Stochastic simulations of remodeling applied to a two-dimensional trabecular bone structure

In this investigation the stochastic dynamics of trabecular bone remodeling (which is a key concept in a range of remodeling models) is verified. For this purpose, two-dimensional binary images of horizontal histological sections of lumbar vertebral bone (L-3) were collected from young and old subjects. Then the stochastic algorithm of remodeling was applied to the images of young trabecular bone. The resulting images of "simulated" old structures and those of "real" old structures were analyzed to evaluate the following architectural parameters: bone volume/trabecular volume; mean area of marrow cavities; mean trabeculae length; marrow space star volume; Euler number density; trabecular thickness; trabecular number; trabecular separation; mean two-point distance along the skeleton of trabecular structure; probability of disconnection; the ratio of marrow space star volume to geometrical volume and dimensionless ratio of mean area of marrow cavities; and mean trabecular length squared (form factor). Using the parametric t-test, the groups of simulated and real old bone images were compared. It is found that the p value of the t-test is never less than 0.20. For eight parameters the p value is >0.45. It is concluded that, as long as the horizontal sections of lumbar vertebrae are considered (sections perpendicular to the direction of the main spine load), the stochastic algorithm of bone remodeling will properly reproduce the architectural properties of trabecular bone.     

Normal postmenopausal women show iliac crest trabecular thickening on vertical sections

Trabecular thickness and the trabecular star volume were estimated in 49 normal individuals (20 males and 29 females) using an unbiased sampling procedure consisting of anisotropic, vertical sections, and a corresponding anisotropic test system. Eight-microns-thick undecalcified stained vertical sections were obtained from iliac crest specimens. Implying a trabecular plate model mean trabecular thickness (Tb.Th., microns) was estimated by different methods: (a) the ratio between bone volume and bone surface (Tb.Th.ratio); (b) the mean of the orthogonal intercepts multiplied with pi/4 (Tb.Th.l0); and (c) the mean of random linear intercepts divided by 2 (Tb.Th.l1). Trabecular star volume (Vtr*, mm3), a true three-dimensional size parameter independent on any model, was estimated using linear intercepts in random directions. Significant (2p less than 0.001) correlations were found between the different methods with coefficients of correlation ranging from 0.71 to 0.89. The Tb.Th. ratio was overall higher (155 +/- 31 microns, (X, SD), 2p less than 0.001) than the other linear estimates, Tb.Th.l0: 144 +/- 26 microns and Tb.Th.l1: 135 +/- 27 microns. Estimates based on orthogonal intercepts (l0) were slightly higher (2p less than 0.001) than those from random linear intercepts (l1). Among all the individuals no significant age-related changes were seen in the measured parameters. However, postmenopausal women (greater than 50 years of age) had more thick trabeculae than younger women (2p = 0.03). Intra-individual distribution analysis disclosed that this finding could not be explained by preferential perforation and loss of thinner trabeculae during bone remodeling. Therefore, normal postmenopausal women do increase trabecular thickness with age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Primary hyperparathyroidism: iliac crest trabecular bone volume, structure, remodeling, and balance evaluated by histomorphometric methods.

Iliac bone biopsies from 69 patients (48 females, 21 males; median age 58 years; range 17-79 years) with primary hyperparathyroidism (PHP) were examined, and static histomorphometric parameters compared to 30 age- and sex-matched normal controls. The control group for the dynamic parameters constituted 20 sex-matched younger normal controls. Fractional volume of trabecular bone was normal, but the trabeculae were thinner (p less than 0.05) in PHP. The structural parameters marrow space star volume, intertrabecular distance, and mean trabecular plate density were not significantly different in PHP patients compared to normal controls, but the age-related increase, for females, in marrow space star volume and decrease, for both sexes, in mean plate density observed in the controls were not noticed in the PHP group. Trabecular bone remodeling was found significantly increased in the PHP patients reflected by increased extension of eroded (p less than 0.001), osteoid (p less than 0.001), and labeled surfaces (p less than 0.05). The activation frequency was increased by approximately 50% (p less than 0.001). Neither PHP patients nor controls showed age-related decrease in trabecular thickness, and accordingly in both groups the bone balance per remodeling cycle was very close to and not significantly different from zero. Normal postmenopausal women (age greater than or equal to 50 yr) had lower trabecular bone volume (p less than 0.001) and higher intertrabecular distance than normal pre-menopausal women (age less than 50 yr).(ABSTRACT TRUNCATED AT 250 WORDS)     

Star volume of marrow space and trabeculae in iliac crest: sampling procedure and correlation to star volume of first lumbar vertebra

The star volume is a new stereologic parameter which can describe structural changes of trabecular bone. It is defined as the mean volume of all the parts of an object which can be seen unobscured in all directions from a particular point inside the object. The mean is over all points inside the object. The parameter is only unbiased when Baddeley's vertical or isotropic sections are used. The star volume of the marrow space (V*m.space) and of the trabeculae (V*tr) was estimated on vertical sections obtained from the first lumbar vertebra and the iliac crest. The continued autopsy material presented was from 7 females and 11 males without malignant or metabolic bone diseases. The mean age was 52 years (range 7-87 years). The sampling procedure was as required for vertical sections and an anisotropic test system was used. A significant age-related increase was shown in marrow space star volume in both lumbar vertebra and iliac crest. The increase in lumbar vertebra marrow space star volume was significantly greater for women than for men. Moreover, a significant correlation was found between the marrow space star volume in lumbar vertebra and iliac crest (r = 0.72, 2p less than 0.01). No such relation was found for trabecular star volume which did not change with age. The results also revealed that marrow space star volume could be measured on conventional bone biopsies preferably, however, on two mutually perpendicular vertical section planes.     

A new method of comprehensive static histomorphometry applied on human lumbar vertebral cancellous bone

The aim of the present study was to assess age-related changes in the human spine by use of established static histomorphometry, and to determine how these static histomorphometric measures are interrelated in human cancellous bone tissue. The material comprised normal human lumbar vertebral bodies (L-2) from 12 women (19-96 years) and 12 men (23-91 years) selected from a larger autopsy material to give an even age and gender distribution. In addition, L-2 from three female subjects (80, 88, and 90 years) with a known vertebral fracture of L-2 were considered. Approximately 9-mm-thick frontal (mediolateral) slices were embedded in methylmetacrylate, stained with aniline blue, and scanned into a computer with a flatbed image scanner at a high resolution (2400 dpi). With a custom-made computer program the following static histomorphometric measures were determined: trabecular bone volume; marrow space star volume; bone space star volume; anisotropy of bone and marrow phase (star length distribution method); node-strut analysis (node:terminus ratio); trabecular thickness; trabecular number; trabecular separation; and trabecular bone pattern factor. In addition, connectivity density was determined (by the ConnEulor method). All 11 histomorphometric measures, except bone space star volume and the two measures of anisotropy, showed a significant correlation with age. Marrow space star volume (r = 0.82) and trabecular bone volume (r = -0.81) showed the highest correlation with age. Furthermore, it was found that all of the histomorphometric measures were correlated, to different degrees. Trabecular bone volume correlated significantly with all ten histomorphometric measures, whereas the two anisotropy measures were poorly correlated to the other measures. Finally, we found the histomorphometric values in this study to be in excellent accordance with various previously published results from studies of human trabecular vertebral bone, the sole exception being marrow space star volume, which was probably due to the small (artificial) region of interest (ROI) that was used in the earlier studies. In conclusion, the new method applied herein allows for easy assessment of age-related changes and also for assessment of relationships between histomorphometric measures in human vertebral cancellous bone.     

Aluminum-induced neo-osteogenesis: a generalized process affecting trabecular networking in the axial skeleton

To determine if aluminum-induced neo-osteogenesis occurs in the axial skeleton, we compared spinal bone density and vertebral histology of beagles treated with aluminum for 8 and 16 weeks to that of untreated normals. Administration of aluminum (1.25 mg/kg) did not alter serum calcium, phosphorus, or creatinine but did result in a significant elevation of vertebral bone density, measured by quantitative computed tomography, after both 8 (286.7 +/- 12.4 mg/ml) and 16 (361.7 +/- 46.5 mg/ml) weeks of treatment compared with controls (212.2 +/- 4.5 mg/ml). In accord with the increased bone density, biopsies from the spine displayed evidence of neo-osteogenesis, including the presence of woven bone, both mineralized and unmineralized, within the marrow space. The genesis of such woven bone units resulted after 16 weeks in a significant increase in trabecular bone volume, woven and lamellar (51.2 +/- 4.4 versus 32.4 +/- 1.2%; p less than 0.05), woven bone volume (9.1 +/- 3.6 versus 0 +/- 0%; p less than 0.05), and trabecular number (4.5 +/- 0.3 versus 3.5 +/- 0.2 per mm; p less than 0.05). In addition, scanning electron microscopic evaluation of the bone biopsies confirmed the existence of new trabecular plates that provided interconnections between existent units. These observations illustrate that aluminum-induced neo-osteogenesis positively influences trabecular networking in the axial skeleton. Such enhancement of bone histogenesis contrasts with the effects of other pharmacologic agents that solely alter the thickness of existing trabecular plates or rods within the vertebral spongiosa.     

Analysis of the influence of image resolution on the discriminating power of trabecular bone architectural parameters

In the study the influence of image resolution on the discriminating power of different parameters quantifying the architecture of trabecular bone is investigated. High-resolution images of 200-microm-thick sections of young and old trabecular bone are collected. Different architectural parameters are evaluated and shown to be statistically different in both groups of images. Then the resolution of the images is artificially degraded to the level comparable with resolution achievable in vivo. It is shown that although the errors of evaluation are quite large for low-resolution images, the statistical difference present in the original data is still observed for parameters that depend only on the global characteristics of trabecular structure or marrow space, or depend only linearly on the number and area of disconnected parts of marrow space or trabecular structure. The parameters that fulfill such conditions are Euler number, mean area of marrow cavities, star volume and trabecular spacing. The statistical difference disappears for parameters that depend on the number and area of disconnected parts of marrow space or trabecular structure in a nonlinear manner, that is, marrow and trabecular disconnection probabilities, two-point distance and second moment of marrow cavities area distribution. It is shown that the mechanism that leads to the losing of discriminating power does not depend crucially on the estimation error or noise level. This mechanism, which involves an interplay between changes of mean values and standard deviations, is the manifestation of partial volume effect amplified with image segmentation.     

Star volume of marrow space and trabeculae of the first lumbar vertebra: sampling efficiency and biological variation

The decrease in the amount of trabecular bone which is seen with age cannot solely be explained by thinning of trabeculae but must also be due to a loss of structural trabecular bone leading to a discontinuity in the trabecular network. Due to the complex architecture and the anisotropy of bone it is difficult to demonstrate this structural change by conventional histomorphometry. Unbiased stereological estimators can however be obtained from anisotropic structures when using vertical sections and a specially designed anisotropic test system. This combined with a new and unbiased stereological parameter for bone structure the star volume can be of major importance in clarifying histological changes of bone structure. The star volume is defined as the mean volume of all the parts of an object which can be seen unobscured in all directions from a particular point with the mean value taken over all points inside the object. It is defined for any type of objects including cavities like marrow space and networks like the trabecular system. Measurements are performed using a frame and a grid with points and lines. The material investigated was the first lumbar vertebra obtained from two females and six males with ages 26 to 75 years without malignant or metabolic bone diseases. The sampling procedure was as required for vertical sections. Results did show a highly significant, five-fold increase in the star volume of the marrow space with age; no such age correlation was found for the star volume of the trabeculae. The only explanation for such an increase in the size of the marrow space is by removing or perforating trabecular bone.(ABSTRACT TRUNCATED AT 250 WORDS)     

Microradiography in the study of trabecular parameters]

The intermethod variation in measurement of trabecular bone volume (VTO) and the indirect estimation of its microstructure according to Parfitt MTPT (microns), MTPD (/mm) and MTPS (microns) were evaluated in seven undecalcified bone biopsies by analyzing the microradiograph of a 100-microns-thick section with an automatic method (IBAS II Zeiss, Munich) in addition to reference methods (manual and semiautomatic) described in the literature and performed on 7-microns-thick stained sections. Three consecutive 7-microns-stained sections and one 100-mu-thick microradiographed section were taken in each specimen. The 100 microns-thick section was also superficially stained. Trabecular bone volume was measured with both a manual integrating eyepiece and an automatic method. The automatic method on the microradiograph underestimated the VTO by 24.42%. There was a correlation (r = 0.75; p < 0.02) between the manual and computerized methods. Mean trabecular plate thickness (microns), mean trabecular plate density (/mm) and mean trabecular separation (microns) were measured with both semiautomatic and automatic methods. The automatic method on the microradiograph underestimated the MTPT (microns) by 18.98% and the MTPD (/mm) by 14.14% and overestimated the MTPS (microns) by 23.17%. For the MTPT (microns) there was a correlation (r = 0.88; p < 0.02), between both methods, and the correlation was good for MTPD (/mm) (r = 0.97; p < 0.001), and MTPS (r = 0.86; p < 0.002).     

Structural and histomorphometric studies of iliac crest trabecular and cortical bone in autosomal dominant osteopetrosis: a study of two radiological types

Cylindrical iliac crest biopsies were obtained from 16 patients with autosomal dominant osteopetrosis after intravital double labeling with tetracycline, and compared with normal age- and sex-matched controls. Ten patients had the radiological type I (5 women, 5 men, aged 17-62 years, mean 42) characterized by diffuse, symmetrical osteosclerosis and enlarged thickness of the cranial vault. Six patients had type II (2 women, 4 men, aged 22-44 years, mean 36), where "Rugger Jersey Spine" and endobone are characteristic findings. Structural studies of cortical and trabecular bone were performed, and trabecular bone resorption and formation rates were studied using dynamic histomorphometry. The total biopsy length (C. Wi) were increased in type I (p less than 0.05), and unchanged in type II. Both types showed increased cortical width (Ct. Wi) (p less than 0.01 and p less than 0.05, respectively), and decreased fractional width of cancellous bone (Cn.Wi/C.Wi) (p less than 0.01 and p less than 0.05). The fractional trabecular bone volume (BV/TV) and trabecular thickness (Tb. Th) were both significantly increased in type I (p less than 0.05), while resorptive and formative indices of trabecular bone remodeling were normal. No difference was found in trabecular bone balance, which was slightly positive in both patients and controls. In type II osteopetrosis the eroded surfaces (OS/BS) were significantly increased (p less than 0.01), as was the total resorptive period RP) (p less than 0.05). The resorption depth (R.D.) was normal, while the resorption rate (MRR) was insignificantly decreased. Many big multinucleated osteoclasts were seen in this type suggesting defective resorptive function.(ABSTRACT TRUNCATED AT 250 WORDS)     

Iliac bone biopsies at the time of periarticular stress fractures during fluoride therapy: comparison with pretreatment biopsies

We attempted to establish whether systemic changes in trabecular bone explain the development of stress fractures in the lower limbs during fluoride therapy for osteoporosis. To this end we compared transiliac bone biopsies obtained before treatment with those taken around the time of stress fractures after 14.3 +/- 10.9 (SD) months of therapy in six patients (group A). Biopsies from a comparable group of six patients without stress fractures at the time of the second biopsy (after 11.9 +/- 2.7 months of treatment) served for comparison (group B). The biopsies were processed undecalcified and examined by routine histomorphometry. The second biopsies did not show any significant improvement in mean bone volume or trabecular architecture. Although the second biopsies in group A had increased erosion surfaces (p less than 0.05) and greater osteoid volume (p less than 0.05), group B biopsies showed no difference in erosion surfaces but an increase in all osteoid parameters: osteoid volume (p less than 0.05), osteoid surface (p less than 0.05), and osteoid seam thickness (p less than 0.01). We reached the following conclusions: (1) the combination of increased erosion and replacement of removed bone by as yet unmineralized osteoid in the stress fracture group must have weakened bone and allowed the development of stress fractures. (2) Stress fracture patients may have mounted a less vigorous osteoblast response to fluoride than non-stress fracture patients. Under these conditions microfractures are likely to heal poorly and propagate to develop into full stress fractures. (3) Renal failure is a contraindication to fluoride therapy.     

Quantitative analysis using the star volume method applied to skeleton patterns extracted with a morphological filter

In this study, a morphological filter was combined with star volume analysis and applied to digital images to determine its potential usefulness in assessing trabecular structure. Three digital "geometric" test patterns (square, rectangle, circle) were created on a CRT (cathode ray tube). Each shape was arranged into five groups by size to yield 15 final "skeletal" patterns that were subsequently assessed with star volume analysis. Also, three digital X-ray images (background, soft tissue, bone block) were processed with a morphological filter to create three sets of 11 skeletal patterns each. These patterns were also assessed with star volume analysis and the ratio of extracted skeletal elements (in pixel numbers) to total pixel numbers was expressed as the pixel percentage. Star volume analysis was then applied to these digital skeletal images to yield the volume of extracted "skeletal" trabecular elements (Vsk) and the volume of nonskeletal (marrow space) elements (Vsp). The Vsk and Vsp were compared for all the different skeletal patterns. The pixel percentages were then compared to the star volume results for the X-ray test patterns. The Vsk decreased and Vsp increased as the number of operations (n) increased for both digital X-ray images and the geometric test patterns when the X-ray images were depicted by pixel percentages. Also, all true bone test patterns were clearly different both visually and quantitatively when compared to the noise skeletons extracted from background and soft tissue. Therefore, as Vsk was increased, so was connectivity. It can be concluded that the application of morphological filters and star volume analysis may be a useful tool in quantitatively determining the characteristics and continuity of trabecular skeletal structures. Further studies involving a larger number of bone samples and using models to compare measurements of calculated versus actual volume should reveal the true potential of this method for evaluating bone structure and its relationship to bone strength and also increase the tools available for evaluating bone diseases such as osteoporosis. Received: October 12, 1999 / Accepted: March 1, 2000     

Age-dependent morphometric alterations in the distal femora of male and female rats.

Morphologic parameters, bone area, bone-to-bone + marrow ratio, periosteal-to-periosteal + endocortical surface ratio, mean trabecular thickness, and surface-to-volume ratio were studied in the epiphysis and metaphysis of the distal femora of male and female rats (Heiligenberg strain) between birth and the end of the lifespan. With increasing age, bone area, bone-to-bone + marrow ratio, and mean trabecular thickness increases, whereas periosteal-to-periosteal + endocortical surface ratio and surface-to-volume ratio decreases in both parts of bone during the first 150 days. Afterwards, periosteal-to-periosteal + endocortical surface ratio, mean trabecular thickness, and surface-to-volume ratio remain constant, whereas the bone area and the bone-to-bone + marrow ratio decrease. Modeling data were measured by use of the vital labeling technique with calcein. From the stained bone area, the bone formation, the bone resorption, and the periosteal mineral apposition rates have been calculated. The bone formation rate, about 13,000%/year in the metaphysis and 2,000%/year in the epiphysis, respectively, is greatest after birth and decreases continuously with increasing age to 3.5%/year for both bone regions. During the first 150 days the bone resorption rate is lower than the bone formation rate, leading to an increase in bone area, but afterwards it is higher so that the area decreases. Likewise the periosteal mineral apposition rate is greater in the metaphysis (24 microns/day at day 50) than in the epiphysis (14 microns/day at day 50), but after 700 days it is comparable for both bone regions (0.07 microns/day). The absolute values of body weight, femur length, and bone area of epiphysis and metaphysis are greater in male rats; only the mean trabecular thickness and the periosteal mineral apposition rate are comparable in both sexes. The relative values of bone-to-bone + marrow ratio, periosteal-to-periosteal + endocortical surface ratio, bone formation rate, and bone resorption rate are comparable for both sexes.     

Age-related changes in vertebral trabecular bone architecture--assessed by a new method

Cylindrical trabecular bone specimens (d = 7 mm) were drilled in a vertical direction from the central part of the third vertebral body (L3) from 23 normal individuals aged 15-87 years (10 males and 13 females). The bone samples were embedded in methylmetacrylate and sawn in 400 microns thick sections with an arbitrary rotation but a fixed vertical axis. The sections were investigated in polarized light at a magnification of x 8. By using this technique, vertical and horizontal trabeculae were clearly separated due to different colors. Photographs were taken. These were magnified, and trabecular thickness and intertrabecular distance were measured, using a Zeiss-integration plate II. A significant age-related decrease was found in the mean horizontal trabecular thickness (r = -0.71, p less than 0.001), while the mean thickness of the vertical trabeculae was unchanged with age (r = 0.06, n.s.). Furthermore, a significant increase was found both for the mean distance between the horizontal trabeculae (r = 0.79, p less than 0.001) and between the vertical trabeculae (r = 0.75, p less than 0.001). The present study gave a clear and striking visual presentation of both the thinning and disappearance of the horizontal supporting struts in the vertebral trabecular lattice and the total removal of some of the vertical trabeculae--leading to the dramatic loss of bone strength previously demonstrated.     

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.     

Microfractures in coxarthrosis

We examined the subchondral bone architecture of the femoral head in relation to trabecular microfracture. Three groups of femoral head specimens were studied. Twenty-eight specimens taken during hip replacement had grade III or IV arthrosis (70 +/- 8 years). From autopsy, 40 femoral heads were obtained, 18 in a group greater than 50 years of age (72 +/- 10 years) and 22 in a group less than 50 years of age (25 +/- 11 years). None of these 40 heads had worse than grade II arthrosis. Coronal slices of the femoral heads were macerated and examined under a dissecting microscope to count trabecular microfractures. For bone histomorphometry, blocks were taken from the subchondral principal compressive and tensile trabeculae. The bone volume, trabecular thickness, and marrow space were quantified. In the subchondral principal compressive region, the arthrotic group had more bone volume, thicker trabeculae, similar trabecular space, and trabecular microfractures when compared with the group greater than 50 years old. In the tensile region, there were no differences except for decreased trabecular microfracture number in the arthrotic group. With the thinnest trabeculae in the compressive region occurring in the greater than 50 years old group, the trabeculae of the younger age group have thinned with age, but with the onset of arthrosis, the thinning is overtaken by pathologic thickening of trabeculae.     

Increased RANK ligand in bone marrow of orchiectomized rats and prevention of their bone loss by the RANK ligand inhibitor osteoprotegerin

Orchiectomized (ORX) rats were used to examine the extent to which their increased bone resorption and decreased bone density might relate to increases in RANKL, an essential cytokine for bone resorption.Serum testosterone declined by > 95% in ORX rats 1 and 2 weeks after surgery (p < 0.05 versus sham controls), with no observed changes in serum RANKL. In contrast, RANKL in bone marrow plasma and bone marrow cell extracts was significantly increased (by  100%) 1 and 2 weeks after ORX. Regression analyses of ORX and sham controls revealed a significant inverse correlation between testosterone and RANKL levels measured in marrow cell extracts (R = − 0.58), while marrow plasma RANKL correlated positively with marrow plasma TRACP-5b, an osteoclast marker (R = 0.63). The effects of RANKL inhibition were then studied by treating ORX rats for 6 weeks with OPG-Fc (10 mg/kg, twice/week SC) or with PBS, beginning immediately after surgery. Sham controls were treated with PBS. Vehicle-treated ORX rats showed significant deficits in BMD of the femur/tibia and lower trabecular bone volume in the distal femur (p < 0.05 versus sham). OPG-Fc treatment of ORX rats increased femur/tibia BMD and trabecular bone volume to levels that significantly exceeded values for ORX or sham controls. OPG-Fc reduced trabecular osteoclast surfaces in ORX rats by 99%, and OPG-Fc also prevented ORX-related increases in endocortical eroded surface and ORX-related reductions in periosteal bone formation rate. Micro-CT of lumbar vertebrae from OPG-Fc-treated ORX rats demonstrated significantly greater cortical and trabecular bone volume and density versus ORX-vehicle controls. In summary, ORX rats exhibited increased RANKL protein in bone marrow plasma and in bone marrow cells, with no changes in serum RANKL. Data from regression analyses were consistent with a potential role for testosterone in suppressing RANKL production in bone marrow, and also suggested that soluble RANKL in bone marrow might promote bone resorption. RANKL inhibition prevented ORX-related deficits in trabecular BMD, trabecular architecture, and periosteal bone formation while increasing cortical and trabecular bone volume and density. These results support the investigation of RANKL inhibition as a strategy for preventing bone loss associated with androgen ablation or deficiency.     

Microfractures in coxarthrosis

We examined the subchondral bone architecture of the femoral head in relation to trabecular microfracture. Three groups of femoral head specimens were studied. Twenty-eight specimens taken during hip replacement had grade III or IV arthrosis (70 × 8 years). From autopsy, 40 femoral heads were obtained, 18 in a group greater than 50 years of age (72 × 10 years) and 22 in a group less than 50 years of age (25 × 11 years). None of these 40 heads had worse than grade II arthrosis. Coronal slices of the femoral heads were macerated and examined under a dissecting microscope to count trabecular microfractures. For bone histomorphometry, blocks were taken from the subchondral principal compressive and tensile trabeculae. The bone volume, trabecular thickness, and marrow space were quantified. In the subchondral principal compressive region, the arthrotic group had more bone volume, thicker trabeculae, similar trabecular space, and trabecular microfractures when compared with the group greater than 50 years old. In the tensile region, there were no differences except for decreased trabecular microfracture number in the arthrotic group. With the thinnest trabeculae in the compressive region occurring in the greater than 50 years old group, the trabeculae of the younger age group have thinned with age, but with the onset of arthrosis, the thinning is overtaken by pathologic thickening of trabeculae.