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.

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.

Trabecular bone microarchitecture after alendronate treatment of osteoporotic women.

OBJECTIVE: To compare the microarchitecture of iliac crest trabecular bone from women treated for two to three years with alendronate versus that of women treated with placebo. RESEARCH DESIGN AND METHODS: Three-dimensional micro-computed tomography (micro-CT; resolution 20 microm) and two-dimensional histomorphometry (resolution 5-7 microm) were used to examine trabecular bone from single transilial biopsies obtained at the completion of clinical trials. MAIN OUTCOME MEASURES: Microarchitectural variables, including bone volume, trabecular number, trabecular thickness, and trabecular spacing in specimens from alendronate- and placebo-treated women were examined. Three-dimensional images of trabecular bone from both groups were constructed from CT images. Correlations among variables and between techniques were also calculated. RESULTS: Eighty-eight specimens were suitable for evaluation by both techniques. As measured by two-dimensional histomorphometry, bone volume fraction (as a proportion of total volume) and trabecular thickness were significantly greater in alendronate specimens, 17.1 +/- 5.5% vs. 13.4 +/- 5.5% (p = 0.0043) and 127 +/- 29 microm vs. 109 +/- 28 microm (p = 0.0090), respectively, and trabecular spacing was significantly smaller, 729 +/- 227 microm vs. 862 +/- 338 microm (p = 0.005). Micro-CT yielded similar findings: bone volume and trabecular number were significantly greater in alendronate specimens: 19.4 +/- 6.2% vs. 16.2 +/- 6.3% (p = 0.0412) and 1.46(+/-) 0.32 vs. 1.31(+/-) 0.33 per mm (p = 0.0346). Two-dimensional and micro-CT measured characteristics correlated strongly with one another, with Pearson product moment correlation coefficients ranging from 0.60 (for trabecular thickness) to 0.83 (for bone volume). CONCLUSIONS: Trabecular microarchitecture of the ilium, whether studied by two- or three-dimensional methods, is better (greater bone volume, greater trabecular thickness, decreased trabecular spacing) after alendronate treatment than after two to three years of treatment with placebo. Bone volume in a trabecular region is strongly correlated to its microarchitecture, suggesting that bone quantity predicts values for these microarchitectural endpoints.

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.

[Progressive limb lengthening with a centromedullary nail versus an external fixator: experimental study in sheep]. / L'allongement progressif des membres par clou mécanique centro-médullaire d'allongement: étude expérimentale chez le mouton.

PURPOSE OF THE STUDY: Progressive limb lengthening with an external fixator often leads to pin-related complications. A new technique allowing progressive lengthening with a centromedullary nail without external fixation has been developed. This original double-locked device consists of matching male and female components fitted with a continuous thread. Lengthening is achieved via a one-way ratchet system. Twelve back-and-forth movements produce 1.25 mm lengthening. MATERIAL AND METHODS: We tested this new device on 20 sheep and compared results with external fixation lengthening in 20 other sheep. The animals were divided into groups for sacrifice on days 5, 10, 20, 45 and 90. Serial x-ray were obtained for all animals. In the 45-day and 90-day groups, histomorphometric (trichrome goldner coloration and polarized light microscopy) and densitometric studies were also performed. Bone mineral density (BMD) was determined and bone trabecular density (BTD) and trabecular bone volume (TBV) were expressed in percent of bone trabecular surface area. RESULTS: Mean lengthening in the 45-day and 90-day groups was 39 mm for the nail and 20 mm for external fixation (1 mm/day). At 90 days, 3 sheep out of 4 had consolidated radiologically with external fixation and 2 out of 4 with the nail. BMD was slightly better for external fixation (0.811 vs 0.695/cm(2)). This difference could probably be attributed to the greater lengthening obtained with the nail. At 45 days, BMD was the same (0.6 g/cm(2)) for both devices. BTD was nearly two-fold higher for the nail compared with external fixation (59.65% vs 32.61% at 90 days), most probably due to primary bone formation. The histomorphometric study allowed an analysis of the osteoid border. Bone quality obtained in the bone regenerate with the nail was superior to that obtained with external fixation. Primary bone formation resulted from membrane ossification with direct transformation of fibroblasts into osteoblasts. CONCLUSION: This work demonstrated that progressive lengthening can be achieved with a specifically designed centromedullary nail without iterative opening of the operative site. Tolerance to this type of device and quality of the bone regenerate are altogether satisfactory.

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.

Lumbar osteoarthritis, bone mineral density, and quantitative ultrasound.

Low bone mass is a major risk factor for osteoporotic fractures. Thus, bone density evaluation, performed by Dual Energy X-ray Absorptiometry (DXA) is important for diagnosis and monitoring treatment of osteoporosis. The accuracy of DXA, particularly at the lumbar spine, can be affected by several factors such as degenerative diseases. To evaluate the effects of vertebral osteophytosis on densitometric measurements, we examined 198 women, aged 32-81 years, who had undergone lateral X-ray of the lumbar spine. We classified patients according to different grades of osteophytosis, and evaluated bone density at the lumbar spine and the proximal femur by DXA. We also performed quantitative ultrasound at the heel (QUS). Patients with severe osteophytosis were significantly older (p < 0.0005), and values were adjusted for this parameter. We observed a significant increase in lumbar bone density with worsening osteophytosis (p < 0.02). On the contrary, no significant differences were found at the femur and QUS. According to bone density at the femoral neck, we subdivided patients into two groups: osteoporotic (group A) and non-osteoporotic (group B). Both groups showed increasingly high bone density at the spine with worsening osteophytosis (A: p < 0.01; B: p < 0.02). No differences were found in all the other evaluations. In conclusion, lumbar spine measurement is dramatically influenced by osteophytosis, particularly in the elderly. Consequently, other strategies should be performed such as evaluation of the hip and also measurement of the heel by ultrasound, which could be an interesting approach in these cases.

Longitudinal study of bone loss in pre- and perimenopausal women: evidence for bone loss in perimenopausal women.

Bone loss before and around the time of menopause is not well characterized by longitudinal studies. We measured bone mineral density at various skeletal sites--total body, femoral neck, trochanter, anteroposterior (AP) and lateral spine, and forearm--with dual-energy X-ray absorptiometry in a large prospective cohort of 272 untreated pre- and perimenopausal women aged 31-59 years, at 1 year intervals for 3 years. Sex steroids and the following markers of bone remodeling were measured: serum osteocalcin (OC), procollagen I carboxyterminal extension peptide, bone alkaline phosphatase (BAP) and urinary crosslinks (CTX and NTX). Seventy-six women were classified as perimenopausal and 196 as premenopausal. Over the 3 years, premenopausal women had no significant bone loss at any site and a small but significant increase in bone mineral density at the trochanter, total hip, AP spine and radius. Perimenopausal women significantly lost bone from cancellous and cortical sites, i.e., the femoral neck, trochanter and lumbar spine. In perimenopausal women with increased follicle stimulating hormone, the rate of bone loss at the femoral neck correlated negatively with OC and BAP. In perimenopausal women, serum estradiol levels decreased during the 3 years of follow-up and bone loss from the trochanter and the AP spine was correlated with serum estradiol after 3 years. In conclusion, among premenopausal women there is no bone loss. In contrast, there is a rapid and diffuse bone loss in perimenopausal women, related to decreased estrogen secretion. Bone markers may be useful to identify these women losing bone.

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.

High-resolution computed tomography for architectural characterization of human lumbar cancellous bone: relationships with histomorphometry and biomechanics.

The aim of the present study on human vertebral cancellous bone was to validate structural parameters measured with high-resolution (150 microm) computed tomography (HRCT) by referring to histomorphometry and to try to predict mechanical properties of bone using HRCT. Two adjacent vertical cores were removed from the central part of human L2 vertebral body taken after necropsy in 22 subjects aged 47-95 years (10 women, 12 men; mean age 79 +/- 14 years). The right core was used for structural analysis performed by both HRCT and histomorphometry. Two cancellous bone specimens were extracted from the left core: a cube for HRCT and a compression test, and a cylinder for a shear test. Significant correlations were found between HRCT and histomorphometric measurements (BV/TV, trabecular thickness, separation and number, and node-strut analysis), but with higher values for most of the tomographic parameters (BV/TV and trabecular thickness determined by HRCT were overestimated by a factor 3.5 and 2.5 respectively, as compared with histomorphometry). The maximum compressive strength and Young's modulus were highly correlated (rho = 0.99, p<0.0005). Significant correlation was obtained between bone mineral density (determined using dual-energy X-ray absorptiometry) and the maximum compressive strength (rho = 0. 64, p = 0.002). In addition the maximum compressive strength and architectural parameters determined by HRCT or histomorphometry showed significant correlations (e.g., for HRCT, BV/TV: rho = 0.88, p<0.0005, N.Nd/TV: rho = 0.73, p<0.001). The shear strength was significantly correlated with BV/TV (rho = 0.62, p = 0.002), Tb.Sp (rho = -0.58, p = 0.004) and TSL (rho = 0.55, p = 0.006) measured by HRCT. In conclusion, an HRCT system with 150 microm resolution is not sufficient to predict the true values of the structural parameters measured by histomorphometry, although high correlations were found between the two methods. However, we showed that a resolution of 150 microm allowed us to predict the mechanical properties of human cancellous bone. In vivo peripheral systems with such a resolution should be of interest and would deliver an acceptable radiation dose to the patient.

Effect of multiple intravenous pamidronate courses in Paget's disease of bone.

BACKGROUND: Pamidronate is a bisphosphonate whose short-term biological efficacy in Paget's disease of bone was convincingly established many years ago. A less well studied area is the efficacy of pamidronate in slowing disease progression and in preventing and treating complications. MATERIAL AND METHODS: We conducted an uncontrolled retrospective study of 79 Paget's disease patients given multiple intravenous pamidronate courses over a mean period of 45 +/- 19 months. The pamidronate dose per course was 180 mg, usually given over three days. The disease was severe and in some cases had proved refractory to other medications. Reasons for pamidronate therapy were pain or other subjective symptoms; established bone, joint, or nervous system complications; or prevention or these complications in patients with involvement of high-risk sites. RESULTS: Bone and joint pain improved under therapy, and in 78% of cases the outcome in terms of complication treatment and/or prevention was favorable. An important finding was waning of the clinical and biological effects of pamidronate as the number of courses increased. Fourteen percent of patients developed resistance to pamidronate, which seemed more closely related to disease extension than to focal lesion activity. CONCLUSION: These data suggest that a prompt return to normal of laboratory markers, most notably total alkaline phosphatase, should be sought, if needed by using higher doses than in our study.

The effects of alendronate on bone turnover and bone quality.

During Phase III clinical trials with alendronate, biochemical and histological studies assessed bone turnover and bone quality in patients treated for 3 years. Patients were randomised in double-blind fashion to receive placebo, alendronate 5 or 10 mg/day for 3 years or 20 mg/day for 2 years followed by 5 mg/day for 1 year. All patients also received 500 mg/day of calcium carbonate. Decreases in bone resorption with alendronate preceded decreases in bone formation. After approximately 6 months of continuous treatment, a new steady state of bone turnover was attained, leading to the increase in bone density. No subsequent decrease in the rate of bone turnover or of frozen bone was noted. Alendronate treatment did not impair bone mineralisation, induce the formation of woven bone or have any other adverse effects on bone quality.

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.

Fractal analysis of bone texture on os calcis radiographs compared with trabecular microarchitecture analyzed by histomorphometry.

Microarchitecture of trabecular bone is an important determinant of bone fragility; to date, its evaluation requires bone biopsy with histomorphometry analysis. Methods of noninvasive characterization of trabecular bone microarchitecture are in development and we have developed and validated a bone texture analysis applied to bone radiographs and based on fractal geometry. The aim of our study was to compare this fractal analysis of trabecular bone texture on radiographs to the trabecular microarchitecture analyzed by bone histomorphometry on os calcis biopsies. Thirty eight ossa calcis from 19 human cadavers were studied. Fractal analysis of the trabecular bone of os calcis radiographs was performed by the maximum likelihood estimator following the fractional brownian motion model. The ossa calcis were dissected, then transcortical biopsy cores focused on the fractal analysis region of interest were obtained. Structural and connectivity parameters were measured with both automatic and semiautomatic analyzers. We have found a significant relationship between the fractal Hmean parameter and structural histomorphometric indices; the best correlation was found with trabecular separation (r = -0.55; P = 0.0004). Based on a stepwise regression analysis, trabecular spacing and trabeculae number together would explain 38% of the variance of the fractal parameter. Although the relationship with connectivity indices was poor, our fractal analysis of os calcis trabecular bone texture on radiographs seemed to partially reflect the trabecular bone microarchitecture.

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.

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.

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.

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)