Discretizing Low-Intensity Whole-Body Vibration Into Bouts With Short Rest Intervals Promotes Bone Defect Repair in Osteoporotic Mice

Continuous administration of low-intensity whole-body vibration (WBV) gradually diminishes bone mechanosensitivity over time, leading to a weakening of its osteogenic effect. We investigated whether discretizing WBV into bouts with short rest intervals was effective in enhancing osteoporotic bone repair. Ten-week-old female mice were ovariectomized and underwent drill-hole defect surgery (Day 0) on the right tibial diaphysis at 11 weeks of age. The mice underwent one of three regimens starting from Day 1 for 5 days/week: continuous WBV at 45 Hz and 0.3 g for 7.5 min/day (cWBV); 3-s bouts of WBV at 45 Hz, 0.3 g followed by 9-s rest intervals, repeated for 30 min/day (repeated bouts of whole-body vibration with short rest intervals [rWBV]); or a sham treatment. Both the cWBV and rWBV groups received a total of 20,250 vibration cycles per day. On either Day 7 or 14 posteuthanasia (n = 6/group/timepoint), the bone and angiogenic vasculature in the defect were computed tomography imaged using synchrotron light. By Day 14, the bone repair was most advanced in the rWBV group, showing a higher bone volume fraction and a more uniform mineral distribution compared with the sham group. The cWBV group exhibited an intermediate level of bone repair between the sham and rWBV groups. The rWBV group had a decrease in large-sized angiogenic vessels, while the cWBV group showed an increase in such vessels. In conclusion, osteoporotic bone repair was enhanced by WBV bouts with short rest intervals, which may potentially be attributed to the improved mechanosensitivity of osteogenic cells and alterations in angiogenic vasculature.


In-Situ Deformation Imaging of Articular Cartilage Using Grating-Based Phase-Contrast X-ray CT at a Synchrotron Light Source

Quantification of deformation behavior of articular cartilage (AC) is crucial for understanding its mechanical function and response to mechanical stimuli. Here, we explored whether grating-based phase-contrast X-ray CT using monochromatic synchrotron light (grating-based msPXCT) enables in-situ quantification of local deformation in AC. Grating-based msPXCT of a porcine AC sample during axial compression test was conducted using a Talbotgrating interferometer. Local displacements and strains were computed using a digital volume correlation method. The magnitude of axial strain decreased from the upper to middle sample zones and reached almost constant over the middle-lower zone, consistent with the depth-dependent density increase with compression. Thus, grating-based msPXCT may be suitable for quantitative analysis of AC deformation.


Effects of Whole-Body Vibration on Breast Cancer Bone Metastasis and Vascularization in Mice

We evaluated whether whole-body vibration (WBV) prevented bone loss induced by breast cancer (BC) metastasis and the involvement of bone marrow vasculature. One day after orthotopic transplantation of mammary 4T1 tumor cells, 8-week-old BALB/c mice were subjected to 0.3 g/90 Hz vertical vibration for 20 min/day for 5 days/week (BC-WBV) or sham-handled (BC-Sham) over 3 weeks. Age-matched intact mice (Intact) were also sham-handled. Both tibiae were harvested from BC-WBV (n=7), BC-Sham (n=9), and Intact (n=5) mice for bone structure imaging by synchrotron radiation-based computed tomography (SRCT) and hematoxylin and eosin staining, whereas right tibiae were harvested from other BC-WBV and BC-Sham (n=6 each) mice for vascular imaging by SRCT. Tumor cells were similarly widespread in the marrow in BC-WBV and BC-Sham mice. In BC-Sham mice, cortical bone volume, trabecular volume fraction, trabecular thickness, trabecular number density, and bone mineral density were smaller, and marrow volume and trabecular separation were larger than in Intact mice. However, although trabecular thickness was smaller in BC-WBV than Intact mice, the others did not differ between the two groups. Serum osteocalcin tended to be higher in BC-WBV than BC-Sham mice. Compared with BC-Sham mice, BC-WBV mice had a smaller vessel diameter, a trend of a larger vessel number density, and smaller vessel diameter heterogeneity. In conclusion, WBV mitigates bone loss in BC bone metastasis, which may be partly due to increased bone anabolism. The alteration of marrow vasculature appears to be favorable for anti-tumor drug delivery. Further studies are needed to clarify the multiple actions of WBV on bone, tumor, and marrow vasculature and how they contribute to bone protection in BC metastasis.


In Vivo Monitoring of Bone Microstructure by Propagation-Based Phase-Contrast Computed Tomography Using Monochromatic Synchrotron Light

Hard X-ray phase-contrast imaging is sensitive to density variation in objects and shows a dose advantage for in vivo observation over absorption-contrast imaging. We examined the capability of propagation-based phase-contrast tomography (PB-PCT) with single-distance phase retrieval for tracking of bone structure and mineral changes using monochromatic synchrotron light. Female mice underwent ovariectomy and drill-hole surgery in the right tibial diaphysis and were divided into two groups: OVX and OVX-E (n = 6 each); the latter group was treated with intraperitoneal administration of 14,15-epoxyeicosatrienoic acid (14,15-EET) for promoting bone repair. Age-matched mice subjected to sham ovariectomy and drill-hole surgery (Sham) were also prepared (n = 6). In vivo CT scans of the drilled defect were acquired 3, 7, and 11 days after surgery, and tomographic images were matched by three-dimensional registration between successive time points for monitoring the process of defect filling. In addition, using absorption-contrast CT as the reference method, the validity of PB-PCT was evaluated in one mouse by comparing images of tibial metaphyseal bone between the two methods in terms of bone geometry as well as the measure of mineralization. Although phase retrieval is strictly valid only for single-material objects, PB-PCT, with its lower radiation dose, could provide a depiction of bone structure similar to that from absorption-contrast CT. There was a significant correlation of linear absorption coefficients between the two methods, indicating the possibility of a rough estimate of the measure of mineralization by PB-PCT. Indeed, delayed bone regeneration (OVX vs. Sham) and the efficacy of 14,15-EET for improving osteoporotic bone repair (OVX-E vs. OVX) could be detected in both bone volume and mineralization by PB-PCT. Thus, in combination with single-distance phase retrieval, PB-PCT would have great potential for providing a valuable tool to track changes in bone structure and mineralization, and for evaluating the effects of therapeutic interventions as well.

Laboratory Investigation; a Journal of Technical Methods and Pathology


The Disuse Effect on Canal Network Structure and Oxygen Supply in the Cortical Bones of Rats

In this study, based on the measurements of intracortical vascular canal structure, we investigated the disuse effect on local O supply in the cortical bones of growing rats. Hindlimb disuse was produced by unilateral sciatic neurectomy (SN) at 4 weeks age. The canal network structures within tibial cortical bone were evaluated in 8- and 12-week-old rats undergoing SN or no treatment (control) by synchrotron radiation micro-CT. Additionally, we developed an intracortical network model by combining the imaged-based canal network with a bone matrix containing theoretical lacunar-canalicular network, and determined the distribution of O concentration in bone tissue numerically. In the control bone, canal network was reduced with growth, resulting in decreased blood flow and averaged O concentration and increased spatial heterogeneity in tissue O concentration. Disuse reduced the canal network, leading to a lower flow rate, lower average O concentration and higher heterogeneity of O concentrations. However, the rarefaction of the canal network with growth was smaller under the disuse condition, and accordingly, the flow rate, the average O concentration and the heterogeneity of O concentrations remained stable. In particular, although the fraction of the canal volume was smaller, the densities of canal segments and bifurcation points under disuse condition tended to be higher than those of the control bone. The heterogeneity of O concentration was lower. Our results indicated that the disuse may lead to more uniformity in the canal network structure and thereby uniform O, possibly contributing to O supply efficiency.

Biomechanics and Modeling in Mechanobiology


Strain Measurements of the Tibial Insert of a Knee Prosthesis Using a Knee Motion Simulator

The longevity of a knee prosthesis is influenced by the wear of the tibial insert due to its posture and movement. In this study, we assumed that the strain on the tibial insert is one of the main reasons for its wear and investigated the influence of the knee varus-valgus angles on the mechanical stress of the tibial insert. Knee prosthesis motion was simulated using a knee motion simulator based on a parallel-link six degrees-of-freedom actuator and the principal strain and pressure distribution of the tibial insert were measured. In particular, the early stance phase obtained from in vivo X-ray images was examined because the knee is applied to the largest load during extension/flexion movement. The knee varus-valgus angles were 0° (neutral alignment), 3°, and 5° malalignment. Under a neutral orientation, the pressure was higher at the middle and posterior condyles. The first and second principal strains were larger at the high and low pressure areas, respectively. Even for a 3° malalignment, the load was concentrated at one condyle and the positive first principal strain increased dramatically at the high pressure area. The negative second principal strain was large at the low pressure area on the other condyle. The maximum equivalent strain was 1.3-2.1 times larger at the high pressure area. For a 5° malalignment, the maximum equivalent strain increased slightly. These strain and pressure measurements can provide the mechanical stress of the tibial insert in detail for determining the longevity of an artificial knee joint.


Effect of Low-Intensity Whole-Body Vibration on Bone Defect Repair and Associated Vascularization in Mice

Medical and Biological Engineering and Computing


Pulmonary Kinematic Analysis With Non-Rigid Deformable Registration for Detecting Localised Emphysema

Computer Methods in Biomechanics and Biomedical Engineering. Imaging & Visualization


Distribution of Nanoparticle Depositions after a Single Breathing in a Murine Pulmonary Acinus Model

International Journal of Heat and Mass Transfer


Effect of Concurrent Use of Whole-Body Vibration and Parathyroid Hormone on Bone Structure and Material Properties of Ovariectomized Mice

This study was designed to determine the effectiveness of whole-body vibration (WBV) and intermittent parathyroid hormone (iPTH) in combination against estrogen deficiency-induced osteoporosis. Female C57BL/6J mice were bilaterally ovariectomized (OVX, n = 40) or sham-operated (sham-OVX, n = 8) at 9 weeks of age. Two weeks later, the OVX mice were randomly divided into four groups (n = 10 each): the control group (c-OVX) and groups treated with iPTH (p-OVX), WBV (w-OVX) and both (pw-OVX). The p-OVX and pw-OVX groups were given human PTH (1-34) at a dose of 30 µg/kg/day. The w-OVX and pw-OVX groups were exposed to WBV at an acceleration of 0.3 g and 45 Hz for 20 min/day. All mice were euthanized after the 18-day treatment, and the left tibiae were harvested. The proximal metaphyseal region was µCT-scanned, and its cortical bone cross-section was analyzed by Fourier transform infrared microspectroscopy and nanoindentation testing. A single application of iPTH or WBV to OVX mice had no effect on bone structure or material properties of cortical bone, which were compromised in comparison to those in sham-OVX mice. The combination of iPTH and WBV improved trabecular bone volume, thickness, and connectivity in OVX mice. Although the combined treatment failed to improve cortical bone structure, its mineral maturity and hardness were restored to the levels observed in sham-OVX mice. There was no evidence of interaction between the two treatments, and the combined effects seemed to be additive. These results suggest combining WBV with iPTH has great potential for treating postmenopausal osteoporosis.


Individual and Combined Effects of Noise-Like Whole Body Vibration and Parathyroid Hormone Treatment on Bone Defect Repair in Ovariectomized Mice

The effectiveness of intermittent administration of parathyroid hormone and exposure to whole-body vibration on osteoporotic fracture healing has been previously investigated, but data on their concurrent use are lacking. Thus, we evaluated the effects of intermittent administration of parathyroid hormone, whole-body vibration, and their combination on bone repair in osteoporotic mice. Noise-like whole-body vibration with a broad frequency range was used instead of conventional sine-wave whole-body vibration at a specific frequency. Mice were ovariectomized at 9 weeks of age and subjected to drill-hole surgery in the right tibial diaphysis at 11 weeks. The animals were divided into four groups (n = 12 each): a control group, and groups treated with intermittent administration of parathyroid hormone, noise-like whole-body vibration, and both. From postoperative day 2, the groups treated with intermittent administration of parathyroid hormone and groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration were subcutaneously administered parathyroid hormone at a dose of 30 µg/kg/day. The groups treated with noise-like whole-body vibration and groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration were exposed to noise-like whole-body vibration at a root mean squared acceleration of 0.3g and frequency components of 45-100 Hz for 20 min/day. Following 18 days of interventions, the right tibiae were harvested, and the regenerated bone was analyzed by micro-computed tomography and nanoindentation testing. Compared with the control group, callus volume fraction was 40% higher in groups treated with intermittent administration of parathyroid hormone and 73% higher in groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration, and callus thickness was 35% wider in groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration. Indentation modulus was 46% higher in groups treated with noise-like whole-body vibration and 43% higher in groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration, and hardness was 31% higher in groups treated with both intermittent administration of parathyroid hormone and noise-like whole-body vibration compared with the control group. There was no interaction between the two treatments for both structure and mechanical indexes. The main effects of intermittent administration of parathyroid hormone and noise-like whole-body vibration on bone repair included increased bone formation and enhanced mechanical function of regenerated bone, respectively. The combined treatment resulted in further regeneration of bone with high indentation modulus and hardness, suggesting the therapeutic potential of the combined use of noise-like whole-body vibration and intermittent administration of parathyroid hormone for enhancing osteoporotic bone healing.

Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine


再生骨の骨質および力学特性に対する全身性微振動および断続的副甲状腺ホルモン(1-34)投与の相乗的作用の解析


Stimulating Angiogenesis Mitigates the Unloading-Induced Reduction in Osteogenesis in Early-Stage Bone Repair in Rats

Accelerating fracture healing during bed rest allows early mobilization and avoids prolonged fracture healing times. We tested the hypothesis that stimulating angiogenesis with deferoxamine (DFO) mitigates the unloading-induced reduction in early-stage bone repair. Rats aged 12 weeks were subjected to cortical drilling on their tibial diaphysis under anesthesia and treated with hindlimb unloading (HU), HU and DFO administration (DFOHU), or weight bearing (WB) for 5 or 10 days (HU5/10, DFOHU5/10, WB5/10; n = 8 per groups) until sacrifice for vascular casting with a zirconium dioxide-based contrast agent. Taking advantage of its absorption discontinuity at the K-absorption edge, vascular and bone images in the drill-hole defects were acquired by synchrotron radiation subtraction CT. Bone repair was reduced in HU rats. The bone volume fraction (B.Vf) was 88% smaller in HU5 and 42% smaller in HU10 than in WB5/10. The bone segment densities (B.Seg) were 97% smaller in HU5 and 141% larger in HU10 than in WB5/10, and bone thickness (B.Th) was 38% smaller in HU10 than in WB10. The vascular volume fraction (V.Vf) was 35% and the mean vessel diameter (V.D) was 13% smaller in HU10 than in WB10. When compared according to categorized vessel sizes, V.Vf in the diameter ranges 20-30, 30-40, and >40 m were smaller in HU10 than in WB10, and V.Seg in the diameter range >40 m was smaller in HU10 than in WB10. In contrast, there was no difference in B.Vf between DFOHU5/10 and WB5/10 and in V.Vf between DFOHU10 and WB10, though B.Seg remained 86% smaller in DFOHU5 and 94% larger in DFOHU10 than in WB5/10, and B.Th and V.D were 23% and 14% lower in DFOHU10 than in WB10. Vessel size-specific V.Vf in the diameter ranges 10-20 and 20-30 m was larger in DFOHU5 than in HU5. In conclusion, the enhanced angiogenic ingrowth mitigates the reduction in bone repair during mechanical unloading.


Second Harmonic Generation Microscopy and Synchrotron Radiation CT for the Determination of Collagen and Mineral Deposition in Early-Stage Bone Repair: Effect of Whole Body Micro-Vibration


Subtraction Micro-Computed Tomography of Angiogenesis and Osteogenesis During Bone Repair Using Synchrotron Radiation With a Novel Contrast Agent

Quantitative three-dimensional (3D) imaging of angiogenesis during bone repair remains an experimental challenge. We developed a novel contrast agent containing 0.07- to 0.1-μm particles of zirconium dioxide (ZrCA) and established subtraction μCT using synchrotron radiation (sSRCT) for quantitative imaging of angiogenesis and bone repair. This method was applied to a rat model of tibial bone repair 3 days (DAY3; n = 2), 5 days (DAY5; n = 8), or 10 days (DAY10; n = 8) after drill-hole injury. Using the same drill-hole defect model, its potential use was illustrated by comparison of bone repair between hindlimbs subjected to mechanical unloading (n = 6) and normal weight bearing (n = 6) for 10 days. Following vascular casting with ZrCA, the defect site was scanned with 17.9- and 18.1-keV X-rays. In the latter, image contrast between ZrCA-filled vasculature and bone was enhanced owing to the sharp absorption jump of zirconium dioxide at 18.0 keV (k-edge). The two scan data sets were reconstructed with 2.74-μm voxel resolution, registered by mutual information, and digitally subtracted to extract the contrast-enhanced vascular image. K2HPO4 phantom solutions were scanned at 17.9 keV for quantitative evaluation of bone mineral. Angiogenesis had already started, but new bone formation was not found on DAY3. New bone emerged near the defect boundary on DAY5 and took the form of trabecular-like structure invaded by microvessels on DAY10. Vascular and bone volume fractions, blood vessel and bone thicknesses, and mineralization were higher on DAY10 than on DAY5. All these parameters were found to be decreased after 10 days of hindlimb unloading, indicating the possible involvement of angiogenesis in bone repair impairment caused by reduced mechanical stimuli. In conclusion, the combined technique of sSRCT and ZrCA vascular casting is suitable for quantitative 3D imaging of angiogenesis and its surrounding bone regeneration. This method will be useful for better understanding the linkage between angiogenesis and bone repair.

Laboratory Investigation; a Journal of Technical Methods and Pathology


Estimation of Change of Bone Structures After Total Hip Replacement Using Bone Remodeling Simulation

The principal cause of femoral stem failure is the loosening of the total hip replacement due to bone resorption in the vicinity of the prosthesis (stress shielding). Bone rebuilds its structure continuously according to the daily mechanical stimuli. Therefore, surgical intervention alters the mechanical condition of bone severely. In this study, we propose a method to predict the change of bone structure after total hip replacement using bone remodeling simulation. The bone-stem complex structure model after total hip replacement was reconstructed based on CT-images used for preoperative planning by orthopedic experts. The bone remodeling simulation was conducted under the daily loading condition using our previous remodeling model, and the average equivalent stresses in the Gruen zone were evaluated. The predicted bone loss relevant to stress shielding was consistent to follow-up clinical data. Moreover, the remodeling simulation when using the stems of different size for the same patient could detect the size-dependent change of stress in the Gruen zone. In particular, the zone under the neck of the stem showed significant changes of stress and large bone loss, accompanying the risk of loosening or fracture. Prediction of bone structure changes after total hip replacement gives us significant information for longevity of prosthesis. Simulation results showed that the present computational framework could be considered to have potential in preoperative planning of total hip replacement.


人工股関節手術計画における力学的評価指標 -メカノスタット理論/リモデリングシミュレーションからの検討-

日本臨床バイオメカニクス会誌

日本臨床バイオメカニクス会誌


Relationship Between Aortic Mineral Elements and Osteodystrophy in Mice With Chronic Kidney Disease

In chronic kidney disease (CKD), osteodystrophy and arterial calcification often coexist. However, arterial alterations have not been addressed in CKD unaccompanied by evidence of calcification. We investigated the association of phosphate (P) and calcium (Ca) accumulation in calcification-free aortas with CKD-induced osteodystrophy. Aortic accumulation of magnesium (Mg), an inhibitor of calcification, was also examined. Male mice aged 26 weeks with CKD characterized by hyperparathyroidism and hyperphosphatemia (Nx, n = 8) and age-matched healthy male mice (shams, n = 8) were sampled for blood, and thoracic vertebrae and aortas were harvested. Bone structure and chemicals were analyzed by microcomputed tomography and infrared microspectroscopy, respectively, and aortic accumulation of P, Ca, and Mg was evaluated by plasma-atomic emission spectrometry. Volume fractions of cortical and trabecular bones were smaller in Nx than in sham animals (P < 0.05), attributed to cortical thinning and reduction in trabecular number, respectively. Bone chemicals were not different between the groups. No calcification was found in either group, but P, Ca, and Mg contents were higher in Nx than in shams (P < 0.05). The mass ratio of Ca/P was lower in Nx than in shams (P < 0.05), but that of Mg/Ca and Mg/P was not different between the groups. Aortic P and Ca contents were inversely correlated with the volume fraction of cortical bone (P < 0.05). In conclusion, the relationship of osteodystrophy with aortic P and Ca accumulation suggests the existence of a bone-vascular axis, even in calcification-free arteries in CKD. The preservation of ratios of Mg/Ca and Mg/P despite CKD development might contribute to calcification resistance.


Identification of Individual Muscle Length Parameters From Measurements of Passive Joint Moment Around the Ankle Joint

Journal of Biomechanical Science and Engineering


Evaluation of Viscoelastic Property of Articular Cartilage Based on Mechanical Model Considering Tissue Microstructure


Effects of Vitamin K on the Morphometrical and Material Properties of Bone in The Tibiae of Growing Rats

Suboptimal vitamin K nutriture is evident during rapid growth. We aimed to determine whether vitamin K(2) (menaquinone-4 [MK-4]) supplementation is beneficial to bone structure and intrinsic bone tissue properties in growing rats. Male Wistar rats (5 weeks old) were assigned to either a control diet (n = 8) or an MK-4-supplemented diet (22 mg d(-1) kg(-1) body weight, n = 8). After a 9-week feeding period, we determined the serum concentration ratio of undercarboxylated osteocalcin to -carboxylated osteocalcin and the urinary deoxypyridinoline level. All rats were then euthanized, and their tibiae were analyzed by micro-computed tomography for trabecular architecture and synchrotron radiation micro-computed tomography for cortical pore structure and mineralization. Fourier transform infrared microspectroscopy and a nanoindentation test were performed on the cortical midlayers of the anterior and posterior cortices to assess bone tissue properties. Neither body weight nor tibia length differed significantly between the 2 groups. Dietary MK-4 supplementation decreased the ratio of undercarboxylated osteocalcin to -carboxylated osteocalcin but did not affect deoxypyridinoline, indicating a positive effect on bone formation but not bone resorption. Trabecular volume fraction and thickness were increased by MK-4 (P < .05). Neither the cortical pore structure nor mineralization was affected by MK-4. On the other hand, MK-4 increased mineral crystallinity, collagen maturity, and hardness in both the anterior and posterior cortices (P < .05). These data indicate the potential benefit of MK-4 supplementation during growth in terms of enhancing bone quality.