Histomorphometric assessment of the long-term effects of alendronate on bone quality and remodeling in patients with osteoporosis.

Treatment effects on bone quality and remodeling was assessed in postmenopausal women with osteoporosis treated with oral alendronate. One transiliac bone biopsy was obtained from 231 women at either 24 mo (n = 11) or 36 mo (n = 120) from the start of treatment with alendronate at doses of between 5 and 20 mg/d, or placebo. 64 biopsies at 24 mo (31 from the placebo group and 33 alendronate-treated patients) and 95 biopsies at 36 mo (40 from the placebo group and 55 alendronate-treated patients) provided adequate cancellous tissue, and were analyzed by histomorphometry. Mineral apposition rate was unaffected by treatment. At 24 and 36 mo, osteoid thickness, volume, and surface significantly decreased. At each of the doses studied, mineralizing surface and activation frequency significantly decreased at each time point (e.g., -92% and -87%, respectively, for the 10 mg daily dose after 2 yr). These diminutions were of the same magnitude for each dose at 24 mo, and for the two highest doses at 36 mo. A significant increase in wall thickness accompanied by a reduction in erosion depth was detected in biopsies obtained at 24 mo. These findings confirm that mineralization is normal, and trabecular bone turnover markedly decreased in patients receiving long-term dosing with alendronate. The findings also suggest that the observed increases in bone mineral density could result both from a reduction in the remodeling space due to a decreased activation frequency and a possible trend to a positive bone balance. In addition, further studies focused on a possible increase in the degree of mineralization of bone are required.

Contribution of the advanced glycation end product pentosidine and of maturation of type I collagen to compressive biomechanical properties of human lumbar vertebrae.

Collagen characteristics contribute to bone biomechanical properties. Yet, few studies have analyzed the independent contributions of bone mineral density (BMD) and post-translational modifications of type I collagen to whole bone strength. Thus, the aim of this study was to determine the relative contributions of BMD and both enzymatic and non-enzymatic collagen crosslink concentration to the biomechanical properties of human vertebrae. Nineteen L3 vertebrae were collected after necropsy (age 26-93; 10 males, 9 females). BMD of the vertebral body was measured by DXA, and the vertebrae were compressed to failure to assess the stiffness, failure load and work to fracture. After mechanical testing, the concentration of both enzymatic crosslinks pyridinoline (PYD), and deoxypyridinoline (DPD) as well as, and the non-enzymatic crosslinks pentosidine (PEN) were analyzed in trabecular and cortical bone by reversed-phase HPLC. The extent of aspartic acid isomerization of type I collagen C telopeptide (CTX) was evaluated by ELISA of native (alpha CTX) and isomerized (beta CTX) forms. BMD was significantly positively related with stiffness (R(2) = 0.74; P < 0.0001), failure load (R(2) = 0.69; P < 0.0001) and work to fracture (R(2) = 0.44; P = 0.002). Bivariate regression analysis showed no association between collagen traits and biomechanical properties. However, in a multiple regression model, BMD and trabecular PEN were both significantly associated with failure load and work to fracture (multiple R(2) = 0.83, P = 0.001 and R(2) = 0.67, P = 0.001, respectively). Similarly, BMD and trabecular alpha/beta CTX ratio were both associated with stiffness (multiple R(2) = 0.83, P = 0.015). These findings indicate that post-translational modifications of type I collagen have an impact on skeletal fragility.

Relationship between compressive properties of human os calcis cancellous bone and microarchitecture assessed from 2D and 3D synchrotron microtomography.

The aim of this study was to determine the contribution of 2D and 3D microarchitectural characteristics in the assessment of the mechanical strength of os calcis cancellous bone. A sample of cancellous bone was removed in a medio-lateral direction from the posterior body of calcaneus, taken at autopsy in 17 subjects aged 61-91 years. The sample was first used for the assessment of morphological parameters from 2D morphometry and 3D synchrotron microtomography (microCT) (spatial resolution=10 microm). The 2D morphometry was obtained from three slices extracted from the 3D microCT images. Very good concordance was shown between 3D microCT slices and the corresponding physical histologic slices. In 2D, the standard histomorphometric parameters, fractal dimension, mean intercept length, and connectivity were computed. In 3D, histomorphometric parameters were computed using both the 3D mean intercept length method and model-independent techniques. The 3D fractal dimension and the 3D connectivity, assessed by Euler density, were also evaluated. The cubic samples were subjected to elastic compressive tests in three orthogonal directions (X, Y, Z) close to the main natural trabecular network directions. A test was performed until collapse of trabecular network in the main direction (Z). The mechanical properties were significantly correlated to most morphological parameters resulting from 2D and 3D analysis. In 2D, the correlation between the mechanical strength and bone volume/tissue volume was not significantly improved by adding structural parameters or connectivity parameter (nodes number/tissue volume). In 3D, one architectural parameter (the trabecular thickness, Tb.Th) permitted to improve the estimation of the compressive strength from the bone volume/tissue volume alone. However, this improvement was minor since the correlation with the BV/TV alone was high (r=0.96). In conclusion, which is in agreement with the statistic's rules, we found, in this study, that the determination of the os calcis bone compressive strength using the 3D bone volume fraction cannot be improved by adding 3D architectural parameters.

Apparent pre- and postmenopausal bone loss evaluated by DXA at different skeletal sites in women: the OFELY cohort.

We measured the bone mineral density (BMD) at various skeletal sites (total body, hip, anteroposterior [AP] and lateral [lat] spine, and forearm) in a large population-based cohort of women aged 31-89 years (the OFELY cohort), and results were analyzed according to age and postmenopausal years. A significant apparent bone loss was found before the menopause in cancellous bone, i.e., at the lat spine and Ward's triangle (-10%; p < 0.05-0.001). Cross-sectional analysis indicated that, after the menopause, apparent bone loss was accelerated within the 10 years following menopause, continued thereafter at all sites except the AP spine, and was again accelerated in elderly menopausal for more than 25 years. Between 30 and 80 years, BMD decreased by 15 to 44% (T score -1.6 to -3.4) according to the site. The amount of apparent bone loss was highest at the Ward's triangle when expressed in percentage (44%) and at the mid- and distal radius when expressed in number of standard deviations from the peak bone mass (-3.4). As a result, the percentage of women classified as osteoporotic according to the World Heath Organization, i.e., with a T score < or = -2.5, varied substantially from site to site and was highest at the radius (37% and 46%) and lateral spine (25-31%), intermediate at the Ward's triangle, AP spine, and whole body BMD, and lowest at the whole body bone mineral content, femoral neck, and trochanter (10-12%). In conclusion, this cross-sectional but large study suggests that there is a moderate apparent premenopausal bone loss that occurs only at cancellous bone sites and that apparent bone loss is accelerated at most skeletal sites after the age of 75 years. Because of the highly variable coefficient of variation of the peak bone mass at various skeletal sites, the percentage of postmenopausal women identified as being osteoporotic varies widely according to the site of measurement.

Vitamin D receptor gene polymorphisms are not related to bone turnover, rate of bone loss, and bone mass in postmenopausal women: the OFELY Study.

Vitamin D receptor (VDR) gene polymorphisms have been reported to account for most of the well established genetic influence on bone mineral density (BMD). However, discordant studies have been published and it is still not clear whether VDR genotypes influence bone mass accretion and/or postmenopausal bone loss. In this study, we analyzed VDR gene polymorphisms, i.e., that of BsmI, ApaI, and TaqI restriction enzymes in 268 untreated postmenopausal women 1-26 years postmenopausal. There were 37 BBAA homozygote (absence of BsmI and ApaI restriction sites on both alleles), 55 bbaa homozygote (presence of restriction sites on both alleles), and 176 heterozygotes. At baseline, women between the three genotypes did not differ significantly in age, years since menopause, body mass index (BMI), nor dietary calcium intake. We found no relationship between VDR genotypes and bone turnover assessed by three serum markers of bone formation and three urinary bone resorption markers, nor with BMD measured at the spine, hip, forearm, and whole body by dual-energy X-ray absorptiometry (DXA). Rates of bone loss assessed by repeated DXA measurements over 2 years were highly significant (p = 0.02-0.0001) at all skeletal sites except for the lumbar spine but did not differ between genotypes at any sites either before or after adjustment for potential confounding factors such as years since menopause, BMI, calcium intake, serum 25 hydroxyvitamin D levels, and baseline BMD. When we restricted the analysis to early postmenopausal women, within 10 years of menopause (n = 128), lumbar spine bone loss became significant, but no significant difference between VDR genotypes in the rate of bone loss measured at any site was found. We conclude that VDR genotypes are not predictive of bone turnover, rate of postmenopausal bone loss, and bone mass in either early or late postmenopausal women. In a subgroup of women with a low calcium intake (below 600 mg/day), we also found no significant differences between genotypes in BMD and the rate of bone loss measured at any site, although the sample size (n = 64) may be too small to detect small differences. In conclusion, these data, along with the absence of relationships between VDR gene polymorphisms and peak bone mass that we recently reported, suggest that the determination of VDR genotypes is probably not a useful clinical test for the risk assessment of osteoporosis.

Iliac trabecular bone formation predicts radial trabecular bone density changes in type 1 osteoporosis.

In 28 patients with idiopathic or postmenopausal type 1 (spinal crush fracture) osteoporosis, resorption indices and dynamic measurements of trabecular bone formation based on in vivo tetracycline labeling in 7.5 mm transiliac biopsies have been compared with trends in radial cortical and trabecular bone density measured with computed tomography. Positive correlations were observed between trabecular bone density trends in the radius and indices of bone formation in the ilium. These were improved when one of the two resorption indices was included with a formation index in bivariate regressions. Marked interindividual variations in radial bone density trends were also seen in cortical bone. These correlated poorly with trends in trabecular bone. Weak negative relationships between cortical bone trends and indices relating to bone formation and resorption were observed, but a positive association was seen with single-labeled surfaces on iliac trabeculae. If, as has been suggested, there are periodic variations in bone formation, the results suggest that axial and peripheral trabecular bone density trends are synchronized in osteoporosis, perhaps in response to systemic factors, such as circulating hormones.

The assessment of bone formation and bone resorption in osteoporosis: a comparison between tetracycline-based iliac histomorphometry and whole body 85Sr kinetics.

Bone formation and resorption have been measured in patients with idiopathic osteoporosis by histomorphometry of 7.5-mm trephine biopsies and in the whole body by 85Sr radiotracer methodology and calcium balances. The studies were synchronized and most were preceded by double in vivo tetracycline labeling. Correlations between histological and kinetic bone formation indices were better when better when based on the extent of double tetracycline labels than on measurements of osteoid by visible light microscopy. Correction of the kinetic data for long-term exchange, using 5 months' serial whole body counting of retained 85Sr, improved the fit of the kinetic to the histological data. A statistical analysis of the measurement uncertainties showed that the residual scatter in the best correlations (between exchange-corrected bone formation rates and double-labeled osteoid surface indices) could be attributed to measurement imprecision alone. The exchange-corrected resorption rate correlated fairly well with iliac trabecular resorption surfaces, and using a volume referent rather than a surface referent for the histological index improved the statistical fit when patients with therapeutically accelerated bone turnover were included. A much better correlation was obtained by including osteoid volume acting as an independent predictor of bone resorption in a bivariate regression with a resorption surface index. The residual errors could then be accounted for by known measurement uncertainties. Whereas osteoid taking a double label closely predicted the kinetic rate of bone formation, further analysis suggested that osteoid that took no label or a single label was more closely related to bone resorption, presumably as a secondary result of the coupling of bone formation to bone resorption. The idea that continued bone loss in some patients is associated with defective osteoblastic bone formation is supported by the low rates found in some patients by both techniques. Heuristically these studies validate both in vivo tetracycline labeling for dynamic histomorphometry and corrections for long-term exchange in kinetic studies of bone formation, providing a quantitative framework for the design and analysis of future studies of bone remodeling in the osteoporoses.

Comparison of trabecular bone microarchitecture and remodeling in glucocorticoid-induced and postmenopausal osteoporosis.

Long-term treatment with glucocorticoids (GCs) leads to a rapid bone loss and to a greater risk of fractures. To evaluate the specific effects of this treatment on cancellous bone remodeling, structure, and microarchitecture, we compared 22 transiliac biopsy specimens taken in postmenopausal women (65 +/- 6 years) receiving GCs (> or = 7.5 mg/day, for at least 6 months) and 22 biopsy specimens taken in age-matched women with postmenopausal osteoporosis (PMOP), all untreated and having either at least one vertebral fracture or a T score < -2.5 SD. On these biopsy specimens, we measured static and dynamic parameters reflecting trabecular bone formation and resorption. Also, we performed the strut analysis and evaluated the trabecular bone pattern factor (TBPf), Euler number/tissue volume (E/TV), interconnectivity index (ICI), and marrow star volume (MaSV). Glucocorticoid-induced osteoporosis (GIOP), when compared with PMOP, was characterized by lower bone volume (BV/TV), trabecular thickness (Tb.Th), wall thickness (W.Th), osteoid thickness (O.Th), bone formation rate/bone surface (BFR/BS), adjusted mineral apposition rate/bone surface (Aj.AR/BS), and higher ICI and resorption parameters. After adjustment for BV/TV, the W.Th remained significantly lower in GIOP (p < 0.0001). The active formation period [FP(a+)] was not different. Patients with GIOP were divided into two groups: high cumulative dose GCs (HGCs; 23.7 +/- 9.7 g) and low cumulative dose GCs (LGCs; 2.7 +/- 1.2 g). HGC when compared with LGC was characterized by lower W.Th (p < 0.05), BV/TV (p < 0.001), Tb.Th (p < 0.05), trabecular number (Tb.N; p < 0.05), FP(a+)(p < 0.05), and nodes (p < 0.05), and higher E/TV (p < 0.05), ICI (p < 0.005), and TBPf (p < 0.05). When HGC was compared with PMOP, the results were similar except for the MaSV, which was significantly higher (p < 0.005). In summary, GIOP was characterized by lower formation and higher resorption than in PMOP, already present after LGC. With HGCs, these changes were associated with a more dramatic bone loss caused by a major loss of trabecular connectivity.

Effects of alendronate on bone quality and remodeling in glucocorticoid-induced osteoporosis: a histomorphometric analysis of transiliac biopsies.

Effects of alendronate (ALN) on bone quality and turnover were assessed in 88 patients (52 women and 36 men aged 22-75 years) who received long-term oral glucocorticoid exposure. Patients were randomized to receive oral placebo or alendronate 2.5, 5, or 10 mg/day for 1 year and stratified according to the duration of their prior glucocorticoid treatment. Transiliac bone biopsies were obtained for qualitative and quantitative analysis after tetracycline double-labeling at the end of 1 year of treatment. As previously reported in glucocorticoid-induced osteoporosis, low cancellous bone volume and wall thickness were noted in the placebo group as compared with normal values. Alendronate treatment was not associated with any qualitative abnormalities. Quantitative comparisons among the four treatment groups were performed after adjustment for age, gender, and steroid exposure. Alendronate did not impair mineralization at any dose as assessed by mineralization rate. Osteoid thickness (O.Th) and volume (OV/BV) were significantly lower in alendronate-treated patients, irrespective of the dose (P = 0.0003 and 0.01, respectively, for O.Th and OV/BV); however, mineral apposition rate was not altered. As anticipated, significant decreases of mineralizing surfaces (76% pooled alendronate group; P = 0.006), activation frequency (-72%; P = 0.004), and bone formation rate (-71%; P = 0.005) were also noted with alendronate treatment. No significant difference was noted between the changes observed with each dose. Absence of tetracycline label in trabecular bone was noted in approximately 4% of biopsies in placebo and alendronate-treated groups. Trabecular bone volume, parameters of microarchitecture, and resorption did not differ significantly between groups. In conclusion, alendronate treatment in patients on glucocorticoids decreased the rate of bone turnover, but did not completely suppress bone remodeling and maintained normal mineralization at all alendronate doses studied. Alendronate treatment did not influence the osteoblastic activity, which is already low in glucocorticoid-induced osteoporosis.

Serum osteocalcin is increased in patients with osteomalacia: correlations with biochemical and histomorphometric findings.

The synthesis of osteocalcin or bone gla protein by osteoblasts is markedly stimulated by 1,25-(OH)2D, a key hormone in the regulation of bone mineralization. The circulating levels of osteocalcin have been shown to reflect both the osteoid matrix production and the formation rate of mineralized bone in several metabolic bone diseases (osteoporosis, thyrotoxicosis, primary hyperparathyroidism) in which both mechanisms are tightly coupled because of the absence of mineralization defect. In this study, we measured in 12 patients (7 women, 5 men, 56 +/- 15 yr old) with untreated osteomalacia serum osteocalcin and vitamin D metabolites (25OHD and 1,25-(OH)2D). The results were correlated with biochemical and histomorphometric assessment of bone remodeling. Osteomalacia was due to vitamin D deficiency (5 cases), to vitamin D malabsorption (6 cases), and to hypophosphataemia in 1 case. When compared to control values, serum osteocalcin was increased in patients with osteomalacia (7.4 +/- 4 vs. 3.7 +/- 1.3 ng/mL; P less than 0.001) and was positively correlated with serum alkaline phosphatase (r = 0.65; P = 0.03) and negatively with 25 OHD (r = -0.61; P = 0.04). Serum osteocalcin was not correlated with 1,25-(OH)2D [r = -0.45; not significant (NS)] even after exclusion of the patient with hypophosphataemia. Serum osteocalcin was positively correlated with the osteoid volume and osteoid perimeter (r = 0.71 and 0.69 respectively; P less than 0.01) but not with any of the tetracycline-based parameter of bone mineralization at the tissue level (r ranging from -0.41 to +0.42, NS). Serum 25 OHD, but not 1,25-(OH)2D, was positively correlated with the mineralization rate (r = 0.59; P less than 0.05 and r = 0.54; NS). We conclude that in patients with osteomalacia, a condition which is characterized by an increased osteoid accumulation due to a decreased mineralization rate, the increased level of serum osteocalcin reflects the increased osteoid synthesis but not the mineralization defect. In this disease, serum osteocalcin is inversely correlated to the severity of vitamin D deficiency reflected by serum 25 OHD, but not to the serum levels of 1,25-(OH)2D.

Intermethod variation in bone histomorphometry: comparison between manual and computerized methods applied to iliac bone biopsies.

The intermethod variation in the measurement of basic bone histomorphometric parameters was evaluated on 100 undecalcified transiliac bone biopsies. Two contiguous samples were taken from 50 patients (33 females; 17 males; mean age: 52 +/- 19 years) for diagnostic purposes. The diagnoses were osteoporosis (n = 38), renal osteodystrophy (n = 18), primary hyperparathyroidism (n = 16), osteomalacia (n = 12), metastatic bone disease (n = 2), thyrotoxic bone (n = 2), fluorosis (n = 2), and 10 biopsies were considered as "normal" bone. Trabecular bone volume (TBV) was measured with both a manual integrating eyepiece and an automatic (QUANTIMET 720-Cambridge Instruments, Cambridge, England) method. Trabecular resorption surfaces (TRS), trabecular osteoid surfaces (TOS), and volume (TOV) were measured with both a manual and a semiautomatic (VIDEOPLAN-Kontron, Munich, West Germany) method. The calcification rate (CR) was measured with both a manual and a semiautomatic method in eight cases after double labeling with tetracycline. Inter- and intraobserver variations were always lower with the automatic and semiautomatic methods than with the manual method, except for TOV. For all the parameters there was a highly significant correlation between manual and computerized methods (0.98 greater than r greater than 0.90). For TBV and CR no significant difference was noted, but for TBV the QUANTIMET appeared more sensitive, that is, better able to detect low values of the structure to be measured. For TRS, the manual method underestimated low values and appeared less sensitive than the semiautomatic method. For the 100 biopsies, the VIDEOPLAN underestimated the osteoid parameters by 13% for TOS and 26% for TOV.(ABSTRACT TRUNCATED AT 250 WORDS)

Automatic-interactive measurement of resorption cavities in transiliac bone biopsies and correlation with deoxypyridinoline.

Measuring bone resorption accurately by histomorphometry of bone biopsies is a challenge. Several techniques have been proposed including the measurement of eroded surfaces and resorption depth, but they have not been compared between themselves nor with biochemical assessment of bone resorption. In addition, there is a need for a rapid method that could be used more routinely. We describe here an automatic interactive method using a color analyzer (Visiolab, BIOCOM, France) with a specific software for the evaluation of erosion depth, eroded volume, eroded surface, osteoclast number, and surface. Thirty transiliac undecalcified bone biopsies stained with Goldner's trichrome were used in this study, taken from subjects suffering from osteoporosis or primary hyperparathyroidism. At the time of the biopsy a 2 h fasting morning urine sample was collected for measurement by HPLC of total deoxypyridinoline, the most sensitive marker of bone resorption. There was a highly significant correlation between maximum erosion depth measured directly and the one calculated according to the count of eroded lamellae (E. F. Eriksen, et al. Metab Bone Dis Relat Res 5:243-252; 1984) (r = 0.76; p = 0.0001). A significant correlation was found between urinary deoxypyridinoline and eroded volume/bone volume in cancellous and endocortical bone measured with the automatic interactive technique (r = 0.48; p = 0.007). In contrast, other histological indexes of bone resorption did not correlate with urinary deoxypyridinoline. The volume of resorption cavities appears to be the most valid index of bone resorption rate as it was correlated with the urinary excretion of total deoxypyridinoline.(ABSTRACT TRUNCATED AT 250 WORDS)

The anabolic effect of human PTH (1-34) on bone formation is blunted when bone resorption is inhibited by the bisphosphonate tiludronate--is activated resorption a prerequisite for the in vivo effect of PTH on formation in a remodeling system?

Parathyroid hormone (PTH) and its (1-34) fragment are stimulators of bone turnover that have an anabolic effect increasing trabecular bone mass when administered intermittently by daily subcutaneous injections. Its clinical use in osteoporosis, however, has been limited by the concomitant increased bone resorption and deleterious effect on cortical bone. To evaluate if a treatment combining PTH and a potent inhibitor of bone resorption would retain the anabolic effect of PTH without increasing bone resorption, we analyzed the effects of PTH (1-34) (500 IU/d) with or without the bisphosphonate tiludronate (1 mg/kg per day) for 3 months on biochemical and histological indices of bone turnover in old female sheep, an animal model which has a slow bone remodeling activity that resembles the one of elderly women. As expected, PTH (1-34) induced a significant increase of urinary pyridinoline and hydroxyproline (reflecting bone resorption), and of serum osteocalcin and alkaline phosphatase (reflecting bone formation), that were consistent with an increase of resorption and tetracycline-based formation of bone measured on iliac crest biopsy. In contrast, all biochemical and histological indices of bone turnover were decreased in sheep receiving tiludronate, a potent inhibitor of bone resorption. Surprisingly, in the combined therapy group, biochemical and histological indices of both resorption and formation did not differ from the control groups. Thus, the model of old sheep, which closely resembles the situation in old human, shows that the anabolic effect of PTH on bone is not maintained when PTH is coadministered with a bisphosphonate, in marked contrast to results noted in the growing rat.(ABSTRACT TRUNCATED AT 250 WORDS)

Mechanical properties of ewe vertebral cancellous bone compared with histomorphometry and high-resolution computed tomography parameters.

The goal of the present study was to determine if a high-resolution computed tomography (HRCT) system with 150 microns resolution was sufficient to predict mechanical properties in ewe lumbar vertebrae. To answer this question, we used a triangular comparison between: HRCT; biomechanics (compression and shear tests); and histomorphometry, which was the reference method for the measurements of morphometric parameters. Two dissected lumbar vertebrae (L-4 and L-5) from 32 ewes were used. Both compressive and shear properties correlated significantly with amount of bone and structural parameters evaluated by histomorphometry (bone volume/tissue volume, trabecular thickness, trabecular separation), but no significant correlation was found with the trabecular number. With our shear test involving the trabecular architecture itself more significant correlations were found with the node-strut analysis parameters than from the compressive test. Significant correlations were also found between HRCT and histological parameters (bone volume/tissue volume, bone surface/bone volume, trabecular separation, trabecular number, total strut length, number of nodes, and number of termini). Correlations between HRCT structural parameters and mechanical properties on L-4 were of the same magnitude as the correlations between the histomorphometric structural parameters and mechanical results on L-5 but with the remarkable advantage the HRCT is a noninvasive method. In spite of the resolution (150 microns) of our HRCT system, which entailed mainly an enlargement of the thinnest trabeculae or their loss during the segmentation process, we obtained coherent relationships between mechanical and tomographic parameters. The thinnest trabeculae probably had little effect on the mechanical strength. Also, this type of resolution allows us to consider the possibility of perfecting an in vivo HRCT system. However, physical density and bone mineral density correlated much better with strength than either classical histomorphometric or tomographic parameters. The current conclusion is fairly negative with respect to the ability of HRCT to assess mechanical properties nondestructively as compared with dual-energy X-ray absorptiometry. But, the noninvasive nature of the imaging modality and the capacity for three-dimensional imaging at arbitrary orientation make HRCT a promising tool in the quantitative assessment of cancellous architecture.

Do ultrasound measurements on the os calcis reflect more the bone microarchitecture than the bone mass?: a two-dimensional histomorphometric study.

Few studies have analyzed the relationship between ultrasound measurements (US) and corresponding histomorphometric parameters of the calcaneus. To address this question we have compared US and histomorphometric parameters in 17 whole human os calcis from amputation or necropsy. Speed of sound (SOS), broadband ultrasound attenuation (BUA), and bone mineral density (BMD) were measured on the whole foot at the calcaneal site using an Achilles device and a DPX-L densitometer (Lunar). The os calcis was dissected and a 1-cm-wide transcortical parallelepiped extracted with a biopsy needle, focused on the center of the measured area. Histomorphometry was performed on undecalcified biopsies. Structural and connectivity parameters were measured on 7-microns-thick sections with both automatic (Biocom) and semiautomatic analyzers (Ibas 1, Kontron). We found that all ultrasonic and densitometric parameters reflected the true amount of bone and were correlated with only some of the parameters reflecting bone microarchitecture. From stepwise regression analysis, we found that 68%, 67%, 72%, and 74% of the variance of SOS, BUA stiffness, and BMD, respectively, were explained significantly by trabeculae thickness only. Ultrasonic measurements appear to reflect bone quantity rather than bone microarchitecture. The current conclusion is fairly negative with respect to the ability of ultrasound to assess structural parameters, but our limited sample size did not give enough power to our study to reach statistically significant correlations. In addition, the calcaneus is anisotropic and the ultrasound interaction in bone is a three-dimensional phenomenon. So, a three-dimensional study rather than a two-dimensional one should be performed.

Bone histology in adults with aseptic necrosis. Histomorphometric evaluation of iliac biopsies in seventy-seven patients.

UNLABELLED: We studied the bone histology by histomorphometric methods in transiliac bone-biopsy specimens from seventy-seven adult patients with aseptic osteonecrosis and normal kidney function. The trabecular bone volume, trabecular osteoid volume, trabecular osteoid surfaces, thickness index of osteoid seams, total resorption surfaces, calcification rate, tetracycline-labeled surfaces, and bone-formation rate at the basic multicellular unit level and at the tissue level were determined. Histological evidence of osteomalacia was found in nine patients, of whom four were alcoholics. In the remaining sixty-eight patients--fifteen treated with corticosteroids, twenty-nine alcoholics, and twenty-four who did not have any detectable etiological factor--a common histomorphometric profile was found. This consisted morphologically of a reduction in trabecular bone volume and in the thickness of osteoid seams, and dynamically of a reduction in calcification rate and in total labeled surfaces. All of these changes suggested a marked decrease in osteoblastic appositional rate and in bone-formation rate at the cell and tissue levels. This could induce a healing defect of microfractures and thus facilitate subchondral fractures. CLINICAL RELEVANCE: This histological study indicated that non-apparent bone disease--either osteoporosis or osteomalacia--may underlie aseptic osteonecrosis in almost all patients, and be found even when blood and urinary biochemical parameters, usually reflecting bone-remodeling, are normal. An iliac-crest bone biopsy with static and dynamic histomorphometric study is the appropriate method for detecting these abnormalities. These results are of importance for understanding the pathophysiological mechanisms underlying osteonecrosis as well as its prevention and treatment.

Mechanical characterization in shear of human femoral cancellous bone: torsion and shear tests.

In order to investigate and compare the mechanical behaviour of human cancellous bone during different shear loading modes, two tests were performed to characterise human femoral cancellous bone in shear: a torsion test until failure and a shear test using a sharpened stainless steel tube. Paired cylindrical samples were core drilled from 12 human femoral heads, symmetrically with respect to the coronal plane and along the primary trabecular direction. The distal part of the sample was assigned to a torsion test and the shear test was performed on the proximal part along two perpendicular anatomical directions. Apparent densities and tissue densities were measured on both torsion and shear specimens. The mean torsion properties were shear modulus G, 289 (183) MPa, ultimate stress tau(torsion), 6.1 (2.7) MPa, ultimate strain gamma(ultimate), 4.6 (1.3)%, yield stress tau(yield), 4.3 (1.9) MPa and yield strain gamma(yield), 1.8 (0.3)%. Strong correlation was obtained between G and tau(torsion) (r'=0.853, p<0.001). These torsion properties were correlated with apparent density of torsion specimens showing, respectively: r'=0.713, p=0.005 and r'=0.671, p=0. 008. Properties from the shear test were invariable with regard to the two tested directions then isotropic ultimate shear stress and isotropic elementary shear stress, which represent the mean values of the two tested directions were, respectively, tau(shear), 10.0 (4. 5) MPa and tau(elem), 18.8 (6.1) MPa. Both shear stresses were correlated with apparent density of shear specimens: tau(shear), r'=0.564, p=0.045 and tau(elem), r'=0.636, p=0.024. Apparent densities for shear specimens were superior than for torsion specimens (p=0.06) and the comparison was the opposite for tissue densities (p=0.028), showing strong density gradients of cancellous bone in the femoral head. These torsion and shear tests which permit the evaluation of cancellous bone behavior under two different types of shear loading, may be performed on different human sites and the measured shear properties may be compared to structural properties of cancellous bone.

Staining procedure for the detection of microcracks: application to ewe bone.

Microcracks are one of the determinants of the bone strength and their accumulation may contribute to increased fracture risk. They are detected after bulk staining with various dyes, including basic fuschin, calcein and xylenol orange. The duration of staining usually varies across types of bone and species. The ewe is a large animal with a bone remodeling similar to humans, used as an animal model in bone histomorphometry studies. The aim of the present study was to determine the optimal conditions for bulk staining with xylenol orange of ewe bone. Xylenol orange 5mM in 70% ethanol was applied to iliac crest and vertebral biopsies for 2 or 15 days or 1, 2 or 3 months. After embedding, sections of 40, 50 and 80 µm thick were cut with either a precision diamond wire saw or a microtome. The staining was not visible after 2 or 15 days and was heterogeneous after 1 or 2 months. The quality of 40 and 50 µm thick sections was not preserved compared with those of 80 µm. Microcracks were suitably observed on ewe bone after bulk staining with xylenol orange for 3 months, in 80 µm thick sections. We conclude that the staining procedures should differ when examining ewe or human bone. This may be due to differences in bone matrix composition.