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A resolution-enhanced Fourier transform infrared spectroscopic study of the environment of the CO3(2-) ion in the mineral phase of enamel during its formation and maturation.

A resolution-enhanced Fourier Transform Infrared (FTIR) Spectroscopic study of the CO3(2-) ion in pig enamel of increasing age and maturity has demonstrated the existence of four different, main carbonate locations. The major CO3(2-) site arises as a result of the substitution of CO3(2-) ions in the positions occupied by PO4(3-) ions in the apatitic lattice. In addition, two minor locations have been identified in positions in which the CO3(2-) ions substitute for OH- ions. The fourth carbonate group appears to be in an unstable location. Its concentration has been found to decrease with aging and maturation, during which there is a progressive increase in the amount of mineral deposited in the enamel. The distribution of the carbonate ions in the different apatitic sites varies randomly during the formation of the mineral phase in enamel and during its maturation. Although these changes have been shown to be related to changes in the composition of the mineral phase, a comparison of the parameters assessing the degree of crystallinity of the mineral phase from upsilon 2CO3(2-) and upsilon 4PO4(3-) infrared absorption data reveals a significant discrepancy related to the nonhomogeneous partition of the CO3(2-) ion in the mineral phase. After maximum mineralization is reached, the composition of the mature mineral phase is decidedly different than that of the initial mineral deposited; the changes affect principally the concentrations of Ca2+, OH-, and HPO4(2-) ions, but not the CO3(2-) ions.

Aging↗

The relationship between bone mineral density and immobilization duration in hemiplegic limbs.

OBJECTIVE: Prolonged immobilization in stroke is known to result in hypercalciuria, hypercalcemia, accelerated bone resorption, and osteoporosis. Furthermore, bone mineral loss accelerated with increasing duration of hemiplegia. Although stroke is a common disease that causes sudden immobilization, relatively few investigations of bone metabolism in stroke have been reported. The aim of this study was to investigate the changes in bone mineral density of the forearms and legs related to duration of hemiplegia-induced immobilization after stroke. METHODS: Forty-one hemiplegic patients with stroke were evaluated. The patients' age, gender and duration of hemiplegia-induced immobilization were recorded. The measurements of bone mineral density (BMD) in all patients were evaluated with DEXA using the Norland apparatus. The BMD values (g/cm2) were determined by measurements made in the lumbar vertebrae, both forearm and legs (femoral neck and trochanter). RESULTS: We found that bone mineral density was decreased in the affected extremities relative to the intact contralateral side on measurements by dual energy x-ray absorptiometry in bones such as forearm, femoral neck and trochanter. There was a significant difference between bone mineral density of paretic and nonparetic forearms and legs. Bone mineral density of the upper limbs was lower than that of the lower limbs. There was a negative correlation between duration of hemiplegia and BMD values. CONCLUSIONS: Bone mineral loss may be related to the duration of hemiplegia-induced immobilization. Bone mineral loss is accelerated when the duration of hemiplegia is prolonged.

Absorptiometry, Photon↗

Interleukin-1beta-induced release of matrix proteins into culture media causes inhibition of mineralization of nodules formed by periodontal ligament cells in vitro.

The mechanism by which interleukin-1beta (IL-1) inhibits the formation of mineralized tissue nodules by periodontal ligament (PDL) cells in vitro was investigated through the processes of morphological analysis, immunoprecipitation, and Northern blot analysis. PDL cells were obtained from a 2-day-old coagulum in tooth socket and cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bone serum (FBS) and antibiotics. Confluent cells were grown for up to 3 weeks in the presence of ascorbic acid (AA), beta-glycerophosphate (GP), and dexamethasone (Dex), or IL-1. PDL cells cultured in the presence of GP and AA did not differentiate, but those treated with Dex, GP, and AA (Dex group) underwent differentiation, showing four stages (confluent, multilayer, nodule, and mineralization) of disparate morphological characteristics. In contrast, the cells treated with IL-1, Dex, GP, and AA (IL-1 group) did form multilayers but failed to form mineralized nodules. Electron microscopy demonstrated that the Dex-induced mineralized nodules contain multilayers of fibroblastic cells, numerous collagen fibrils, and dense globular as well as fused electron dense patches that are associated with numerous apatite crystals. The nodule-like structures in the IL-1 group were also comprised of multilayered fibroblastic cells, but they contained only a small number of collagen fibrils, and no dense globular or fused patches. Von Kossa staining confirmed the presence of numerous mineralized nodules in the Dex group and their scarceness in the IL-1 group. Northern blot analysis of IL-1-treated cells, however, revealed the presence of mRNAs for type I collagen (Col I), secreted protein, acidic and rich in cysteine (SPARC), osteopontin (OPN), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OC), whose expression patterns and levels were comparable to those of the Dex group. Immunoprecipitation analysis of OPN and BSP in the cell/matrix layers and the culture media after [35S]-methionine labeling showed their deposition primarily in the mineralized nodules of the Dex group, and their release into the media in the IL-1 group. Immunogold labeling demonstrated the location of OPN and BSP in mineralized nodules of the Dex group, but no significant labeling occurred in the nodule-like structures from the IL-1 group. Interestingly, IL-1 treatment increased the expression of collagenase mRNA by sevenfold, compared with that of the Dex group. These data suggest that the IL-1-induced formation of unmineralized nodules by PDL cells results not so much from the downregulated formation of matrix proteins, which plays a crucial role in the mineralization process, as from their release into the culture media. Finally, collagenase synthesis upregulated by IL-1 may be involved in this process.

Animals↗

Comparison of mineral quality and quantity in iliac crest biopsies from high- and low-turnover osteoporosis: an FT-IR microspectroscopic investigation.

Fourier-transform infrared microspectroscopy (FTIRM) allows analysis of mineral content, mineral crystal maturity and mineral composition at approximately 10-micron spatial resolution. Previous FTIRM analyses comparing 4-micron thick sections from non-decalcified iliac crest biopsies from women with post-menopausal osteoporosis, as contrasted with iliac crest tissue from individuals without evidence of metabolic bone disease, demonstrated significant differences in average mineral content (decreased in osteoporosis) and mineral crystal size/perfection (increased in osteoporosis). More importantly, these parameters, which vary throughout the tissue in relation to the tissue age in healthy bone, showed no such variation in bone biopsies from patients with osteoporosis. The present study compares the spatial and temporal variation in mineral quantity and properties in trabecular bone in high- and low-turnover osteoporosis. Specifically, six biopsies from women (n=5) and one man with high-turnover osteoporosis (age range 39-77) and four women and two men with low turnover osteoporosis (age range 37-63) were compared to ten "normal" biopsies from three men and seven woman (age range: 27-69). "High turnover" was defined as the presence of increased resorptive surface, higher than normal numbers of osteoclasts and greater than or equal to normal osteoblastic activity. "Low turnover" was defined as lower than normal resorptive surface, decreased osteoclast number and less than normal osteoblastic activity. Comparing variations in FTIR-derived values for each of the parameters measured at the surfaces of the trabecular bone to the maximum value observed in multiple trabeculae from each person, the high-turnover samples showed little change in the mineral: matrix ratio, carbonate: amide I ratio, crystallinity and acid phosphate content. The low-turnover samples also showed little change in these parameters, but in contrast to the high-turnover samples, the low-turnover samples showed a slight increase in these parameters, indicative of retarded, but existent resorption and formation. These data indicate that FTIR microspectroscopy can provide quantitative information on mineral changes in osteoporosis that are consistent with proposed mechanisms of bone loss.

Adult↗

Reductions in degree of mineralization and enzymatic collagen cross-links and increases in glycation-induced pentosidine in the femoral neck cortex in cases of femoral neck fracture.

INTRODUCTION: Enzymatic and glycation-induced nonenzymatic cross-links play important roles in the expression of bone strength. The cross-link pattern is affected by tissue maturation and senescence. The aim of our study was to understand the distinctive posttranslational modifications of collagen in areas with different degrees of mineralization with and without hip fracture. METHODS: Sixteen female cases of intracapsular hip fracture (78+/-6 years) and 16 age- and gender-matched postmortem controls (76+/-6 years) were included in this study. A sample of each femoral neck cortex was fractionated into low (1.7 to 2.0 g/ml) and high (>2.0 g/ml) density portions. The contents of enzymatic cross-links (dihydroxylysinonorleucine, hydroxylysinonorleucine, lysinonorleucine, pyridinoline, and deoxypyridinoline) and nonenzymatic cross-links (pentosidine) and the extent of lysine (Lys) hydroxylation were determined in each fraction. RESULTS: In the controls, there was no significant difference in the contents of enzymatic cross-links between low- and high-mineralized bone fractions whereas pentosidine content was significantly higher in high-mineralized bone compared with low-mineralized bone (p=0.0014). When comparing enzymatic cross-link contents between controls and fracture cases, a trend toward lower (p=0.0961) cross-link content in low-mineralized bone and a significant reduction (p<0.0001) in high-mineralized bone were observed. Pentosidine content of low-mineralized bone was significantly higher in fracture cases than in controls (p<0.0001). The extent of Lys hydroxylation was significantly higher in fracture cases than in controls (p<0.001). The higher hydroxylation of Lys in collagen from fracture cases relative to controls was associated with significantly higher values of hydroxylysine-derived cross-link such that the enzymatic cross-link patterns correlated with the extent of Lys hydroxylation in the collagen molecules. CONCLUSIONS: These results suggest that reductions in the degree of mineralization and enzymatic cross-links and excessive formation of pentosidine may play an important role in explaining poor bone quality in osteoporosis.

Aged↗

The degree of mineralization of bone tissue measured by computerized quantitative contact microradiography.

Strength of bones depends on bone matrix volume (BMV), bone microarchitecture, but also bone mineralization, and we have recently shown in osteoporotic patients treated with alendronate that fracture risk and bone mineral density (BMD) were changed without modifications of BMV or bone microarchitecture. Mineralization of bone matrix implies two successive steps: a rapid primary mineralization on the calcification front followed by a slow process of secondary mineralization progressively adding about one-half of the mineral content on bone matrix. These two steps are clearly illustrated by microradiographs of compact and cancellous bone tissue from transiliac human biopsies. Our working hypothesis is based on the impact of changes in bone remodeling rate on the degree of mineralization of bone, i.e., on the BMD measured at the tissue level. Contact quantitative microradiography using a computerized microdensitometric method, is described and allows the measurement of the mean degree of mineralization of bone (MDMB). This parameter may be quantitatively evaluated by exposing an aluminum calibration step-wedge and a plane-parallel calcified tissue section simultaneously to the same beam of X-rays, then determining, from the resulting microradiograph, the thickness of aluminum that produces the same X-ray absorption as a given region of the bone tissue section. To be used as a control group, iliac bone samples were taken at necropsy from 43 subjects (30 women aged 48.4 +/- 3.7 years and 13 men aged 66.0 +/- 4.4 years) who died suddenly showing no apparent bone disease. A control MDMB, which does not change with age, and a control distribution of these values are thus established. These control values are necessary for interpreting the changes in MDMB observed in bone conditions untreated or treated.

Absorptiometry, Photon↗

Mineralization of basement membrane mediates dentogingival adhesion in mammalian and nonmammalian vertebrates.

In order to elucidate the mechanism of adhesion between the gingiva and the tooth, detailed comparative ultrastructural studies of the dentogingival border were done in the monkey and shark. The tissues were prepared with or without demineralization for the ultrastructural observations. At the border, the internal basement membrane, which is firmly bound to the junctional epithelium through hemidesmosomes, was specialized differently in these species. In the monkey, the lamina densa was closely associated at its enamel side with an additional layer which had characteristics of the lamina densa and was referred to as the supplementary lamina densa. In the shark, the lamina densa showed a unique, hemidesmosome-related specialization in the form of the intermittent occurrence of bulges along its surface facing the epithelium. In nondemineralized tissues a part of the basement membrane, that is, the supplementary lamina densa (monkey) and the main lamina densa but not bulges (shark), was preferentially mineralized. The mineral deposit was continuous with that in the enamel and enameloid/dentine, thus constituting an advancing edge of mineralization. The network arrangement of the mineral crystals in the monkey basement membrane resembled the pattern of the cord network of the basement membrane, suggesting the presence of a delicate mutual basement membrane-mineral interaction. Thus, the organic phase and the mineral phase are allowed to make contact at this mineralized area of the basement membrane and firmly bind to one another. Therefore, strong gingiva-tooth adhesion is established by partial mineralization of the internal basement membrane, in a way similar to that found in the previously reported association of maturation stage ameloblasts with the enamel.

Animals↗

Matrix vesicles promote mineralization in a gelatin gel.

Extracellular matrix vesicles (MVs) are associated with initial calcification in a variety of tissues, but the mechanisms by which they promote mineralization are not certain. In this study, MVs isolated from fourth passage rat growth plate chondrocyte cultures were included within a gelatin gel into which calcium and phosphate ions diffused from opposite ends. In this gel, apatite formation occurs by 3.5 days in the absence of mineralization promoters, allowing measurement of the ability of different factors to "nucleate" apatite before this time or to assess the effects of molecules which modulate the rate and extent of mineral deposition. Mineral ion accumulation and crystal type are assayed at 5 days. In this study, MV protein content in the central band of a 10% gelatin gel was varied by including 100 microliters of a Tris-buffered solution containing 0-300 micrograms/ml MV protein. There was a concentration-dependent increase in mineral accretion. Whereas 10 micrograms MV protein in the gel did not significantly promote apatite formation as compared with vesicle-free gels, 20 and 30 micrograms MV protein in the gel did promote apatite deposition. Inclusion of 10 mM beta-glycerophosphate in the gels, along with MVs, did not significantly increase apatite formation despite the demonstrable alkaline phosphatase activity of the MVs. In contrast, MVs at all concentrations significantly increased apatite accumulation when proteoglycan aggregates or ATP, inhibitors of apatite formation and proliferation, were included in the gel. Slight increases in calcium, but not phosphate accumulation, were also noted when an ionophore was included with the MVs to facilitate Ca ion transport into the vesicles. FT-IR analysis of the mineral formed in the vesicle-containing gels revealed the presence of a bone-like apatite. These data suggest that MVs facilitate mineralization by providing enzymes that modify inhibitory factors in the extracellular matrix, as well as by providing a protected environment in which mineral ions can accumulate.

Adenosine Triphosphate↗

Differing lumbar vertebral mineralization rates in ambulatory pediatric patients with osteogenesis imperfecta.

The purpose of this study was to evaluate serial changes in bone mineral density (BMD) of the lumbar spine in individual children and adolescents with untreated osteogenesis imperfecta (OI) using dual X-ray absorptiometry (DXA). Twenty-seven pediatric patients with OI who had no historical or radiographic evidence of lumbar fracture, required no assistive device for mobility, and were taking no medications known to affect skeletal mineralization during the study period comprised the investigational group. Absolute BMD and age- and gender-matched BMD (Z-scores) were assessed relative to standard parameters of growth (height, weight, age, height adjusted for age and gender and body surface area) and severity of disease (lifetime fracture rate). The spinal mineralization rate (SMR) between examinations for 15 patients with more than one measurement (n = 20 intervals) was expressed as the magnitude of the change in BMD Z-score per year. Both BMD and BMD Z-score were closely correlated with height, height Z-score, weight and body surface area and were inversely related to fracture rate (P < 0.001 for all comparisons). BMD was also highly correlated with patient age (P < 0.001). Stepwise regression analysis showed that together height Z-score and lifetime fracture rate improved the prediction of BMD Z-score (r = 0. 71; P < 0.001). SMRs ranged from -0.5 to 3.5. The average change in SMR between sequential measurements was 168% for the five children who had more than two DXA examinations. Linear regression showed a significant negative correlation between SMR and height Z-score (r = -0.79, P < 0.001). We conclude that vertebral body size is a critical determinant of BMD and BMD Z-score in OI because DXA results are expressed per unit area, not per unit volume. Pediatric patients with OI mineralize their lumbar vertebrae at rates similar to healthy children but tend to lag behind in overall mineralization. The rate of mineralization at any age appears to be related to the patient's height (adjusted for age- and gender-matched controls) and inversely related to the patient's lifetime rate of fractures. Our data suggest that vertebral mineralization in children with OI is related primarily to rapid increases in vertebral volume and only secondarily to increases in vertebral mineral density.

Adolescent↗

Vascularization and cartilage mineralization of the thyroid cartilage of Munich minipigs and domestic pigs.

Thyroid cartilages of Munich minipigs and domestic pigs were investigated by polychrome sequential labeling, radiography, intravascular injections, histologic examination and scanning electron microscopy in order to gain further insight into the process of vascularization and cartilage mineralization. The relationship between vascularization and cartilage mineralization has only been studied in chondroepiphyses of long bones. Vessels branch off the perichondrial vascular network and enter parts of the thyroid cartilage with a large transverse diameter. Cartilage canals, which are perichondral invaginations, contain an arteriole, a venule, a capillary network and connective tissue. The capillaries form a glomerulus-like structure deep in the matrix of the cartilage. Neighbouring cartilage canals do not display any anastomoses. Cartilage mineralization occurs in large areas of the thyroid cartilage. It is only found in the interterritorial extracellular matrix. Mineralization of the cartilage is evident in areas supplied with cartilage canals as well as in non-supplied areas. Mineralized interterritorial matrix is composed of circular structures of different sizes fusing to form plaques. In scanning electron microscopy circular structures appear as globules. It is possible to visualize the dynamic process of cartilage mineralization with polychrome sequential labeling; it proceeds up to 4 microm per week. Distribution of cartilage canals reveals their nutritional role for the cartilage. According to investigations in chondroepiphyses, cartilage mineralization starts adjacent to the glomerular end of cartilage canals. In contrast, no correlation between cartilage vascularization and the beginning of cartilage mineralization of the thyroid cartilage of Munich minipigs and of domestic pigs has been found.

Animals↗

The appearance of matrix vesicles and mineralization during tooth development in three teleost fishes with well-developed enameloid and orthodentine.

Ultra-thin resin sections of tooth bearing parts from three species were examined by electron microscopy. During enameloid matrix formation, matrix vesicles (MV) and fine aggregations of crystal-like structures probably derived from the MV were found in the enameloid matrix. In enameloid, however, no additional mineralization occurred along the collagen fibrils until mineralization had begun initially at the junction between the enameloid and orthodentine areas. Advance of mineralization along collagen fibrils was inhibited in the enameloid area before initial mineralization despite the presence of MV. Many MV were observed at the initial mineralization site; they appeared in the predentine during odontogenesis after the initial mineralization and were still visible during the stage of basal dentine formation. The odontoblasts continued to produce MV from the early stage of enameloid matrix formation until the late stage of basal dentine formation. However, despite their long appearance, MV are probably involved directly with the mineralization only when the initial mineralization starts.

Animals↗

In vivo and in vitro study of the effects of chlorpromazine on tooth mineralization in rats and mice.

The effects of chlorpromazine (CPZ) on tooth mineralization were examined using incisor dentine in adult rats and cultured tooth germs of mandibular first molars dissected from mouse embryos. CPZ (10, 50 and 250 mg/kg, s.c.) substantially inhibited dentine mineralization as evaluated by contact microradiographs. Plasma calcium and phosphorus concentrations were not decreased by CPZ (10 and 50 mg/kg). Physicochemical effects were not involved in the action of CPZ on the mineralization. In vitro experiments showed that CPZ (1 and 10 microM) inhibited mineralization and alkaline phosphatase (ALP) activity in the tooth germs. As CPZ has the properties of a calmodulin antagonist, the calmodulin antagonists W-7 and W-5 were also examined. Both inhibited mineralization and ALP activity in tooth germs; W-5 had less effect than W-7. These in vivo and in vitro findings suggest that CPZ inhibited cell-mediated mineralization in dentine without affecting the calcium-regulating system and physicochemical mineral deposition. In addition, calmodulin could be involved in cell-mediated mineralization.

Alkaline Phosphatase↗

Morphometric analysis of the deposition and mineralization of enamel and dentine from rat incisor during the recovery phase following a low-calcium regimen.

The administration of a low-calcium diet to pups nursed by a mother on the same diet has been shown to induce a slowing of growth. A reduction of the apposition rate of dentine, which was normally mineralized, and a dramatic reduction of the extent of mineralization of enamel, whose organic matrix was otherwise produced in an almost normal amount, was observed in the incisors of these animals. Modifications of the mineral apposition rate of dentine, measured after administration of tetracycline (10 mg/kg), and the thickness and the microhardness of the two tissues, the latter being an expression of the degree of mineralization, were now investigated when hypocalcic pups were restored to a normal-calcium diet for 10 or 60 days. Enamel microhardness was increased by more than 60% after only 10 days of restored diet and had become the same as in the control tissue after 60 days, without any significant increase in thickness. Dentine thickness and mineral apposition rate increased significantly, to become similar to those of the controls after 60 days of restored diet. In dentine there was no significant variation of microhardness between experimental pups and controls, either during the low-calcium diet or the restorative period. These results indicate that the deposition of the organic matrix of enamel is a process independent from that of its mineralization, and that the mineralization of the organic matrix happens by its replacement even a long time after its deposition. In contrast, the deposition and mineralization of dentine are strictly interdependent processes, at least in these experimental conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Bone mineral measurement: Skylab experiment M-078.

The observation that bone mineral is lost in patients who are either immobilized or remain in bed for extended periods of time formed the basis for the concern that large amounts of bone mineral may be lost during long periods of weightlessness. This concern was magnified when early X-ray densitometry studies suggested that rather large amounts of mineral could be lost during rather short periods of weightlessness (4-14 days). Even though these Gemini results have recently been modified, they still reflect substantial losses in the upper extremity. This led to a series of prolonged bed-rest studies (30-36 weeks) which, in addition to careful calcium balance, also employed a newer, more precise method of estimating bone mineral in the radius, ulna, and os calcis. It employed an essentially monoenergetic photon source (125I) and a scintillation detector operating in a rectilinear scanning mode to measure bone mineral by the absorptiometric technique. Bed-rest studies revealed variable mineral losses but suggested that little if any is lost during 4-6 weeks, with variable amounts being lost in 8 weeks. Losses up to 40% were noted in the os calcis after 9 months, with essentially none in the radius and ulna. When this technique was employed during the Apollo 14, 15, and 16 missions, only one crewman (CMP Apollo 15) showed significant losses in the os calcis and none in the radius or ulna. These results were, therefore, in concert with the bed-rest data but at variance with the earlier Gemini data. The variability observed during bed rest was reconciled when it was observed that the rate of loss could be correlated with the initial 24-hour urinary hydroxyproline excretion and the initial os calcis mineral content. Prediction terms were established. Measurements of the SL-II crew after 28 days of weightlessness revealed no significant bone mineral losses. The Skylab data lie within the predicted limits obtained from the bed-rest data. The relevance of the prediction terms to the Skylab and longer missions discussed.

Absorptiometry, Photon↗

Lipid levels and bone mineral density.

PURPOSE: There has been considerable debate about the potential relationship between the use of statin lipid-lowering drugs and fracture risk; several observational studies suggest a protective effect but no randomized controlled trials have confirmed such a benefit. Because statins are given preferentially to persons with hyperlipidemia, if lipid levels were associated with bone mineral density, this could explain the discrepancy between epidemiological observations and randomized controlled trials. The aim of this study was to examine the relationship between lipid levels and bone mineral density. SUBJECTS AND METHODS: We included the 13592 participants in the National Health and Nutritional Examination Survey (NHANES) III who had bone mineral density and lipid levels measured; participants who reported the use of a lipid-lowering therapy were excluded. We examined the unadjusted bone mineral density across quintiles of total cholesterol, low-density lipoprotein (LDL), and high-density lipoprotein (HDL). We then constructed multivariable models, including age, sex, body mass index, and other potential confounders. RESULTS: In crude analyses, higher total cholesterol and LDL levels were associated with lower bone mineral densities (both P values for trend <.001), whereas higher HDL levels were associated with higher bone mineral densities (P value for trend <.001). However, in fully adjusted models, there was no significant relationship between total cholesterol, LDL, or HDL levels and bone mineral density (all P values for trend >.1). CONCLUSIONS: These results do not support a relationship between lipid levels and bone mineral density.

Adolescent↗

Bone mineral density in subjects using central nervous system-active medications.

PURPOSE: Decreased bone mineral density defines osteoporosis according to the World Health Organization and is an important predictor of future fractures. The use of several types of central nervous system-active drugs, including benzodiazepines, anticonvulsants, antidepressants, and opioids, have all been associated with increased risk of fracture. However, it is unclear whether such an increase in risk is related to an effect of bone mineral density or to other factors, such as increased risk of falls. We sought to examine the relationship between bone mineral density and the use of benzodiazepines, anticonvulsants, antidepressants, and opioids in a representative US population-based sample. SUBJECTS AND METHODS: We analyzed data on adults aged 17 years and older from the Third National Health and Nutrition Examination Survey (NHANES III, 1988-1994). Total femoral bone mineral density of 7114 male and 7532 female participants was measured by dual-energy x-ray absorptiometry. Multivariable linear regression models were used to quantify the relation between central nervous system medication exposure and total femoral bone mineral density. Models controlled for relevant covariates, including age, sex, and body mass index. RESULTS: In linear regression models, significantly reduced bone mineral density was found in subjects taking anticonvulsants (0.92 g/cm2; 95% confidence interval [CI]: 0.89 to 0.94) and opioids (0.92 g/cm2; 95% CI: 0.88 to 0.95) compared with nonusers (0.95 g/cm2; 95% CI: 0.95 to 0.95) after adjusting for several potential confounders. The other central nervous system-active drugs--benzodiazepines or antidepressants--were not associated with significantly reduced bone mineral density. CONCLUSION: In cross-sectional analysis of NHANES III, anticonvulsants and opioids (but not benzodiazepines or antidepressants) were associated with significantly reduced bone mineral density. These findings have implications for fracture-prevention strategies.

Adolescent↗

Skeletal tissue growth, differentiation and mineralization in the NASA rotating wall vessel.

The NASA Rotating Wall Vessel (RWV) is a device that creates a unique environment that supports three-dimensional tissue growth, a heightened level of cell differentiation and randomizes the position of the downward gravitational force on cells. Embryonic bone formation encompasses a cascade of chondrogenic and osteogenic events, which can be altered by changes in gravitational loading. These studies were conducted to determine if the chondrogenic cascade in bone formation would be enhanced or hindered in the unique culture environment of the RWV. Embryonic mouse pre-bone tissues were placed in the RWV at one of four different stages of chondrogenesis, ranging from undifferentiated mesenchyme cells to chondrocytes on the verge of undergoing terminal chondrocyte differentiation. After culture, tissues were analyzed for their size, the amount of alkaline phosphatase (ALP) activity and their ability to form a mineralized matrix. Tissue consisting of cells at the early phase of chondrogenesis grew very little and did not differentiate or mineralize when cultured in the RWV. Some tissues were cultured for short periods in the RWV then cultured in standard culture dishes (SCD). Following this culture regime, the cartilage grew only a small amount, but alkaline phosphatase activity increased, and some mineralized regions formed. The pattern of mineralization was abnormal, with two mineralized zones at each end of the cartilage instead of a single central zone. Tissues that were at the three more advanced stages of chondrogenesis when placed in the RWV showed substantial growth, differentiation and mineralization. Mineralization patterns in these older tissues was normal. Tissues at the oldest stage of chondrogenesis showed more growth and as much or more mineralization as tissue cultured only in SCD. These data suggest that exposure to the RWV at early stages of chondrogenesis severely limits the ability for cartilage growth and yields abnormal downstream morphogenesis. However, at later stages of chondrogenesis, the RWV environment may be beneficial and enhance growth and development. Future studies to characterize intercellular signaling molecules and gene expression activities of chondrocytes in the RWV will be valuable for understanding the mechanism of skeletogenesis.

Animals↗

Long-term risedronate treatment normalizes mineralization and continues to preserve trabecular architecture: sequential triple biopsy studies with micro-computed tomography.

The objective of the study was to assess the time course of changes in bone mineralization and architecture using sequential triple biopsies from women with postmenopausal osteoporosis (PMO) who received long-term treatment with risedronate. Transiliac biopsies were obtained from the same subjects (n = 7) at baseline and after 3 and 5 years of treatment with 5 mg daily risedronate. Mineralization was measured using 3-dimensional (3D) micro-computed tomography (CT) with synchrotron radiation and was compared to levels in healthy premenopausal women (n = 12). Compared to the untreated PMO women at baseline, the premenopausal women had higher average mineralization (Avg-MIN) and peak mineralization (Peak-MIN) by 5.8% (P = 0.003) and 8.0% (P = 0.003), respectively, and lower ratio of low to high-mineralized bone volume (BMR-V) and surface area (BMR-S) by 73.3% (P = 0.005) and 61.7% (P = 0.003), respectively. Relative to baseline, 3 years of risedronate treatment significantly increased Avg-MIN (4.9 +/- 1.1%, P = 0.016) and Peak-MIN (6.2 +/- 1.5%, P = 0.016), and significantly decreased BMR-V (-68.4 +/- 7.3%, P = 0.016) and BMR-S (-50.2 +/- 5.7%, P = 0.016) in the PMO women. The changes were maintained at the same level when treatment was continued up to 5 years. These results are consistent with the significant reduction of turnover observed after 3 years of treatment and which was similarly maintained through 5 years of treatment. Risedronate restored the degree of mineralization and the ratios of low- to high-mineralized bone to premenopausal levels after 3 years of treatment, suggesting that treatment reduced bone turnover in PMO women to healthy premenopausal levels. Conventional micro-CT analysis further demonstrated that bone volume (BV/TV) and trabecular architecture did not change from baseline up to 5 years of treatment, suggesting that risedronate provided long-term preservation of trabecular architecture in the PMO women. Overall, risedronate provided sustained benefits on mineralization and architecture, two key determinants of bone strength, over 5 years lending support for its long-term efficacy in fracture risk reduction.

Adult↗