Short-TE projection reconstruction NMR microscopy of trabecular bone.

The aim of this study was to assess the potential of projection reconstruction (PR) NMR microscopy in the quantitative evaluation of trabecular bone architecture. Short-TE PR spin-echo microimages were acquired at 7.05 T on normal bone explants. The main structural parameters such as bone volume fraction (BVF), trabecular thickness (Tb.Th.) and trabecular separation (Tb.Sp.) were obtained from the 3D microimages using the method of directed secants. Quantitative structural data were then compared with those derived from conventional spin-echo microimages. Our study indicates that projection reconstruction NMR microscopy promises to be more accurate than the conventional FTI method in the analysis of trabecular bone.     

Characterization of trabecular bone structure from high-resolution magnetic resonance images using fuzzy logic

The purpose of this work was to apply fuzzy logic image processing techniques to characterize the trabecular bone structure with high-resolution magnetic resonance images. Fifteen ex vivo high-resolution magnetic resonance images of specimens of human radii at 1.5 T and 12 in vivo high-resolution magnetic resonance images of the calcanei of peri- and postmenopausal women at 3 T were obtained. Soft segmentation using fuzzy clustering was applied to MR data to obtain fuzzy bone volume fraction maps, which were then analyzed with three-dimensional (3D) fuzzy geometrical parameters and measures of fuzziness. Geometrical parameters included fuzzy perimeter and fuzzy compactness, while measures of fuzziness included linear index of fuzziness, quadratic index of fuzziness, logarithmic fuzzy entropy, and exponential fuzzy entropy. Fuzzy parameters were validated at 1.5 T with 3D structural parameters computed from microcomputed tomography images, which allow the observation of true trabecular bone structure and with apparent MR structural indexes at 1.5 T and 3 T. The validation was statistically performed with the Pearson correlation coefficient as well as with the Bland-Altman method. Bone volume fraction correlation values (r) were up to .99 (P<.001) with good agreements based on Bland-Altman analysis showing that fuzzy clustering is a valid technique to quantify this parameter. Measures of fuzziness also showed consistent correlations to trabecular number parameters (r>.85; P<.001) and good agreements based on Bland-Altman analysis, suggesting that the level of fuzziness in high-resolution magnetic resonance images could be related to the trabecular bone structure.     

Interrelationships between 3-T-MRI-derived cortical and trabecular bone structure parameters and quantitative-computed-tomography-derivedbone mineral density

Recently, 3-T magnetic resonance imaging (MRI) has been introduced for bone imaging. Through higher signal-to-noise ratios, as compared to 1.5-T MRI, it promises to be a more powerful tool for the assessment of cortical and trabecular bone measures. The goal of our study was to compare MRI-derived cortical and trabecular bone measures to quantitative computed tomography (QCT)-derived bone mineral density (BMD). Using 3-T MRI in 51 postmenopausal women, apparent (app.) measures of bone volume/total volume, trabecular number (Tb.N), trabecular thickness (Tb.Th) and trabecular separation were derived at the distal radius, distal tibia and calcaneus. Cortical thickness (Ct.Th) was calculated at the distal radius and distal tibia. These measures were compared to QCT-derived BMD of the spine, hip and radius. Significant correlations (^?P<.05; ^??P<.001; ^???P<.0001) were found between spine BMD- and MRI-derived Ct.Th (r_radius=.55, ^?P<.05; r_tibia=.67, ^???P<.0001) and app. Tb.N (r_radius=.33, ^?P<.05; r_tibia=.35, ^?P<.05) at the radius and tibia. Furthermore, within the first 10 mm at the radius, an inverse correlation for Ct.Th and app. BV/TV (r_6mm=−.56, P<.001; r_10mm=−.36, P<.05) and app. Tb.Th (r_6mm=−.54, P<.001; r_10mm=−.41, P<.05) was found.     

The influence of the resolution and contrast on measuring the articular cartilage volume in magnetic resonance images

The progression of OA in patients may be followed by measuring the volume of articular cartilage from MR images. We attempted to determine the reproducibility of volume measurements of articular cartilage made from magnetic resonance images of the knees and the dependence of the reproducibility on image resolution and contrast-to-noise. A fat-suppressed 3D technique was used to generate four image sets with different image resolution. Each patient was imaged twice to obtain image pairs at each resolution. To assess the dependence of reproducibility on noise we generated six image sets for each patient by adding noise to the original images and repeating the comparison. On each image set, the femoral, tibial, and patellar cartilage were outlined by a combination of computer and manual methods, and the images were used to calculate the volume of each cartilage plate. Comparing the coefficient of variance between the volume measurements made from the two visits, the volume measurements made from images with the highest resolution (0.275 x 0.275 x 1.0 mm) had the highest reproducibility. The high resolution images of the tibia and femur had the least partial-volume averaging and, as a result, better defined the boundaries between cartilage and adjacent tissues. A different trend was evident for the patella. For studies of osteoarthritis therapies, we recommend using MR images with the highest possible in-plane spatial resolution to provide the most reproducible volume measurements of knee cartilage.     

X射线显微CT用于大鼠骨小梁结构分析的研究

应用X射线显微CT(X-μCT)对正常及骨质疏松大鼠的骨小梁结构进行了分析,并与骨组织形态计量法的测量值进行了比较,探讨了X射线光谱技术在骨结构分析中的应用。实验对大鼠骨样品进行X-μCT扫描,扫描条件为 80 kVp,80 μA,360°旋转,帧平均4帧,角度增益 0.4°,分辨率14 μm。三维重建并分析了骨小梁结构,结构参数包括骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)以及骨小梁间隔(Tb.Sp)。结果表明,采用X-μCT分析不同组大鼠的骨小梁结构参数值之间存在显著差异(P<0.05),测定值与传统骨组织形态计量法的测定值显著相关,其中胫骨骨小梁BV/TV,Tb.Th,Tb.N,Tb.Sp的相关系数r分别为0.984,0.960,0.995,0.988,腰椎骨小梁各结构参数的相关系数分别为0.938,0.968,0.877,0.951。因此,X-μCT可以较好地呈现并区分正常骨组织、骨质疏松骨组织以及经雌激素治疗后骨组织的微观结构,可以实现对骨小梁结构参数的分析测定,与骨组织形态计量法相比是一种更精确、立体、快速且无损测量骨微结构和评价骨质量的方法。     

Wavelet-based characterization of vertebral trabecular bone structure from magnetic resonance images at 3 T compared with micro-computed tomographic measurements

Trabecular bone structure and bone density contribute to the strength of bone and are important in the study of osteoporosis. Wavelets are a powerful tool in characterizing and quantifying texture in an image. The purpose of this study was to validate wavelets as a tool in computing trabecular bone thickness directly from gray-level images. To this end, eight cylindrical cores of vertebral trabecular bone were imaged using 3-T magnetic resonance imaging (MRI) and micro-computed tomography (microCT). Thickness measurements of the trabecular bone from the wavelet-based analysis were compared with standard 2D structural parameters analogous to bone histomorphometry (MR images) and direct 3D distance transformation methods (microCT images). Additionally, bone volume fraction was determined using each method. The average difference in trabecular thickness between the wavelet and standard methods was less than the size of 1 pixel size for both MRI and microCT analysis. A correlation (R) of .94 for microCT measurements and that of .52 for MRI were found for the bone volume fraction. Based on these results, we conclude that wavelet-based methods deliver results comparable with those from established MR histomorphometric measurements. Because the wavelet transform is more robust with respect to image noise and operates directly on gray-level images, it could be a powerful tool for computing structural bone parameters from MR images acquired using high resolution and thus limited signal scenarios.     

Comparison of high-resolution MRI, optical microscopy and SEM for quantitation of trabecular architecture in the rat femur

Magnetic resonance imaging (MRI) has been used to analyze trabecular bone architecture in femur heads taken from adult Wistar rats. The aim of this study was to validate the use of MRI in assessing trabecular structure and morphology by comparing standard measures of bone morphology in the rat femur obtained from high resolution MRI with those obtained by conventional optical microscopy and by scanning electron microscopy (SEM). MR images were obtained on a Bruker 4.7 T micro-imaging system using a three-dimensional spin echo sequence with spatial resolution of 23 microm in-plane and a slice thickness of 39 microm. Optical images were obtained by de-calcifying the bone in EDTA and then sectioning 5-microm-thick slices. SEM images were obtained from bone embedded in epoxy resin with surface preparation by diamond polishing. Values of standard bone morphological parameters were compared and correlation coefficients between the MRI and the optical- and SEM-derived measures of morphology were calculated. Partial volume effects in MRI were minimized in this study by the use of very thin slices, yielding better agreement with optical- and SEM-derived measures of trabecular bone morphology than have been obtained in previous studies. Correlations between the MRI and optical data were significantly lower than those between the MRI and SEM data. Effects of de-calcification were also investigated. The results indicate that comparison of MRI with thin (de-calcified) optical images may be inherently flawed due to the destructive de-calcification and sectioning process used to prepare samples for the optical imaging.     

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.     

In vitro ultrasonic characterization of human cancellous femoral bone using transmission and backscatter measurements: relationships to bone mineral density

Thirty-eight slices of pure trabecular bone 1-cm thickness were extracted from human proximal femurs. A pair of 1-MHz central frequency transducers was used to measure quantitative ultrasound (QUS) parameters in transmission [normalized broadband ultrasound attenuation (nBUA), speed of sound (SOS)] and in backscatter [broadband ultrasound backscatter (BUB)]. Bone mineral density (BMD) was measured using clinical x-ray quantitative computed tomography. Site-matched identical region of interest (ROIs) of 7 x 7 mm2 were positioned on QUS and QCT images. This procedure resulted in 605 ROIs for all the specimens data pooled together. The short-term precision of the technique expressed in terms of CV was found to be 2.3% for nBUA, 0.3% for SOS and 4.5% for BUB. Significant linear correlation between QUS and BMD were found for all the 605 ROIs pooled, with r2 values of 0.73, 0.77, and 0.58 for nBUA, SOS, and BUB, respectively (all p < 0.05). For the BUB, the best regression was obtained with a polynomial fit of second order (r2 = 0.63). An analysis of measurements errors was developed. It showed that the residual variability of SOS is almost completely predicted by measurements errors, which is not the case for BUA and BUB, suggesting a role for micro-architecture in the determination of BUA and BUB.     

Surface bioactivity modification of titanium by CO2 plasma treatment and induction of hydroxyapatite: In vitro and in vivo studies

Since metallic biomaterials used for orthopedic and dental implants possess a paucity of reactive functional groups, bioactivity modification of these materials is challenging. In the present work, the titanium discs and rods were treated with carbon dioxide plasma and then incubated in a modified simulated body fluid 1.5SBF to obtain a hydroxyapatite layer. Surface hydrophilicity of samples, changes of surface chemistry, surface morphologies of samples, and structural analysis of formed hydroxyapatite were investigated by contact angle to water, X-ray photoelectron spectrometer (XPS), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and X-ray diffraction (XRD). The results demonstrated that hydrophilicity of titanium surface was improved and hydroxyl groups increased after modification with carbon dioxide plasma treatment. The hydroxyl groups on the surface of titanium were the richest after carbon dioxide plasma treatment under the condition of 20 W for less than 30 s. The hydroxyapatite formability of titanium surface was enhanced by carbon dioxide plasma pretreatment, which was attributed to the surface chemistry. MC3T3-E1 cell as a model cell was cultured on the Ti, CPT-Ti and CPT/SBF-Ti discs in vitro, and the results of the morphology and differentiation of the cell showed that CPT/SBF-Ti was the highest bioactive. The relative parameters of the new bone around the Ti and CPT/SBF-Ti rods including bone mineral density (BMD), a ratio of bone volume to total volume (BV/TV), trabecular thickness (Tb.Th.) and trabecular number (Tb.N.) were analyzed by a micro-computed tomography (micro-CT) after 4-, 8- and 12-week implantation periods in vivo. The results indicated that the CPT/SBF-Ti was more advantageous for new bone formation.     

MR phase imaging to quantify bone volume fraction: computer simulations and in vivo measurements

Magnetic resonance phase images can be used to assess trabecular bone by measuring the standard deviation of the phases in a region of interest. The standard deviation of regional phase measurements reflects the degree of magnetic field inhomogeneity caused by susceptibility differences between bone and marrow. A 3D computer model of trabecular bone was developed and then used to explore the influence of bone volume fraction and imaging parameters such as pixel size and slice width on the standard deviation of regional phase measurements. The results from these tests show that with appropriate selection of these parameters, phase spread strongly reflects variations in trabecular bone density (a correlation of R(2) = 0.98 with bone volume fraction between 0 and 10%). The technique was then applied in vivo on the radius of 25 patients who already had a bone density scan with peripheral quantitative tomography and a correlation between phase standard deviation and trabecular bone density was found (R(2) = 0.46).     

Inter-rater and intra-rater reliability of subchondral bone and cartilage thickness measurement from MRI

MRI is often used to visualize and quantify the articular cartilage layer of load bearing joints affected by degenerative diseases, such as osteoarthritis (OA). Although the role played by the subchondral bone in the etiology and/or progression of OA may be important, the ability to visualize and quantify subchondral bone with MRI has received little attention. In this report we examined the inter-rater and intra-rater reliability of subchondral bone and cartilage thickness measurements from MR images of cadaver femoral head specimens. A 3D-SPGR pulse sequence tuned to eliminate chemical shift artifact through phase cancellation was used to image the specimens. Three raters manually segmented four specimens on two different occasions. Subchondral bone and cartilage thickness measurements were calculated from the segmented images. Inter-rater and intra-rater reliabilities were very high (>.98) for both cartilage and subchondral bone thickness measurements. We conclude that subchondral bone thickness can be measured as reliably as cartilage thickness from MR images.     

Effects of risedronate in a rat model of osteopenia due to orchidectomy and disuse: densitometric, histomorphometric and microtomographic studies

Dual energy X-ray absorptiometry (DXA), histomorphometry and X-ray microtomography (microCT) were used to assess effects of risedronate and testosterone in a combined rat model of orchidectomy (ORX) and local paralysis induced by botulinum neurotoxin (BTX). Four groups of mature rats were studied for 1 month: SHAM operated; ORX and right hindlimb immobilization (BTX); ORX+BTX+risedronate or testosterone. Changes in bone and body composition were measured by DXA (BMC, lean and fat mass), histomorphometry (BV/TV(2D), Tb.Th and microarchitectural parameters) and microCT (BV/TV(3D), SMI and cortical parameters). ORX and BTX had additive effects on bone loss since differences were maximized on the immobilized bone. The decrease in BMC on the tibial metaphysis reached -33.6% vs. -11.3% in the non-immobilized limb. BV/TV and Tb.N decreased and Tb.Sp increased in both hindlimbs whereas Tb.Th was significantly lower only in the immobilized limb. Decrease of tibial cortical area and thickness was greater in the immobilized limb. Risedronate prevented BMC, BV/TV and architecture loss but not reduction in Tb.Th. Cortical bone was preserved only in the non-immobilized limb. Testosterone was unable to prevent trabecular and cortical bone loss, but it prevents loss of whole body lean mass. In conclusion, ORX and BTX resulted in additive effects on bone loss. Risedronate had protective effects on trabecular bone loss but was less effective on cortical 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.     

Helmholtz-pair transmit coil with integrated receive array for high-resolution MRI of trabecular bone in the distal tibia at 7T

A Helmholtz-pair local transmit RF coil with an integrated four-element receive array RF coil and foot immobilization platform was designed and constructed for imaging the distal tibia in a whole-body 7T MRI scanner. Simulations and measurements of the B(1) field distribution of the transmit coil are described, along with SAR considerations for operation at 7T. Results of imaging the trabecular bone of three volunteers at 1.5T, 3T and 7T are presented, using identical 1.5T and 3T versions of the 7T four-element receive array. The spatially registered images reveal improved visibility for individual trabeculae and show average gains in SNR of 2.8× and 4.9× for imaging at 7T compared to 3T and 1.5T, respectively. The results thus display an approximately linear dependence of SNR with field strength and enable the practical utility of 7T scanners for micro-MRI of trabecular bone.     

Interobserver reproducibility of quantitative cartilage measurements: comparison of B-spline snakes and manual segmentation.

The objective of this work was to develop a segmentation technique for thickness measurements of the articular cartilage in MR images and to assess the interobserver reproducibility of the method in comparison with manual segmentation. The algorithm is based on a B-spline snakes approach and is able to delineate the cartilage boundaries in real time and with minimal user interaction. The interobserver reproducibility of the method, ranging from 3.3 to 13.6% for various section orientations and joint surfaces, proved to be significantly superior to manual segmentation.     

Relationship between cancellous bone induced magnetic field and ultrastructure in a rat ovariectomy model

The site-dependent variations in trabecular bone morphology were studied in the rat tibia by magnitude and phase difference three-dimensional nuclear magnetic resonance microscopy and image processing, and the implications of ovariectomy were evaluated. Specimens excised from the proximal tibial metaphysis in ovariectomized (n = 7) and intact control (n = 4) rats were imaged at 9.4T with their anatomic axes parallel to the direction of the magnetic field. An echo-offset 3D rapid spin-echo excitation pulse sequence was used to generate phase difference maps, from which the standard deviation of the phase difference, sigma(delta psi), was calculated. In addition, a fictitious rate constant, R2', was calculated from the slope of the exponential portion of the Fourier transform of the phase difference histogram. Trabecular bone volume fraction was also determined in the same volume of interest. The results show strong correlations between bone volume fraction and both sigma(delta psi) and R2', suggesting that these parameters could be useful for nondestructive assessment of trabecular bone volume.     

Optimization and validation of a rapid high-resolution T1-w 3D FLASH water excitation MRI sequence for the quantitative assessment of articular cartilage volume and thickness

In view of follow up, survey and development of therapeutic strategies for osteoarthritis where cartilage deterioration plays an important role, a non invasive, reliable and quantitative assessment of the articular cartilage is desirable. The currently available high resolution T(1)-weighted (T1-w) 3D FLASH pulse sequences with frequency selective fat suppression are very time consuming. We have 1) optimized a high resolution T1-w 3D FLASH water excitation (WE) sequence for short acquisition time and cartilage visualization, and 2) validated this sequence for cartilage volume and thickness quantification. The spectral fat presaturation was replaced by selective water excitation. The flip angle of the WE sequence was optimized for the contrast to noise (C/N(cart)) ratio of cartilage. Sagittal datasets (voxel size: 0.31 x 0.31 x 2 mm(3)) of the knees of nine healthy volunteers were acquired both, with the 3D FLASH WE (17.2/6.6/30 degrees ) sequence (WE) and a previously validated 3D FLASH fat saturated (42/11/30 degrees ) sequence (FS). For validation of the WE sequence, cartilage volume, mean and maximal cartilage thickness of the two sequences were compared. Reproducibility was assessed by calculating the coefficient of variation (COV %) of 4 consecutive WE data sets in the volunteers. The acquisition time was reduced from 16'30" (FS) down to 7'14" for the WE sequence. Image contrast and visualization of the cartilage was very similar, but delineation of the basal layer of the cartilage was slightly improved with the WE sequence. A flip angle of 30 degrees provided the best C/N(cart) ratios (WE). Reproducibility (COV) was between 1.9 and 5.9%. Cartilage volume and thickness agreed within 4% between FS and WE sequence. The WE sequence allows for rapid, valid and reproducible quantification of articular cartilage volume and thickness, prerequisites for follow-up examinations. The reduced acquisition time (50% of FS) enables routine clinical application and thus may contribute to a broader assessment of osteoarthritis.     

Dependence of threshold variations on the assessment of histomorphometric indices from computed microtomography using synchrotron radiation

In this work, the variation of threshold level for segmentation of tomographic image in a micron scale accomplished with synchrotron radiation was studied. Three different sites of rat femora were evaluated from computed microtomography performed at SYRMEP beamline of ELETTRA Synchrotron Laboratory with a resolution of 9 μm. An optimal threshold level was selected and then varied in two points up and down in steps of one point, corresponding to 0.39% per point. Histomorphometric indices were calculated directly from the three dimensional segmented images of trabecular microstructure. The results showed that histomorphometric indices presented small variations for all indices in all femur sites. The greater variation occurred for trabecular thickness (Tb.Th) [from ?2.84% to 2.99%] while the smaller one occurred for trabecular number (Tb.N) [?0.26% to 0.35].     

Influence of the precision of spectral backscatter measurements on the estimation of scatterers size in cancellous bone

The goal of this study is to propose a model for the ultrasonic frequency-dependent backscatter coefficient in femoral cancellous bone. This model has been developed with success to predict backscatter in human calcaneal bone [Jenson, Ultr. Med. Biol. 2003]. A weak scattering model is used and the backscatter coefficient is expressed in terms of a Gaussian autocorrelation function of the medium. The backscatter coefficient is computed and comparison is made with experimental data for 37 specimens and for frequency ranging from 0.4 to 1.2 MHz. An excellent agreement between experimental data and predictions is found for both the magnitude and the frequency-dependence of the backscatter coefficient. Then, a nonlinear regression is performed for each specimen, and the mean trabecular thickness is estimated. Experimental data and theoretical predictions are averaged over the 37 specimens. We also find a close agreement between theoretical predictions obtained using the Gaussian autocorrelation function (scatterer size=134+/-15 microm) and the mean trabecular thickness (Tb.Th=132+/-12 microm) derived from the analysis of bone 3-D micro-architecture using high-resolution micro-tomography. However, the correlation between individual experimental and estimated Tb.Th values is moderate (R(2)=0.44). The performance of the estimator are limited mainly by two factors: interference noise due to random positioning of the scatterers and attenuation. We show that the fundamental limitation of our estimator due to the speckle noise is around 5 microm for trabecular thickness estimation. This limitation is lower than the observed biological variability which is around 30 microm and should not be a limiting factor for individual prediction. A second limitation is the tremendous attenuation encountered in highly scattering media such as cancellous bone, which results in highly damped backscatter signals. The compensation for attenuation is difficult to perform, and it may be a critical point that limits the precision of the estimator.