Guidelines for the assessment of bone density and microarchitecture in vivo using high-resolution peripheral quantitative computed tomography.

INTRODUCTION: The application of high-resolution peripheral quantitative computed tomography (HR-pQCT) to assess bone microarchitecture has grown rapidly since its introduction in 2005. As the use of HR-pQCT for clinical research continues to grow, there is an urgent need to form a consensus on imaging and analysis methodologies so that studies can be appropriately compared. In addition, with the recent introduction of the second-generation HrpQCT, which differs from the first-generation HR-pQCT in scan region, resolution, and morphological measurement techniques, there is a need for guidelines on appropriate reporting of results and considerations as the field adopts newer systems. METHODS: A joint working group between the International Osteoporosis Foundation, American Society of Bone and Mineral Research, and European Calcified Tissue Society convened in person and by teleconference over several years to produce the guidelines and recommendations presented in this document. RESULTS: An overview and discussion is provided for (1) standardized protocol for imaging distal radius and tibia sites using HR-pQCT, with the importance of quality control and operator training discussed; (2) standardized terminology and recommendations on reporting results; (3) factors influencing accuracy and precision error, with considerations for longitudinal and multi-center study designs; and finally (4) comparison between scanner generations and other high-resolution CT systems. CONCLUSION: This article addresses the need for standardization of HR-pQCT imaging techniques and terminology, provides guidance on interpretation and reporting of results, and discusses unresolved issues in the field.

Systemic mastocytosis identified in two women developing fragility fractures during lactation.

Two women presenting with fragility fractures during lactation had bone mineral density (BMD) reduced more greatly than usually associated with lactation. The first woman was 29 years old with a BMD T-score of - 3.2 SD at the spine and- 2.0 SD at the femoral neck. The second woman was 35 years old with a BMD T-score of - 4.5 SD at the spine and - 2.8 SD at the femoral neck. Both women had increased cortical porosity and reduced trabecular density. Investigation identified an elevated serum tryptase, and marrow biopsy confirmed the diagnosis of mastocytosis. Lactation causes bone loss, but the occurrence of fractures in the setting of severe deficits in BMD and microstructural deterioration signals the need to consider additional causes of bone loss.

Operator variability in scan positioning is a major component of HR-pQCT precision error and is reduced by standardized training.

In this study, we determined that operator positioning precision contributes significant measurement error in high-resolution peripheral quantitative computed tomography (HR-pQCT). Moreover, we developed software to quantify intra- and inter-operator variability and demonstrated that standard positioning training (now available as a web-based application) can significantly reduce inter-operator variability. INTRODUCTION: HR-pQCT is increasingly used to assess bone quality, fracture risk, and anti-fracture interventions. The contribution of the operator has not been adequately accounted in measurement precision. Operators acquire a 2D projection ("scout view image") and define the region to be scanned by positioning a "reference line" on a standard anatomical landmark. In this study, we (i) evaluated the contribution of positioning variability to in vivo measurement precision, (ii) measured intra- and inter-operator positioning variability, and (iii) tested if custom training software led to superior reproducibility in new operators compared to experienced operators. METHODS: To evaluate the operator in vivo measurement precision, we compared precision errors calculated in 64 co-registered and non-co-registered scan-rescan images. To quantify operator variability, we developed software that simulates the positioning process of the scanner's software. Eight experienced operators positioned reference lines on scout view images designed to test intra- and inter-operator reproducibility. Finally, we developed modules for training and evaluation of reference line positioning. We enrolled six new operators to participate in a common training, followed by the same reproducibility experiments performed by the experienced group. RESULTS: In vivo precision errors were up to threefold greater (Tt.BMD and Ct.Th) when variability in scan positioning was included. The inter-operator precision errors were significantly greater than the short-term intra-operator precision (p < 0.001). New trained operators achieved comparable intra-operator reproducibility to experienced operators and lower inter-operator reproducibility (p < 0.001). Precision errors were significantly greater for the radius than for the tibia. CONCLUSION: Operator reference line positioning contributes significantly to in vivo measurement precision and is significantly greater for multi-operator datasets. Inter-operator variability can be significantly reduced using a systematic training platform, now available online ( http://webapps.radiology.ucsf.edu/refline/ ).

Measurement of cortical porosity of the proximal femur improves identification of women with nonvertebral fragility fractures.

UNLABELLED: We tested whether cortical porosity of the proximal femur measured using StrAx1.0 software provides additional information to areal bone mineral density (aBMD) or Fracture Risk Assessment Tool (FRAX) in differentiating women with and without fracture. Porosity was associated with fracture independent of aBMD and FRAX and identified additional women with fractures than by osteoporosis or FRAX thresholds. INTRODUCTION: Neither aBMD nor the FRAX captures cortical porosity, a major determinant of bone strength. We therefore tested whether combining porosity with aBMD or FRAX improves identification of women with fractures. METHODS: We quantified femoral neck (FN) aBMD using dual-energy X-ray absorptiometry, FRAX score, and femoral subtrochanteric cortical porosity using StrAx1.0 software in 211 postmenopausal women aged 54-94 years with nonvertebral fractures and 232 controls in Tromsø, Norway. Odds ratios (ORs) were calculated using logistic regression analysis. RESULTS: Women with fractures had lower FN aBMD, higher FRAX score, and higher cortical porosity than controls (all p < 0.001). Each standard deviation higher porosity was associated with fracture independent of FN aBMD (OR 1.39; 95% confidence interval 1.11-1.74) and FRAX score (OR 1.58; 1.27-1.97) in all women combined. Porosity was also associated with fracture independent of FRAX score in subgroups with normal FN aBMD (OR 1.88; 1.21-2.94), osteopenia (OR 1.40; 1.06-1.85), but not significantly in those with osteoporosis (OR 1.48; 0.68-3.23). Of the 211 fracture cases, only 18 women (9%) were identified using FN aBMD T-score < -2.5, 45 women (21%) using FRAX threshold >20%, whereas porosity >80th percentile identified 61 women (29%). Porosity identified 26% additional women with fractures than identified by the osteoporosis threshold and 21% additional women with fractures than by this FRAX threshold. CONCLUSIONS: Cortical porosity is a risk factor for fracture independent of aBMD and FRAX and improves identification of women with fracture.

Women and men with hip fractures have a longer femoral neck moment arm and greater impact load in a sideways fall.

INTRODUCTION: In a case control study, we report that women and men with hip fractures have a longer moment arm of the force applied on the proximal femur during a sideways fall, a structural feature that may contribute to fracture risk. The impact load and its direction during a sideways fall onto the greater trochanter are partly determined by the geometry of the proximal femur. We hypothesized that the hip geometry in elderly with hip fractures produces a greater impact on the hip during a sideways fall. METHODS: We studied 41 female (77.2 +/- 9.9 years) and 22 male (76.2 +/- 12.1 years) patients with hip fractures and 40 female (85.7 +/- 6.0 years) and 17 male (84.3 +/- 10.1 years) controls. Hip geometry was analyzed on the nonfracture hip in patients and left hip in controls using dual-energy X-ray absorptiometry. RESULTS: There was no difference in areal bone mineral density (aBMD), hip axis length, femoral neck axis length, or neck-shaft angle between cases and controls. However, the moment arm of the force on the hip during a sideways fall was 7.3% and 9.5% longer resulting in 5.6% and 9.1% greater moment in such a fall in female and male cases relative to their respective controls independent of height and weight (all p < 0.056). In multivariate logistic regression analysis, only the moment arm length in a sideways fall was associated with increased risk of hip fracture in females (odds ratio = 1.91, 95%CI: 1.14-3.20 for each SD increase in moment arm length of sideways fall, p = 0.02) and males (odds ratio = 2.69, 95% CI, 1.19-6.09, p = 0.01). CONCLUSIONS: A longer moment arm in the sideways fall increases the resultant force applied to the hip predisposing to hip fracture.

Role of subchondral bone remodelling in collapse of the articular surface of Thoroughbred racehorses with palmar osteochondral disease.

REASONS FOR PERFORMING STUDY: To gain a better understanding of the aetiology of articular surface collapse in horses with palmar osteochondral disease. OBJECTIVES: To determine whether acceleration of focal bone resorption associated with reduced physical activity contributes to articular surface collapse in racehorses with palmar osteochondral disease. STUDY DESIGN: Cross-sectional study comparing metacarpal bones from horses at varying stages of race training. METHODS: Metacarpal bones from 36 racing Thoroughbred horses were examined with high-resolution peripheral quantitative computed tomography to determine the proportion of the articular surface that had collapsed and with backscattered scanning electron microscopy to quantify porosity and eroded bone surface. Racing and training histories were obtained for comparison with imaging data. RESULTS: In 21 cases, inward collapse of the calcified cartilage layer was observed on backscattered scanning electron microscopy. An increased extent of articular surface collapse was associated with greater numbers of microfractures in the calcified cartilage and superficial subchondral bone (Spearman's correlation [rs ] = 0.62, P<0.001). In the deeper bone (6-10 mm), porosity was lower with a greater extent of articular surface collapse (rs = -0.38, P = 0.023), whereas in the superficial bone (0-4 mm) there was no association between articular surface collapse and porosity (rs = 0.19, P = 0.26). Both porosity (median 14, range 3.8-26 vs. 3.8, 1.6-17%, P = 0.008) and eroded surface (1.1, 0.74-4.5 vs. 0.64, 0.11-4.7 mm(-1) , P = 0.016) of the superficial subchondral bone were higher in resting than in training horses, and in some resting horses subchondral bone voids were highly concentrated, resulting in an apparent loss of support for the overlying calcified cartilage layer. CONCLUSIONS: Articular surface collapse is common in cases of palmar osteochondral disease and is likely to be a sequel to fatigue injury of subchondral bone. Focal subchondral bone resorption appears to contribute to collapse of the calcified cartilage and is potentiated by a reduced-intensity exercise regimen.

Can high-resolution peripheral quantitative computed tomography imaging of subchondral and cortical bone predict condylar fracture in Thoroughbred racehorses?

REASONS FOR PERFORMING STUDY: High-resolution 3D imaging may improve the prediction and/or early identification of condylar fractures of the distal metacarpus/tarsus and reduce the frequency of breakdown injury in racehorses. OBJECTIVES: To test the hypotheses that horses suffering condylar fractures have higher bone volume fraction (BV/TV) of the distal metacarpal epiphysis, greater subchondral bone thickness at the fracture site and higher second moment of inertia in the metacarpal midshaft as identified with high-resolution 3D imaging. STUDY DESIGN: Cross-sectional study using cadaver material. METHODS: Thoroughbreds that died on racetracks were grouped as: 1) horses with third metacarpal (McIII) fractures with a condylar component (cases, n = 13); 2) horses with no limb fracture (controls, n = 8); 3) horses with fractures in other bones or suspensory apparatus disruption (other fatal injuries, n = 16). The palmar condyles of McIII and the midshaft were examined with high resolution peripheral quantitative computed tomography (HR-pQCT). Statistical analysis included logistic regression and Spearman's correlation. RESULTS: There were no significant differences in BV/TV of distal McIII and second moment of inertia of the midshaft between cases and controls. Epiphyseal bone BV/TV was greater in injured limbs of horses with any fatal limb injury (Groups 1 and 3 combined) compared with controls (odds ratio = 1.20, 95% confidence interval 1.01-1.42, P = 0.034). An epiphyseal BV/TV>0.742 resulted in a sensitivity of 82.8% and specificity of 62.5% in identifying horses with fatal limb injury. In horses without condylar fracture, increased subchondral bone thickness was associated with palmar osteochondral disease lesions in the adjacent condyle (rs = 0.65, P<0.001). CONCLUSIONS: Increased BV/TV of the distal metacarpus may have some value for identifying horses at risk of any fatal breakdown injury but not metacarpal condylar fractures. Measurement of parasagittal groove subchondral bone thickness is complicated by adjacent palmar osteochondral disease lesions. Thus, high-resolution imaging of the distal metacarpus appears to have limited ability to identify horses at risk of condylar fractures.

A new method of segmentation of compact-appearing, transitional and trabecular compartments and quantification of cortical porosity from high resolution peripheral quantitative computed tomographic images.

A transitional or cortico-trabecular junctional zone exists at any location composed of both cortical and trabecular bones such as the metaphyses of tubular bones and short bones like the femoral neck. The transitional zone comprises the inner cortex adjacent to the medullary canal and trabeculae abutting against the cortex contiguous with the endocortical surface. This is a site of vigorous remodeling. Intracortical remodeling cavitates the inner cortex expanding this transitional zone at the price of compact-appearing cortex so that it contains porosity, cortical fragments that resemble trabeculae, and trabeculae abutting the eroding cortex. The porosity of the transitional zone is an important source of bone loss. It reduces bone strength exponentially and is a quantifiable `fingerprint' of structural deterioration. A new automated method of segmentation of bone from background and bone into its compact-appearing cortex, transitional zone, and trabecular compartment is described, with a new approach to quantification of cortical porosity. Segmentation is achieved by automatically selecting attenuation profile curves perpendicular to the periosteal surface. Local bone edges are identified as the beginning and the end of the rising and falling S-shaped portions of the curve enabling the delineation of the compartments. Analyzing ~3600 consecutive overlapping profiles around the perimeter of each cross-sectional slice segments the compartments. Porosity is quantified as the average void volume fraction of all voxels within each compartment. To assess accuracy at the distal radius and tibia, µCT images of cadaveric specimens imaged at 19 µm voxel size served as the gold standard. To assess accuracy at the proximal femur, scanning electron microscopy (SEM) images of specimens collected at 2.5 µm resolution served as the gold standard. Agreement between HRpQCT and the gold standards for segmentation and quantification of porosity at the distal radius and tibia ranged from R(2)=0.87 to 0.99, and for the proximal femur ranged from 0.93 to 0.99. The precision error in vivo for segmentation and quantification of porosity in HRpQCT images at the distal radius, given by the root mean square error of the coefficient of variation, ranged from 0.54% for porosity of the transitional zone to 3.98% for area of the compact-appearing cortex. Segmentation of the transitional zone minimizes errors in apportioning cortical fragments and cortical porosity to the medullary compartment and so is likely to allow accurate assessment of fracture risk and the morphological effects of growth, aging, diseases and therapies.

Proteinase-activated receptor-2 is required for normal osteoblast and osteoclast differentiation during skeletal growth and repair.

Proteinase-activated receptor-2 (PAR(2)) is a G-protein coupled receptor expressed by osteoblasts and monocytes. PAR(2) is activated by a number of proteinases including coagulation factors and proteinases released by inflammatory cells. The aim of the current study was to investigate the role of PAR(2) in skeletal growth and repair using wild type (WT) and PAR(2) knockout (KO) mice. Micro computed tomography and histomorphometry were used to examine the structure of tibias isolated from uninjured mice at 50 and 90 days of age, and from 98-day-old mice in a bone repair model in which a hole had been drilled through the tibias. Bone marrow was cultured and investigated for the presence of osteoblast precursors (alkaline phosphatase-positive fibroblastic colonies), and osteoclasts were counted in cultures treated with M-CSF and RANKL. Polymerase chain reaction (PCR) was used to determine which proteinases that activate PAR(2) are expressed in bone marrow. Regulation of PAR(2) expression in primary calvarial osteoblasts from WT mice was investigated by quantitative PCR. Cortical and trabecular bone volumes were significantly greater in the tibias of PAR(2) KO mice than in those of WT mice at 50 days of age. In trabecular bone, osteoclast surface, osteoblast surface and osteoid volume were significantly lower in KO than in WT mice. Bone marrow cultures from KO mice showed significantly fewer alkaline phosphatase-positive colony-forming units and osteoclasts compared to cultures from WT mice. Significantly less new bone and significantly fewer osteoclasts were observed in the drill sites of PAR(2) KO mice compared to WT mice 7 days post-surgery. A number of activators of PAR(2), including matriptase and kallikrein 4, were found to be expressed by normal bone marrow. Parathyroid hormone, 1,25 dihydroxyvitamin D(3), or interleukin-6 in combination with its soluble receptor down-regulated PAR(2) mRNA expression, and fibroblast growth factor-2 or thrombin stimulated PAR(2) expression. These results suggest that PAR(2) activation contributes to determination of cells of both osteoblast and osteoclast lineages within bone marrow, and thereby participates in the regulation of skeletal growth and bone repair.

Structural decay of bone microarchitecture in men with prostate cancer treated with androgen deprivation therapy.

CONTEXT: Androgen deprivation therapy (ADT) used in the treatment of prostate cancer reduces bone mineral density (BMD) and predisposes to fractures. The structural basis of the BMD deficit and bone fragility is uncertain. OBJECTIVE AND PATIENTS: We investigated changes in bone microarchitecture in 26 men (70.6±6.8 yr) with nonmetastatic prostate cancer during the first year of ADT using the new technique of high-resolution peripheral quantitative computed tomography. DESIGN AND SETTING: We conducted a 12-month prospective observational study in the setting of a tertiary referral center. RESULTS: After 12 months of ADT, total volumetric density decreased by 5.2±5.4% at the distal radius and 4.2±2.7% at the distal tibia (both P<0.001). This was due to a decrease in cortical volumetric BMD (by 11.3±8.6% for radius and 6.0±4.2% for tibia, all P<0.001) and trabecular density (by 3.5±6.0% for radius and 1.5±2.3% for tibia, all P<0.01), after correcting for trabecularization of cortical bone. Trabecular density decreased due to a decrease in trabecular number at both sites (P<0.05). Total testosterone, but not estradiol, levels were independently associated with total and corrected cortical volumetric BMD at the tibia. CONCLUSIONS: Sex steroid deficiency induced by ADT for prostate cancer results in microarchitectural decay. Bone fragility in these men may be more closely linked to testosterone than estradiol deficiency.

Heparin-flush associated thrombocytopenia--induced hemorrhage: a case report.

Thrombocytopenia is a well known complication of heparin. It occurs when heparin is used in full therapeutic dose or in subcutaneous mini-doses for prophylaxis. This has also been reported with minuscule dose of heparin used in heparin-flushes to keep vascular access catheters patient. We are reporting for the first time, a case of severe hemorrhage due to thrombocytopenia associated with heparin flushes.