Cancellous Bone Microstructure Research Articles

Passive bicycle training stimulates epiphyseal bone formation and restores bone integrity independent of locomotor recovery in a rat spinal cord injury model.

It is unknown whether activity-based physical therapy (ABPT) modalities that mobilize the paralyzed limbs improve bone integrity at the highly fracture-prone epiphyseal regions of the distal femur and proximal tibia following severe spinal cord injury (SCI). In this study, 4-mo-old skeletally mature littermate-matched male Sprague-Dawley rats received either SHAM surgery or severe contusion SCI. At 1 wk postsurgery, SCI rats were stratified to undergo no-ABPT, two 20-min bouts/day of quadrupedal bodyweight-supported treadmill training (qBWSTT), or hindlimb passive isokinetic bicycle (cycle) training, 5 days/wk for another 3 wk. We assessed locomotor recovery and plantar flexor muscle mass, tracked cancellous and cortical bone microstructure at the distal femoral and proximal tibial epiphyses using in vivo microcomputed tomography (microCT), and evaluated bone turnover at the tibial epiphysis with histomorphometry. All SCI animals displayed persistent hindlimb paralysis and pervasive muscle atrophy. Over the initial 2 wk, which included 1 wk of no exercise and 1 wk of ABPT acclimation, a similar magnitude of bone loss developed in all SCI groups. Thereafter, cancellous bone loss and cortical bone decrements increased in the SCI no-ABPT group. qBWSTT attenuated this trabecular bone loss but did not prevent the ongoing cortical bone deficits. In comparison, twice-daily cycle training increased the number and activity of osteoblasts versus other SCI groups and restored all bone microstructural parameters to SHAM levels at both epiphyseal sites. These data indicate that a novel passive isokinetic cycle training regimen reversed cancellous and cortical bone deterioration at key epiphyseal sites after experimental SCI via osteoblast-mediated bone anabolic mechanisms, independent of locomotor recovery or increased muscle mass.NEW & NOTEWORTHY This study was the first to assess how quadrupedal bodyweight-supported treadmill training or passive isokinetic bicycle (cycle) training impacts bone recovery at the distal femoral and proximal tibial epiphyses in a rat model of severe contusion spinal cord injury. Our results demonstrate that passive isokinetic cycle training completely restored cancellous and cortical bone microstructural parameters at these sites via osteoblast-mediated bone anabolic actions, independent of locomotor recovery or increased plantar flexor muscle mass.

A numerical simulation method for ultrasonic transmission in cancellous bone based on a four-parameter random growth method

Abstract Quantitative ultrasound (QUS) has been widely used in non-destructive evaluation of bone health in research and clinical practice. To make a more accurate bone evaluation, the transmission characteristics of ultrasound in the bone need to be understood in detail. In the two-dimensional finite element model, cancellous bone is usually simulated by a non-porous structure solid or by approximating bone trabeculae as ellipses, which is different from real bone. However, although the error of the model constructed by bone CT images is small, it needs to be based on real bone samples, and the samples are limited. Therefore, a modeling method of cancellous bone based on four-parameter random growth method was proposed in this paper, and on this basis, numerical simulation of ultrasonic transmission was carried out. Firstly, based on the four-parameter random growth method, the aggregation algorithm is used to concentrate discrete pixels and smooth the edge of pores. Meanwhile, the built-in algorithm ensures the same porosity before and after processing to reduce the discrete pore structure. Secondly, based on COMSOL to establish the simulation model of ultrasonic propagation in cancellous bone, we analyzed the change of acoustic field distribution, discussed the correlation between the porosity of cancellous bone and backscattering coefficient (BSC) based on the ultrasonic backscattering method, and compared the experimental results of CT scan images of bone samples. The experimental results show that the cancellous bone modeling method in this paper has the same conclusion as the method based on CT images, which verifies the feasibility of this method. This method can generate a geometric model of the cancellous bone microstructure with specified porosity and different bone trabecular distribution, which is similar to the real bone structure, and can be directly imported into the finite element software to facilitate the study of bone microstructure related problems. It provides a more convenient method to study the mechanism of ultrasonic propagation in cancellous bone and has important significance in solving the inverse problem of recovering effective bone parameters from the received ultrasonic signals.

Gaussian random field-based characterization and reconstruction of cancellous bone microstructure considering the constraint of correlation structure

The macro scale physical properties of cancellous bone materials are governed by the microstructural features, which is of great significance for the multi-scale research of cancellous bone and the inverse design of bone-mimicking materials. Therefore, it is essential to characterize the natural cancellous bone samples, and reconstruct the microstructures with the biomimetic osteointegration and mechanical properties. In this research, a novel approach for the characterization and reconstruction of cancellous bone was proposed, based on the medical image analysis and anisotropic three-dimensional Gaussian random field (GRF). The geometric similarity, i.e. the interface curvature distribution (ISD), was meticulously studied, which is important to the osteointegration ability. And the mechanical properties were validated by the stress-strain curves under the large compressive strain simulated by the smoothed particle hydrodynamic (SPH) method. In addition, the effects of the generation parameters of GRF-based biomimetic microstructures on the apparent properties were analyzed. The ISD results demonstrated that both GRF and micro-CT groups had the similar columnar morphological properties, while the latter had more hyperbolic features. And it was found that the GRF-based biomimetic microstructures and the natural bone samples based on micro-CT (MCT) had the similar failure mode. The concordance correlation coefficient between MCT and GRF pairs was 0.8685, with a Pearson ρ value of 0.8804, and significance level p<0.0001. The Bland-Altman LoA was 0.1647 MPa with 95 % (1.96SD) lower and upper bound value between −0.2892 and 0.6185 MPa. The two groups had almost the same elastic modulus with the mean absolute percentage error (MAPE) of 7.84 %. While the yield stress and total conversion energy of the GRF-based samples were lower than those of the natural bone samples, and the MAPE were 16.99 % and 16.27 %, respectively. Although it meant the lower structural efficiency, the huge design space of this approach and advanced 3D printing technology can provide great potential for the design of orthopedic implants.

SURGICAL TECHNIQUE AND COMPARISON OF AUTOLOGOUS CANCELLOUS BONE GRAFTS FROM VARIOUS DONOR SITES IN RATS

Autologous cancellous bone graft is the gold standard in large bone defect repair. However, studies using autologous bone grafting in rats are rare and donor sites as well as harvesting techniques vary. The aim of this study was to determine the feasibility of autologous cancellous bone graft harvest from 5 different anatomical sites in rats and compare their suitability as donor sites for autologous bone graft.13 freshly euthanised rats were used to describe the surgical approaches for autologous bone graft harvest from the humerus, iliac crest, femur, tibia and tail vertebrae (n=4), determine the cancellous bone volume and microstructure of those five donor sites using µCT (n=5), and compare their cancellous bone collected qualitatively by looking at cell outgrowth and osteogenic differentiation using an ALP assay and Alizarin Red S staining (n=4).It was feasible to harvest cancellous bone graft from all 5 anatomical sites with the humerus and tail being more surgically challenging. The microstructural analysis showed a significantly lower bone volume fraction, bone mineral density, and trabecular thickness of the humerus and iliac crest compared to the femur, tibia, and tail vertebrae. The harvested volume did not differ between the donor sites. All donor sites apart from the femur yielded primary osteogenic cells confirmed by the presence of ALP and Alizarin Red S stain. Bone samples from the iliac crest showed the most consistent outgrowth of osteoprogenitor cells.The tibia and iliac crest may be the most favourable donor sites considering the surgical approach. However, due to the differences in microstructure of the cancellous bone and the consistency of outgrowth of osteoprogenitor cells, the donor sites may have different healing properties, that need further investigation in an in vivo study.

Microstructural Analysis of Cancellous Bone in Fluorosis Rats.

Skeletal fluorosis likely alters bone structural properties on the cortical and cancellous tissue levels in view that fluorine ion replaces bone mineral composition. Our previous study showed high bone turnover occurred in cortical bone of skeletal fluorosis. Therefore, this study further analyzed the microstructure of cancellous bone in fluorosis rats. Rats were randomly assigned into three groups: the control, low-dose fluoride group (10 mgF-/kg·day), and high-dose fluoride group (20 mgF-/kg·day). Rats were orally administered with fluoride for 1, 2, and 3 months of periods. The trabecular bone parameters of tibia were detected with micro CT and analyzed with software. The activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), and the content of malondialdehyde (MDA) in serum were measured. Results showed that severity of dental fluorosis rose with the increase of dose and prolongation of fluoride exposure. Meantime, the poorer connectivity and less trabecular bone network were observed in cancellous bone of rats treated with fluoride. Data analysis indicated that fluoride treatment significantly decreased bone volume and connectivity degree, but amplified trabecular space in 1 and 2 months of periods. Intriguingly, trabecular thickness significantly decreased in 1-month high-dose fluoride group, but returned to the control in 3 months of period. Fluoride treatment mainly inhibited the GPX activity and increased the MDA level to activate oxidative stress. This study confirmed that excessive fluoride impaired cancellous bone and caused redox imbalance.

Exploring the relationship of brown adipose tissue to bone microarchitecture using 7T MRI and micro-CT.

Brown adipose tissue (BAT) is involved both in energy production and bone metabolism. The purpose of this study was to analyze the relationship between BAT and microarchitecture at cancellous and cortical bone using Kunming mice and the methods of 7T magnetic resonance imaging (MRI) combined with micro-CT. Twenty-four female Kunming mice were examined by 7T MRI and measured T2* relaxation time on the deep and superficial interscapular BAT (iBAT) and subcutaneous white adipose tissue (sWAT). Cancellous bone microarchitecture of the distal femur and cortical bone of the middle femur were examined by micro-CT. A paired t-test was used to analyze the differences in T2* values between iBAT and sWAT. The correlation between BAT T2* values and bone microstructure parameters were analyzed using Pearson's correlation. T2* values of the deep and superficial iBAT (6.36±3.31 ms and 6.23±2.61 ms) were significantly shorter than those of sWAT (16.30±3.05 ms, t(deep) iBAT=-10.816), t(superficial) iBAT =-12.276, p<0.01). Deep iBAT T2* values were significantly and negatively correlated with bone volume, cancellous thickness, and bone thickness (Th) and trabecular thickness (Tb.Th) of the cancellous bone of femur. Deep iBAT T2* values were significantly and positively correlated with the structural model index of cancellous bone of femur. Deep iBAT T2* values were significantly and negatively correlated with bone mineral density of the cortical bone of femur. MRI can distinguish the two adipose tissues from each other. T2* values of BAT were lower than WAT on MRI. BAT related bone remodeling was more correlated with the microstructure of cancellous bone than that of cortical bone.

Influence of Chinese Herbal Formula on Bone Characteristics of Cobb Broiler Chickens.

To evaluate the prevention and treatment effect of a Chinese herbal formula (CHF) on the bone disease of Cobb broiler chickens, compare its efficacy with Bisphosphonates (BPs), and provide a theoretical basis for studying the nutritional regulation technology of CHF to improve the bone characteristics of broiler chickens. In this study, 560 one-day-old Cobb broiler chickens were examined for the influence of Chinese herbal formula (CHF) and Bisphosphonates (BPs). Different doses of CHF and BPs were added to the diet, and the 30- and 60-day-old live weight, tibial bone strength, the microstructure of the distal femur cancellous bone, blood biochemical indexes related to bone metabolism, and genes related to bone metabolism were determined and analyzed. The results showed that the live weight of Cobb broilers fed with CHF and BPs in the diet was as follows: The live weight of the CHF group was higher than that of the normal control (NC) group, while the live weight of the BPs group was lower than that of the NC group; the CHF and BPs improved the bone strength of Cobb broilers and increased the elastic modulus, yield strength, and maximum stress of the tibia. CHF and BPs increased the cancellous bone mineral density (BMD), bone tissue ratio (BV/TV), bone surface area tissue volume ratio (BS/TV), bone trabecular thickness (Tb.Th), and bone trabecular number (Tb.N) in the distal femur, and decreased the bone surface area bone volume ratio (BS/BV) and bone trabecular separation (Tb.Sp). Thus, the microstructure of the bone tissue of the distal femur was improved to a certain extent. Both the CHF and the BPs also increased the serum levels of the vitamin D receptor (VDR), osteoprotegerin (OPG), and alkaline phosphatase (ALP), and decreased the content of osteocalcin (OT). Meanwhile, CHF and BPs upregulated the expression of osteogenic genes (BMP-2, OPG, Runx-2) to promote bone formation and downregulated the expression of osteoclastic genes (RANK, RANKL, TNF-α) to inhibit bone resorption, thus affecting bone metabolism. Conclusion: The CHF could improve the skeletal characteristics of Cobb broilers by upregulating the expression of bone-forming-related genes and downregulating the expression of bone-breaking-related genes, thus preventing and controlling skeletal diseases in Cobb broilers. Its effect was comparable to that of BPs. Meanwhile, the CHF-H group achieved the best results in promoting the growth and improvement of the skeletal characteristics of Cobb broilers based on the live weight and skeletal-characteristics-related indexes.

Surgical technique and comparison of autologous cancellous bone grafts from various donor sites in rats.

Autologous cancellous bone graft is the gold standard in large bone defect repair. However, studies using autologous bone grafting in rats are rare. To determine the feasibility of autologous cancellous bone graft harvest from different anatomical donor sites (humerus, ilium, femur, tibia, and tail vertebrae) in rats and compare their suitability as donor sites, a total of 13 freshly euthanized rats were used to describe the surgical technique, determine the cancellous bone volume and microstructure, and compare the cancellous bone collected quantitatively and qualitatively. It was feasible to harvest cancellous bone grafts from all five anatomical sites with the humerus and tail being more surgically challenging. The microstructural analysis using micro-computed tomographyshowed a significantly lower bone volume fraction, bone mineral density, and trabecular thickness of the humerus and iliac crest compared to the femur, tibia, and tail vertebrae. The harvested weight and volume did not differ between the donor sites. All donor sites apart from the femur yielded primary osteogenic cells confirmed by the presence of alkaline phosphatase and Alizarin Red S stain. Bone samples from the iliac crest showed the most consistent outgrowth of osteoprogenitor cells. In conclusion, the tibia and iliac crest may be the most favorable donor sites considering the surgical approach. However, due to the differences in microstructure of the cancellous bone and the consistency of outgrowth of osteoprogenitor cells, the donor sites may have different healing properties, that need further investigation in an in vivo study.

Determination of E-modulus of cancellous bone derived from human humeri and validation of plotted single trabeculae: Development of a standardized humerus bone model

BackgroundEvaluation of the mechanical behavior of the microstructure of cancellous bone seems important for the understanding of the mechanical behavior of bone. Prevention and treatment of fragility fractures due to osteoporosis is a major challenge according to ageing population. A bone model might help to assess fracture risk. Measurement of single trabeculae of bone should give further information compared with bone densitometry alone. This study measures the mechanical properties of single cancellous trabeculae derived from human proximal humerus. Methods34 single trabeculae dissected from human humeral heads were measured and evaluated mechanically. Trabeculae were fixed on microscope slides and geometrical data were reported during axial rotation of the specimens to measure the transverse section using computer aided design (CAD). The samples were subjected to a two-point bending test and were loaded with a measure-stamp at a defined distance. Force and deflection were measured by high-resolution sensors. The E-modulus was then calculated in combination with finite elements method simulation (FEM), using the previously obtained CAD-Data. ResultsThe average E-modulus from 34 valid measurements of human humeral trabeculae was 1678 MPa with a range from 829 to 3396 MPa, which is consistent with existing literature. The planned additional validation of the measurement method using manufactured three-dimensional synthetic trabeculae with known mechanical properties showed an average elastic modulus of single trabeculae of 51.5 MPa, being two dimensions lower than the value reported in the datasheet of the plastic. ConclusionThis newly developed, time and cost-efficient procedure allows the measurement of E-modulus in single trabeculae. Measurement of mechanic parameters of single trabeculae might give insights on mechanic behavior of bone and be relevant for the research of systemic bone diseases, complementing the existing data on bone-mineral-density. Further examination of single trabeculae of human cancellous bone should give an insight on the mechanical behavior of bone also considering systemic bone diseases.

Ultrasonic Bone Assessment: Ability of Apparent Backscatter Techniques to Detect Changes in the Microstructure of Human Cancellous Bone.

Ultrasonic backscatter techniques may offer a useful approach for detecting changes in bone caused by osteoporosis. The goal of this study was to investigate how bone mineral density (BMD) and the microstructure of human cancellous bone affect three ultrasonic backscatter parameters that have been identified as potentially useful for ultrasonic bone assessment purposes: the apparent integrated backscatter (AIB), the frequency slope of apparent backscatter (FSAB), and the frequency intercept of apparent backscatter (FIAB). Ultrasonic measurements were performed with a 3.5-MHz broadband transducer on 54 specimens of human cancellous bone prepared from the proximal femur. Microstructural parameters and BMD were measured using X-ray microcomputed tomography (micro-CT). Relationships between AIB, FSAB, FIAB, and the micro-CT parameters were investigated using univariate and multivariate statistical analysis techniques. Moderate-to-strong univariate correlations were observed between the backscatter parameters and microstructure and BMD in many cases. The partial correlation analysis indicated that the backscatter parameters are dependent on microstructure independently of BMD in some cases. Multiple stepwise linear regression analysis used to generate multivariate models found that microstructure was a significant predictor of the backscatter parameters in most cases.

Assessment of Intravertebral Mechanical Strains and Cancellous Bone Texture Under Load Using a Clinically Available Digital Tomosynthesis Modality.

Vertebral fractures are the most common osteoporotic fractures, but clinical means for assessment of vertebral bone integrity are limited in accuracy, as they typically use surrogate measures that are indirectly related to mechanics. The objective of this study was to examine the extent to which intravertebral strain distributions and changes in cancellous bone texture generated by a load of physiological magnitude can be characterized using a clinically available imaging modality. We hypothesized that digital tomosynthesis-based digital volume correlation (DTS-DVC) and image texture-based metrics of cancellous bone microstructure can detect development of mechanical strains under load. Isolated cadaveric T11 vertebrae and L2-L4 vertebral segments were DTS imaged in a nonloaded state and under physiological load levels. Axial strain, maximum principal strain, maximum compressive and tensile principal strains, and von Mises equivalent strain were calculated using the DVC technique. The change in textural parameters (line fraction deviation, anisotropy, and fractal parameters) under load was calculated within the cancellous centrum. The effect of load on measured strains and texture variables was tested using mixed model analysis of variance, and relationships of strain and texture variables with donor age, bone density parameters, and bone size were examined using regression models. Magnitudes and heterogeneity of intravertebral strain measures correlated with applied loading and were significantly different from background noise. Image texture parameters were found to change with applied loading, but these changes were not observed in the second experiment testing L2-L4 segments. DTS-DVC-derived strains correlated with age more strongly than did bone mineral density (BMD) for T11.

Biomimetic Ti–6Al–4V alloy/gelatin methacrylate hybrid scaffold with enhanced osteogenic and angiogenic capabilities for large bone defect restoration

Titanium-based scaffolds are widely used implant materials for bone defect treatment. However, the unmatched biomechanics and poor bioactivities of conventional titanium-based implants usually lead to insufficient bone integration. To tackle these challenges, it is critical to develop novel titanium-based scaffolds that meet the bioadaptive requirements for load-bearing critical bone defects. Herein, inspired by the microstructure and mechanical properties of natural bone tissue, we developed a Ti–6Al–4V alloy (TC4)/gelatin methacrylate (GelMA) hybrid scaffold with dual bionic features (GMPT) for bone defect repair. GMPT is composed of a hard 3D-printed porous TC4 metal scaffold (PT) backbone, which mimics the microstructure and mechanical properties of natural cancellous bone, and a soft GelMA hydrogel matrix infiltrated into the pores of PT that mimics the microenvironment of the extracellular matrix. Ascribed to the unique dual bionic design, the resultant GMPT demonstrates better osteogenic and angiogenic capabilities than PT, as confirmed by the in vitro and rabbit radius bone defect experimental results. Moreover, controlling the concentration of GelMA (10%) in GMPT can further improve the osteogenesis and angiogenesis of GMPT. The fundamental mechanisms were revealed by RNA-Seq analysis, which showed that the concentration of GelMA significantly influenced the expression of osteogenesis- and angiogenesis-related genes via the Pi3K/Akt/mTOR pathway. The results of this work indicate that our dual bionic implant design represents a promising strategy for the restoration of large bone defects.

Study on mechanical properties and permeability of elliptical porous scaffold based on the SLM manufactured medical Ti6Al4V.

In this paper, we take the elliptical pore structure which is similar to the microstructure of cancellous bone as the research object, four groups of bone scaffolds were designed from the perspective of pore size, porosity and pore distribution. The size of the all scaffolds were uniformly designed as 10 × 10 × 12 mm. Four groups of model samples were prepared by selective laser melting (SLM) and Ti6Al4V materials. The statics performance of the scaffolds was comprehensively evaluated by mechanical compression simulation and mechanical compression test, the manufacturing error of the scaffold samples were evaluated by scanning electron microscope (SEM), and the permeability of the scaffolds were predicted and evaluated by simulation analysis of computational fluid dynamics (CFD). The results show that the different distribution of porosity, pore size and pores of the elliptical scaffold have a certain influence on the mechanical properties and permeability of the scaffold, and the reasonable size and angle distribution of the elliptical pore can match the mechanical properties and permeability of the elliptical pore scaffold with human cancellous bone, which has great potential for research and application in the field of artificial bone scaffold.

Negligible Effect of Estrogen Deficiency on Development of Skeletal Changes Induced by Type 1 Diabetes in Experimental Rat Models.

Although postmenopausal osteoporosis often occurs concurrently with diabetes, little is known about interactions between estrogen deficiency and hyperglycemia in the skeletal system. In the present study, the effects of estrogen deficiency on the development of biochemical, microstructural, and mechanical changes induced by streptozotocin-induced diabetes mellitus (DM) in the rat skeletal system were investigated. The experiments were carried out on nonovariectomized (NOVX) and ovariectomized (OVX) control and diabetic mature female Wistar rats. Serum levels of bone turnover markers (CTX-I and osteocalcin) and 23 cytokines, bone mass and mineralization, histomorphometric parameters, and mechanical properties of cancellous and compact bone were determined. The results were subjected to two-way ANOVA and principal component analysis (PCA). Estrogen deficiency induced osteoporotic changes, with increased bone resorption and formation, and worsening of microstructure (femoral metaphyseal BV/TV decreased by 13.0%) and mechanical properties of cancellous bone (the maximum load in the proximal tibial metaphysis decreased by 34.2%). DM in both the NOVX and OVX rats decreased bone mass, increased bone resorption and decreased bone formation, and worsened cancellous bone microarchitecture (for example, the femoral metaphyseal BV/TV decreased by 17.3% and 18.1%, respectively, in relation to the NOVX controls) and strength (the maximum load in the proximal tibial metaphysis decreased by 35.4% and 48.1%, respectively, in relation to the NOVX controls). Only in the diabetic rats, profound increases in some cytokine levels were noted. In conclusion, the changes induced by DM in female rats were only slightly intensified by estrogen deficiency. Despite similar effects on bone microstructure and strength, the influence of DM on the skeletal system was based on more profound systemic homeostasis changes than those induced by estrogen deficiency.

A review on artificial bone modelling: Materials and manufacturing techniques

Artificial bone modelling has been done in recent times in order to replicate the mechanical properties and microstructure of the human cancellous and cortical bone. The applications of artificial bone models include bone grafting and testing of orthopaedic instruments. The recent developments in this field make the use of natural polymers and bio-compatible metals to manufacture porous structures replicating bone properties. The aim of this review is to consolidate the research efforts done in the field of making artificial bone models of the human trabecular bone by studying the material and manufacturing aspects of the process. The article reviews the bone models under the categories of polymers and composites and metal scaffolds. A combination of natural polymers along with a bio-compatible synthetic polymer and bio-ceramic reinforcement has proven to be a suitable candidate in replicating the mechanical properties of the bone while ensuring bio-compatibility. Three manufacturing techniques were reviewed with respect to the ability to replicate the bone microstructure. SLM is a potential candidate in terms of design flexibility and manufacturing accuracy. A direction to future research in extending the application of artificial bone models to the testing of dental implants has also been provided.

Ultrasonic Bone Assessment Using the Backscatter Amplitude Decay Constant

Ultrasonic backscatter techniques are being developed to detect changes in bone caused by osteoporosis. The present study introduces a new technique that measures the exponential decay in the amplitude of the backscatter signal quantified by a parameter called the backscatter amplitude decay constant (BADC). Measurements were performed on 54 specimens of cancellous bone from 14 human femurs using a 3.5-MHz transducer. Six methods were tested to determine BADC. The recommended method measures the time slope of the natural log of the rectified signal. Measured values of BADC ranged from approximately 0.1 μs-1 to 0.6 μs-1. Moderate to strong correlations (Spearman's ρ >0.7) were found between BADC and the density and microstructural characteristics of the specimens determined using X-ray microcomputed tomography. The results of this study suggest that BADC may be able to detect changes in the density and microstructure of cancellous bone caused by osteoporosis and other diseases.

Microcomputed Tomography (microCT)‐based Three‐Dimensional (3D) Image Registration to Assess Bone Loss in Spinal Cord Injured Rats

BACKGROUND3D image registration is a technique where in‐vivo microCT scans are collected at different timepoints and regions of interest (ROI) are constructed and aligned to improve the precision of determining bone microstructure. In the rodent spinal cord injury (SCI) model, the rapid bone loss occurring at the distal femur precludes the use of standard 3D registration strategies.PURPOSETo (1) adapt a microCT‐based 3D registration protocol to our rodent SCI model, (2) determine the degree of cancellous bone loss at the distal femoral epiphysis after SCI, and (3) assess the effects of bodyweight‐supported treadmill training (TM) or passive bicycle training (PBT) on bone loss after SCI.METHODS16‐wk old male Sprague‐Dawley rats were stratified to receive: 1) T9 laminectomy (SHAM) (n=9), 2) severe T9 contusion (SCI) (n=10), 3) SCI+TM (n=10), or 4) SCI+PBT (n=14). TM and PBT began 1‐wk post‐surgery (post‐sx, two 20‐min bouts/day, 5‐d/wk for 3‐wks). In‐vivo microCT scans were performed pre‐sx and 2‐ and 4‐wks post‐sx. Images were aligned with a 3D registration protocol. ROIs were developed to assess cancellous bone microstructure at the distal femoral epiphysis using two separate protocols that either included or excluded new bone formed by periosteal bone expansion over the 4‐wk experiment.RESULTSDifferences were noted between the ROI protocols, with the ROI that included periosteal bone growth underestimating SCI‐induced bone loss. As such, the results reported hereafter were derived from the ROI that excluded new bone resulting from periosteal bone expansion. No differences in bone outcomes were present in SHAMs at any timepoint except for a slightly higher trabecular separation (Tb.Sp) at 4‐wks (p&lt;.05). At 2‐wks, SCI displayed 14% lower cancellous bone volume (BV/TV) than pre‐sx (p&lt;.01), characterized by 13% lower trabecular number (Tb.N) (p&lt;.05) and 7% higher Tb.Sp (p&lt;.05). Bone loss was more pronounced at 4‐wks after SCI, evidenced by lower trabecular thickness (Tb.Th) and higher Tb.Sp vs 2‐wks (both p&lt;.01). SCI+TM and SCI+PBT displayed a similar magnitude of bone loss to SCI at 2‐wks (1‐wk after starting exercise). Thereafter, SCI+TM displayed no further bone loss, resulting in 9% less BV/TV loss than SCI (p&lt;.01). In comparison, PBT increased BV/TV 15% from 2‐ to 4‐wks (p&lt;.01), due to 5% higher Tb.Th (p&lt;.01) and 12% higher Tb.N (p&lt;.05), ultimately restoring BV/TV to pre‐sx levels. Structural model index (SMI) and trabecular pattern factor (Tb.Pf) increased in SCI (p&lt;0.05) and SCI+TM at 4‐wks (p&lt;.05 to &lt;.01), signifying transition from rod‐like to weaker plate‐like trabeculae and a less connected trabecular network, respectively. In comparison, SMI (p&lt;.05) and Tb.Pf (p&lt;.01) increased in SCI+PBT at 2‐wks before returning to pre‐sx levels by 4‐wks.CONCLUSIONUsing our 3D registration protocol, we determined that SCI causes severe cancellous bone loss and changes indicative of an overall weakening of bone architecture at the distal femoral epiphysis. TM attenuated bone loss at this skeletal site, while PBT promoted new bone formation and restored BV/TV to pre‐sx levels.Support or Funding InformationThis work was supported by the APS UGSRF.

Locomotor training with adjuvant testosterone preserves cancellous bone and promotes muscle plasticity in male rats after severe spinal cord injury.

Loading and testosterone may influence musculoskeletal recovery after spinal cord injury (SCI). Our objectives were to determine (a) the acute effects of bodyweight-supported treadmill training (TM) on hindlimb cancellous bone microstructure and muscle mass in adult rats after severe contusion SCI and (b) whether longer-term TM with adjuvant testosterone enanthate (TE) delivers musculoskeletal benefit. In Study 1, TM (40 min/day, 5 days/week, beginning 1 week postsurgery) did not prevent SCI-induced hindlimb cancellous bone loss after 3 weeks. In Study 2, TM did not attenuate SCI-induced plantar flexor muscles atrophy nor improve locomotor recovery after 4 weeks. In our main study, SCI produced extensive distal femur and proximal tibia cancellous bone deficits, a deleterious slow-to-fast fiber-type transition in soleus, lower muscle fiber cross-sectional area (fCSA), impaired muscle force production, and levator ani/bulbocavernosus (LABC) muscle atrophy after 8 weeks. TE alone (7.0 mg/week) suppressed bone resorption, attenuated cancellous bone loss, constrained the soleus fiber-type transition, and prevented LABC atrophy. In comparison, TE+TM concomitantly suppressed bone resorption and stimulated bone formation after SCI, produced near-complete cancellous bone preservation, prevented the soleus fiber-type transition, attenuated soleus fCSA atrophy, maintained soleus force production, and increased LABC mass. 75% of SCI+TE+TM animals recovered voluntary over-ground hindlimb stepping, while no SCI and only 20% of SCI+TE animals regained stepping ability. Positive associations between testosterone and locomotor function suggest that TE influenced locomotor recovery. In conclusion, short-term TM alone did not improve bone, muscle, or locomotor recovery in adult rats after severe SCI, while longer-term TE+TM provided more comprehensive musculoskeletal benefit than TE alone.

Influence of cancellous bone microstructure on ultrasonic attenuation: a theoretical prediction

BackgroundQuantitative ultrasound has been used for the assessment of cancellous bone status. The attenuation mechanisms of cancellous bone, however, have not been well understood, because the microstructure of cancellous bone is significantly inhomogeneous and the interaction between ultrasound and the microstructure of cancellous bone is complex. In this study, a theoretical approach was applied to investigate the influence of the microstructure of cancellous bone on ultrasonic attenuation.ResultsThe scattering from a trabecular cylinder was significantly angle dependent. The dependencies of the ultrasonic attenuation on frequency, scatterer size, and porosity were explored from the theoretical calculation. Prediction results showed that the ultrasonic attenuation increased with the increase of frequency and decreased linearly with the increase in porosity, and the broadband ultrasound attenuation decreased with the increase in porosity. All these predicted trends were consistent with published experimental data. In addition, our model successfully explained the principle of broadband ultrasound attenuation measurement (i.e., the attenuation over the frequency range 0.3–0.65 MHz was approximately linearly proportional to frequency) by considering the contributions of scattering and absorption to attenuation.ConclusionThe proposed theoretical model may be a potentially valuable tool for understanding the interaction of ultrasound with cancellous bone.

Compression failure characterization of cancellous bone combining experimental testing, digital image correlation and finite element modeling

Cancellous bone yield strain has been reported in the literature to be relatively constant and independent from microstructure and apparent density, while fracture strain shows higher scattering. The objective of this work is to assess this hypothesis, characterizing the compression fracture in cancellous bone from a numerical approach and relating it to morphological parameters. Quasi-static compression fractures of cancellous bone samples are modeled using high-resolution image-based finite elements, correlating the numerical models and experimental results. The yield strain and the strain at fracture are inferred from the micro-CT-based finite element models by inverse analysis. The validation of the fracture models is carried out through digital image correlation (DIC). To develop this work, cancellous bone parallelepiped-shaped specimens were prepared and micro-CT scanned at 22 μm spatial resolution. A morphometric analysis was carried out for each specimen in order to characterize its microstructure. Quasi-static compression tests were conducted, recording the force-displacement response and a sequence of images during testing for the application of the DIC technique. This was applied without the need of a speckle pattern benefiting from the irregular microstructure of cancellous bone. The finite element models are also used to simulate the local fracture of trabeculae at the micro level using a combination of continuum damage mechanics and the element deletion technique. Equivalent strain, computed both from DIC and micro-FE, was the best predictor of the compression fracture pattern. The procedure followed in this work permits the estimation of failure parameters that are difficult to measure experimentally, which can be used in numerical models.