Distal skeletal tibia assessed by HR-pQCT is highly correlated with femoral and lumbar vertebra failure loads.

Dual energy X-ray absorptiometry (DXA) is the standard for assessing fragility fracture risk using areal bone mineral density (aBMD), but only explains 60-70% of the variation in bone strength. High-resolution peripheral quantitative computed tomography (HR-pQCT) provides 3D-measures of bone microarchitecture and volumetric bone mineral density (vBMD), but only at the wrist and ankle. Finite element (FE) models can estimate bone strength with 86-95% precision. The purpose of this study is to determine how well vBMD and FE bone strength at the wrist and ankle relate to fracture strength at the hip and spine, and to compare these relationships with DXA measured directly at those axial sites. Cadaveric samples (radius, tibia, femur and L4 vertebra) were compared within the same body. The radius and tibia specimens were assessed using HR-pQCT to determine vBMD and FE failure load. aBMD from DXA was measured at the femur and L4 vertebra. The femur and L4 vertebra specimens were biomechanically tested to determine failure load. aBMD measures of the axial skeletal sites strongly correlated with the biomechanical strength for the L4 vertebra (r=0.77) and proximal femur (r=0.89). The radius correlated significantly with biomechanical strength of the L4 vertebra for vBMD (r=0.85) and FE-derived strength (r=0.72), but not with femur strength. vBMD at the tibia correlated significantly with femoral biomechanical strength (r=0.74) and FE-estimated strength (r=0.83), and vertebral biomechanical strength for vBMD (r=0.97) and FE-estimated strength (r=0.91). The higher correlations at the tibia compared to radius are likely due to the tibia's weight-bearing function.

Mechanical stimuli of trabecular bone in osteoporosis: A numerical simulation by finite element analysis of microarchitecture.

Mechanical stimuli are one of the factors that influence bone cell activity and therefore the remodeling of bone. These stimuli are dependent on the microarchitecture of the tissue and can be altered by changes in the bone that occur typically with osteoporosis. The objective of this study was to quantify the variation in the mechanical stimuli of trabecular bone due to changes in the microarchitecture. The morphology of 76 cubes of trabecular bone from human tibia were obtained from microcomputed tomography images and estimated possibilities for mechanical stimuli were determined using poro-viscoelastic finite element models based on the three-dimensional images. The distributions of Von Mises stress, octahedral strain, strain energy density, fluid velocity and pore pressure were predicted for the solid and the marrow phases of bone. We predicted that with variations in the morphology of the trabecular bone, such as an increase of 30% porosity, there is a significant decrease in the mechanical stimuli of the tissue when subjected to constant strain. The average stress and strain in the bone phase may reduce 50% and the fluid velocity in the marrow phase 88%. These decreases may intrinsically affect the mechanoregulation of bone regeneration that contributes to the etiology of osteoporosis.

The poro-viscoelastic properties of trabecular bone: a micro computed tomography-based finite element study.

Bone is a porous structure with a solid phase that contains hydroxyapatite and collagen. Due to its composition, bone is often represented either as a poroelastic or as a viscoelastic material; however, the poro-viscoelastic formulation that allows integrating the effect of both the fluid flow and the collagen on the mechanical response of the tissue, has not been applied yet. The objective of this study was to develop a micro computed tomography (µCT)-based finite element (FE) model of trabecular bone that includes both the poroelastic and the viscoelastic nature of the tissue. Cubes of trabecular bone (N=25) from human distal tibia were scanned with µCT and stress relaxation experiments were conducted. The µCT images were the basis for sample specific FE models, and the stress relaxation experiments were simulated applying a poro-viscoelastic formulation. The model considers two scales of the tissue: the intertrabecular pore and the lacunar-canalicular pore scales. Independent viscoelastic and poroelastic models were also developed to determine their contribution to the poro-viscoelastic model. All the experiments exhibited a similar relaxation trend. The average reaction force before relaxation was 9.28 × 10(2)N (SD ± 5.11 × 10(2)N), and after relaxation was 4.69 × 10(2)N (SD ± 2.88 × 10(2)N). The slope of the regression line between the force before and after relaxation was 1.92 (R(2)=0.96). The poro-viscoelastic models captured 49% of the variability of the experimental data before relaxation and 33% after relaxation. The relaxation predicted with viscoelastic models was similar to the poro-viscoelastic ones; however, the poroelastic formulation underestimated the reaction force before relaxation. These data suggest that the contribution of viscoelasticity (fluid flow-independent mechanism) to the mechanical response of the tissue is significantly greater than the contribution of the poroelasticity (fluid flow-dependent mechanism).

A comparison of methods for in vivo assessment of cortical porosity in the human appendicular skeleton.

The recent advent of high-resolution peripheral quantitative computed tomography (HR-pQCT) provides new opportunities to measure in vivo human bone microarchitecture. Increasingly, cortical porosity (CtPo) is of particular interest due to its relationship with bone quality and turnover. The two approaches that have emerged to measure CtPo from HR-pQCT are threshold-based and density-based methods, and the purpose of this work was to compare the performance of each against a gold-standard synchrotron radiation micro-computed tomography (SRµCT) measurement. Human cadaveric cortical bone specimens (N=23) were measured by SRµCT and HR-pQCT, and high correlations were found for both methods. The density-based approach had an r2=0.939 (95% confidence interval (CI) of +6.17% to +20.99%) and consistently overestimated porosity as measured by SRµCT, while the threshold-based approach had an r2=0.977 and consistently underestimated porosity (95% CI of -2.60% to -10.76%). The density-based approach is prone to beam hardening artifacts and susceptible to natural variations of tissue mineral density (TMD), but is less affected by motion artifacts that may occur in in vivo scans. The threshold-based method has the advantage that it provides structural information that complements the cortical porosity measure, such as number of pores and connectivity, and can accurately detect the larger pores which are the most relevant to bone biomechanical strength. With the first generation HR-pQCT systems the accuracy of detecting pores larger than 140 µm diameter is excellent (r2=0.983; 95% CI of -4.88% to +2.45%). The accuracy of the threshold-based method will improve as new HR-pQCT systems emerge and provide a robust quantitative approach to measure cortical porosity.

Assessing the local mechanical environment in medial opening wedge high tibial osteotomy using finite element analysis.

High-tibial osteotomy (HTO) is a surgical technique aimed at shifting load away from one tibiofemoral compartment, in order the reduce pain and progression of osteoarthritis (OA). Various implants have been designed to stabilize the osteotomy and previous studies have been focused on determining primary stability (a global measure) that these designs provide. It has been shown that the local mechanical environment, characterized by bone strains and segment micromotion, is important in understanding healing and these data are not currently available. Finite element (FE) modeling was utilized to assess the local mechanical environment provided by three different fixation plate designs: short plate with spacer, long plate with spacer and long plate without spacer. Image-based FE models of the knee were constructed from healthy individuals (N = 5) with normal knee alignment. An HTO gap was virtually added without changing the knee alignment and HTO implants were inserted. Subsequently, the local mechanical environment, defined by bone compressive strain and wedge micromotion, was assessed. Furthermore, implant stresses were calculated. Values were computed under vertical compression in zero-degree knee extension with loads set at 1 and 2 times the subject-specific body weight (1 BW, 2 BW). All studied HTO implant designs provide an environment for successful healing at 1 BW and 2 BW loading. Implant von Mises stresses (99th percentile) were below 60 MPa in all experiments, below the material yield strength and significantly lower in long spacer plates. Volume fraction of high compressive strain ( > 3000 microstrain) was below 5% in all experiments and no significant difference between implants was detected. Maximum vertical micromotion between bone segments was below 200 µm in all experiments and significantly larger in the implant without a tooth. Differences between plate designs generally became apparent only at 2 BW loading. Results suggest that with compressive loading of 2 BW, long spacer plates experience the lowest implant stresses, and spacer plates (long or short) result in smaller wedge micromotion, potentially beneficial for healing. Values are sensitive to subject bone geometry, highlighting the need for subject-specific modeling. This study demonstrates the benefits of using image-based FE modeling and bone theory to fine-tune HTO implant design.

Predicting the permeability of trabecular bone by micro-computed tomography and finite element modeling.

The intrinsic permeability of bone plays an important role in the transport of nutrients and minerals within the tissue, and affects the mechanical stimuli that are related to the fate of the stem cells. The objective of this study was to establish a method to assess trabecular bone permeability using experimental and finite element (FE) modeling approaches based on micro computed tomography (µCT) images. Human cadaveric tibia cube specimens (N=23) were scanned with µCT. The permeability was measured experimentally using a custom-developed constant-head permeameter, and computationally by a poroelastic formulation to simulate the fluid flow within the discretized bone matrix and pore phase. The average of the experimentally measured permeability was 4.84 × 10(-10)m(2) with a standard deviation of 3.70 × 10(-10)m(2). A regression model of the µCT determined that the maximum bone area to total area ratio (maxBA/TA) for all slices that are perpendicular to the direction of fluid flow explained 84% of the variability of the natural logarithm of the experimentally measured permeability. The 2D measure of maxBA/TA performed better than 3D measures in general, although some parameters were reasonably well associated with permeability such as bone volume ratio (BV/TV, r=-0.71), the bone surface/bone volume (BS/BV, r=0.73), and the trabecular thickness (TbTh, r=-0.71). The correlation between the permeability predicted with FE models and experimentally measured permeability was reasonable (r=0.69), but the FE approach did not accurately represent the wide variability of permeability measured experimentally. The results of this study suggest that the changes in the trabecular bone microarchitecture have an exponential relationship with permeability, and the use of µCT-based 2D measurement of maxBA/TA performs well at predicting permeability, thus providing a convenient approach to measure this important aspect affecting biomechanical functions in the tissue.

An in vivo investigation of the initiation and progression of subchondral cysts in a rodent model of secondary osteoarthritis.

INTRODUCTION: Subchondral bone cysts (SBC) have been identified in patients with knee osteoarthritis (OA) as a cause of greater pain, loss of cartilage and increased chance of joint replacement surgery. Few studies monitor SBC longitudinally, and clinical research using three-dimensional imaging techniques, such as magnetic resonance imaging (MRI), is limited to retrospective analyses as SBC are identified within an OA patient cohort. The purpose of this study was to use dual-modality, preclinical imaging to monitor the initiation and progression of SBC occurring within an established rodent model of knee OA. METHODS: Eight rodents underwent anterior cruciate ligament transection and partial medial meniscectomy (ACLX) of the right knee. In vivo 9.4 T MRI and micro-computed tomography (micro-CT) scans were performed consecutively prior to ACLX and 4, 8, and 12 weeks post-ACLX. Resultant images were co-registered using anatomical landmarks, which allowed for precise tracking of SBC size and composition throughout the study. The diameter of the SBC was measured, and the volumetric bone mineral density (vBMD) was calculated within the bone adjacent to SBC. At 12 weeks, the ACLX and contralateral knees were processed for histological analysis, immunohistochemistry, and Osteoarthritis Research Society International (OARSI) pathological scoring. RESULTS: At 4 weeks post-ACLX, 75% of the rodent knees had at least 1 cyst that formed in the medial tibial plateau; by 12 weeks all ACLX knees contained SBC. Imaging data revealed that the SBC originate in the presence of a subchondral bone plate breach, with evolving composition over time. The diameter of the SBC increased significantly over time (P = 0.0033) and the vBMD significantly decreased at 8 weeks post-ACLX (P = 0.033). Histological analysis demonstrated positive staining for bone resorption and formation surrounding the SBC, which were consistently located beneath the joint surface with the greatest cartilage damage. Trabecular bone adjacent the SBC lacked viable osteocytes and, combined with bone marrow changes, indicated osteonecrosis. CONCLUSIONS: This study provides insight into the mechanisms leading to SBC formation in knee OA. The expansion of these lesions is due to stress-induced bone resorption from the incurred mechanical instability. Therefore, we suggest these lesions can be more accurately described as a form of OA-induced osteonecrosis, rather than 'subchondral cysts'.

Subchondral cysts create increased intra-osseous stress in early knee OA: A finite element analysis using simulated lesions.

AIM OF STUDY: To investigate the role of intra-osseous lesions in advancing the pathogenesis of Osteoarthritis (OA) of the knee, using Finite Element Modeling (FEM) in conjunction with high-resolution imaging techniques. METHODS: Twenty early stage OA patients (≤ Grade 2 radiographic score) were scanned with a prototype, cone-beam CT system. Scans encompassed the mid-shaft of the femur to the diaphysis of the proximal tibia. Individual bones were segmented to create 3D geometric models that were transferred to FE software for loading experiments. Patient-specific, inhomogeneous material properties were derived from the CT images and mapped directly to the FE models. Duplicate models were also created, with a 3D sphere (range 3-12 mm) introduced into a weight-bearing region of the joint, mimicking the size, location, and composition of a subchondral bone cyst (SBC). A spherical shell extending 1mm radially around the SBC served as the sample volume for measurements of von Mises equivalent stress. Both models were vertically loaded with 750 N, or approximately 1 body weight during a single-leg stance. RESULTS: All FE models exhibited a physiologically realistic weight-bearing distribution of stress, which initiated at the joint surface and extended to the cortical bone. Models that contained the SBC experienced a nearly two-fold increase in stress (0.934 ± 0.073 and 1.69 ± 0.159 MPa, for the non-SBC and SBC models, respectively) within the bone adjacent to the SBC. In addition, there was a positive correlation found between the diameter of the SBC and the resultant intra-osseous stress under load (p = 0.004). CONCLUSIONS: Our results provide insights into the mechanism by which SBC may accelerate OA, leading to greater pain and disability. Based on these findings, we feel that patient-derived FE models of the OA knee - utilizing in vivo imaging data - present a tremendous potential for monitoring joint mechanics under physiological loads.

In vitro quantification of wear in tibial inserts using microcomputed tomography.

BACKGROUND: Wear of polyethylene tibial inserts can decrease the longevity of total knee arthroplasty. Wear is currently assessed using laboratory methods that may not permit backside wear measurements or do not quantify surface deviation. QUESTIONS/PURPOSES: We developed and validated a technique to quantify polyethylene wear in tibial inserts using microcomputed tomography (micro-CT), a nondestructive high-resolution imaging technique that provides detailed images of surface geometry in addition to volumetric measurements. METHODS: Six unworn and six wear-simulated polyethylene tibial inserts were evaluated. Each insert was scanned three times using micro-CT at a resolution of 50 µm. The insert surface was reconstructed for each scan and the insert volume was calculated. Gravimetric analysis was performed for all inserts, and the micro-CT and gravimetric volumes were compared to determine accuracy. We created three-dimensional surface deviation maps. RESULTS: Micro-CT generated high-quality three-dimensional renderings of the insert surface geometry. Between-scan precision was 0.07%; we observed no difference between micro-CT and gravimetric volume measurements. CONCLUSIONS: Micro-CT can provide precise and accurate volumetric measurements in addition to quantifiable three-dimensional surface deviation maps for the entire insert surface. The technique has the potential to evaluate wear in wear simulator trials and retrieval studies. CLINICAL RELEVANCE: This micro-CT technique combines the benefits of volumetric and surface scanning methods to quantify wear across all surfaces of polyethylene components with a single tool. When applied in wear simulator and retrieval studies, these measurements can be used to evaluate and predict the wear properties of the components.

Integration and evaluation of a needle-positioning robot with volumetric microcomputed tomography image guidance for small animal stereotactic interventions.

PURPOSE: Preclinical research protocols often require insertion of needles to specific targets within small animal brains. To target biologically relevant locations in rodent brains more effectively, a robotic device has been developed that is capable of positioning a needle along oblique trajectories through a single burr hole in the skull under volumetric microcomputed tomography (micro-CT) guidance. METHODS: An x-ray compatible stereotactic frame secures the head throughout the procedure using a bite bar, nose clamp, and ear bars. CT-to-robot registration enables structures identified in the image to be mapped to physical coordinates in the brain. Registration is accomplished by injecting a barium sulfate contrast agent as the robot withdraws the needle from predefined points in a phantom. Registration accuracy is affected by the robot-positioning error and is assessed by measuring the surface registration error for the fiducial and target needle tracks (FRE and TRE). This system was demonstrated in situ by injecting 200 microm tungsten beads into rat brains along oblique trajectories through a single burr hole on the top of the skull under micro-CT image guidance. Postintervention micro-CT images of each skull were registered with preintervention high-field magnetic resonance images of the brain to infer the anatomical locations of the beads. RESULTS: Registration using four fiducial needle tracks and one target track produced a FRE and a TRE of 96 and 210 microm, respectively. Evaluation with tissue-mimicking gelatin phantoms showed that locations could be targeted with a mean error of 154 +/- 113 microm. CONCLUSIONS: The integration of a robotic needle-positioning device with volumetric micro-CT image guidance should increase the accuracy and reduce the invasiveness of stereotactic needle interventions in small animals.

A bony connection signals laryngeal echolocation in bats.

Echolocation is an active form of orientation in which animals emit sounds and then listen to reflected echoes of those sounds to form images of their surroundings in their brains. Although echolocation is usually associated with bats, it is not characteristic of all bats. Most echolocating bats produce signals in the larynx, but within one family of mainly non-echolocating species (Pteropodidae), a few species use echolocation sounds produced by tongue clicks. Here we demonstrate, using data obtained from micro-computed tomography scans of 26 species (n = 35 fluid-preserved bats), that proximal articulation of the stylohyal bone (part of the mammalian hyoid apparatus) with the tympanic bone always distinguishes laryngeally echolocating bats from all other bats (that is, non-echolocating pteropodids and those that echolocate with tongue clicks). In laryngeally echolocating bats, the proximal end of the stylohyal bone directly articulates with the tympanic bone and is often fused with it. Previous research on the morphology of the stylohyal bone in the oldest known fossil bat (Onychonycteris finneyi) suggested that it did not echolocate, but our findings suggest that O. finneyi may have used laryngeal echolocation because its stylohyal bones may have articulated with its tympanic bones. The present findings reopen basic questions about the timing and the origin of flight and echolocation in the early evolution of bats. Our data also provide an independent anatomical character by which to distinguish laryngeally echolocating bats from other bats.

The role of Akt1 in terminal stages of endochondral bone formation: angiogenesis and ossification.

Longitudinal bone growth is the result of endochondral bone formation which takes place in the growth plate. The rate of chondrocyte proliferation and hypertrophy, vascular invasion with the formation of primary ossification centers and cartilage replacement by bone tissue are all important processes required for normal growth. We have shown a role for the PI3K signaling pathway in chondrocyte hypertrophy and bone growth in tibia explant cultures. In this current study, we aimed to investigate the role of Akt1, an important target of PI3K, in endochondral ossification. Akt1 KO mice showed reduced size compared to their littermates throughout life, but the largest difference in body size was observed around 1 week of age. Focusing on this specific developmental stage, we discovered delayed secondary ossification in the long bones of Akt1 KO mice. A delay in formation of a structure resembling a secondary ossification center was also seen in tibia organ cultures treated with the PI3K inhibitor LY294002. The expression of matrix metalloproteinase-14 (MMP-14), the main protease responsible for development of secondary ossification centers, was decreased in the epiphysis of Akt1 KO mice, possibly explaining the delay in secondary ossification centers seen in the Akt1 KO mice. Bone mineral density (BMD) and bone mineral content (BMC) measured in the proximal tibia of 1-year-old mice were decreased in Akt1 KO mice, suggesting that the original delay in ossification might affect bone quality in older animals.

Molecular and histological analysis of a new rat model of experimental knee osteoarthritis.

Articular cartilage degeneration is the most consistently observed feature of osteoarthritis (OA). Animal and human studies have shown that various forms of exercise influence the course of the disease in different ways. In addition, early changes in articular cartilage that influence the progression of OA, such as the expression of cytokines, require further investigation. We have used a surgically induced experimental model of knee OA to address these questions. Here, we discuss our recent studies investigating the effects of an exercise paradigm in surgically induced OA, which determined that the destabilized knee joint is susceptible to enhanced degeneration when subjected to low-intensity, low-impact exercise. Further, we investigated early global changes in gene expression in articular chondrocytes from degenerating cartilage. Identified candidate genes including genes involved in chemokine, endothelin, and transforming growth factor-alpha signaling are discussed in the context of articular cartilage degeneration in early OA.

Forced mobilization accelerates pathogenesis: characterization of a preclinical surgical model of osteoarthritis.

Preclinical osteoarthritis (OA) models are often employed in studies investigating disease-modifying OA drugs (DMOADs). In this study we present a comprehensive, longitudinal evaluation of OA pathogenesis in a rat model of OA, including histologic and biochemical analyses of articular cartilage degradation and assessment of subchondral bone sclerosis. Male Sprague-Dawley rats underwent joint destabilization surgery by anterior cruciate ligament transection and partial medial meniscectomy. The contralateral joint was evaluated as a secondary treatment, and sham surgery was performed in a separate group of animals (controls). Furthermore, the effects of walking on a rotating cylinder (to force mobilization of the joint) on OA pathogenesis were assessed. Destabilization-induced OA was investigated at several time points up to 20 weeks after surgery using Osteoarthritis Research Society International histopathology scores, in vivo micro-computed tomography (CT) volumetric bone mineral density analysis, and biochemical analysis of type II collagen breakdown using the CTX II biomarker. Expression of hypertrophic chondrocyte markers was also assessed in articular cartilage. Cartilage degradation, subchondral changes, and subchondral bone loss were observed as early as 2 weeks after surgery, with considerable correlation to that seen in human OA. We found excellent correlation between histologic changes and micro-CT analysis of underlying bone, which reflected properties of human OA, and identified additional molecular changes that enhance our understanding of OA pathogenesis. Interestingly, forced mobilization exercise accelerated OA progression. Minor OA activity was also observed in the contralateral joint, including proteoglycan loss. Finally, we observed increased chondrocyte hypertrophy during pathogenesis. We conclude that forced mobilization accelerates OA damage in the destabilized joint. This surgical model of OA with forced mobilization is suitable for longitudinal preclinical studies, and it is well adapted for investigation of both early and late stages of OA. The time course of OA progression can be modulated through the use of forced mobilization.

Micro-computed tomography of a 500-year-old tooth: technical note.

OBJECTIVE: To determine whether micro-computed tomography (micro-CT) could be used to reconstruct ancient dental anatomy accurately and differentiate the enamel from the dentin, as well as to verify whether micro-CT could detect tooth disorders such as attrition or caries accurately. METHODS: Micro-CT imaging was performed, using a cone-beam micro-CT specimen scanner, on a 500-year-old human tooth found in a burial jar in the Cardomom Mountains in southwestern Cambodia. RESULTS: The occlusal surface of the tooth showed marked attrition, with the dentin extending close to the enamel layer on the crown. In addition to this, micro-CT images depicted calculus on the buccal surface and a cervical root caries lesion present on the distal surface. The sclerotic zone of the carious lesion (located deep in the destroyed dentin) and the dentin were effectively differentiated through excellent resolution and superior tissue contrast of the volume data set. Axial slices from apical to coronal show the carious lesion extending vertically along the dentin-enamel junction with an intact outer enamel surface. CONCLUSION: Micro-CT is a reproducible, nondestructive and highly accurate technique that can be successfully applied to the study of ancient teeth.