Ultrasonic characterization of orthotropic elastic bovine bones

The aim of the present study was to determine the mechanical properties of bovine bones. An ultrasonic method was used to determine acoustical parameters such as the longitudinal and transverse velocities in the longitudinal and two radial directions of compact bone, i.e., in all directions of the plane. Waves propagating through bovine femoral bones were studied using an ultrasonic scanner for linear and sectorial scanning. The mechanical parameters of compact bone, such the Young's modulus and Poisson's ratio in the orthotropic case, were then determined from the measured velocities. The results are in line with those in the literature.     

Quantification of guided mode propagation in fractured long bones

Guided modes propagation in intact, fractured and healing long bone has drawn significant research interests. However, mode quantifications for the direct comparison are still necessary to address. The aim of the study is to analyze the mode interaction with a notch-fracture in the long bone and find quantitative ultrasound parameters sensitive to depth and width variation of the fracture. We analyzed the impacts of the partially and completely diaphyseal osteotomy on fundamental guided modes propagation using the two-dimension finite-difference time-domain (2D-FDTD) simulations. The long bones were built as three layer models by a cortical plate embedded between overlying soft tissue and inner-coated marrow. Narrowband low-frequency sinusoids (100 kHz) were employed to only excite two fundamental guided modes. The mode amplitude variations were investigated as functions of the gap-breakage width and depth. It is found that the transverse fractures have strong influences on the anti-symmetric mode A0 transmission and reflection, whereas amplitudes of the symmetric mode S0 are not sensitive to the fracture degree. The quantitative results consistently indicate that reflection energy and transmission coefficients of the S0 and A0 modes can be used to quantify the mode interaction in the fractured long bone and further to evaluate long bone fracture status. Future study is needed to investigate the physical experiments on realistic fractured long bone and to insure that the proposed ultrasound parameters can be used to quantitatively evaluate the long bone fracture in clinical application.     

Mineralogical changes of bioapatite in femoral bones of mice during pregnancy

Mineralogical changes and solubility of bioapatite in mice cortical bones (2- and 5 months old) were investigated by Raman spectroscopy and inductively coupled plasma spectrometry in this study. The Raman analyses show that the full width at half maximum values of the 960?cm^?1 peak exhibit a slight increase during mouse pregnancy, which suggests slightly decreasing crystallinity. Additionally, aging significantly increases the solubility of bone bioapatite. The 1070?cm^?1 peak indicates the enriched carbonate incorporation during aging from 2 to 5 months. The elevated solubility of bioapatite and low bone density are consistent with the low crystallinity of bioapatite.     

Skeletal muscle contraction in protecting joints and bones by absorbing mechanical impacts

We have previously hypothesized that the dissipation of mechanical energy of external impact is a fundamental function of skeletal muscle in addition to its primary function to convert chemical energy into mechanical energy. In this paper, a mathematical justification of this hypothesis is presented. First, a simple mechanical model, in which the muscle is considered as a simple Hookean spring, is considered. This analysis serves as an introduction to the consideration of a biomechanical model taking into account the molecular mechanism of muscle contraction, kinetics of myosin bridges, sarcomere dynamics, and tension of muscle fibers. It is shown that a muscle behaves like a nonlinear and adaptive spring tempering the force of impact and increasing the duration of the collision. The temporal profiles of muscle reaction to the impact as functions of the levels of muscle contraction, durations of the impact front, and the time constants of myosin bridges closing, are obtained. The absorption of mechanical shock energy is achieved due to the increased viscoelasticity of the contracting skeletal muscle. Controlling the contraction level allows for the optimization of the stiffness and viscosity of the muscle necessary for the protection of the joints and bones.     

Detection of long bones based on Barker code excited ultrasonic guided waves

Single pulse excited ultrasonic guided wave surfers high attenuation during the propagation in long bones.This results in small amplitude and low signal-to-noise ratio(SNR)of measured signals.Thus,the Barker code excitation is introduced into long bone detection to improve the quality of received signals,due to its efficiency in increasing amplitude and SNR.Both simulation and in vitro experiment were performed,and the results were decoded by the weighted match filter(WMF) and the finite impulse response- least squares inverse filter(FIRLSIF),respectively.The comparison between the results of Barker code excitation and sine pulse excitation was presented.For 13-bit Barker code excitation,WMF produced 13 times larger amplitude than sine pulse excitation,while FIR-LSIF achieved higher peak-sidelobe-level(PSL) of —63.59 dB and better performance in noise suppression.The results show that the Barker code excited guided waves have the potential to be applied to the long bone detection.     

The spectrum entropy estimation method for ultrasonic backscattering signal in cancellous bones

In the application of cancellous bone ultrasound diagnosis based on backscattering method,it is of great importance to estimate fast and accurately whether the valid backscattering signal exists in the received signal.We propose a fast estimation method based on spectrum entropy method.With 984 records of adult calcaneus clinical data,we estimate the validity of the backscatter signal using this method.The results of the proposed method and the results of experience-base judgement were compared and analyzed.And two key parameters,the signal range length and the segment number of the spectrum entropy,were analyzed.The results show when the signal range length is 13 us and the segment number is 15-20,this method can get the best result(accuracy95%,sensitivity99%,specificity87%),while taking little calculation time(1.5 ms).Therefore,this spectrum entropy method can satisfy the accuracy and real-time requirements in the ultrasonic estimation for cancellous bone.     

X-ray imaging characterization of femoral bones in aging mice with osteopetrotic disorder

Aging mice with a rare osteopetrotic disorder in which the entire space of femoral bones are filled with trabecular bones are used as our research platform. A complete study is conducted with a micro computed tomography (CT) system to characterize the bone abnormality. Technical assessment of femoral bones includes geometric structure, biomechanical strength, bone mineral density (BMD), and bone mineral content (BMC). Normal aging mice of similar ages are included for comparisons. In our imaging work, we model the trabecular bone as a cylindrical rod and new quantitative which are not previously discussed are developed for advanced analysis, including trabecular segment length, trabecular segment radius, connecting node number, and distribution of trabecular segment radius. We then identified a geometric characteristic in which there are local maximums (0.0049, 0.0119, and 0.0147 mm) in the structure of trabecular segment radius. Our calculations show 343% higher in percent trabecular bone volume at distal-metaphysis; 38% higher in cortical thickness at mid-diaphysis; 11% higher in cortical cross-sectional moment of inertia at mid-diaphysis; 42% higher in cortical thickness at femur neck; 26% higher in cortical cross-sectional moment of inertia at femur neck; 31% and 395% higher in trabecular BMD and BMC at distal-metaphysis; 17% and 27% higher in cortical BMD and BMC at distal-metaphysis; 9% and 53% higher in cortical BMD and BMC at mid-diaphysis; 25% and 64% higher in cortical BMD and BMC at femur neck. Our new quantitative parameters and findings may be extended to evaluate the treatment response for other similar bone disorders.     

Experimental aspect of solid-state nuclear magnetic resonance studies of biomaterials such as bones

Solid-state nuclear magnetic resonance (SSNMR) spectroscopy is increasingly becoming a popular technique to probe micro-structural details of biomaterial such as bone with pico-meter resolution. Due to high-resolution structural details probed by SSNMR methods, handling of bone samples and experimental protocol are very crucial aspects of study. We present here first report of the effect of various experimental protocols and handling methods of bone samples on measured SSNMR parameters. Various popular SSNMR experiments were performed on intact cortical bone sample collected from fresh animal, immediately after removal from animal systems, and results were compared with bone samples preserved in different conditions. We find that the best experimental conditions for SSNMR parameters of bones correspond to preservation at −20 °C and in 70% ethanol solution. Various other SSNMR parameters were compared corresponding to different experimental conditions. Our study has helped in finding best experimental protocol for SSNMR studies of bone. This study will be of further help in the application of SSNMR studies on large bone disease related animal model systems for statistically significant results.     

3D histomorphometric quantification of trabecular bones by computed microtomography using synchrotron radiation

Conventional bone histomorphometry is an important method for quantitative evaluation of bone microstructure. X-ray computed microtomography is a non-invasive technique, which can be used to evaluate histomorphometric indices in trabecular bones (BV/TV, BS/BV, Tb.N, Tb.Th, Tb.Sp). In this technique, 3D images are used to quantify the whole sample, differently from the conventional one, in which the quantification is performed in 2D slices and extrapolated for 3D case. In this work, histomorphometric quantification using synchrotron 3D X-ray computed microtomography was performed to quantify the bone structure at different skeletal sites as well as to investigate the effects of bone diseases on quantitative understanding of bone architecture. The images were obtained at Synchrotron Radiation for MEdical Physics (SYRMEP) beamline, at ELETTRA synchrotron radiation facility, Italy. Concerning the obtained results for normal and pathological bones from same skeletal sites and individuals, from our results, a certain declining bone volume fraction was achieved. The results obtained could be used in forming the basis for comparison of the bone microarchitecture and can be a valuable tool for predicting bone fragility.     

Effect of selected signals of interest on ultrasonic backscattering measurement in cancellous bones

This study examined how the signals of interest (SOI) effect on the backscattering measurement numerically based on 3-D finite-difference time-domain (FDTD) method. High resolution microstructure mappings of bovine cancellous bones provided by micro-CT were used as the input geometry for simulations. Backscatter coefficient (BSC), integrated backscatter coefficient (IBC) and apparent integrated backscatter (AIB) were calculated with changing the start (L1) and duration (L2) of the SOI. The results demonstrated that BSC and IBC decrease as L1 increases, and AIB decreases more rapidly as L1 increases. The backscattering parameters increase with fluctuations as a function of L2 when L2 is less than 6 mm. However, BSC and IBC change little as L2 continues to increase, while AIB slowly decreases as L2 continues to increase. The results showed how the selections of the SOI effect on the backscattering measurement. An explicit standard for SOI selection was proposed in this study and short L1 (about 1.5 mm) and appropriate L2 (6 mm-12 mm) were recommended for the calculations of backscattering parameters.     

MR imaging of fatigue stress injuries to bones: intra- and interobserver agreement

The aim of this study was to determine the validity of MR imaging (MRI) in the assessment of stress-related injuries to bone.MR images of 50 military recruits (8 females and 42 males; 18-27 (mean 20) years of age) were retrospectively evaluated twice for stress injuries to bone by 4 radiologists (2 musculoskeletal radiologists, 2 radiology residents). Coronal T1-weighed (T1W) and STIR images, as well as axial and coronal T2-weighted (T2W) fat-suppressed images were taken using a 1.0T scanner. Rates for sensitivity, specificity, and accuracy of MRI of the stress-related injuries were calculated. Intraobserver and interobserver agreement was determined with kappa statistics.Rates for MRI sensitivity were 27-96%, for specificity 65-100%, and for diagnostic accuracy 58-97%. Lowest rates were seen when reading T1W images and highest when reading STIR images. Readers showed moderate to excellent intraobserver agreement (kappa 0.75-0.95). Interobserver agreement was fair to excellent (kappa 0.41-0.91), and the lowest values were seen in the interpretation of T1W images. Normal findings could be differentiated from various grades of stress injury to bone.MRI is a valid means of revealing the presence of stress injuries to bone and their staging. Observer agreement is good to excellent when using T2W images and STIR images, while T1W images are of lesser value.     

Ultrasonically determined thickness of long cortical bones: two-dimensional simulations of in vitro experiments

Previously it has been demonstrated that cortical bone thickness can be estimated from ultrasonic guided-wave measurements, in an axial transmission configuration, together with an appropriate analytical model. This study considers the impact of bone thickness variation within the measurement region on the ultrasonically determined thickness (UTh). To this end, wave velocities and UTh were determined from experiments and from time-domain finite-difference simulations of wave propagation, both performed on a set of ten human radius specimens (29 measurement sites). A two-dimensional numerical bone model was developed with tunable material properties and individualized geometry based on x-ray computed-tomography reconstructions of human radius. Cortical thickness (CTh) was determined from the latter. UTh data for simulations were indeed in a excellent accordance (root-mean-square error was 0.26 mm; r2=0.94, p<0.001) with average CTh within the measurement region. These results indicate that despite variations in cortical thickness along the propagation path, the measured phase velocity can be satisfactorily modeled by a simple analytical model (the A(0) plate mode in this case). Most of the variability (up to 85% when sites were carefully matched) observed in the in vitro ultrasound data was explained through simulations by variability in the cortical thickness alone.     

Ultrasonically determined thickness of long cortical bones: Three-dimensional simulations of in vitro experiments

It was reported in a previous study that simulated guided wave axial transmission velocities on two-dimensional (2D) numerically reproduced geometry of long bones predicted moderately real in vitro ultrasound data on the same bone samples. It was also shown that fitting of ultrasound velocity with simple analytical model yielded a precise estimate (UTh) for true cortical bone thickness. This current study expands the 2D bone model into three dimensions (3D). To this end, wave velocities and UTh were determined from experiments and from time-domain finite-difference simulations of wave propagation, both performed on a collection of 10 human radii (29 measurement sites). A 3D numerical bone model was developed with tuneable fixed material properties and individualized geometry based on X-ray computed tomography reconstructions of real bones. Simulated UTh data were in good accordance (root-mean-square error was 0.40 mm; r(2)=0.79, p<0.001) with true cortical thickness, and hence the measured phase velocity can be well estimated by using a simple analytical inversion model also in 3D. Prediction of in vitro data was improved significantly (by 10% units) and the upgraded bone model thus explained most of the variability (up to 95% when sites were carefully matched) observed in in vitro ultrasound data.     

Use of multiple acoustic wave modes for assessment of long bones: model study

Multiple acoustic wave mode method has been proposed as a new modality in axial bone QUS. The new method is based on measurement of ultrasound velocity at different ratio of wavelength to the bone thickness, and taking into account both bulk and guided waves. It allows assessment of changes in both the material properties related to porosity and mineralization as well as the cortical thickness influenced by resorption from inner layers, which are equally important in diagnostics of osteoporosis and other bone osteopenia. Developed method was validated in model studies using a dual-frequency (100 and 500 kHz) ultrasound device. Three types of bone phantoms for long bones were developed and tested: (1) tubular specimens from polymer materials to model combined changes of material stiffness and cortical wall thickness; (2) layered specimens to model porosity in compact bone progressing from endosteum towards periosteum; (3) animal bone specimens with both cortical and trabecular components. Observed changes of the ultrasound velocity of guided waves at 100 kHz followed gradual changes in the thickness of the intact cortical layer. On the other hand, the bulk velocity at 500 kHz remained nearly constant at the different cortical layer thickness but was affected by the material stiffness. Similar trends were observed in phantoms and in fragments of animal bones.     

Prediction of backscatter coefficient in trabecular bones using a numerical model of three-dimensional microstructure

A model of ultrasonic backscattering for cancellous bone saturated by water is proposed. This model assumes that scattering is caused by the solid trabeculae and describes the cancellous bone as a weak scattering medium. The backscatter coefficient is related to the spatial Fourier transform of bone microarchitecture and to the density and compressibility fluctuations between the solid trabeculae and the saturating fluid. The computations of the model make use of three-dimensional numerical images of bone microarchitecture, obtained by tomographic reconstructions with a 10 microm spatial resolution. With this model, the predictions of the frequency dependence and of the magnitude of the backscatter coefficient are reasonably accurate. The theoretical predictions are compared to experimental data obtained on 19 specimens. An accuracy error of approximately 1 dB was found (difference between the averaged experimental values and theoretical predictions). One limit of the model may come from inaccurate values of trabecular bone characteristics needed for the computations (density and longitudinal velocity), which are yet to be precisely determined for human trabecular bone. However, the model is only slightly sensitive to variations of bone material properties. It was found that an accuracy error of 2.2 dB at maximum resulted from inaccurate a priori values of bone material properties. A computation of the elastic mean free path in the medium suggests that multiple scattering plays a minor role in the working frequency bandwidth (0.4-1.2 MHz). It follows from these results that a weak scattering medium model may be appropriate to describe scattering from trabecular bone.     

Joint spectrogram segmentation and ridge-extraction method for separating multimodal guided waves in long bones

Ultrasonic guided waves (GWs) can be used to evaluate long bones effectively because of the ability to provide the information of the whole bone. In this study, a joint spectrogram segmentation and ridge-extraction (JSSRE) method was proposed to separate multiple modes in long bones. First, the Gabor time-frequency transform was applied to obtain the spectrogram of multimodal signals. Then, a multi-class image segmentation algorithm was used to find the corresponding region of each mode in the spectrogram, including an improved watershed transform and a region growing procedure. Finally, the ridges were extracted and the time domain signals representing individual modes were reconstructed from these ridges in each region. The validations of this method were discussed by simulated multimodal signals with different signal-to-noise ratios (SNR). The correlation coefficients between the original signals without noise and the reconstructed signals were calculated to analyze the results quantitatively. The results showed that the extracted ridges were in good agreement with generated theoretical dispersion curves, and the reconstructed signals were highly related to the original signals, even under the SNR=3 dB situation.     

Middle ear cavity and ear canal pressure-driven stapes velocity responses in human cadaveric temporal bones

Drive pressure to stapes velocity (V(st)) transfer function measurements are collected and compared for human cadaveric temporal bones with the drive pressure alternately on the ear canal (EC) and middle ear cavity (MEC) sides of the tympanic membrane (TM), in order to predict the performance of proposed middle-ear implantable acoustic hearing aids, as well as provide additional data for examining human middle ear mechanics. The chief finding is that, in terms of the V(st) response, MEC stimulation performs at least as well as EC stimulation below 8 kHz, provided that the EC is unplugged. Plugging the EC causes a reduced response for MEC drive below 2 kHz, due to a corresponding reduction of the pressure difference between the two sides of the TM. Between 8 and 11 kHz, the MEC drive transfer functions feature an approximately 17 dB drop in magnitude below the EC drive case, the cause of which remains unknown. The EC drive transfer functions reported here feature significantly less magnitude roll-off above 1 kHz than previous studies [with a slope of -2.3 vs -6.7 dB/octave for Aibara et al., Hear. Res. 152, 100-109 (2001)], and significantly more phase group delay (134 vs 62 micros for Aibara et al.).     

Application of an effective medium theory for modeling ultrasound wave propagation in healing long bones

Quantitative ultrasound has recently drawn significant interest in the monitoring of the bone healing process. Several research groups have studied ultrasound propagation in healing bones numerically, assuming callus to be a homogeneous and isotropic medium, thus neglecting the multiple scattering phenomena that occur due to the porous nature of callus. In this study, we model ultrasound wave propagation in healing long bones using an iterative effective medium approximation (IEMA), which has been shown to be significantly accurate for highly concentrated elastic mixtures. First, the effectiveness of IEMA in bone characterization is examined: (a) by comparing the theoretical phase velocities with experimental measurements in cancellous bone mimicking phantoms, and (b) by simulating wave propagation in complex healing bone geometries by using IEMA. The original material properties of cortical bone and callus were derived using serial scanning acoustic microscopy (SAM) images from previous animal studies. Guided wave analysis is performed for different healing stages and the results clearly indicate that IEMA predictions could provide supplementary information for bone assessment during the healing process. This methodology could potentially be applied in numerical studies dealing with wave propagation in composite media such as healing or osteoporotic bones in order to reduce the simulation time and simplify the study of complicated geometries with a significant porous nature.     

Three-dimensional finite element modeling of guided ultrasound wave propagation in intact and healing long bones

The use of guided waves has recently drawn significant interest in the ultrasonic characterization of bone aiming at supplementing the information provided by traditional velocity measurements. This work presents a three-dimensional finite element study of guided wave propagation in intact and healing bones. A model of the fracture callus was constructed and the healing course was simulated as a three-stage process. The dispersion of guided modes generated by a broadband 1-MHz excitation was represented in the time-frequency domain. Wave propagation in the intact bone model was first investigated and comparisons were then made with a simplified geometry using analytical dispersion curves of the tube modes. Then, the effect of callus consolidation on the propagation characteristics was examined. It was shown that the dispersion of guided waves was significantly influenced by the irregularity and anisotropy of the bone. Also, guided waves were sensitive to material and geometrical changes that take place during healing. Conversely, when the first-arriving signal at the receiver corresponded to a nondispersive lateral wave, its propagation velocity was almost unaffected by the elastic symmetry and geometry of the bone and also could not characterize the callus tissue throughout its thickness. In conclusion, guided waves can enhance the capabilities of ultrasonic evaluation.     

Study of fossil bones by synchrotron radiation micro‐spectroscopic techniques and scanning electron microscopy

Earlymost Villafranchian fossil bones of an artiodactyl and a perissodactyl from the Milia excavation site in Grevena, Greece, were studied in order to evaluate diagenetic effects. Optical microscopy revealed the different bone types (fibro‐lamellar and Haversian, respectively) of the two fragments and their good preservation state. The spatial distribution of bone apatite and soil‐originating elements was studied using micro‐X‐ray fluorescence (µ‐XRF) mapping and scanning electron microscopy. The approximate value of the Ca/P ratio was 2.2, as determined from scanning electron microscopy measurements. Bacterial boring was detected close to the periosteal region and Fe bearing oxides were found to fill bone cavities, e.g. Haversian canals and osteocyte lacunae. In the perissodactyl bone considerable amounts of Mn were detected close to cracks (the Mn/Fe weight ratio takes values up to 3.5). Goethite and pyrite were detected in both samples by means of metallographic microscopy. The local Ca/P ratio determined with µ‐XRF varied significantly in metal‐poor spots indicating spatial inhomogeneities in the ionic substitutions. XRF line scans that span the bone cross sections revealed that Fe and Mn contaminate the bones from both the periosteum and medullar cavity and aggregate around local maxima. The formation of goethite, irrespective of the local Fe concentration, was verified by the Fe K‐edge X‐ray absorption fine structure (XAFS) spectra. Finally, Sr K‐edge extended XAFS (EXAFS) revealed that Sr substitutes for Ca in bone apatite without obvious preference to the Ca₁ or Ca₂ unit‐cell site occupation.