A degenerative medial meniscus retains some protective effect against osteoarthritis-induced subchondral bone changes.

OBJECTIVES: The objective was to estimate the impact of the meniscus on cartilage and subchondral bone in knee osteoarthritis (OA). METHODS: In a sample of 46 knee specimens (26 females), 13 (7 females) were classified as OA according to the Kellgren-Lawrence classification. Outerbridge and meniscal grading were performed. Using micro-computed tomography images, we analyzed the cartilage thickness, subchondral plate thickness and micro-architecture of trabecular subchondral bone at different depths and in two different locations of the medial tibial plateau: one peripheral (PER) covered by the meniscus and one central (CENT) uncovered by the meniscus. RESULTS: Uncoverage by the meniscus was associated with bone sclerosis, defined as higher bone volume to total volume (BV/TV), higher trabecular number, thicker trabeculae with lower spacing, and anisotropy and a more plate-like architecture. The protective effect of meniscal coverage was observed in the uppermost 5 to 6 mm of the subchondral bone. As compared with normal knees, knees with OA showed significantly higher bone sclerosis (P <0.05­0.001) at the PER location, but only BV/TV (P=0.03) and trabecular number (P=0.02) differed between OA and non-OA knees at the CENT location uncovered by meniscus. CONCLUSIONS: OA results showed a partial dedifferentiation of the subchondral bone micro-architecture between PER and CENT locations probably due to menisci that still retain some of their protective effects on the subchondral bone.

Microcracks in subchondral bone plate is linked to less cartilage damage.

OBJECTIVES: Osteoarthritis (OA) is a disease of the whole joint characterized by cartilage loss and subchondral bone remodeling. The role of microcracks in cartilage integrity and subchondral bone homeostasis is not fully understood. The main goal of this work was to evaluate microcrack density in both calcified cartilage and subchondral bone plate in relation to cartilage damage in humans and to better define the association of microcracks and osteocyte density in subchondral bone. METHODS: We investigated 18 bone cores from cadaveric human knees that were stained with En-Bloc Basic Fuchsin. We quantified microcrack density, osteocyte density, cartilage surfaces and cartilage damage. The presence of microcracks was confirmed for each bone core by scanning electron microscopy. Finally, trabecular subchondral bone parameters were measured by micro-CT. RESULTS: Microcracks were detected in both calcified cartilage and subchondral bone plate. The density of microcracks in both calcified cartilage (CC) and subchondral bone plate (SBP) was negatively correlated with cartilage damage (r = -0.45, p < 0.05). The presence of microcracks in SBP was associated with a lower histological OA score. Osteocytes formed a dendrite network that abruptly stopped at the border of calcified cartilage. Osteocyte density in subchondral bone plate was increased in the presence of microcracks in calcified cartilage. CONCLUSIONS: Subchondral bone plate microcracks might be required for maintaining cartilage homeostasis. Microcracks in calcified cartilage may trigger osteocyte density in subchondral bone plate with subsequent regulation of subchondral bone remodeling to prevent cartilage damage.

Multiscale and multimodality computed tomography for cortical bone analysis.

In clinical studies, high resolution peripheral quantitative computed tomography (HR-pQCT) is used to separately evaluate cortical bone and trabecular bone with an isotropic voxel of 82 µm3, and typical cortical parameters are cortical density (D.comp), thickness (Ct.Th), and porosity (Ct.Po). In vitro, micro-computed tomography (micro-CT) is used to explore the internal cortical bone micro-structure with isotropic voxels and high resolution synchrotron radiation (SR); micro-CT is considered the 'gold standard'. In 16 tibias and 8 femurs, HR-pQCT measurements were compared to conventional micro-CT measurements. To test modality effects, conventional micro-CT measurements were compared to SR micro-CT measurements at 7.5 µm3; SR micro-CT measurements were also tested at different voxel sizes for the femurs, specifically, 7.5 µm3 versus 2.8 µm3. D.comp (r = -0.88, p < 10-3) was the parameter best correlated with porosity (Po.V/TV). The correlation was not affected by the removal of pores under 130 µm. Ct.Th was also significantly highly correlated (r = -0.89 p < 10-3), while Ct.Po was correlated with its counterpart Po.V/TV (r = 0.74, p < 10-3). From SR micro-CT and conventional micro-CT at 7.5 µm3 in matching areas, Po.V/TV and pore diameter were underestimated in conventional micro-CT with mean ± standard deviation (SD) biases of -2.5 ± 1.9% and -0.08 ± 0.08 mm, respectively. In contrast, pore number (Po.N) and pore separation (Po.Sp) were overestimated with mean ± SD biases of +0.03 ± 0.04 mm-1 and +0.02 ± 0.04 mm, respectively. The results from the tibia and femur were similar when the results of SR micro-CT at 7.5 µm3 and 2.8 µm3 were compared. Po.V/TV, specific surface of pores (Po.S/Po.V), and Po.N were underestimated with mean biases of -1.7 ± 0.9%, -4.6 ± 4.4 mm-1, and -0.26 ± 0.15 mm-1, respectively. In contrast, pore spacing was overestimated at 7.5 µm3 compared to 2.8 µm3 with mean biases of 0.05 ± 0.03 mm. Cortical bone measurements from HR-pQCT images provided consistent results compared to those obtained using conventional micro-CT at the distal tibia. D.comp was highly correlated to Po.V/TV because it considers both the micro-porosity (Haversian systems) and macro-porosity (resorption lacunae) of cortical bone. The complexity of canal organization, (including shape, connectivity, and surface) are not fully considered in conventional micro-CT in relation to beam hardening and cone beam reconstruction artifacts. With the exception of Po.V/TV measurements, morphological and topological measurements depend on the characteristics of the x-ray beam, and to a lesser extent, on image resolution.

Cartilage morphology assessed by high resolution micro-computed tomography in non OA knees.

OBJECTIVE: At the microscopic level, there is no established 3D method to measure cartilage volume and thickness. An easy, precise and accurate volume and thickness measurements of human cartilage has been developed from micro-computed tomography (micro-CT) images. DESIGN: In the medial tibial plateau of twenty nine left cadaveric knees classified as normal, vertical cores (7 mm in diameter) were extracted in three locations: lateral (LAT), medial posterior (MP) and medial anterior (MA). The cores were imaged in mineral oil with micro-CT (voxel size 10.2 µm) and were measured cartilage volume (Cart.Vol, mm(3)) and cartilage thickness (Cart.Th, mm) using a thickness plugin. Short term reproducibility and standard deviation (ST_RMSCV%, ST_RMSSD, mm) were determined. To assess accuracy, holes with diameters of 2 mm, 3 mm, and 4 mm were artificially generated and nominal and measured hole sizes were compared. RESULTS: Precision of Cart.Vol_ST_RMSCV% was 1.35%. The mean biases between nominal hole volume and measurements were -0.49 ± 1.5 mm(3) (2 mm), -0.41 ± 4.2 mm(3) (3 mm) and +0.34 ± 4.4 mm(3) (4 mm). ST_RMSSD was 100 times lower than the biological variation and the highest bias to measure volume was 24 times lower. Cart.Th results statistically differed among core extraction locations: 1.75 ± 0.28 mm (LAT), 1.84 ± 0.40 mm (MP) and 2.32 ± 0.41 mm (MA). CONCLUSION: With a standard laboratory micro-CT system, it was possible to measure cartilage volume and thickness with good precision and accuracy relative to the biological variation of the specimen cohort. Cartilage measurements from micro-CT probably will improve the knowledge of the relationship between cartilage and subchondral bone and may allow to better understand the OA process.

3D characterization of pores in the cortical bone of human femur in the elderly at different locations as determined by synchrotron micro-computed tomography images.

UNLABELLED: Diaphysis, inferior, and lateral superior regions of the femoral neck are subjected to diverse mechanical loads. Using micro-CT based on synchrotron radiation, three-dimensional morphology and connectivity of the pore network are location dependent, underlying different remodeling mechanisms. INTRODUCTION: The three-dimensional (3D) morphology and connectivity of the pore network at various locations in human femurs subjected to diverse mechanical loads were assessed using micro-CT based on synchrotron radiation. METHODS: The cortex from 20 human femurs (mean age, 78.3 ± 12.4 years) was taken from the diaphysis (D), the inferior (IN), and the lateral superior (LS) regions of the femoral neck. The voxel size of the 3D reconstructed image was 7.5 µm. Cortical thickness and pore volume/tissue volume (Po.V/TV), pore diameter (Po.Dm) and spacing (Po.Sp) were determined. The pore surface/pore volume ratio (Po.S/Po.V), the number of pores (Po.N), the degrees of anisotropy (DA), and the connectivity density (ConnD), the degree of mineralization (DMB) were also determined. RESULTS: The characteristics of the pore network in femoral cortical bone were found to be location dependent. There was greater porosity, Po.Dm, and Po.N, and more large (180-270 µm), extra-large (270-360 µm) and giant pores (>360 µm) in the LS compared to the IN and D. The difference in porosity in between the periosteal and endosteal layers was mostly due to an increase of Po.Dm rather than Po.N. There was a lower DMB of bone in the LS, which is consistent with a higher remodeling rate. CONCLUSION: The results provide evidence for large variations in the structure of the internal pore network in cortical bone. These variations could involve different underlying remodeling mechanisms.

Comparison of radiograph-based texture analysis and bone mineral density with three-dimensional microarchitecture of trabecular bone.

PURPOSE: Hip fracture is a serious health problem and textural methods are being developed to assess bone quality. The authors aimed to perform textural analysis at femur on high-resolution digital radiographs compared to three-dimensional (3D) microarchitecture comparatively to bone mineral density. METHODS: Sixteen cadaveric femurs were imaged with an x-ray device using a C-MOS sensor. One 17 mm square region of interest (ROI) was selected in the femoral head (FH) and one in the great trochanter (GT). Two-dimensional (2D) textural features from the co-occurrence matrices were extracted. Site-matched measurements of bone mineral density were performed. Inside each ROI, a 16 mm diameter core was extracted. Apparent density (Dapp) and bone volume proportion (BV/TV(Arch)) were measured from a defatted bone core using Archimedes' principle. Microcomputed tomography images of the entire length of the core were obtained (Skyscan 1072) at 19.8 microm of resolution and usual 3D morphometric parameters were computed on the binary volume after calibration from BV/TV(Arch). Then, bone surface/bone volume, trabecular thickness, trabecular separation, and trabecular number were obtained by direct methods without model assumption and the structure model index was calculated. RESULTS: In univariate analysis, the correlation coefficients between 2D textural features and 3D morphological parameters reached 0.83 at the FH and 0.79 at the GT. In multivariate canonical correlation analysis, coefficients of the first component reached 0.95 at the FH and 0.88 at the GT. CONCLUSIONS: Digital radiographs, widely available and economically viable, are an alternative method for evaluating bone microarchitectural structure.

Trabecular and cortical bone separately assessed at radius with a new ultrasound device, in a young adult population with various physical activities.

The aim was to evaluate a new ultrasound device in a young adult population and to assess its reproducibility via comparison to DXA measurements and geometrical measurements from high-resolution radiographs. Ninety-three subjects aged between 20 and 51 years were recruited and divided into four groups according to their gender and physical activity status: 22 male athletes, 19 male controls, 21 female athletes, and 31 female controls. Ultrasonic measurements were assessed by the prototype LD-100 (Oyo Electric Co., Kyoto, Japan) on the dominant distal radius. Attenuation in the radius (dB), cortical bone thickness (mm), radius thickness (mm), mass density of cancellous bone (mg/cm(3)), and elasticity (GPa) of cancellous bone were obtained. BMD was measured by DXA at the dominant distal radius. Radius images were obtained with a direct high-resolution digital X-ray device (BMA, D(3)A Medical Systems), and radius and cortical thicknesses were estimated using a specific software (ImageJ, Bethesda, USA), in an area site-matched with LD-100. There was a significant positive correlation between site-matched BMD measurement and LD-100 parameters (p<0.004), X-ray radius thickness, and LD-100 parameters except elasticity (p<0.05, r>0.32), X-ray cortical thickness and LD-100 attenuation and cortical thickness (p<0.01). A significantly higher attenuation, cortical and radius thicknesses were found in athletes compared to controls (p<0.05). The radius thickness measured on radiographs was significantly higher in athletes versus controls in both sexes, and cortical thickness was significantly higher in male athletes versus controls. These data suggest a positive influence of physical activity on bone cortical measurements. This study also confirmed the particular interest of bone assessment by ultrasound.

Hypoleptinaemia in extreme body mass models: the case of international rugby players.

Leptinaemia has been poorly studied in athlete populations with the consequences of athletic hypoleptinaemia yet to be examined. Our aim was to determine if systemic leptinaemia is influenced in high body mass athletes. We recruited 24 rugby players (21.5+/-4.7 years; 11.8+/-2.9h/week) and 26 controls (22.3+/-3.1 years; 1.9+/-1.4h/week). BMD (whole body (WB), limbs, lumbar spine and non-dominant femur) and body composition (WB Lean Mass (LM) and FM) were measured by Dual X-ray Absorptiometry. Circulating levels of serum leptin (ng/ml), adiponectin (microg/ml), insulin (ng/ml), osteocalcin (ng/ml) and CTx (ng/ml) were assessed by ELISA assays. BMD were significantly higher in rugby players vs controls, at all bone sites, yet after adjustments for body mass index. They had significantly higher LM and FM but no differences in %FM. They had significantly higher osteocalcin but lower CTx, insulin and leptin concentrations. Leptin levels were strongly correlated to %FM (r=0.85, p<0.0001), as well as to absolute FM (r=0.77, p=0.0002), in the rugbymen group. Rugby practice was associated to a bone remodelling process in favour of bone formation. There was a significant hypoleptinaemia in our rugby players, while their percent FM was equivalent and absolute FM significantly higher than the control levels. These data suggest that leptin is under control of physical activity and not just fat mass.

Side-to-side and within-side variability of 3D bone microarchitecture by conventional micro-computed tomography of paired iliac crest biopsies.

Bone microarchitecture in osteoporosis can be characterized by examining iliac bone biopsies and treatment effects assessed by comparing a baseline biopsy from one side to a posttreatment biopsy from the other side, a method that assumes limited side-to-side variability. New techniques based on micro-computed tomography (microCT) provide information on the three-dimensional (3D) microarchitecture of bone. We used microCT to measure side-to-side and within-side variability of 3D microarchitectural parameters of trabecular and cortical bone in paired iliac-crest biopsies, one from each side. A Bordier needle trephine was used to collect biopsies from 30 postmenopausal female cadavers (mean age, 73.7+/-10.7 years; range, 55-96 years). Biopsies were chemically defatted then imaged using a desktop microCT scanner (voxel size, 10.77 microm). Parameters measured in trabecular bone consisted of bone volume/tissue volume (BV/TV, %), direct trabecular thickness and trabecular spacing (Tb.Th and Tb.Sp, microm) using the sphere method, bone surface/bone volume (BS/BV, mm(-1)), trabecular number (Tb.N, mm(-1)), structure model index (SMI), trabecular pattern factor (Tb.Pf), and degree of anisotropy (DA). In cortical bone, we measured cortical thickness (Cort.Th), porosity (Cort.Porosity), and pore diameter (Po.Dm). For trabecular bone parameters, reproducibility as assessed from two microCT acquisitions ranged from 4.1% to 6.9%. To assess side-to-side variability, we matched the volumes of interest selected in the right and left iliac crests. The mean difference in absolute individual percent variation (mAbsDelta(ind)) between the two sides ranged from 10.8% to 14.8% for all trabecular parameters except Tb.Pf (74%) and SMI (84%). In cortical bone, mAbsDelta(ind) were 11.6% for Po.Dm, 15.1% for Cort.Porosity, and 27.6% for Cort.Th. To assess within-side variability, we divided the trabecular iliac crest volume into three equal parts, one adjacent to each cortex and one in the middle. Values of mAbsDelta(ind) versus the middle part were ranging from 7.6% for Tb.Sp to 26.2% for BV/TV. Thus, within-side variability was similar in magnitude to side-to-side variability. The considerable differences in robustness across trabecular parameters indicate a need for selecting the most stable parameters, most notably for longitudinal studies of small numbers of patients. Acquisition by microCT and image analysis must comply with stringent quality criteria, especially the distance from the cortices must be standardized.

Laws' masks descriptors applied to bone texture analysis: an innovative and discriminant tool in osteoporosis.

OBJECTIVE: The objective of this study was to explore Laws' masks analysis to describe structural variations of trabecular bone due to osteoporosis on high-resolution digital radiographs and to check its dependence on the spatial resolution. Laws' masks are well established as one of the best methods for texture analysis in image processing and are used in various applications, but not in bone tissue characterisation. This method is based on masks that aim to filter the images. From each mask, five classical statistical parameters can be calculated. MATERIALS AND METHODS: The study was performed on 182 healthy postmenopausal women with no fractures and 114 age-matched women with fractures [26 hip fractures (HFs), 29 vertebrae fractures (VFs), 29 wrist fractures (WFs) and 30 other fractures (OFs)]. For all subjects radiographs were obtained of the calcaneus with a new high-resolution X-ray device with direct digitisation (BMA, D3A, France). The lumbar spine, femoral neck, and total hip bone mineral density (BMD) were assessed by dual-energy X-ray absorptiometry. RESULTS: In terms of reproducibility, the best results were obtained with the TRE5E5 mask, especially for three parameters: "mean", "standard deviation" and "entropy" with, respectively, in vivo mid-term root mean square average coefficient of variation (RMSCV)%= 1.79, 4.24 and 2.05. The "mean" and "entropy" parameters had a better reproducibility but "standard deviation" showed a better discriminant power. Thus, for univariate analysis, the difference between subjects with fractures and controls was significant (P<10(-3)) and significant for each fracture group independently (P<10(-4) for HF, P=0.025 for VF and P< 10(-3) for OF). After multivariate analysis with adjustment for age and total hip BMD, the difference concerning the "standard deviation" parameter remained statistically significant between the control group and the HF and VF groups (P<5 x 10(-5), and P=0.04, respectively). No significant correlation between these Laws' masks parameters and BMD was obtained. In addition, this study showed the dependence of Laws' masks parameters on image resolution, which confirms the necessity to perform Laws' textural measurement on high-resolution images. CONCLUSION: The reproducibility and discriminant power of the Laws' masks analysis has been demonstrated on bone images; thus, this method constitutes a promising routine technique for the determination of osteoporosis fracture risk from radiographs.

Effect of specimen conditioning on the microarchitectural parameters of trabecular bone assessed by micro-computed tomography.

The best way to preserve the mechanical properties of bone specimens is hydration in NaCl, whereas the reference process in microCT analysis is defatting. However, for finite element modelling (FEM) it is necessary to use the same bone specimens for biomechanical testing and 3D imaging. This study aimed to evaluate the effect of sample conditioning on trabecular bone microarchitectural parameters. Trabecular bones were analysed by microCT under three successive conditions: first, the fatted samples were analysed immersed in NaCl (process N); second, they were hydrated for 24 h then imaged without immersion (process H); third, the samples were defatted before analysis (process D). The microarchitectural parameters bone volume/tissue volume (BV/TV), trabecular spacing (Tb.Sp), number (Tb.N) and thickness (Tb.Th) were calculated. Except for BV/TV, there was no significant difference between the processes N and D. In process H, BV/TV, Tb.Th and Tb.N were higher and BS/BV and Tb.Sp were lower than in process D. Results showed that the process D may be replaced by the process N. The process H induced significant differences in microarchitectural parameters when compared to process D. Nevertheless, this sample conditioning should be used to develop FEM when microCT images are to be acquired during compressive testing.

Subchondral bone micro-architectural alterations in osteoarthritis: a synchrotron micro-computed tomography study.

OBJECTIVES: We evaluated the three-dimensional (3D) micro-architecture of subchondral trabecular (Tb) bone in osteoarthritis (OA). Due to high signal-to-noise ratio and high resolution, micro-computed tomography (micro-CT) by synchrotron radiation is considered as the gold standard for bone micro-architecture imaging. DESIGN: Subchondral bone were extracted from femoral heads in OA cases in areas without cartilage (OAc-; n=6) and in adjacent areas with cartilage (OAc+; n=6) and compared to eight subchondral bone cores from osteoporosis cases (OP). The voxel size of images was 10.13 microm. We measured the bone volume fraction (BV/TV) and morphological parameters: Tb thickness (TbTh), Tb spacing (TbSp), Tb number (TbN), and bone surface/bone volume (BS/BV). The degree of anisotropy (DA), the connectivity by the Euler number and the degree of mineralization (DM) were equally assessed. RESULTS: BV/TV and morphological parameters showed significant differences between OAc- and OP samples (P<0.01 except TbTh: P<0.05) and between OAc- and OAc+ (0.05

Bone status in primary hyperparathyroidism.

Traditional bone involvement, such as osteoitis fibrosa, has become very rare (< 1%) in primary hyperparathyroidism (PHPT); nevertheless, fractures seem more frequent than in controls, with a predilection for fractures of the distal extremity of the radius, pelvis, ribs and vertebrae, and a relative modest incidence of fractures of the upper extremity of the femur. Histo-morphometric studies have stressed a discrepancy between cortical and trabecular bone with an increase of bone remodeling. The cortical width is constantly diminished and the cortical porosity is increased whereas trabecular volume is normal and micro-architecture preserved. Bone mineral density (BMD) allows an early diagnosis of bone disease and takes a growing place in the management of patients. Since the consensus conference in 1991, the measurement of BMD has been incorporated in the surgical decision with a threshold: Z-score < -2. The demineralisation predominates on sites rich in cortical bone (1/3 proximal of the distal radius); the radius, which was the first site evaluated for technical reasons, is also the most discriminating one. Spine demineralisation is met in more severe forms and BMD measurement of the whole body is promising but requires more studies. In the absence of a radical processing, moderate forms remain stable, whereas more severe forms have a tendency to deteriorate. The evaluation of spine and femoral BMD is useful for the follow-up because the bone gain after parathyroidectomy is significant early on at these sites (rich in trabecular bone with high bone turnover), whereas the BMD of radius is relatively stable.

Evaluation of error bounds on calcaneal speed of sound caused by surrounding soft tissue.

For absorptiometry measurements, soft tissue may have an impact on quantitative ultrasound (QUS) measurements. In the present study, we focused primarily on the quantification of measurement error on speed of sound (SOS) caused by surrounding soft tissue. The relevant soft tissue parameters affecting the inherent SOS inaccuracies are thickness and sound velocity. To meet our goal, SOS measurements were taken at the right heel using a QUS imaging device in 21 healthy subjects. Site-matched measurements of soft tissue thickness (STT) and bone width were performed using magnetic resonance imaging of the heel. Several bone velocities were calculated either by accounting for bone width (SOSBW) only or by taking into account the exact path lengths of all major components traversed by ultrasound &lapr;V(b)). Given that soft tissue composition is difficult to determine in vivo, we chose to estimate lower and upper error bounds on bone velocity (V(b lower) and V(b upper)) by spanning the full range of available values in the literature. The mean BW was 30.7 +/- 2.7 mm and the mean medial and external STTs were 8.8 +/- 1.7 and 8.5 +/- 1.5 mm, respectively. Accounting for true BW only resulted in no significant difference between SOS (1533 +/- 37) and SOSBW (1531 +/- 33). By contrast, accounting for both true BW and surrounding soft tissue resulted in an increase in the calculated bone velocity and statistically significant differences between SOS and V(b upper) (1568 +/- 36) and V(b lower) (1542 +/- 34). Root mean square errors between SOS and the calculated velocities were 0.34, 2. 32, and 0.70% for SOSBW, V(b upper), and V(b lower), respectively. We report here measurement errors caused by soft tissue to be 3 to 20 times higher than the SOS short-term precision (SOS coefficient of variation of 0.1%). Our results suggest that inaccuracies in SOS measurement caused by overlying soft tissue cannot be neglected. Overlying soft tissues may influence outcomes of longitudinal studies, especially if variations in tissue thickness and composition occur during the longitudinal follow-up. A practical way of minimizing the measurement error could be to perform an adequate correction for the overlying soft tissue. However, ideally, this should require knowing both the thickness and sound velocity in soft tissue. One might preferably conduct experimental investigations that directly control soft tissue thickness and composition to resolve this problem.

Ultrasound measurement on the calcaneus: influence of immersion time and rotation of the foot.

The aim of this study was to evaluate the influence of immersion time and rotation of the heel around the leg axis on the reproducibility of measurements using an ultrasound bone imaging scanner (UBIS) with a temperature-controlled water bath. Measurements were obtained in 10 men, 11 premenopausal women and 10 postmenopausal women. The right foot of all subjects was scanned 12 times with an interval of 3 min between each scan. The first 10 measurements, performed to study the effect of immersion time, were taken without intermediate repositioning. Measurements 11 and 12 were also taken without removing the foot, but the calcaneus position was varied by an angle of +/-2.5 degrees with respect to the reference position used during the first 10 measurements. Prolonged immersion of the heel led to a decrease in the variations of quantitative ultrasound (QUS) parameters between successive measurements. Following rapid variations with immersion time, the QUS parameters reached a plateau. The effect of immersion time on measurement error remained significant until the first two or three scans for broadband ultrasound attenuation (BUA) and until the sixth or seventh scans for speed of sound (SOS). The variation in BUA was more pronounced (p<0.05) for the group of postmenopausal women (20. 7% change in BUA between the first and tenth scans; p<0.005) than for the group of premenopausal women (6.8% change in BUA between the first and tenth scans; p<0.005). The variations in SOS were similar in the two groups (0.8% variation; p<0.005). The impact of immersion time was smaller for men than for women [2.5%, (p<0.01) and 0.4% (p<0.005) of the change between the first and tenth scans for BUA and SOS respectively]. On the whole, the measurement errors due to rotation of the heel were lower than those caused by immersion time. The variations were significant only in men and premenopausal women. Both immersion time and rotation of the heel may play a role in the precision of QUS and should be carefully standardized, particularly for longitudinal studies. In addition, following these results we have adopted a standardized protocol to derive the technique reproducibility in groups of premenopausal and postmenopausal women. The coefficients of variation were 1.1% for BUA and 0.1% for SOS in premenopausal women, and 1.4% for BUA and 0.13% for SOS in postmenopausal women.

Clinical evaluation of spine morphometric X-ray absorptiometry.

A new method for vertebral height measurements, morphometric X-ray absorptiometry (MXA) based on dual-energy X-ray absorptiometry, has been proposed. This technique overcomes some limitations of morphometric radiography (MRX): the effective radiation dose is low, some sources of geometric distortion are eliminated, such as dependence on patient position, magnification gradient, and the effect of scoliosis is minimized. The purpose of this study was to compare morphometric parameters obtained by both methods (MXA and MRX), and to evaluate the agreement between morphometric evaluations and qualitative reading for vertebral fracture diagnosis. The evaluation was performed with an Hologic QDR 2000 device in 67 women without vertebral fractures and 31 women with vertebral fractures (according to a qualitative assessment). The reproducibility of the image analysis was <4% and comparable to MRX. The estimated bias between the two methods was on average 10 mm, and was a function of the vertebral height, according to the Bland and Altman method. The agreement between MXA and other methods for vertebral fracture diagnosis was poor for the thoracic level above T7, due to a lack of resolution and rib interposition. Agreement was also low for T10, due to the motion of the diaphragm muscle. MXA cannot currently be used for the diagnosis of thoracic vertebral fracture in clinical practice. Technological improvements are necessary to make this promising method useful as a screening tool to evaluate the presence of thoracic vertebral fractures.

Assessment of the relationship between broadband ultrasound attenuation and bone mineral density at the calcaneus using BUA imaging and DXA.

The purposes of this study was to determine the relationship between broadband ultrasound attenuation (BUA) and bone mineral density (BMD) measured at different regions of the calcaneus with identical site-matched regions of interest (ROIs). Dual-energy X-ray absorptiometry (DXA) measurements of the calcaneus and BUA imaging were performed in 30 women (15 premenopausal and 15 postmenopausal). Four square ROIs were located in the great tuberosity and one square ROI in the foramen calcaneus. A ROI adapted to the shape and size of the whole calcaneus was also considered. All ROIs were analyzed three times with both techniques to minimize intra-observer variability. The correlation coefficient between attenuation and frequency was used as an index of BUA measurement error. Before accepting a measurement of BUA in inhomogeneous material, it could be useful to map the spatial variations of the measurement error. In all ROIs we found the BUA and BMD were strongly related (r = 0.78-0.91, p < 0.001). The correlation between BUA and BMD was slightly higher in the inferior part of the posterior tuberosity than in the superior part and in the foramen calcaneus. The very high correlation between attenuation and frequency found in all ROIs (r = 0.99) suggests that measurement errors of propagation were probably not significant. Ultrasound imaging yields the opportunity for studying the spatial acoustic properties in the calcaneus and their relation to bone mass or structural parameters provided by independent imaging techniques. BUA measured with current transmission techniques reflects mainly bone mass, and microarchitecture to a smaller extent.

Quantitative ultrasound imaging at the calcaneus using an automatic region of interest.

A new approach to measuring bone properties at the calcaneus using ultrasound parametric imaging has recently emerged. However, an additional source of observer-related error is the substantial regional variations in the pattern of ultrasound parameters. The contribution of intra-observer and inter-observer variability to the coefficient of variation can be eliminated using an algorithm which selects the region of interest (ROI) completely automatically. The objective of the present study was the clinical assessment of an automatic ROI for both broadband ultrasonic attenuation (BUA) and speed of sound (SOS) measurement using ultrasound parametric imaging. The automatic ROI was defined as the circular region of lowest attenuation in the posterior tuberosity of the calcaneus. We have tested this algorithm using clinical images of the calcaneus from 265 women. Mean coefficients of variation were 1.6% (95% confidence interval 1.4%-1.9%) and 0.26% (95% confidence interval 0.23%-0.32%) for BUA and SOS respectively (standardized CV was 2.1% for BUA and 2.6% for SOS). Z-scores in an osteoporotic group were -0.61 and -0.52 for BUA and SOS respectively. In healthy women, the age-related decline was -0.50 dB/ MHz per year (0.7%/year) for BUA and -1.2 m/s per year (0.08%/year) for SOS. In the subgroup of healthy postmenopausal women, using stepwise multiple regression, we found that BUA was predicted best by years since menopause (YSM) and weight, with overall model r2 = 0.28; SOS was predicted best by YSM only (r2 = 0.21). Neither the range of biological variation of ultrasound parameters nor the clinical value were affected by the choice of the region of lowest attenuation for measurement. The automatic procedure was totally independent of operator interaction, therefore excluding loss of precision due to intra- or inter-observer variability. The results showed the high precision and robustness of the procedure. These factors make this approach viable for routine clinical use.

Bone effects of dydrogesterone in ovariectomized rats: a biologic, histomorphometric, and densitometric study.

It has been suggested that progesterone might affect bone metabolism. Dydrogesterone (DD) has a chemical structure very close to that of natural progesterone, without androgenic effects. We compared the effect of a daily treatment with DD and estradiol (E2, 40 micrograms/kg) on 70 female rats (8 weeks old), divided in seven groups: controls; ovariectomized (OVX); OVX + E2, OVX + DD 2.5 mg/kg; OVX + DD 5 mg/kg; OVX + E2 + DD 2.5 mg/kg; and OVX + E2 + DD 5 mg/kg. Two months later we studied estradiol, osteocalcin, and acid phosphatases (TRAP) levels; histomorphometric parameters at the trabecular part of the femur; and bone mineral density of the tibiae (trabecular and cortical areas) and caudal vertebrae (cortical bone) by dual energy X-ray absorptiometry (Hologic QDR 2000). In the OVX group, we found a nonsignificant increase of osteocalcin and TRAP, a decrease in the trabecular bone volume, and an increase in the intertrabecular distance. These variations were not seen in the groups treated with E2 (with or without DD). DD had no effect on trabecular architecture parameters. It induced a dose-dependent increase in the osteoclast number (TRAP + cells), without increase in trabecular separation. There was no difference in bone density of caudal vertebrae between the different groups, nor of the tibial density between the OVX and OVX + DD groups. The bone densities were not different in the control and E2-treated groups, with or without DD. During dydrogesterone treatment, there is an increase in osteoclast number, without an apparent increase in osteoclast function. Dydrogesterone did not prevent bone loss due to ovariectomy in rats, and it did not affect the protective effect of estradiol.