Double lamellae in trabecular osteons: towards a new method for age estimation by bone microscopy.

The study of anatomical ageing has a dual purpose in biological anthropology. On the one hand, it can provide insights into age associated changes in the body and thus widen the understanding of human senescence; and on the other hand it can provide means of estimation of age at death. This paper explores normal ageing in the pattern of remodelling of trabecular bone in humans. The material consists of necropsies of bone from the ilium of 25 males. A 1 cm2 prism extending from the outer to the inner surface of the iliac bone was removed from men who had died with no clinical signs of diseases, which would usually affect bone structure and metabolism. The samples were cut, and studied by light microscopy at a magnification of 100 x. New trabecular bone is formed in disk-shaped osteons with a clear double lamellar structure. In each sample, the number of double lamellae in a mean of 21 complete osteons was counted. The mean number of lamellae was taken as the measurement of interest. The log of the mean counts was found to regress linearly and with no evidence for heteroscedacity on age. The correlation between the two was high and negative (r=-0.83, p<0.001). The material is too limited to provide a useful basis for age estimation as such, but the study demonstrates the potential for palaeodemographic application of the method.

Plasma 25-hydroxyvitamin D and not 1,25-dihydroxyvitamin D is associated with parathyroid adenoma secretion in primary hyperparathyroidism: a cross-sectional study.

BACKGROUND: Primary hyperparathyroidism (PHPT) is associated with reduced plasma 25-hydroxyvitamin D (P-25OHD) and usually increased plasma 1alpha,25-dihydroxyvitamin D (P-1,25(OH)2D). Parathyroid tissue expresses the vitamin D receptor and it is thought that circulating 1,25(OH)2D participate in the regulation of parathyroid cell proliferation, differentiation and secretion. AIM: To investigate the relations between circulating levels of 1,25(OH)2D and 25OHD respectively and parathyroid adenoma weight (AW), plasma-parathyroid hormone (P-PTH) and PTH secretion expressed as P-PTH/AW. DESIGN: Cross-sectional study. MATERIAL: One hundred and seventy-one consecutive hypercalcaemic caucasian patients aged 19-87 years (median 63, 84% females) with surgically proven parathyroid adenoma. RESULTS: A weak positive correlation was found between P-25OHD and P-1,25(OH)2D (r=0.24, P<0.005). AW depended on sex and body mass index. Following adjustment, it was correlated positively to P-PTH, calcium (Ca) and alkaline phosphatase (AP) and inversely to plasma phosphate in a multiple regression model. AW was not associated with vitamin D metabolites. Preoperative P-PTH correlated positively to plasma levels of Ca and AP, but inversely to phosphate and 25OHD (P<0.001) levels. P-PTH was not associated with P-1,25(OH)2D (P=0.65). The P-PTH:AW ratio correlated inversely to P-25OHD (P<0.05), but showed no relations to plasma levels of Ca, phosphate or 1,25(OH)2D (P=0.22). CONCLUSION: In this material, low levels of 25OHD were related to higher levels of P-PTH and higher PTH:AW ratios in patients with PHPT suggesting that vitamin D deficiency increase PTH secretion activity. Neither PTH secretion nor AW was associated with circulating levels of 1,25(OH)2D.

Changes in bone histomorphometry after long-term treatment with intermittent, cyclic etidronate for postmenopausal osteoporosis.

Intermittent, cyclic etidronate therapy (400 mg/day for 2 weeks followed by 13 weeks free from study drug administration) resulted in a significant increase in lumbar bone mineral content and a significant decrease in the rate of new vertebral fractures in patients with postmenopausal osteoporosis. To investigate the effect of the treatment on bone histomorphometry, transiliac crest bone biopsy samples were obtained in this study before treatment and after 60 and 150 weeks of treatment with either intermittent, cyclic etidronate (n = 33) or placebo (n = 33). After 60 weeks of etidronate therapy, significant decreases in activation frequency (from 0.55 to 0.09 year,-1 P < 0.01) and resorption depth (from 53.2 to 37.8 microns, P < 0.05) were observed, leading to a positive balance per remodeling cycle. In the placebo group, no significant changes were seen. The 150 week bone biopsy samples were suboptimal for analysis, probably as a result of a regional acceleratory phenomenon. Our results suggest that, as a result of reductions in both activation frequency and resorption depth, intermittent, cyclic etidronate therapy may protect the trabecular network against fortuitous perforations and thereby maintain the strength of the bony tissue.

The influence of growth hormone on cancellous and cortical bone of the vertebral body in aged rats.

The influence of growth hormone administration on cancellous and cortical bone of the vertebral body in 2-year-old male rats has been investigated. All rats were injected for 80 days, then killed. Controls were given saline, and three recombinant human growth hormone (rhGH) injected groups were given either rhGH (2.7 mg/kg/day) for the first 20 or 40 days, followed by saline injection, or rhGH for all 80 days. Tetracycline labeling was performed on days 41 and 69. In all groups given rhGH, an increase in the cortical bone volume was found. In the rhGH 40-day group, single labeling corresponding to injection on day 41 was seen all around the anterior surface of the vertebral body wall (toward the abdominal cavity). In the rhGH 80-day group, double labeling was seen all around the anterior surface of the vertebral body, and a substantial increase in the mineralizing surface/total surface, mineral apposition rate, and mineralized bone formation rate was found. In the cortical bone of the anterior wall, cavities had developed in the rhGH 40- and 80-day groups. In the cancellous bone, no differences in bone volume, bone volume/total volume, or bone surface/bone volume were seen, but in the middle part of the vertebral body a decrease in the mineralizing surface/total surface was found in the rhGH 80-day group. The height of the vertebral body was not influenced by rhGH administration, whereas the transversal and midsaggital diameters were increased in the rhGH 80-day group. The compressive mechanical strength of the vertebral body specimens was increased in the rhGH 80-day group, and this increase most likely could be explained by formation and deposition of cortical bone.

Serum markers of type I collagen formation and degradation in metabolic bone disease: correlation with bone histomorphometry.

Type I collagen makes up more than 90% of bone matrix. Therefore, analysis of antigens related to collagen formation and degradation in bone should provide good and specific estimates of both bone resorption and bone formation rates. In this study we measured serum levels of the pyridinoline cross-linked telopeptide domain of type I collagen (ICTP) as a marker of bone resorption and serum carboxy-terminal propeptide of type I procollagen (PICP) as a marker of bone formation. Serum levels of the two antigens were correlated to histomorphometric indices of bone resorption and bone formation calculated from iliac crest bone biopsies in a group of 18 individuals with high- and low-turnover bone disease (myxedema, primary hyperparathyroidism, and thyrotoxicosis). After logarithmic transformation the regression of S-ICTP on volume-referent resorption rate (BRs/R/BV) was significant (r = 0.61, p < 0.01, SEM/Y = 56%). S-ICTP also showed a significant regression on the volume-referent cancellous bone balance (r = -0.45, p < 0.05, SEM/Y = 412%). S-PICP was significantly correlated to the mineral appositional rate (r = 0.53, p < 0.05) and volume-referent bone formation rate (r = 0.61, p < 0.01, SEM/Y = 48%). The correlation to bone turnover as expressed in the activation frequency was also highly significant (r = 0.61, p < 0.01, SEM/Y = 51%). No significant correlation with wall thickness or bone balance was demonstrable per remodeling cycle. Thus, assays employing antigens that reflect collagen formation and degradation are useful instruments for the evaluation of rates of bone remodeling in metabolic bone disease.

Cancellous bone remodeling occurs in specialized compartments lined by cells expressing osteoblastic markers.

We describe a sinus, referred to as a bone remodeling compartment (BRC), which is intimately associated with cancellous bone remodeling. The compartment is lined on its marrow side by flattened cells and on its osseous side by the remodeling bone surface, resembling a roof of flattened cells covering the bone surface. The flat marrow lining cells are in continuity with the bone lining cells at the margins of the BRC. We examined a large number of diagnostic bone biopsy specimens received during recent years in the department. Furthermore, 10 patients (8 women and 2 men, median age 56 [40-69] years) with the high turnover disease of primary hyperparathyroidism who were treated with parathyroidectomy and followed for 3 years were included in the histomorphometric study. Bone samples for the immuno-enzyme staining were obtained from an amputated extremity of child. The total cancellous bone surface covered by BRC decreases by 50% (p < 0.05) following normalization of turnover and is paralleled by a similar 50% decrease in remodeling surface (p < 0.05). The entire eroded surface and two-thirds of the osteoid surface are covered by a BRC. BRC-covered uncompleted walls are 30% (p < 0.05) thinner than those without a BRC. This indicates that the BRC is invariably associated with the early phases of bone remodeling, that is, bone resorption, whereas it closes during the late part of bone formation. Immuno-enzyme staining shows that the flat marrow lining cells are positive for alkaline phosphatase, osteocalcin, and osteonectin, suggesting that they are bone cells. The first step in cancellous bone remodeling is thought to be the lining cells digesting the unmineralized matrix membrane followed by their disappearance and the arrival of the bone multicellular unit (BMU). We suggest that the lining cell barrier persists during bone remodeling; that the old lining cells become the marrow lining cells, allowing bone resorption and bone formation to proceed under a common roof of lining cells; that, at the end of bone formation, new bone lining cells derived from the flattened osteoblasts replace the marrow lining cells thereby closing the BRC; and that the two layers of lining cells eventually becomes a single layer. The integrity of the osteocyte-lining cell system is reestablished by the new generation of lining cells. The BRC most likely serves multiple purposes, including efficient exchange of matrix constituents and minerals, routing, monitoring, or modulating bone cell recruitment, and possibly the anatomical basis for the coupling of bone remodeling.

Growth hormone treatment in adults with adult-onset growth hormone deficiency increases iliac crest trabecular bone turnover: a 1-year, double-blind, randomized, placebo-controlled study.

The effects of growth hormone (GH) substitution on bone metabolism were evaluated by dynamic histomorphometry on iliac crest bone biopsies. Twenty-nine patients, aged 21-61 years (mean 45.5 years), with adult-onset GH deficiency (GHD) were randomized to receive subcutaneous injections with GH (2 IU/m2/day = 0.67 mg/m2/day) or placebo for 12 months. Serum insulin-like growth factor I (IGF-I) levels increased 263 +/- 98% (mean +/- SD) during GH treatment (p < 0.0001). In the GH group, osteoid surface increased during treatment from 11% (3-15%) (median [25-75 percentiles]) to 21% (10-27%) (p = 0.01) and mineralizing surface from 4% (1-8%) to 11%(7-16%) (p = 0.04). Moreover, erosion surface tended to increase in the GH group from 2% (1-3%) to 4% (3-5%) (p = 0.07). The quiescent surface decreased in the GH group from 87% (83-96%) to 74% (68-87%) (p = 0.01). The adjusted appositional rate, mineral apposition rate, bone formation rate, bone erosion rate, mineralization lag time, and osteoid thickness remained unchanged during treatment. Erosion depth showed a trend toward increase in the GH group (p = 0.09), whereas wall thickness was unchanged. Bone balance at the remodeling unit level and activation frequency were unchanged. At the tissue level, bone erosion rate increased significantly from 26% (17-36%)/year to 39% (23-72%)/year (p = 0.03). Similarly, the bone formation rate at the tissue level tended to increase, from 24% (15-31%)/year to 36% (17%-63%)%/year (p = 0.06). Finally, bone balance at the tissue level decreased significantly from 1% (-2-2%)/year to -5% (-13-1%)/year (p = 0.01). No significant difference in change was seen in the cancellous bone volume. We conclude that 12 months of GH substitution therapy increases trabecular bone turnover. Moreover, our data suggest that bone balance at the bone multicellular unit level is not changed to positive.

Acute effects of parathyroid hormone on vitamin D metabolism in patients with the bone loss of aging.

This study was carried out to evaluate the effects of an iv injection of parathyroid extract on serum levels of 1,25-dihydroxyvitamin D [1,25-(OH)2D3] in elderly osteopenic patients and age-matched nonosteopenic controls. Serum concentrations of 1,25-(OH)2D were reduced in elderly osteopenic subjects (mean +/- SEM, 20 +/- 3 pg/ml) compared with values in age-matched nonosteopenic controls (35 +/- 3 pg/ml), whereas no differences were found in serum 24,25-dihydroxyvitamin D levels (1.5 +/- 0.3 and 2.2 +/- 0.5 ng/ml, respectively). An iv injection of parathyroid extract was followed by a significant increase in serum 1,25-(OH)2D levels in both osteopenic patients (16 +/- 6 pg/ml) and controls (15 +/- 5 pg/ml). The mean 4-h increases in serum 1,25-(OH)2D of 11-18 pg/ml were not significantly different in the two groups. The results indicate that the reduced 1,25-(OH)2D concentrations in the osteopenic patients are secondary to changes in factors that normally stimulate this enzyme system.

Differences in static cortical bone remodeling parameters in human mandible and iliac crest.

Knowledge of the baseline turnover characteristics, and of possible general and local factors influencing alveolar bone responses, is particularly important in the planning of oral rehabilitation. The conventional tool used to obtain information on bone turnover is the iliac crest biopsy, but it is not clear whether it mirrors the situation involving the jaws. The aim of this study was to compare static bone remodeling parameters in the mandible and in the iliac crest to obtain baseline values for the mandible and to test the hypothesis of site specificity of bone remodeling. Bone specimens were obtained from 50 subjects (mean age 64 +/- 17) at autopsy. Three sites were sampled: iliac crest; jaw angle; and foramen mentalis area. In addition, occlusal status was recorded. On undecalcified thin sections, cortical porosity (Ct.Po), eroded sites (ESi), formative sites (FSi), osteonal diameter (On.Dm), Haversian canal diameter (H.Ca.Dm), and wall width (W.Wi) were measured. Ct.Po in the jaw angle and in the foramen mentalis area was lower (48% and 50%, respectively) than in the iliac crest, as was ESi and FSi (80% in the jaw angle and 74% in the foramen mentalis area). In the foramen mentalis area, Ct.Po was greater in subjects with occlusion. On.Dm, H.Ca.Dm, and W.Wi were significantly larger and mutually correlated within the mandible, whereas no correlation was found between mandibular sites and iliac crest. Static cortical bone remodeling parameters are different in the mandible and the iliac crest, thus confirming the hypothesis of site specificity of bone remodeling. Within the mandible, the parameters were correlated, whereas there was no correlation between the mandible and the iliac crest. This could be ascribed to the different functional demands to the mandible and the iliac crest, which was also reflected in the observed influence of functional occlusion on bone remodeling in the mandible. It can thus be concluded that bone reaction to dental intervention is more dependent on the local environment than on general bone turnover as reflected by the iliac crest.

Missing observations in bone histomorphometry on osteoporosis: implications and suggestions for an approach.

Crucial bone histomorphometric indices, i.e., turnover-related indices, are based on tetracycline double labeling. However, these indices are particularly exposed to loss of information because of missing readings on double labels. If the failure to make the observation is related to its magnitude, then selection bias may invalidate the conclusions. Therefore, ignoring missing double labels may lead to a selection of high-turnover patients. The aim of this study was to analyze the dimension and the impact of excluding iliac crest bone biopsies with missing readings in women with spinal crush fracture osteoporosis (n = 158, median 68 years, range 49-80 years). Furthermore, two different lower limits of the mineral apposition rate (MAR) were examined to explore their usefulness as a biological minimum that can be used for cases with missing readings, i.e., recoding of missing values. The average MAR (calculated as the mean of all interlabel widths measured in each individual) shows a lower limit of 0.3 microm/day, suggesting an apparent minimum for the interlabel width (Ir.L.Wi) of 3-4 microm. Identifying the smallest interlabel width measured in each individual and calculating the minimal MAR shows that 77% of the minimal MAR values are below 0.3 microm/day and reach a minimum of 0.1 microm/day, corresponding to an interlabel width of about 1 microm. Therefore, the minimal MAR presents a biological minimum of 0.1 microm/day. This value is used for our recoding: if no labels are sampled (2% of our population), Ir.L.Wi is assigned the value 0; if none or an insufficient number of double labels are sampled (29% of our population), then Ir.L.Wi is assigned the value 1 microm. Excluding cases with missing readings on any dependent variable increases the mineralizing surface (MS/BS) by 60% (2p < 0.01); other indices show no significant change. The suggested recoding decreases the average MAR by 4% (2p < 0.01), prolongs the remodeling period by 19% (2p < 0.01), and tends to decrease the activation frequency (2p = 0.09). Furthermore, the number of excluded biopsies tends to be larger among the older (2p = 0.09) and more severely osteopenic individuals (2p = 0.09). We conclude that ignoring missing double labels leads to selection bias; therefore, specific measures such as recoding procedures are needed to allow proper representation of low turnover patients. There is also a risk of bias caused by the exclusion of the older, osteopenic patients in bone histomorphometric osteoporosis trials.

Reconstruction of cortical bone remodeling in untreated primary hyperparathyroidism and following surgery.

Iliac crest bone biopsies from 39 patients (23 women, 16 men) with untreated primary hyperparathyroidism (PHP) were examined. Static histomorphometric parameters for cortical bone was compared with values obtained from 39 age- and sex-matched normal controls. Thirty-five of the patients were double-labeled with tetracycline before biopsy. The cortical remodeling cycle was reconstructed in these patients and compared with 24 sex-matched but younger normal controls. In 9 patients, a second intact bone biopsy was obtained 6-12 months after successful surgery for PHP. In these patients, pre- and post-treatment biopsies were compared. Cortical porosity was increased in PHP (p < 0.001) due to a high turnover state with expansion of the remodeling space (p < 0.001). The net balance of the remodeling cycle on the haversian surface was found unchanged in PHP, since the average haversian canal diameter remained unchanged. In the female PHP-patients, a significant increase in the final resorption depth (p < 0.01) was balanced by a proportional increase in the wall width (p < 0.001). Among all PHP patients, no significant decrease in cortical width was found. However, a significant reduction in absolute (p < 0.05) and fractional cortical width (p < 0.05) was seen in postmenopausal PHP women giving rise to a cortical bone loss. Reconstruction of the cortical remodeling cycle revealed a doubling in activation frequency from 0.6#/year in normals to 1.2#/year in PHP (p < 0.001), giving rise to a high turnover state. Surgical cure of PHP was accompanied by marked changes in cortical bone metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)

Trabecular bone remodeling and bone balance in hyperthyroidism.

In vivo tetracycline double-labeled iliac crest bone biopsies from 15 hyperthyroid patients were used for the reconstruction of curves describing the variation of resorption depth and formation thickness with time. The curves emerging were compared to curves reconstructed from 13 age- and sex-matched normal individuals (mean age 44 years). The median function period for resorptive cells in hyperthyroid patients (16 days) was about one-third the resorptive period in normals (51 days). No significant difference between the osteoclast-, mononuclear-, or preosteoblast-like cell resorption depths could be demonstrated between the two groups. Consequently, the median resorption rate in hyperthyroid patients (3.8 microns/day) was more than 3 times higher than the value in the control group (1.1 micron/day). Median Sigma, was shorter in the hyperthyroid group (109 days) than in the control group (151 days, P less than 0.05), as was the median initial mineralization lag time (5 and 16 days, respectively, P less than 0.01). No significant difference between the measured mean completed wall thickness (mcwT) values in the hyperthyroid groups and the control group could be demonstrated (58.1 and 60.5 micron respectively). Median initial mineralization rate in the hyperthyroid group (1.2 micron3/micron2 per day) was not significantly higher than the value calculated in the control group (0.9 micron3/micron2 per day), but median initial matrix appositional rate in hyperthyroid (4.8 microns3/micron2 per day) was 3 times higher than the value calculated for normals (1.6 micron3/micron2 per day) (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Kinetics of trabecular bone resorption and formation in hypothyroidism: evidence for a positive balance per remodeling cycle.

Using a recently described model for reconstruction of the entire remodeling sequence in trabecular bone after tetracycline double labeling, bone remodeling was investigated in 18 hypothyroid female patients and 11 age- and sex-matched controls. In the hypothyroid group, the final resorption depth was reduced to 42 microns vs 53 microns in the control group (P less than 0.01). The total resorption period was increased to 76 days vs 32 days in the control group (P less than 0.001), and the resorption rate was lowered to 0.8 micron/day vs 2.1 microns/day in the control group (P less than 0.001). The function period for preosteoblast-like cells was 22 days compared to 14 days in normals (P less than 0.05). The total bone formation period was prolonged to 620 days in the hypothyroid group compared to 151 days in the control group (P less than 0.001), and the matrix and mineral appositional rates were lowered to 0.7 and 0.5 micron3/micron2 per day compared to 1.8 (P less than 0.001) and 1.4 (P less than 0.001) micron3/micron2 per day, respectively. The mineralization lag time in the hypothyroid group was 48 days vs 25 days in the control group (P less than 0.001), and at the start of bone formation this difference was even more pronounced (63 and 16 days, respectively, P less than 0.001). The mean completed wall thickness was increased in the hypothyroid group to 59.5 microns compared to 54 micron in the control group (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Trabecular bone remodeling and balance in primary hyperparathyroidism.

The total remodeling sequences in 19 primary hyperparathyroid patients and 16 approximately age-matched and sex-matched controls were reconstructed from histomorphometric analyses of bone specimens obtained after intravital tetracycline double labeling. In the primary hyperparathyroid group the total amount of work performed by resorptive cells was reduced, as indicated by the significantly lower three-dimensional mononuclear and preosteoblast-like cell resorption depths (35.8 microns vs 44.5 microns in normals, P less than 0.01 and 45.3 microns vs 56.6 microns in normals, P less than 0.01, respectively). The active resorption period (i.e., the function period for osteoclasts and mononuclear cells) was reduced to 19 days compared to 29 days in normals (P less than 0.05), but no difference with respect to bone resorption rates could be demonstrated between the two groups. The median bone formation period (Sigmaf) in primary hyperparathyroid patients was not different from the value obtained in normals (172 days vs 134 days, respectively), and the matrix appositional rate (Ama), as well as the mineralization lag time (tm), were also unchanged. The initial mineralization rate (Ami(i)) was not significantly different from the value obtained in normals, but averaged over the total bone formation period, a reduced mineralization rate could be demonstrated (0.32 micron3/micron2 per day vs 0.46 micron3/micron2 per day in normals, P less than 0.01). The measured final three-dimensional thickness of bone formed during Sigmaf (mcwTm) was reduced in the primary hyperparathyroid group (51.1 microns vs 55.9 microns in normals, P less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Trabecular bone resorption depth decreases with age: differences between normal males and females.

The resorption depth below osteoclasts, mononuclear cells, and preosteoblast-like cells can be estimated by counting the number of lamellae of known thickness eroded below the three cell types. In a previous study on bone resorption in young normal individuals, we demonstrated that the mean depth below osteoclasts was smaller than the mean depth below mononuclear cells, which again was smaller than the mean depth below preosteoblast-like cells, the last taken as the final depth reached. In order to investigate the variation in resorption depth with age in both sexes, we examined bone biopsy specimens from 42 normal females and 34 normal males aged 17-90 years. For each patient the mean osteoclastic, mononuclear, and preosteoblast-like cell resorption depths were calculated, and the surface extensions of the three types of resorption lacunae were determined. The surface extent of preosteoblast-like cell lacunae increased with age in females (p less than 0.05). Osteoclastic resorption depth was constant throughout the ages in females as well as in males. Mononuclear and preosteoblast-like cell resorption depths decreased significantly with age in females (p less than 0.01 and p less than 0.05, respectively) as well as in males (p less than 0.05 and p less than 0.01, respectively). The mean depth of lacunae where resorption had terminated (i.e., preosteoblast-like cell resorption depth) was larger in females aged 30-60 years than in men of the same age (p less than 0.05). The reduction in final resorption depth with age is parallel to the decrease in mean thickness of completed walls previously described.(ABSTRACT TRUNCATED AT 250 WORDS)

Cortical bone remodeling in autosomal dominant osteopetrosis: a study of two different phenotypes.

Previous analyses of cancellous bone remodeling in autosomal dominant osteopetrosis (ADO) have suggested a diminished osteoclastic resorption at the endocortical surface. Consequently, cortical width increases with age in both ADO type I and II. By means of histomorphometric methods, the remodeling cycle of the Haversian surface in the iliac crest has recently been reconstructed in normal individuals. The aim of the present investigation was to study cortical bone remodeling in ADO type I and II. Transcortical iliac crest bone biopsies obtained from 21 ADO patients were studied. Of these, 14 had ADO type I and seven had ADO type II. Twenty of the patients had received intravital double labeling with tetracycline before the biopsies were obtained. The static histomorphometric parameters for cortical bone mass and structure were compared with a control group of 21 sex- and age-matched individuals. Regarding the dynamic histomorphometric parameters for reconstruction of the cortical bone remodeling cycle, the ADO type I material was compared with control material from 14 sex- and age-matched individuals, whereas the ADO type II material was compared with control material from six sex-matched but younger individuals. Significant increases were seen in absolute as well as fractional cortical widths in both ADO types. Furthermore, an age-related increase was observed in both absolute and fractional cortical widths in ADO type I, whereas ADO type II was nonsignificant with regard to age. The fractional cancellous width was reduced in both type I and type II. However, only in ADO type I did the fractional cancellous width significantly correlate inversely with age. In the control group, neither cortical dimensions nor cancellous width correlated significantly with age. No significant differences were observed between patients and the control group in osteon dimensions; fractional resorptive, formative, or quiescent sites; resorptive or formative remodeling rates; remodeling periods; or activation frequency. An age-related increase in cortical porosity was seen in the group of normal individuals, but not in the two patient groups. The fractional remodeling space was increased in the ADO type II group. In conclusion, cortical dimensions are increased in both ADO type I and II and positively correlated to age in type I. However, cortical remodeling at the Haversian surface is essentially normal in both ADO type I and II. This could be explained by a defective endocortical bone resorption, as no periosteal accretion was observed, and because cancellous bone remodeling previously has been described to be normal.

Unbiased stereological estimation of osteoid and resorption fractional surfaces in trabecular bone using vertical sections: sampling efficiency and biological variation.

The conventional stereological procedure for estimating bone surface densities in iliac crest biopsies provides biased estimates because bone substructure is anisotropic and the sectioning of the biopsies is intended to be parallel to the cylindrical axis of the iliac crest biopsy. It has recently been shown that random anisotropic sections with an identifiable axis or direction which is arbitrary but fixed can be used for unbiased estimates of surface areas. The arbitrary axis is called "vertical," a denomination which does not imply anything about its relation to directions in the world's field of gravity. Sampling efficiency and biological variation of fractional osteoid and resorptive surfaces were investigated in 14 paired bone specimens obtained from two females and five males (mean age 64) using an integral anisotropic test system constructed of cycloids. Results obtained at direct microscopy were compared to results obtained with projection onto the table. Mean fractional osteoid and resorption surfaces were 6.2 and 1.4% using direct microscopy and 5.6 and 1.7% using projection. Estimation of variance at each level of sampling showed that the majority of the total observed variance was due to true variance between individuals; for fractional osteoid surface the unavoidable biological variation was 80.5% of the observed variation and for fractional resorption surfaces it was 57%. The efficiency of the estimation procedure is therefore close to optimal and the only way to improve the precision in groups of patients and controls would be to include more individuals in the study.(ABSTRACT TRUNCATED AT 250 WORDS)

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)

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)

Age- and sex-related changes in iliac cortical bone mass and remodeling.

Iliac crest bone biopsies were obtained from 64 normal individuals (41 women and 23 men) aged 19-90 (mean 48.2) years. Thirty-four were double-labeled with tetracycline before biopsy. The following variables were measured in all biopsies: biopsy core width (C.Wi), absolute (A.Ct.Wi) and fractional (F.Ct.Wi) cortical width, absolute (A.Cn.Wi) and fractional (F.Cn.Wi) cancellous width, cortical porosity (Ct.Po), osteon diameter (On.Dm), Haversian canal diameter (Ha.Ca.Dm), and wall thickness (W.Th). In the tetracycline-labeled biopsies the typical cortical remodeling cycle was reconstructed and the activation frequency was estimated. A negative cortical bone balance with aging was found in both sexes. In females F.Ct.Wi decreased (p < 0.02) with aging because of marrow expansion (p < 0.05). Furthermore, Ct.Po increased (p < 0.001) because of a decrease in W.Th (p < 0.01) and an increase in H.Ca.Dm (p < 0.001). In males the negative cortical bone balance with aging was exclusively caused by an increase in Ct.Po (p < 0.001) partially explained by an expanding H.Ca.Dm (p < 0.01). The On.Dm increased with aging (p < 0.01), but surprisingly, no fall in W.Th was observed. Reconstruction of the remodeling cycle did not reveal any significant difference between younger women and men. However, the activation frequency rose from 0.5 per year in premenopausal to 1.0 per year in postmenopausal women (p < 0.001), giving rise to a high turnover state, an increase in the remodeling space, and thereby the porosity in the cortical bone immediately after menopause. The present study has shown a reduction in cortical bone mass in elderly people, compared with younger, which may be explained by an age-related remodeling imbalance. This reduction is further increased in women in the postmenopausal state because of a postmenopausal accelerated bone turnover.