Loss of trabeculae by mechano-biological means may explain rapid bone loss in osteoporosis.

Osteoporosis is characterized by rapid and irreversible loss of trabecular bone tissue leading to increased bone fragility. In this study, we hypothesize two causes for rapid loss of bone trabeculae; firstly, the perforation of trabeculae is caused by osteoclasts resorbing a cavity so deep that it cannot be refilled and, secondly, the increases in bone tissue elastic modulus lead to increased propensity for trabecular perforation. These hypotheses were tested using an algorithm that was based on two premises: (i) bone remodelling is a turnover process that repairs damaged bone tissue by resorbing and returning it to a homeostatic strain level and (ii) osteoblast attachment is under biochemical control. It was found that a mechano-biological algorithm based on these premises can simulate the remodelling cycle in a trabecular strut where damaged bone is resorbed to form a pit that is subsequently refilled with new bone. Furthermore, the simulation predicts that there is a depth of resorption cavity deeper than which refilling of the resorption pits is impossible and perforation inevitably occurs. However, perforation does not occur by a single fracture event but by continual removal of microdamage after it forms beneath the resorption pit. The simulation also predicts that perforations would occur more easily in trabeculae that are more highly mineralized (stiffer). Since both increased osteoclast activation rates and increased mineralization have been measured in osteoporotic bone, either or both may contribute to the rapid loss of trabecular bone mass observed in osteoporotic patients.     

Multiscale modeling of elastic properties of cortical bone

We model cortical bone as a composite material with hierarchical structure. At a nanostructural level, bone is composed of cross-linked collagen molecules, containing water and non-collagenous proteins in their gaps, reinforced with hydroxyapatite-like nanocrystals. Such a nanocomposite structure represents a mineralized collagen fibril, which serves as a primary building block of bone. At a sub-microstructural level (few microns), the mineralized collagen fibrils are embedded in an extrafibrillar hydroxyapatite matrix to form a single lamella, which also contains the lacunar cavities. At a microstructural level (hundreds of microns) one can distinguish two lamellar structures in the mature cortical bone: osteons, made of concentric layers of lamellae surrounding long hollow Haversian canals, and interstitial lamellae made of remnants of old osteons. At a mesostructural level (several millimeters), the cortical bone is represented by a random collection of osteons and resorption cavities in the interstitial lamellae. A macrostructural level is the whole bone level containing both the cortical (compact) and trabecular (spongy) bone types. In this paper, we predict analytically the effective elastic constants of cortical bone by modeling its elastic response at these different scales, spanning from the nanostructural to mesostructural levels, using micromechanics methods and composite materials laminate theories. The results obtained at a lower scale serve as inputs for the modeling at a higher scale. The predictions are in good agreement with the experimental data reported in literature.     

Nanostructural analysis of trabecular bone

The mechanical properties of bone are dictated by the size, shape and organization of the mineral and matrix phases at multiple levels of hierarchy. While much is known about structure–function relations at the macroscopic level, less is known at the nanoscale, especially for trabecular bone. In this study, high resolution transmission electron microscopy (HRTEM) was carried out to analyze shape and orientation of apatite crystals in murine femoral trabecular bone. The distribution and orientation of mineral apatites in trabecular bone were different from lamellar bone and the c-axis of the tablet-like mineral apatite crystals in trabecular bone was arranged with no preferred orientation. The difference in the orientation distribution of apatite crystals of trabecular bone in the present study compared with that of lamellar bone found in the literature can be attributed to the more complex local stress state in trabecular bone. Apatite crystals were also found to be multi-crystalline, not single crystalline, from dark field image analysis. From the observations of this study, it is suggested that Wolff’s law can be applicable to the nanostructural orientation and distribution of apatite crystals in trabecular bone. It was also found that small round crystalline particles observed adjacent to collagen fibrils were similar in size and shape to the apatite crystals in biomimetically nucleated synthetic amorphous calcium phosphate, which suggests that they are bone mineral apatite nuclei.     

A scalable multi‐level preconditioner for matrix‐free µ‐finite element analysis of human bone structures

The recent advances in microarchitectural bone imaging disclose the possibility to assess both the apparent density and the trabecular microstructure of intact bones in a single measurement. Coupling these imaging possibilities with microstructural finite element (µFE) analysis offers a powerful tool to improve bone stiffness and strength assessment for individual fracture risk prediction. Many elements are needed to accurately represent the intricate microarchitectural structure of bone; hence, the resulting µFE models possess a very large number of degrees of freedom. In order to be solved quickly and reliably on state‐of‐the‐art parallel computers, the µFE analyses require advanced solution techniques. In this paper, we investigate the solution of the resulting systems of linear equations by the conjugate gradient algorithm, preconditioned by aggregation‐based multigrid methods. We introduce a variant of the preconditioner that does not need assembling the system matrix but uses element‐by‐element techniques to build the multilevel hierarchy. The preconditioner exploits the voxel approach that is common in bone structure analysis, and it has modest memory requirements, at the same time robust and scalable. Using the proposed methods, we have solved in 12min a model of trabecular bone composed of 247 734 272 elements, yielding a matrix with 1 178 736 360 rows, using 1024 CRAY XT3 processors. The ability to solve, for the first time, large biomedical problems with over 1 billion degrees of freedom on a routine basis will help us improve our understanding of the influence of densitometric, morphological, and loading factors in the etiology of osteoporotic fractures such as commonly experienced at the hip, spine, and wrist. Copyright © 2007 John Wiley & Sons, Ltd.     

Kinetic characterization of the deproteinization of trabecular and cortical bovine femur bones

The present study proposes an interpretation of the mechanism of bone deproteinization. Cortical and trabecular bovine femur bones were deproteinized using 6% NaOCl (37, 50, 60 °C). The kinetic parameters (rate constant and activation energy) were calculated, and the surface area of each type of bone was considered. A statistical analysis of the rate constants shows that cortical bone deproteinizes at a lower rate than trabecular. The activation energy is higher for trabecular than cortical bone, and no significant differences are found in the protein concentration values for both bones. Therefore, although trabecular bone deproteinizes at a higher rate than cortical, trabecular bone requires more energy for the deproteinization reaction to take place. Considering that both types of bones are constituted by mineral, protein, and water; the present work shows that the individual inner matrix architecture of trabecular and cortical bones, along with characteristics such as the mineral concentration and its bonding with collagen fibers, may be the responsible factors that control protein depletion.     

Influence of density, elasticity, and structure on ultrasound transmission through trabecular bone cylinders

The aim of this in vitro study is to evaluate the potentiality of quantitative ultrasound (QUS) to separate information on density, elasticity, and structure on specimens of trabecular bone. Fifteen cylinders of spongy bone extracted from equine vertebrae were progressively demineralized and subjected to QUS, micro computed tomography (muCT), Dual energy X-ray absorptiometry (DXA) at various mineralization levels. Eventually all cylinders underwent a compression test to calculate the Young's modulus. Correlation analysis shows that speed of sound (SOS) is strictly associated to bone mineral density (BMD), Young's modulus, and all muCT parameters except for degree of anisotropy (DA). Fast wave amplitude (FWA) is directly correlated with bone surface and total volume ratio (BS/TV) and trabecular separation (Tb Sp), and inversely correlated with trabecular number (Tb N). Because muCT parameters were strictly correlated to BMD and Young's modulus data, partial correlation analysis was performed between SOS, FWA, and structural and elastic data in order to eliminate the effect of density. SOS was significantly correlated to bone volume and total volume ratio (BV/TV), BS/TV, and Young's modulus, and FWA was significantly correlated to Tb Sp only. These results show that SOS is strongly influenced by volumetric mineral bone density and elastic modulus of the specimen, and FWA is mainly affected by trabecular separation independently on density. Therefore, SOS and FWA are able to provide different and complementary information, at least on trabecular bone samples.     

Insights into the effects of tensile and compressive loadings on microstructure dependent fracture of trabecular bone

The micro-scale finite element models used in the past to understand yielding failure of trabecular bone have not addressed the microcrack formation and its effect on microstructure dependent fracture. An understanding of microcrack based failure mechanisms can be important to develop insights into response of trabecular bone to external loading before final failure. With this goal, we analyze tensile and compressive fracture failure at two different ages in two trabecular bone micrographs obtained from an ovine femur using a recently developed cohesive finite element method (CFEM) framework. The results and analyses indicate that examined trabecular microstructures are optimally designed for resisting compressive loading. Under tensile loading, initial damage in a microstructure is localized in a single random trabecula. Final microstructure failure occurs immediately after the failure of the trabecula. However, under compressive loading, failure of the first trabecula does not precede immediate complete failure of microstructure. Under compression the propagation fracture toughness (characterized by change in energy release rate as a function of crack density) increases with increase in crack density. However, under tension the propagation fracture toughness decreases with increasing crack density. The fracture mechanism remains unaffected by age variation. Effect of tissue property random variation on the variation in fracture strength diminishes under tension and increases under compression with increase in the age. Overall, results indicate that structural arrangement of the trabecular bone (besides the hierarchical chemical composition) can be an important contributor to its unique fracture resistance properties.     

Modeling trabecular bone adaptation to local bending load regulated by mechanosensing osteocytes

Cancellous bone has a complicated three-dimensional porous microstructure that consists of strut-like or plate-like trabeculae. The arrangement of the trabeculae is remodeled throughout the organism’s lifetime to functionally adapt to the surrounding mechanical environment. During bone remodeling, osteocytes buried in the bone matrix are believed to play a pivotal role as mechanosensory cells and help regulate the coupling of osteoclastic bone resorption and osteoblastic bone formation according to the mechanical stimuli. Previously, we constructed a mathematical model of trabecular bone remodeling incorporating cellular mechanosensing and intercellular signal transmission, in which osteocytes are assumed to sense the flow of interstitial fluid as a mechanical stimulus that regulates bone remodeling. Our remodeling simulation could describe the reorientation of a single strut-like trabecula under uniaxial loading. In the present study, to investigate the effects of a bending load on trabecular bone remodeling, we simulated the morphological change in a single trabecula under a cyclic bending load based on our mathematical model. The simulation results showed that the application of the bending load influences not only the formation of the plate-like trabecula but also the changes in trabecular topology. These results suggest the possibility that the characteristic trabecular morphology, such as the strut-like or plate-like form, is determined depending on the local mechanical environment.     

Ultrasonic scanner for in vivo measurement of cancellous bone properties from backscattered data

A dedicated ultrasonic scanner for acquiring RF echoes backscattered from the trabecular bone was developed. The design of device is based on the goal of minimizing of custom electronics and computations executed solely on the main computer processor and the graphics card. The electronic encoder-digitizer module executing all of the transmission and reception functions is based on a single low-cost field programmable gate array (FPGA). The scanner is equipped with a mechanical sector-scan probe with a concave transducer with 50 mm focal length, center frequency of 1.5 MHz and 60% bandwidth at -6 dB. The example of femoral neck bone examination shows that the scanner can provide ultrasonic data from deeply located bones with the ultrasound penetrating the trabecular bone up to a depth of 20 mm. It is also shown that the RF echo data acquired with the scanner allow for the estimation of attenuation coefficient and frequency dependence of backscattering coefficient of trabecular bone. The values of the calculated parameters are in the range of corresponding in vitro data from the literature but their variation is relatively high.     

Development of a numerical cancellous bone model for finite-difference time-domain simulations of ultrasound propagation

The trabecular frame in cancellous bone has numerous porous spaces of various sizes and shapes. Their continual arrangement changes with position in the bone. Assuming that the complicated pore space is the aggregation of spherical pores, in this study, the trabecular structure was analyzed using a three-dimensional (3-D) X-ray microcomputed tomography (muCT) image. Analysis involved a 3-D cancellous bone model developed for numerical simulations of ultrasound propagation. In this model, the trabecular structure was simplified by regularly arranging spherical pores in a solid bone. Using a viscoelastic, finite-difference, time-domain (FDTD) method with the simplified cancellous bone model, ultrasound pulse waveforms propagating through cancellous bone were simulated in two cases of the propagations parallel and perpendicular to the main trabecular orientation. The porosity dependences of the propagation properties, attenuation, and propagation speed were derived from the simulated waveforms. Comparisons with simulated results using the realistic cancellous bone model reconstructed from a 3-D muCT image, assisted to further validate this simplified model.     

Evaluation of trabecular bone formation in a canine model surrounding a dental implant fixture immobilized with an antimicrobial peptide derived from histatin

JH8194 induces osteoblast differentiation, although it was originally designed to improve antifungal activity. This suggests that JH8194 is useful for implant treatment. Therefore, the aim of this study was to evaluate the osseointegration capacity of JH8194-modified titanium dental implant fixtures (JH8194-Fi). The implants were randomly implanted into the edentulous ridge of dog mandibles. Healing abutments were inserted immediately after implant placement. Three weeks later, peri-implant bone levels, the first bone-to-implant contact points, and trabecular bone formation surrounding the implants were assessed by histological and digital image analyses based on microcomputed tomography (microCT). The histological analysis revealed an enhancement of mature trabecular bone around the JH8194-Fi compared with untreated fixtures (control-Fi). Similarly, microCT combined with analysis by Zed View? also showed increased trabecular bone formation surrounding the JH8194-Fi compared with the control-Fi (Student's t-test, P < 0.05). JH8194 may offer an alternative biological modification of titanium surfaces to enhance trabecular bone formation around dental implants, which may contribute to the transient acquirement of osseointegration and the long-term success of implant therapy.     

Numerical analysis of variability in ultrasound propagation properties induced by trabecular microstructure in cancellous bone

The manner by which the trabecular microstructure affects the propagation of ultrasound waves through cancellous bone is numerically investigated by finite difference time-domain (FDTD) simulation. Sixteen 3-D numerical models of 6.45times6.45times6.45 mm with a voxel size of 64.5 mum are reconstructed using a 3-D microcomputed tomographic (muCT) image taken from a bovine cancellous bone specimen of approximately 20times20times9 mm. All cancellous bone models have an oriented trabecular structure, and their trabecular elements are gradually eroded to increase the porosity using an image processing technique. Three erosion procedures are presented to realize various changes in the trabecular microstructure with increasing porosity. FDTD simulations of the ultrasound pulse waves propagating through the cancellous bone models at each eroded step are performed in 2 cases of the propagations parallel and perpendicular to the major trabecular orientation. The propagation properties of the wave amplitudes and propagation speeds are derived as a function of the porosity, and their variability due to the trabecular microstructure is revealed. To elucidate an effect of the microstructure, the mean intercept length (MIL), which is a microstructural parameter, is introduced, and the correlations of the propagation properties with the MILs of the trabecular elements and pore spaces are investigated.     

The correlation between the SOS in trabecular bone and stiffness and density studied by finite-element analysis

For the clinical assessment of osteoporosis (i.e., a degenerative bone disease associated with increased fracture risk), ultrasound has been proposed as an alternative or supplement to the dual-energy X-ray absorptiometry (DEXA) technique. However, the interaction of ultrasound waves with (trabecular) bone remains relatively poorly understood. The present study aimed to improve this understanding by simulating ultrasound wave propagation in 15 trabecular bone samples from the human lumbar spine, using microcomputed tomography-based finite-element modeling. The model included only the solid bone, without the bone marrow. Two structural parameters were calculated: the bone volume fraction (BV/TV) and the structural (apparent) elastic modulus (E_s), and the ultrasound propagation parameter speed of sound (SOS). Relations between BV/TV and E_s were similar to published experimental relations. At 1 MHz, correlations between SOS and the structural parameters BV/TV and E_s were rather weak, but the results can be explained from the specific features of the trabecular structure and the intrinsic material elastic modulus E_i. In particular, the systematic differences between the three main directions provide information on the trabecular structure. In addition, at 1 MHz the correlation found between the simulated SOS values and those calculated from the simple bar equation was poor when the three directions are considered separately. Hence, under these conditions, the homogenization approach - including the bar equation - is not valid. However, at lower frequencies (50-300 kHz) this correlation significantly improved. It is concluded that detailed analysis of ultrasound wave propagation through the solid structure in various directions and with various frequencies, can yield much information on the structural and mechanical properties of trabecular bone.     

Exploration of trabecular bone nonlinear elasticity using time-of-flight modulation

Bone tissue contains microcracks that may affect its mechanical properties as well as the whole trabecular structure. The relationship between crack density and bone strength is nevertheless poorly understood. Linear ultrasound techniques being almost insensitive to the level of damage, we propose a method to measure acoustic non- linearity in trabecular bone using time-of-flight modulation (TOFM) measurements. Ultrasonic short bursts times-of- flight (TOF) are modulated as a result of nonlinear interaction with a low-frequency (LF) wave in the medium. TOF variations are directly related to elastic modulus variations. Classical and nonclassical nonlinear parameters beta, delta, and alpha can be derived from these measurements. The method was validated in materials with classical, quadratic, nonlinear elasticity. In dense trabecular bone region, TOFM related to classical, quadratic, nonlinear elasticity as a function of the LF pressure exhibits tension-compression asymmetry. The TOFM amplitude measured in dense areas of trabecular bone is almost one order of magnitude higher than in a low-density area, but the linear parameters show much smaller variations: 5% for ultrasound propagation velocity and 100% for broadband ultrasonic attenuation (BUA). In high-density trabecular bone regions, beta depends on the LF pressure amplitude and can reach 400 at 50 kPa.     

Severe Cortical and Trabecular Osteopenia in Secondary Hyperparathyroidism

Background: Peripheral quantitative computed tomography (pQCT) provides real volumetric bone density values, not only of the total, but also of trabecular and cortical bone, separately. In addition, it provides data on bone geometry that can be related to the risk of fracture. Methods: Total, cortical, and trabecular volumetric bone mineral densities (BMD), as well as the main geometric parameters (cross‐sectional area, cortical area, trabecular area, and cortical thickness) were assessed by pQCT at the distal radius in 24 hemodialysis patients affected by severe secondary hyperparathyroidism (PTH, mean ± SD: 1444 ± 695 pg/mL). The strength‐strain index (SSI), a biomechanical parameter describing bone fragility, was also determined. Results: Compared with a control group of 64 healthy age‐matched subjects, volumetric BMD (mg/cm³) was significantly reduced in all patients (total BMD: 243 ± 87 vs. 405 ± 138, cortical BMD: 605 ± 218 vs. 856 ± 204, trabecular BMD: 95 ± 51 vs. 182 ± 75). Cortical area and cortical thickness showed significant modifications, while cross‐sectional area did not. SSI was significantly reduced (547 ± 125 vs. 927 ± 306 mm³). PTH levels showed a significant inverse correlation with cortical BMD (r = ?0.56), cortical thickness (r = ?0.46), cortical area (r = ?0.61), and SSI (r = ?0.54). Quantitative analysis of bone demonstrated cortical porosity. Conclusions: In dialysis patients with severe secondary hyperparathyroidism, pQCT showed a significant cortical osteopenia, associated with geometric and mechanical bone impairment. Interestingly, we also found a comparable deficit of trabecular bone, which may be related to the very high PTH levels. Generalized cortical thinning, intracortical porosity and cortical‐endosteal resorption (“trabecularization” of the cortical bone) are major determinants of reduced bone strength, which may be quantitated by pQCT.     

基于视觉模型的苏木精—伊红染色法染色病理切片骨小梁提取

为了提取HE染色骨组织切片的骨小梁,实现骨小梁面积定量检测,以极坐标扩散方式创建视觉模型,划分骨小梁与骨细胞、骨皮质的边界。提取背景某一像素的RGB值,把病理切片所有像素的RGB值对背景像素求取三维欧式距离,获得阈值,对图像二值化分割,通过区域填充获得骨小梁。结果表明视觉模型能较好地获得感兴趣区域的边界,计量的骨小梁面积精度高于常规软件,检测结果鲁棒性相对于常规软件提高了95%以上,人机交互少,降低了检测的盲目性,提高了检测速度。     

基于多特征发现的生理切片骨小梁识别技术研究

目标区域计算机自动识别是骨组织形态计量学的发展趋势。研究图像多特征信息,识别苏木素尹红染色病理切片中的骨小梁。分析骨组织病理切片HSL颜色空间通道的直方图特征,进行基于颜色通道的阈值分割,采用Lazy Snapping算法获得骨小梁深层次特征信息,改进扫描种子填充算法,使其融合目标区域多特征信息。实验结果表明改进的扫描种子填充算法将复杂图像骨小梁提取时间从常规软件的10多分钟提速到257ms。改进的扫描线算法,能快速获得稳健的目标区域。     

Stress analysis in bone tissue around single implants with different diameters and veneering materials: A 3-D finite element study

The aim of this study was to evaluate the stress distribution on bone tissue with a single prosthesis supported by implants of large and conventional diameter and presenting different veneering materials using the 3-D finite element method. Sixteen models were fabricated to reproduce a bone block with implants, using two diameters (3.75 × 10 mm and 5.00 × 10 mm), four different veneering materials (composite resin, acrylic resin, porcelain, and NiCr crown), and two loads (axial (200 N) and oblique (100 N)). For data analysis, the maximum principal stress and von Mises criterion were used. For the axial load, the cortical bone in all models did not exhibit significant differences, and the trabecular bone presented higher tensile stress with reduced implant diameter. For the oblique load, the cortical bone presented a significant increase in tensile stress on the same side as the loading for smaller implant diameters. The trabecular bone showed a similar but more discreet trend. There was no difference in bone tissue with different veneering materials. The veneering material did not influence the stress distribution in the supporting tissues of single implant-supported prostheses. The large-diameter implants improved the transference of occlusal loads to bone tissue and decreased stress mainly under oblique loads. Oblique loading was more detrimental to distribution stresses than axial loading.     

Hierarchical analysis and multi-scale modelling of rat cortical and trabecular bone

The aim of this study was to explore the hierarchical arrangement of structural properties in cortical and trabecular bone and to determine a mathematical model that accurately predicts the tissue''s mechanical properties as a function of these indices. By using a variety of analytical techniques, we were able to characterize the structural and compositional properties of cortical and trabecular bones, as well as to determine the suitable mathematical model to predict the tissue''s mechanical properties using a continuum micromechanics approach. Our hierarchical analysis demonstrated that the differences between cortical and trabecular bone reside mainly at the micro- and ultrastructural levels. By gaining a better appreciation of the similarities and differences between the two bone types, we would be able to provide a better assessment and understanding of their individual roles, as well as their contribution to bone health overall.     

Capillary action: enrichment of retention and habitation of cells via micro-channeled scaffolds for massive bone defect regeneration

The development of a biomaterial substitute that can promote bone regeneration in massive defects has remained as a significant clinical challenge even using bone marrow cells or growth factors. Without an active, thriving cell population present throughout and stable anchored to the construct, exceptional bone regeneration does not occur. An engineered micro-channel structures scaffold within each trabecular has been designed to overcome some current limitations involving the cultivation and habitation of cells in large, volumetric scaffolds to repair massive skeletal defect. We created a scaffold with a superior fluid retention capacity that also may absorb bone marrow cells and provide growth factor-containing body fluids such as blood clots and/or serum under physiological conditions. The scaffold is composed of 3 basic structures (1) porous trabecular network (300–400 μm) similar to that of human trabecular bones, (2) micro-size channels (25–70 μm) within each trabecular septum which mimic intra-osseous channels such as Haversian canals and Volkmann’s canals with body fluid access, diffusion, nutritional supply and gas exchange, and (3) nano-size pores (100–400 nm) on the surface of each septum that allow immobilized cells to anchor. Combinatorial effects of these internal structures result in a host-adapting construct that enhances cell retention and habitation throughout the 3 cm-height and 4 cm-length bridge-shaped scaffold.