicular system (LCS) serves as a mechanotransductive core where external loading applied to the skeleton is transduced into mechanical signals (e.g., fluid shear) that can be sensed by mechanosensors (osteocytes). The fluid velocity and shear stress within the LCS are affected by various loading parameters. However, the interactive effect of distinct loading parameters on the velocity and shear stress in the LCS remains unclear. To address this issue, we developed a multiscale modeling approach, combining a poroelastic finite element (FE) model with a single osteocytic LCS unit model to calculate the flow velocity and shear stress within the LCS. Next, a sensitivity analysis was performed to investigate individual and interactive effects of strain magnitude, strain rate, number of cycles, and intervening short rests between loading cycles on the velocity and shear stress around the osteocyte. Lastly, we developed a relatively simple regression model to predict those outcomes. Our results demonstrated that the strain magnitude or rate alone were the main factors affecting the velocity and shear stress; however, the combination of these two was not directly additive, and addition of a short rest between cycles could enhance the combination of these two related factors. These results show highly interactive effects of distinct loading parameters on fluid velocity and shear stress in the LCS. Specifically, our results suggest that an enhanced fluid dynamics environment in the LCS can be achieved with a brief number of load cycles combined with short rest insertion and high strain magnitude and rate.

Osteocyte β1 integrin loss causes low bone mass and impairs bone mechanotransduction in mice.

BackgroundThe key focal adhesion protein β1 integrin plays an essential role in early skeletal development. However, roles of β1 integrin expression in osteocytes during the regulation of bone homeostasis and mechanotransduction are incompletely understood.Materials and methodsTo study the in vivo function of osteocyte β1 integrin in bone, we utilized the 10-kb Dmp1 (Dentin matrix acidic phosphoprotein 1)-Cre to generate mice with β1 integrin deletion in this cell type. Micro-computerized tomography, bone histomorphometry and immunohistochemistry were performed to determine the effects of osteocyte β1 integrin loss on bone mass accrual and biomechanical properties. In vivo tibial loading model was applied to study the possible involvement of osteocyte β1 integrin in bone mechanotransduction.ResultsLoss of β1 integrin expression in osteocytes resulted in a severe low bone mass and impaired biomechanical properties in load-bearing long bones and spines, but not in non-weight-bearing calvariae, in mice. The loss of β1 integrin led to enlarged size of lacunar-canalicular system, abnormal cell morphology, and disorientated nuclei in osteocytes. Furthermore, β1 integrin loss caused shortening and disorientated collagen I fibers in long bones. Osteocyte β1 integrin loss did not impact the osteoclast activities, but significantly reduced the osteoblast bone formation rate and, in the meantime, enhanced the adipogenic differentiation of the bone marrow stromal cells in the bone microenvironment. In addition, tibial loading failed to accelerate the anabolic bone formation and improve collagen I fiber integrity in mutant mice.ConclusionsOur studies demonstrate an essential role of osteocyte β1 integrin in regulating bone homeostasis and mechanotransduction.The transnational potential of this article: This study reveals the regulatory roles of osteocyte β1 integrin in vivo for the maintenance of bone mass accrual, biomechanical properties, extracellular matrix integrity as well as bone mechanobiology, which defines β1 integrin a potential therapeutic target for skeletal diseases, such as osteoporosis.

Perilacunar bone tissue exhibits sub-micrometer modulus gradation which depends on the recency of osteocyte bone formation in both young adult and early-old-age female C57Bl/6 mice.

Osteocytes resorb and replace bone local to the lacunar-canalicular system (LCS). However, whether osteocyte remodeling impacts bone quality adjacent to the LCS is not understood. Further, while aging is well-established to decrease osteocyte viability and truncate LCS geometry, it is unclear if aging also decreases perilacunar bone quality. To address these questions, we employed atomic force microscopy (AFM) to generate nanoscale-resolution modulus maps for cortical femur osteocyte lacunae from young (5-month) and early-old-age (22-month) female C57Bl/6 mice. AFM-mapped lacunae were also imaged with confocal laser scanning microscopy to determine which osteocytes recently deposited bone as determined by the presence of fluorochrome labels administered 2d and 8d before euthanasia. Modulus gradation with distance from the lacunar wall was compared for labeled (i.e., bone forming) and non-labeled lacunae in both young and aged mice. All mapped lacunae showed sub-microscale modulus gradation, with peak modulus values 200-400nm from the lacunar wall. Perilacunar modulus gradations depended on the recency of osteocyte bone formation (i.e., the presence of labels). For both ages, 2d-labeled perilacunar bone had lower peak and bulk modulus compared to non-labeled perilacunar bone. Lacunar length reduced with age, but lacunar shape and size were not strong predictors of modulus gradation. Our findings demonstrate for the first time that osteocyte perilacunar remodeling impacts bone tissue modulus, one contributor to bone quality. Given the immense scale of the LCS, differences in perilacunar modulus resulting from osteocyte remodeling activity may affect the quality of a substantial amount of bone tissue.

Simulation study on the effect of resistance exercise on the hydrodynamic microenvironment of osteocytes in microgravity

Osteoporosis occurs in astronauts after long-term space flight owing to the lack of gravity. The mechanical microenvironment of osteocytes in load-bearing bone are changed during resistance exercise, which prevents massive bone loss in the human body. A cylindrical fluid-structure coupling finite element model for osteons with a two-stage pore structure (i.e., Haversian canal, lacunar-canalicular system) was established with the software COMSOL. In the Earth’s gravity field and in microgravity, considering the effects of pulsating pressure of arterioles, a comparative study was performed on the changes in hydrodynamic microenvironment of osteocytes during human body high-intensity exercise at different frequencies (defined as causing bone to produce 3000 με) and the body is at rest. Positive and negative liquid pressure (with respect to one atmosphere pressure) alternately acted on osteocytes during human exercising, but only positive pressure acted on osteocytes during human resting. The variation range of liquid pressure acted on osteocytes during human exercising was significantly higher than that during resting. The liquid flow velocity around osteocytes during body exercise was about four orders of magnitude higher than that during resting. In microgravity, moderate physical exercise can obviously improve the hydrodynamic microenvironment of osteocytes in load-bearing bone, which could compensate for the lack of mechanical stimulation to osteocytes caused by the lack of gravity, thereby promoting the normal physiological function of osteocytes. To a certain extent, these results revealed the biomechanical mechanism by which exercise has an effect in fighting osteoporosis in astronauts.

A Potential Participant in Type 2 Diabetes Bone Fragility: TIMP-1 at Sites of Osteocyte Lacunar-Canalicular System.

Type 2 diabetes mellitus (T2DM) is associated with an increased risk of bone fracture, but the bone mineral density (BMD) is typically normal or higher in such patients. Because the fracture risk is independent of reduced BMD, bone fragility in T2DM may be partially due to poor bone quality. The mechanisms triggering bone quality abnormalities in T2DM are complex, and include the accumulation of advanced glycation end-products, the increased inflammation, and low bone turnover. Matrix metalloproteinases (MMPs) in bone can hydrolyze the bone matrix. Tissue inhibitors of MMPs (TIMPs) can inhibit the activity of MMPs. Both MMPs and TIMPs participate in mediating bone quality. Among all types of TIMPs, TIMP-1 is mostly reportedly increased in the serum of T2DM patients. Because osteocytes can express TIMP-1, and osteocyte pericellular matrix influences bone quality partially regulated by perilacunar/canalicular remodeling, we hypothesized that TIMP-1 at sites of osteocyte lacunar-canalicular system is involved in T2DM bone fragility.

Tunnels in the rock: Dynamics of osteocyte morphogenesis.

Osteocytes are dynamic, bone matrix-remodeling cells that form an intricate network of interconnected projections through the bone matrix, called the lacunar-canalicular system. Osteocytes are the dominant mechanosensory cells in bone and their mechanosensory and mechanotransductive functions follow their morphological form. During osteocytogenesis and development of the osteocyte lacunar-canalicular network, osteocytes must dramatically remodel both their cytoskeleton and their extracellular matrix. In this review, we summarize our current understanding of the mechanisms that govern osteocyte differentiation, cytoskeletal morphogenesis, mechanotransduction, and matrix remodeling. We postulate that the physiologic activation of matrix remodeling in adult osteocytes, known as perilacunar/canalicular remodeling (PLR) represents a re-activation of the developmental program by which the osteocyte network is first established. While much of osteocyte biology remains unclear, new tools and approaches make the present moment a particularly fruitful and exciting time to study the development of these remarkable cells.