Ricardo Francisco Capozza

Mechanical validation of a tomographic (pQCT) index for noninvasive estimation of rat femur bending strength

Cross-sectional moment of inertia (CSMI) and volumetric cortical bone mineral density (vCtBMD) were assessed by peripheral quantitative computed tomography (pQCT) at femur midshafts from 103 Wistar female rats receiving 0 (n = 12) or 15-1000 mu g/kg/day sc of dexamethasone (n = 46) from 5 to 9 weeks of age, or 0 or 80 mg/kg 3/wk of AI(OH)(3) IP (n = 23,22) from 4 to 10 months of age. A bone strength index (BSI), calculated as the product CSMI x vCtBMD, was found to closely correlate (r = 0.94, R(2) = 0.89, p < 0.001) with the actual, mechanically tested bending breaking force of all bones. Correlation and determination coefficients obtained were higher than those usually reported employing different long-bone strength predictive formulae. The curve approached the origin and was linear throughout the wide range of CSMI, vCtBMD and BSI achieved because of age- and treatment-induced differences, showing a very low standard error of the estimate. Instead, different curve slopes and/or intercepts were found in separate analysis between data from each of the experiments when breaking force was correlated with CSMI or vCtBMD alone, or with the DEXA-assessed BMD of the mechanically assayed bone portion. Results suggest that noninvasive assessment of the BSI by means of pQCT technology provides an original tool for a precise and accurate estimation of long-bone bending strength that can be advantageously applied in crosssectional as well as longitudinal, in vivo studies employing animal models.

Dexamethasone effects on mechanical, geometric and densitometric properties of rat femur diaphyses as described by peripheral quantitative computerized tomography and bending tests

In previous studies with cortisol, betamethasone and oxazacort we attributed glucocorticoid effects on bone biomechanics to changes in bone mass and geometry rather than to an action on bone material properties. In this experiment, groups of 7 rats each received subcutaneous doses of 15.6, 31.2, 62.5, 125, 250, 500 or 1000 micrograms/kg per day of dexamethasone (DMS) and an additional 14 animals were controlled untreated for 4 weeks. Their fresh femurs were then scanned by peripheral quantitative computerized tomography (pQCT; XCT-960, Stratec, Germany) at the midshaft and submitted to three-point bending tests. In consonance with our earlier investigations, a significant, log-dose-related reduction in bone load-bearing capacity was observed, associated with an impairment in bone geometric properties (cross-sectional area and moment of inertia) and in body weight gain. However, the pQCT-assessed volumetric mineral density of cortical bone (vCtBMD; regarded as a material quality indicator in terms of mineralization) was significantly reduced by DMS following a dose-response relationship. Furthermore, a direct association was detected between vCtBMD and diaphyseal load-bearing capacity and stiffness. In contrast with our previous approach, data suggests that, apart from changes in bone geometric properties, glucocorticoid effects on bone material quality--as assessed by vCtBMD changes in this study--seem also to play a significant role in the determination of their biomechanical consequences.

Bone Quality Parameters of the Distal Radius as Assessed by pQCT in Normal and Fractured Women

Abstract. The aim of this study was to test the ability of some indicators of different aspects of bone quality (assessed by peripheral quantitative computed tomography in the distal radius) to discriminate between fractured and nonfractured individuals. The study compared 214 women aged 45–85 years, free of any bone-affecting treatment, of whom 107 had suffered a Colles” fracture in the previous 6 months and 107 did not. The determinations included bone tissue or mineral “mass” indicators (trabecular, cortical and total volumetric mineral content, cortical bone area); bone “density” estimates (trabecular, cortical and total volumetric mineral density), and the Cartesian (rectangular) and polar moments of inertia as influences of cross-sectional architecture on resistance to bending and torsional loads, respectively. The influences of body height, weight and age on the tomographic indicators were minimized by adjusting the data according to the partial coefficients of multiple stepwise regressions. The adjusted values of all the indicators were lower in fractured than in nonfractured groups. The prevalence of fractures was directly related to the actual values of the indicators, rather than the age or body habitus of the individuals. The significance of these differences between the assessed indicators decreased in the following order: trabecular “mass” > trabecular “density” > cortical or total “mass” > cortical architecture > total or cortical “density” indicators. Within the same type of bone, the tissue or mineral “mass” indicators performed better than the “density” indicators. The cortical bone density did not give useful information, probably because of technical difficulties. Odds-ratios and receiver-operating characteristic (ROC) analyses confirmed those features. The selected “cut-off” values of the indicators as determined by the ROC curves (very close to those determined by the inflexion points of the logistic reression curves) may indicate reference limits to detect persons at risk of fracture according to the type of information provided by each variable. These results show that these tomographic indicators discriminate well between fractured and nonfractured individuals, and should be suitable to assess how total, cortical and trabecular bone strength in the distal radius could affect different kinds of strength regardless of the age or body habitus of the individual. Their ability to estimate fracture risk from different biomechanical points of view should be assessed by adequately designed, prospective studies.

Determination of femur structural properties by geometric and material variables as a function of body weight in rats. Evidence of a sexual dimorphism.

Femur diaphyses of male and female Wistar rats were densitometrically and biomechanically assayed. The BMD-dependent material properties were better in female than in male bones, but cross-section geometric properties were better in male femurs. As a result, mechanical properties of the integrated diaphyses were better in males, but differences disappeared after statistical adjustment of data to a common body weight. Results evidence a feed-back mechanism locally controlling the strain-dependent bone modelling and the corresponding cross-sectional design as related to bone stiffness, with a set-point adjusted to animal biomass. A sexual dimorphism of bone biomechanics is also described for the species.

Structural analysis of the human tibia by tomographic (pQCT) serial scans

This study analyses the evaluation of tomographic indicators of tibia structure, assuming that the usual loading pattern shifts from uniaxial compression close to the heel to a combined compression, torsion and bending scheme towards the knee. To this end, pQCT scans were obtained at 5% intervals of the tibia length (S5–S95 sites from heel to knee) in healthy men and women (10/10) aged 20–40 years. Indicators of bone mass [cortical area, cortical/total bone mineral content (BMC)], diaphyseal design (peri/endosteal perimeters, cortical thickness, circularity, bending/torsion moments of inertia – CSMIs), and material quality [(cortical vBMD (bone mineral density)] were determined. The longitudinal patterns of variation of these measures were similar between genders, but male values were always higher except for cortical vBMD. Expression of BMC data as percentages of the minimal values obtained along the bone eliminated those differences. The correlative variations in cortical area, BMC and thickness, periosteal perimeter and CSMIs along the bone showed that cortical bone mass was predominantly associated with cortical thickness toward the mid‐diaphysis, and with bone diameter and CSMIs moving more proximally. Positive relationships between CSMIs (y) and total BMC (x) showed men’s values shifting to the upper‐right region of the graph and women’s values shifting to the lower‐left region. Total BMC decayed about 33% from S5 to S15 (where minimum total BMC and CSMI values and variances and maximum circularity were observed) and increased until S45, reaching the original S5 value at S40. The observed gender‐related differences reflected the natural allometric relationships. However, the data also suggested that men distribute their available cortical mass more efficiently than women. The minimum amount and variance of mass indicators and CSMIs, and the largest circularity observed at S15 reflected the assumed adaptation to compression pattern at that level. The increase in CSMIs (successively for torsion, A–P bending, and lateral bending), the decrease in circularity values and the changes in cortical thickness and periosteal perimeter toward the knee described the progressive adaptation to increasing torsion and bending stresses. In agreement with the biomechanical background, the described relationships: (i) identify the sites at which some changes in tibial stresses and diaphyseal structure take place, possibly associated with fracture incidence; (ii) allow prediction of mass indicators at any site from single determinations; (iii) establish the proportionality between the total bone mass at regions with highly predominant trabecular and cortical bone of the same individual, suitable for a specific evaluation of changes in trabecular mass; and (iv) evaluate the ability of bone tissue to self‐distribute the available cortical bone according to specific stress patterns, avoiding many anthropometric and gender‐derived influences.

Association between low lean body mass and osteoporotic fractures after menopause

Objective: This study evaluated dual-energy x-ray absorptiometry-assessed whole-body bone-muscle relationship (bone mineral content/lean mass [BMC/LM]) as an indicator of its nonmechanical perturbations (ie, systemic) in pre- and postmenopausal women. A total of 3,205 women were studied, either healthy (no fracture [No Fx] groups, 1,035 premenopausal, 1,556 postmenopausal) or with recent fractures (Fx groups, 139 premenopausal, 475 postmenopausal) located at osteoporotic sites (hip, spine, long-bone metaphyses; Type II Fx, n = 386) or at other skeletal sites (Type I Fx, n = 228) to evaluate the impact of decreased muscle mass on fracture incidence before and after menopause. Design: SD-scored graphs of BMC/LM proportionality were obtained from the No Fx groups as normal references. Based on the reference BMC versus LM curves obtained from their respective No Fx pre- and postmenopausal controls, BMC-LM SD scores were calculated for all women with fractures. Results: BMC-LM SD scores in all premenopausal women with fractures and in Type I Fx postmenopausal women were similar to the reference. In contrast, SD scores in Type II Fx postmenopausal women were lower than the reference, especially in those with hip fractures. Except for Type II Fx postmenopausal women, all groups showed linear and similar BMC versus LM curves. Type II Fx postmenopausal women showed nonlinear relationships, with progressively decreasing BMC and BMC-LM SD scores as their LM decreased. Conclusions: Results suggest that both LM and BMC-LM SD scores can help to differentiate between systemic and mechanical (disuse-related) osteopenia/osteoporosis after menopause. Low LM values or BMC-LM SD scores seem to constitute additional fracture risk factors beyond those usually detected in premenopausal women or in women with other types of fractures. This application of dual-energy x-ray absorptiometry technology may lead to more effective diagnosis and treatment at low cost.

Biomechanical impact of aluminum accumulation on the pre- and post-yield behavior of rat cortical bone.

In order to analyze the effects on whole-bone behavior of aluminum accumulation, 14 rats, aged 90 days, received i.p. doses of 27 mg/day of elemental Al, as Al(OH)3, during 26 weeks, while 14 rats remained as controls. Their femur diaphyses were studied tomographically by peripheral quantitative computed tomography (pQCT) and mechanically tested in bending. The load/deformation curves obtained allowed distinction between effects observed during the linearly elastic (Hookean) and nonlinear (non-Hookean, or plastic) behaviors of bones before and after the yield point, respectively. Treatment reduced the cortical bone mineralization (volumetric bone mineral density [BMD], −2%; P < 0.01), with a negative impact on the bending stiffness (Young’s elastic modulus) and the yield stress of cortical bone (−18% and −13%; P < 0.05). Despite the absence of any cortical mass increase (cross-sectional area), improved spatial distribution of cortical tissue concerning anterior-posterior bending stress (cross-sectional moment of inertia, 10%; P < 0.05) occurred through a modulation of modeling drifts. Up to the yield point, neither the structural strength (load supported), the strain, nor the structural stiffness (load/deformation ratio) of the diaphyses were affected. This suggests an adaptive response of bone geometry to the impairment of bone material stiffness. However, Al intoxication significantly reduced the ultimate load and the post-yield fraction of that load (−6% and −27%; P < 0.05). This suggests that the proposed, adaptive response could have improved bone design so as to make it adequate for maintaining a normal pre-yield diaphyseal stiffness and strength according to the bone mechanostat theory, but not so as to provide complete protection of the diaphyseal post-yield (and ultimate) strength. Although a relative inhibition of bone formation could not be discarded, an Al-induced impairment of the bone ability to resist loads beyond the yield point could have caused the unusual disparity observed between effects on bone (elastic) stiffness and (ultimate) strength. In addition, to explain the unusual finding, these results suggest that little-studied microstructural factors (spatial arrangement of elements within the mineralized tissue) affecting the post-yield behavior of cortical bone, regardless of bone mineralization in these and other conditions, ought to be further investigated in specifically designed studies as a novel, promising resource in skeletal research.

Effects of large doses of olpadronate (dimethyl-pamidronate) on mineral density, cross-sectional architecture, and mechanical properties of rat femurs

As part of a safety-assessment study, doses of 8, 40, and 200 mg/kg per day, 6 days per week, of sodium olpadronate (dimethyl-APD, Me2-APD) were given by gavage to 10-week-old male and female rats during 27 weeks. Only the 200 mg/kg per day dose provoked toxic effects and a meaningful growth depression, regardless of the animal gender. In male animals, doses of 40 or 200 mg/kg per day improved strength, stiffness, and cross-sectional moment of inertia (CSMI) of femur diaphyses despite the toxic effects observed at the highest dose. Changes in bone mechanical properties were a consequence of those induced in CSMI. Regression analyses showed a treatment-induced improvement in bone modeling (as assessed by CSMI) for the same level of bone material stiffness (as expressed by calculated values of elastic modulus). The high dependency of results on body mass bearing suggested that these effects were exerted through an increase in the efficiency of bone mechanostat. Strikingly, they were not evident in female rats. If not related to a lower bone bioavailability of bisphosphonates in female rats as described by others, this phenomenon may have reflected: (1) their a smaller biomass; and/or (2) a less effective mechanostatic regulation of bone architecture derived from a higher bone material stiffness related to male animals. An increase of BMD with a predominance toward the distal region was observed in all femurs studied. This effect, unrelated to the observed changes in mechanical properties, seems to express a lack of remodeling of primary cartilage or bone tissue.

Absorptiometric assessment of muscle-bone relationships in humans: reference, validation, and application studies.

This report summarizes some preliminary absorptiometric (DXA, QCT/pQCT) studies from our laboratory, supporting the following assumptions. 1. InHomo sapiens at all ages, natural proportionality between DXA-assessed bone mineral mass (bone mineral content, BMC) and muscle mass (lean mass, LM) of the whole body or limbs is specific for ethnicity, gender, and reproductive status, but not for body weight, height, or body mass index. 2. This proportionality is sensitive to many kinds of endocrine-metabolic perturbations. 3. Percentilized or Z-scored charts of the BMC/LM correlations as determined in large samples of healthy individuals can provide a diagnostic reference for evaluating proportionality in different conditions. 4. Employing exclusively DXA, this methodology can be applied to discriminate between “disuse-related” and “metabolic” osteopenias based on the finding of normal or low BMC/LM percentiles or Z-scores respectively, with important therapeutic and monitoring implications.

Novel experimental effects on bone material properties and the pre- and postyield behavior of bones may be independent of bone mineralization

In this article, we summarize the results of six different tomographic/biomechanical rat studies involving hypophysectomy (Hx), ovariectomy, treatment with rhGH, olpadronate, alendronate, and toxic doses of aluminum and the development of a genetic diabetes in theeSS strain. All these conditions induced some interesting and rarely reported effects on postyield bone strength. These effects were generally related neither to the degree of mineralization or the elastic modulus of the bone tissue nor to the preyield behavior of the bones. In two particular cases (Hx,eSS), the elastic modulus of bone tissue varied independently of its degree of mineralization. These results suggest the involvement of some microstructural factor(s) of bone tissue resistance to crack progression (a postyield feature of bone behavior), rather than to crack initiation (the yield-determining factor) in the corresponding mechanism. Changes in collagen or crystal structure may play that role. These changes are relevant to the mechanism of fracture production during plastic deformation, a feature of bone strength that might be independent from mineralization. Therefore, these changes might help to explain some effects of novel treatments on bone strength unrelated to bone mineralization. This questions the belief that the remaining bone mass in metabolic osteopenias is biologically and mechanically normal.