bluuebunny/crossref_metadata_2025 huggingface dataset:
Growth hormone stimulates bone formation and strength of cortical bone in aged rats
Authors: Andreassen Troels T. , Jørgensen Peter H. , Oxlund Hans , Flyvbjerg Allan , Ørskov Hans
Published: February 2010

The influence of growth hormone on bone formation, mechanical strength, and composition has been investigated in femur middiaphyseal cortical bone from 2-year-old male rats. The rats were given biosynthetic human growth hormone (bhGH) at 2.7 mg/kg/day in two daily injections for 20, 40, or 80 days, and all animals were killed 80 days after the start of bhGH administration. Control animals were given saline. All animals were labeled with tetracycline on days 41 and 69. Only in the bhGH-80-day group was subperiosteal tetracycline double labeling seen all around the femur diaphysis, and this pattern was found in all animals of the group. Double labeling subperiosteally at the posteromedial aspect was found in all animals of the experiment, but compared with the control group, a 400% and an 800% increase in mineral apposition rate was seen in the bhGH-40-day and bhGH-80-day groups, respectively. Light microscopy and polarization microscopy showed that this newly deposited bone was organized in the same concentric lammellae and had the same direction of the collagen fibers when compared with the surrounding bone formed before the start of bhGH injections. The cortical bone cross-sectional area was increased in the bhGH-40-day and bhGH-80-day groups. At the endosteum, scattered labeling was found in animals from all groups, and no differences in medullary cross-sectional areas were seen. The mechanical analysis revealed an increased mechanical strength of the whole diaphyseal bone after bhGH administration. When the data were corrected for dimensions of the diaphyseal bone, no differences in intrinsic mechanical properties of the bone tissue were found. No differences in apparent density of dry defatted bone, ash, and collagen were seen, whereas apparent density of dry defatted bone minus ash was decreased in all groups given bhGH. Correspondingly, a slight increase in ash concentrations of the bhGH-injected animals was seen. bhGH administration also increased the body weight, muscle mass, and total serum IGF-I and thyroxine concentrations. (
Validation of accuracy and precision of dual energy X-ray absorptiometry for infants
Authors: Koo Winston W. K. , Massom Larry R. , Walters Jocelyn
Published: February 2010

There is limited information on the validity of bone mineral content measurement by dual energy X-ray absorptiometry (DXA BMC) for use in subjects with low body mass. We evaluated the accuracy and precision of DXA in piglets (body weight 886–5526 g, median 2096 g). Stepwise multiple regression analyses showed that ash weight is the major determinant of DXA BMC (adjusted r2 = 0.98, RMS residual = 3.61 g). The intercept was not significantly different from zero. DXA BMC measurements of other piglets under various clinical situations showed no significant effect from the use of cotton blanket, diaper, or positioning (prone, supine, lateral). In vivo replication of DXA BMC measurements of infants at a postnatal age of from 1–350 days showed a slope of 0.99 and high correlation coefficient (r2 = 0.99, RMS residual = 3.59 g). The intercept was not significantly different from zero, and the average coefficient of variation of duplicate DXA BMC in infants was 2.8%. We conclude that DXA BMC reliably but proportionately underestimates ash weight and is a highly precise method for measuring bone mineral status in young pediatric subjects.
Calvarial and limb bone cells in organ and monolayer culture do not show the same early responses to dynamic mechanical strain
Authors: Rawlinson Simon C. F. , Mosley John R. , Suswillo Rosemary F. L. , Pitsillides Andrew A. , Lanyon Lance E.
Published: February 2010

Responses to mechanical strain in calvaria and limb bone organ cultures were compared by measuring cellular glucose 6-phosphate dehydrogenase (G6PD) activity in situ and prostaglandin release. Normal functional strains were recorded in the ulnae (1000 μϵ) and calvarium (30 μϵ) in vivo in 110 g rats. Organ cultures of ulnae and calvaria from similar animals were loaded to produce dynamic strains (600 cycles, 1 Hz) of 1000 μϵ in the ulna, and 100 or 1000 μϵ in calvaria. In ulnae, both PGE2 and PGI2 were released and resident osteocytes and osteoblasts showed increased G6PD activity. Neither response was seen in calvaria. However, exogenous PGI2 (10−5−10−9 M) stimulated G6PD activity in osteocytes and osteoblasts in organ cultures of both calvaria and ulnae. In ulnar cells the response was linear, in calvarial cells it was biphasic with maximum activity at 10−7 M. Osteoblasts derived from ulnae and cultured on plastic plates subjected to dynamic strain (600 cycles, 1 Hz, 4000 μϵ) showed increased G6PD activity. There was no such response in similarly treated calvarial-derived cells. Calvarial bone cells differ from those of the ulna in that they do not respond to physiological strains in their locality with increased prostanoid release or G6PD activity either in situ or when seeded onto dynamically strained plastic plates. Cells from both sites in organ culture show increased G6PD activity in response to exogenous PGI2, but their dose: Responses differ in shape. These differences may reflect the extent to which functional loading influences bone architecture in these two sites.
Factors contributing to skeletal health of postmenopausal Mexican-American women
Authors: Villa Marie Luz , Marcus Robert , Delay Romelia Ramírez , Kelsey Jennifer L.
Published: February 2010

Incidence rates of hip fracture are lower in Hispanic (HC) than non-Hispanic Caucasians (NHC). To investigate factors that may affect skeletal health of Hispanics, we recruited 152 healthy community-dwelling Mexican-American Caucasian women into a 4-year longitudinal study that evaluates bone mass, nutritional status, muscle strength, mobility, falls, and other factors that may contribute to fracture risk. Results from the baseline component of the study are reported herein. Average bone mineral densities (BMD) evaluated by dual-energy X-ray absorptiometry (DXA) in this study group did not differ from BMDs in healthy, NHC women of similar age. Hip axis length (HAL), however, was significantly shorter than that reported for nonosteoporotic NHC. Factors independently associated with greater BMD and BMC at certain skeletal sites were lean body mass, fat mass, acculturation, years of estrogen use, sun exposure, hip adductor strength, grip strength, erythrocyte folate, and serum glucose concentrations. Factors independently associated with lower BMD and BMC at certain skeletal sites were age, parity, and vertebral deformities (all p < 0.05). Thus, the decreased risk of hip fracture in HC compared with NHC does not appear to be due to high bone mass. However, other factors such as HAL and body composition may play a role in maintenance of skeletal integrity.
Effects of differentiation and transforming growth factor β1 on PTH/PTHrP receptor mRNA levels in MC3T3-E1 cells
Authors: Mccauley Laurie K. , Koh Amy J. , Beecher Christopher A. , Cui Yingqi , Decker James D. , Franceschi Renny T.
Published: February 2010

TGFβ has opposing effects on osteoblasts which are thought to be differentiation stage dependent; however, little is known concerning the effects of TGFβ on osteoblastic characteristics at different stages of maturation. The purpose of this study was to characterize the pattern of mRNA expression for the PTH/PTHrP receptor during normal osteoblastic differentiation in vitro, and evaluate the effects of TGFβ1 on PTH/PTHrP receptor and osteocalcin (OCN) steady-state mRNA at different stages of osteoblastic differentiation. MC3T3-E1 preosteoblasts were plated at low density and induced to differentiate with ascorbic acid and β-glycerophosphate. The first group served as a vehicle control and the remaining five groups received a single 48 h TGFβ1 (3.0 ng/ml)-pulse staggered on a weekly basis for 30 days. Cell cultures were harvested weekly and evaluated for: steady-state PTH/PTHrP receptor and OCN mRNA levels via northern analysis, calcium and phosphorous levels, bone nodules via Von Kossa staining, alkaline phosphatase enzyme levels, and hydroxyproline levels. Group 1 (control) samples followed a normal pattern of proliferation, extracellular matrix deposition, and mineralization. PTH/PTHrP receptor and OCN mRNA expression increased 8-fold and 10-fold respectively, over the collection periods. When TGFβ1 was administered during the first 48 h period (group 2) while cells were rapidly proliferating, there was a persistent inhibition of PTH/PTHrP receptor expression and a striking reduction in OCN mRNA expression at all time points. There was also a down-regulation of PTH/PTHrP receptor and OCN expression when TGFβ1 was administered later during osteoblast differentiation (groups 3–6); however, these effects were not persistent. In addition there was a total lack of bone nodule formation in group two cultures, whereas groups 3–6 had increasing bone nodule formation because the TGFβ1 was administered later in the culture period. These studies indicate that expression of the PTH/PTHrP receptor increases with osteoblastic differentiation and suggest that TGFβ1 inhibits osteoblastic maturation with more persistent effects found in less differentiated osteoblastic cells.
Droloxifene prevents ovariectomy-induced bone loss in tibiae and femora of aged female rats: A dual-energy X-ray absorptiometric and histomorphometric study
Authors: Chen Hong Ka , Ke Hua Zhu , Jee Webster S. S. , Ma Yan Fei , Pirie Christine M. , Simmons Hollis A. , Thompson David D.
Published: February 2010

Our previous studies indicated that droloxifene (DRO), a tissue-specific estrogen antagonist/agonist, prevented bone loss without causing uterine hypertrophy in growing ovariectomized (OVX) rats. Using dual-energy X-ray absorptiometry (DXA) and bone histomorphometry, the current study compared the efficacy of DRO to 17β-estradiol (E2) in preventing OVX-induced bone loss in tibiae and femora of 19-month-old rats to determine whether DRO had similar skeletal effects as E2 in aged female rats. Sprague-Dawley female rats were OVX or sham-operated (sham) at 19 months of age. The sham-operated rats were treated with vehicle (oral), while the OVX rats were treated with vehicle (oral), E2 at 30 μg/kg/day (sc), or DRO at 2.5, 5, or 10 mg/kg/day (oral) for 8 weeks. Bone mineral density (BMD) of whole femora (WF), distal femoral metaphyses (DFM), femoral shafts (FS), and proximal femora (PF) was determined using DXA. Static and dynamic cancellous bone histomorphometric analyses were performed in double-labeled undecalcified longitudinal sections from proximal tibial metaphyses. Ovariectomy for 8 weeks significantly reduced the BMD of WF, DFM, FS, and PF (from −6 to −15%). Treatment with E2 completely prevented the decreases in BMD of WF and DFM and had no significant effects in BMD of FS and PF in aged OVX rats. The decrease in BMD of DFM induced by OVX was prevented by treatment with DRO at all dose levels. In addition, DRO at 10 mg/kg/day prevented OVX-induced decreases in BMD of WF, FS, and PF. Furthermore, proximal tibial cancellous bone histomorphometric results showed that OVX significantly decreased the trabecular bone volume by 34% and increased the activation frequency by 104% while it nonsignificantly increased other indices including percent eroded perimeter, mineral apposition rate, and bone formation rate per bone volume compared with sham-operated controls. Treatment with E2 or DRO at all dose levels completely prevented the OVX-induced decreases in trabecular bone volume and increases in bone turnover, indicating that DRO is an estrogen agonist in bone in aged OVX rats. Together with the previous findings that DRO inhibited body weight gain, reduced total serum cholesterol, and had no effect on uterine weight, we conclude that DRO is as efficacious as E2 in preventing OVX-induced bone loss and inhibiting bone turnover but without estrogenic uterine effects in aged OVX rats. These data suggest that DRO may be superior to E2 for the treatment of postmenopausal and senile osteoporoses.
Vitamin D receptor gene polymorphisms do not predict bone turnover and bone mass in healthy premenopausal women
Authors: Garnero Patrick , Borel Olivier , Sornay-Rendu Elizabeth , Dr. Delmas Pierre D.
Published: February 2010

Bone mineral density (BMD) is under strong genetic control. Polymorphisms at the vitamin D receptor (VDR) gene have been recently suggested to account for up to 75% of this genetic effect. We analyzed these polymorphisms, i.e., that of BsmI, TaqI, and ApaI restriction enzymes by PCR of the DNA in 189 healthy premenopausal women aged 31 to 57 years. For the BsmI polymorphism they were 17% BB homozygotes, 51% Bb heterozygotes, and 32% bb homozygotes, genotype frequencies that are very similar to those previously reported in other Caucasian populations of north European ancestry. Women in the three genotypes for any of the three polymorphisms were matched for age and did not differ in body weight, height, physical activity, nor smoking habits. We found no relationship between the genotype for any of the three polymorphisms nor bone formation and resorption rate assessed by five specific biochemical markers of bone turnover nor with BMD measured at the spine, proximal femur, forearm, and whole body by dual-energy X-ray absorptiometry (DXA). We concluded that these polymorphisms are not predictive of bone turnover nor BMD in a sample of healthy premenopausal women drawn from the French population.
Triiodothyronine potentiates the stimulatory effects of interleukin-1β on bone resorption and medium interleukin-6 content in fetal rat limb bone cultures
Authors: Tarjan Gabor , Dr. Stern Paula H.
Published: February 2010

It has been demonstrated that thyroid hormones stimulate osteoclasts indirectly and that this effect is mediated by products of other cell types present in bone. To determine if interleukin-6 (IL-6) could be a mediator of thyroid hormone action, we investigated the effect of 3,5,3′-triiodothyronine (T3) on bone resorption (45Ca release) and on the IL-6 concentration in medium from cultured 19-day-old fetal rat limb bones. T3 alone increased 45Ca release significantly only at a fairly high concentration (10−6 M) under the conditions used. T3 alone, over a 10−11–10−6 M concentration range, failed to elicit a detectable effect on the medium IL-6 content. However, T3 potentiated the stimulatory effect of interleukin-1β (IL-1β) on IL-6 production in a dose-dependent manner. T3, 10−8 M, also significantly increased IL-1β-stimulated calcium release. Inhibition of IL-1β with 1 μM interleukin-1 receptor antagonist (IL-Ira) abrogated the potentiating effects of T3 on IL-1β-stimulated IL-6 production and blocked the combined effect of T3 and IL-1β on 45Ca release. One micromolar indomethacin significantly, but not completely, inhibited the effect of IL-1β, as well as the combined effect of IL-1β and T3 on resorption and IL-6 production, indicating the involvement of prostaglandins in these actions. Consistent with this, 1 μM prostaglandin E1 (PGE1) significantly increased both the IL-6 production and the calcium release. By potentiating the effect of IL-1β, T3 increased bone resorption at much lower concentrations. We therefore speculate that the enhancement of IL-1β effects may be a biologically relevant mechanism of thyroid hormone action on bone.
The potential role of fibroblasts in periprosthetic osteolysis: Fibroblast response to titanium particles
Authors: Yao Jianling , Dr. Glant Tibor T. , Lark Michael W. , Mikecz Katalin , Jacobs Joshua J. , Hutchinson Nancy I. , Hoerrner Lori A. , Kuettner Klaus E. , Galante Jorge O.
Published: February 2010

Periprosthetic osteolysis with or without aseptic loosening is a major clinical problem in total hip arthroplasty. While the macrophage response to prosthetic wear debris and its role in periprosthetic osteolysis has been extensively studied, information regarding other cell types (fibroblasts, osteoblasts) is limited. This study explored the response of fibroblasts to particulate wear debris. Fibroblasts isolated from interfacial membranes of patients with failed total hip replacements and normal synovial tissue, when challenged with small-sized (<3 μm) titanium (Ti) particles, responded with significantly enhanced expressions of collagenase, stromelysin and, to a much lesser extent, their tissue inhibitor of metalloproteinases (TIMP). These “regulated” expressions at both mRNA and protein levels were correlated with the size and composition of particles. De novo protein synthesis was required for the regulation of these mRNAs. A similar effect could be induced by the treatment of the cells with particle-free conditioned medium from Ti particle-stimulated fibroblasts. Furthermore, this conditioned medium significantly suppressed the mRNA levels of procollagen α1(I) and α1(III) in osteoblast-like MG-63 cells. It is concluded that fibroblasts stimulated with certain particle debris may play an important role in periprosthetic osteolysis by releasing bone-resorbing metalloproteinases and mediator(s) which resulted in suppressed collagen synthesis in osteoblasts.
Intermittent treatment with human parathyroid hormone (hPTH[1-34]) increased trabecular bone volume but not connectivity in osteopenic rats
Authors: Dr. Lane Nancy E. , Thompson J.M. , Strewler Gordon J. , Kinney J.H.
Published: February 2010

Previous studies have determined that intermittent parathyroid hormone (PTH) therapy increases bone mass and improves biomechanical strength in osteopenic animal models. The purpose of this investigation was to determine if intermittent human parathyroid hormone (hPTH[1–34]) therapy increased trabecular bone volume and connectivity in a rat model of established osteopenia using three-dimensional (3D) ex vivo in situ morphometry by X-ray tomographic methods (XTM). Six-month-old retired Sprague-Dawley breeder rats were used. Thirty animals were ovariectomized (OVX) and six were Sham operated. On day 56, post-OVX, a prePTH-treatment OVX group was sacrificed. The remaining OVX animals were randomized into four groups of six animals each, given injections 5 out of every 7 days for 28 days of either vehicle or hPTH(1-34) at 4, 40, or 400 μg/kg of body weight (BW)/day and were sacrificed on day 84 post-OVX. At sacrifice, the left proximal tibias were harvested for XTM scans. hPTH(1-34) at medium and high doses significantly increased trabecular bone volume and trabecular thickness compared with ovariectomized animals treated with vehicle (p < 0.05). The trabecular bone volume was equal to or greater than the Sham-operated animals in both hPTH(1–34) 40 and 400 μg/kg of BW treatment groups. Trabecular bone connectivity decreased by nearly 50% compared to the Sham-operated group at day 84 post-OVX and did not increase with any of the hPTH(1-34) treatments. Intermittent hPTH(1-34) treatment in osteopenic OVX rats increased trabecular bone volume to control levels or higher by thickening existing trabecule. Human PTH(1–34) did not re-establish connectivity when therapy was started after 50% of the trabecular connectivity was lost. We hypothesize that to re-establish trabecular connectivity, a therapeutic intervention would have to be given before a significant distance between trabeculae has developed. Further studies will need to be done to refute or confirm our hypothesis.
Bisphosphonates promote apoptosis in murine osteoclasts in vitro and in vivo
Authors: Hughes David E. , Wright Kenneth R. , Uy Harry L. , Sasaki Akira , Yoneda Toshiyuki , Roodman David G. , Mundy Gregory R. , Dr. Boyce Brendan F.
Published: February 2010

Bisphosphonates inhibit bone resorption and are therapeutically effective in diseases of increased bone turnover, such as Paget's disease and hypercalcemia of malignancy. The mechanisms by which they act remain unclear. Proposed mechanisms include inhibition of osteoclast formation from precursors and inhibitory or toxic effects on mature osteoclasts. We have developed a new in vitro model to study osteoclast survival and in this paper present in vitro and in vivo evidence that may explain both the observed reduction in osteoclast numbers and in bone resorption by mature osteoclasts, namely that bisphosphonates induce programmed cell death (apoptosis). Three bisphosphonates (risedronate, pamidronate, and clodronate) caused a 4- to 24-fold increase in the proportion of osteoclasts showing the characteristic morphology of apoptosis in vitro. This observation was confirmed in vivo in normal mice, in mice with increased bone resorption, and in nude mice with osteolytic cancer metastases, with similar-fold increases to those observed in vitro. Of the three compounds, risedronate, the most potent inhibitor of bone resorption in vivo, was the strongest inducer of osteoclast apoptosis in vitro. Osteoclast apoptosis may therefore be a major mechanism whereby bisphosphonates reduce osteoclast numbers and activity, and induction of apoptosis could be a therapeutic goal for new antiosteoclast drugs.
Pretreatment with low doses of norethindrone potentiates the osteogenic effects of fluoride on human osteosarcoma cells
Authors: Takada Junichi , Baylink David J. , Lau William K.-H.
Published: February 2010

We recently reported that picomolar doses of norethindrone (NET), a synthetic analog of 19-nortestosterone, significantly stimulated human TE85 osteosarcoma cell proliferation, differentiation, and activity in vitro. In the present study, we investigated the possibility that NET interacts with another osteogenic agent, i.e., fluoride, to stimulate human TE85 osteosarcoma cell proliferation, differentiation, and activities. Bone cell proliferation was measured by the stimulation in [3H]thymidine incorporation. Differentiation was monitored by the increase in alkaline phosphatase-specific activity. Osteoblastic activity was assessed by the stimulations in collagen synthesis and in osteocalcin secretion (in the presence of 1 nM 1,25-dihydroxyvitamin D3). When the human TE85 cells were incubated with mitogenic doses of NET and fluoride concurrently, the stimulatory effects of the two agents on these parameters exhibited no significant interaction. The enhancing effect of NET on the osteogenic effect of fluoride was not due to a shift of the fluoride dose response curve. Pretreatment with NET for 24 h followed by a treatment with a mitogenic dose (i.e., 100 μM) of fluoride for an additional 24 h significantly and synergistically potentiated the effects of fluoride on the [3H]thymidine incorporation, alkaline phosphatase-specific activity, collagen synthesis, and osteocalcin secretion, compared with those with the subsequent vehicle (0.05% ethanol) treatments. In contrast, pretreatment with fluoride for 24 h before the addition of NET for 24 h did not produce significant synergistic stimulations in the test parameters. Pretreatment of TE85 cells with the same doses of dihydrotestosterone or progesterone prior to treatment with fluoride under the same conditions did not induce synergistic potentiation of fluoride in [3H]thymidine incorporation, suggesting that the synergistic interaction with fluoride is probably not a common property of anabolic sex steroids. In summary, we found that: (1) the osteogenic effects of fluoride and NET were additive when cells were treated with both agents concurrently; (2) a 24-h pretreatment with picomolar doses of NET potentiated the osteogenic actions of fluoride in human TE85 osteosarcoma cells; and (3) pretreatment with NET produced a subsequent fluoride response that was synergistic. In conclusion, these findings led us to speculate that the osteogenic actions of NET and fluoride act through different mechanisms, and that NET at low doses has a permissive effect on the osteogenic effects of fluoride, and as such NET may be used in concert with fluoride to increase osteoblast proliferation, differentiation, and activity.
Evidence for interrupted bone resorption in human iliac cancellous bone
Authors: Croucher P.I. , Gilks W.R. , Compston J.E.
Published: February 2010

Bone resorption and formation are coupled both in time and space and may occur simultaneously in the same remodeling unit. A number of studies have shown that the formative phase of the remodeling sequence may undergo temporary interruptions prior to completion and it is possible that bone resorption may be subject to similar interruptions. We have investigated this hypothesis by studying the distribution of eroded depth in resorption cavities in human cancellous bone. Eroded depth was assessed in iliac crest cancellous bone from 41 normal healthy subjects using a cubic spline curve fitting technique. The distribution of mean eroded depths was skewed to the right. Comparison of the observed distribution with an expected distribution, which was calculated from previously published data and assumes resorption begins rapidly and slows as it approaches completion, showed a significantly greater proportion of shallower cavities than expected (p < 0.0001). Similarly, comparison of observed and uniform distributions, which assumes a constant rate of resorption throughout the erosion period, also showed a significantly greater proportion of smaller cavities (p < 0.01). In subjects aged less than 39 years, there were fewer small cavities than in those aged 40-59 years. In addition, there was some evidence that females of 40-59 years had a proportionately greater number of smaller cavities than males; however, there were no differences in other age groups. Our results demonstrate a significantly greater proportion of smaller resorption cavities than would be expected from current models of bone remodeling and are consistent with the hypothesis that resorption undergoes temporary interruptions and/or permanent arrest during the process of bone remodeling. (J Bone Miner Res 1995;10:1537-1543)
Assessment of the skeletal status by peripheral quantitative computed tomography of the forearm: Short-term precision in vivo and comparison to dual X-ray absorptiometry
Authors: Grampp Stephan , Lang Philipp , Jergas Michael , Glüer Claus C. , Mathur Ashwini , Engelke Klaus , Genant Harry K.
Published: February 2010

In order to assess precision of peripheral quantitative computed tomography (pQCT), duplicate bone mineral density (BMD) measurements at the radius were performed in 20 healthy premenopausal, 20 healthy postmenopausal, and 20 osteoporotic postmenopausal women using a Stratec XCT-960 system. The short-term reproducibility in vivo for the total, trabecular, and cortical regions of interest (ROI) was expressed as the absolute precision error (standard deviation, SD) and as the relative precision error (SD/mean x 100, or coefficient of variation, CV, in %). Reproducibility in vivo was good in all volunteers but was influenced by the study group and the ROI. The precision error for trabecular BMD was 3 mg/cm3, or about 1.6%. This is large relative to the aging decrease of 0.22%/year, or to the difference (12 mg/cm3 or 7%) between osteoporotic women and postmenopausal controls. In order to compare pQCT to dual X-ray absorptiometry (DXA) at the forearm and at the lumbar spine (L1–L4), 40 premenopausal healthy controls, 40 postmenopausal healthy controls, and 35 postmenopausal osteoporotic women were assessed. DXA measurements performed at the ultradistal, middistal, 1/3, and total ROI of the radius showed only moderate correlations between r = 0.38–0.75, r = 0.27–0.64, and r = 0.38–0.53 for the comparison versus pQCT total BMD, versus pQCT trabecular BMD, and versus pQCT cortical BMD, respectively. Correlations of DXA at the lumbar spine and pQCT were between r = 0.18 and 0.44. DXA at radius and spine was able to discriminate between postmenopausal controls and osteoporotic women (p = 0.001–0.004), but BMD measurements by pQCT did not show this ability (p = 0.15–0.52). However, two nonstandard pQCT parameters, namely the surface area of the cortical bone and the cortical BMC were factors that discriminated well between these two groups (p = 0.002, p = 0.005, respectively). These pQCT parameters also yielded the highest relative annual changes in pre- and postmenopausal control subjects. The measurement of cortical bone in the distal radius proved to be a good predictor of vertebral fracture status and was a good indicator of age-related skeletal change. Our data emphasize the importance of cortical measurements when using pQCT of the radius to assess osteoporosis.
Bone density determinants in elderly women: A twin study
Authors: Dr. Flicker Leon , Hopper John L. , Rodgers Lisa , Kaymakci Bahtiyar , Green Robyn M. , Wark John D.
Published: February 2010

This cross-sectional twin study examined the influence of constitutional, lifestyle, and genetic factors on bone mineral density (BMD) in elderly women. BMD, at the lumbar spine, femoral neck, Ward's triangle, total hip, and total forearm, total body bone mineral content (BMC), and lean mass and fat mass were measured using dual energy X-ray absorptiometry in 69 volunteer female twin pairs (37 monozygotic [MZ], 32 dizygotic [DZ]) aged 60–89 years. Height and weight were measured. Medical history and lifetime tobacco and alcohol use were determined by questionnaire. In terms of within-pair differences, lean mass was independently associated with BMD at all sites. In contrast, fat mass was not associated with BMD at any site once allowance had been made for lean mass. Lifetime tobacco use was independently associated with BMD at the lumbar spine, total hip, and forearm. Total body BMC was independently predicted by lean mass, fat mass, tobacco use, and alcohol consumption. Age and the above independently predictive body composition and lifestyle factors accounted for 20–33% of variation in BMD. After allowing for these covariates, MZ and DZ correlations were consistent with about 75% of residual variation in BMD at the nonforearm sites being determined by genetic factors. For total body BMC, the covariates explained 75% of total variation, and genetic factors 76% of the residual variation. Therefore, at the proximal femur and lumbar spine, after taking into account the relation of BMD with lean mass and smoking, genetic factors appear to play a substantial role in explaining variation in BMD in elderly women.
Enhanced expression of type I receptors for bone morphogenetic proteins during bone formation
Authors: Ishidou Yasuhiro , Kitajima Isao , Obama Hiroya , Maruyama Ikuro , Murata Fusayoshi , Imamura Takeshi , Yamada Naoshi , Ten Duke Peter , Miyazono Kohei , Sakou Takashi
Published: February 2010

Type I receptors for bone morphogenetic proteins (BMPs), i.e., BMPR-IA and BMPR-IB, are transmembrane serine/threonine kinases, that bind osteogenic protein-1 (OP-1, also termed BMP-7) and BMP-4. Using antibodies specific to BMPR-IA and -IB, we have studied the expression of BMP type I receptors in the bone formation process during embryonic development and fracture healing. In the mouse embryo, both BMPR-IA and -IB were expressed in condensing mesenchymal cells at 13.5 days post coitum (p.c.). At 15.5 days p.c., expression of BMPR-IB, but not of BMPR-IA, was observed in the cells in perichondrium of developing cartilage. At 17.5 and 19.5 days p.c., expression of both receptors was observed in chondrocytes and in osteoblasts. In normal rat adult bone, expression of BMPR-IA, but not of BMPR-IB, was observed in osteoblasts in the periosteum. Three days after the femoral fracture, expression of BMPR-IA and -IB was up-regulated in cells at the proliferating osteogenic layer of the periosteum. On day 7, both receptors were found in fibroblast-like spindle cells and chondrocytes in the endochondral ossification sites, and osteoblasts in the newly formed trabecular bone. Expression of BMPR-IA was higher than that of BMPR-IB in osteogenic layer on day 3 and in osteoblasts in the trabecular bone on day 7. On day 14, expression of BMP type I receptors was observed at similar sites, albeit with lower expression levels than were observed on day 7. The present data suggest that expression of BMP type I receptors is up-regulated during bone formation, and that they may play important roles in bone morphogenesis.
Human osteoclasts, not osteoblasts, deposit osteopontin onto resorption surfaces: An in vitro and ex vivo study of remodeling bone
Authors: Dr. Dodds Robert A. , Connor Janice R. , James Ian E. , Lee Rykaczewski Elizabeth , Appelbaum Edward , Dul Edward , Gowen Maxine
Published: February 2010

Osteopontin is a phosphorylated glycoprotein believed to be secreted by osteoblasts and deposited into the bone matrix to facilitate osteoclast adhesion or to initiate osteoid mineralization. Previously we have presented contradictory evidence that osteoclasts express osteopontin mRNA in human remodeling bone. The aim of this study was to ascertain whether osteoclasts synthesize and deposit osteopontin in resorption lacunae. We characterized expression of osteopontin mRNA and protein expression in both intramembranous and endochondral ossification, as well as remodeling bone, in the human osteophyte. Osteopontin mRNA was expressed in osteoclasts with tartrate-resistant acid phosphatase (TRAP) positivity within resorption lacunae. The osteoclasts and immediate resorption surfaces also expressed osteopontin. However, osteopontin mRNA and protein were weak (transient) or undetectable in osteoblasts at adjacent bone formation sites; no osteopontin expression was observed in the osteoid, although occasional reactivity was observed in osteocytes and the mineral-osteoid interface. In contrast, osteopontin was highly expressed in the osteoblasts and matrix of woven bone during intramembranous and endochondral ossification. The matrix expression correlated with mineralization; however, in some instances osteopontin deposition was observed prior to mineralization. Similarly, osteopontin expression was evident in cartilage matrix, solely at foci of mineralization. Chondroclasts expressed osteopontin mRNA and protein; the surfaces of resorbed calcified cartilage also expressed osteopontin. Abnormal, unmineralized matrices apparently lacked deposited osteopontin, but were nevertheless resorbed by osteoclasts; the osteoclasts and resorbed surfaces expressed no osteopontin protein. That osteoclasts are responsible for the deposition of osteopontin was confirmed in vitro, whereby resorption pits in whale dentine and bovine bone slices, produced by isolated human osteoclasts, contained deposited osteopontin. Osteopontin may facilitate the adhesion (or detachment) of the osteoclast to the bone surface. Alternatively, the possibility that osteopontin may act as a postresorptive signal to recruit osteoblasts, or to polarize and direct the mineralization of the formed osteoid, is discussed.
Cell proliferation of human fibroblasts and osteoblasts in osteogenesis imperfecta: Influence of age
Authors: Fedarko N.S. , D'Avis P. , Frazier C.R. , Burrill M.J. , Fergusson V. , Tayback M. , Sponseller P.D. , Shapiro J.R.
Published: February 2010

Clinical studies indicate that as a group, osteogenesis imperfecta (OI) subjects are shorter than age- and sex-matched controls. Not only somatic growth, but also cellular growth appears to be impaired, and these may be related to defects in extracellular matrix common to this disorder. We have investigated the growth characteristics of dermal fibroblasts and trabecular osteoblasts isolated from patients with OI and control subjects of various ages. Cell growth curves and cell doubling times were determined by measuring cell number using crystal violet dye binding. Growth curves were modeled by a modified logistic function, the three parameters of which are markers for biologically relevant growth parameters: the plateau value or upper asymptote, which reflects the maximum cell density upon confluence; the maximal growth rate (μm); and the lag time. Both normal human fibroblasts and osteoblasts showed an age-dependent decrease in μm. Normal fibroblasts exhibited no age-dependence to their upper asymptote or lag time. Fibroblasts derived from patients with OI did not have significantly different upper asymptote values, μm, or lag times when compared with normal fibroblasts. Normal osteoblasts had a decrease in upper asymptote, decrease in μm, but a relatively constant lag time with increasing age. In contrast, OI osteoblast μm was decreased relative to that of normal subjects. For osteoblasts from OI patients, decreased μm appeared unrelated to the age of the subject, whereas OI fibroblasts did exhibit an age-dependent decrease in μm. The percentage of collagenase-digestible protein (a measure of collagen synthesis) produced by normal human fibroblasts correlated well with μm. Treating normal human osteoblasts with the proline analogue 3,4-dehydroproline, which destabilizes collagen triple helix formation and alters collagen synthesis, secretion, and turnover, also decreased μm. A dose response to varying concentrations of 3,4-dehydroproline was observed for normal human bone cell μm. These data suggest a link between type I collagen synthesis and cellular proliferation.
Intact parathyroid hormone levels are not elevated in glucocorticoid-treated subjects
Authors: Paz-Pacheco Elizabeth , El-Hajj Fuleihan Ghada , Leboff Meryl S.
Published: February 2010

To assess whether chronic glucocorticoid therapy results in a compensatory increase in parathyroid hormone (PTH), we measured intact PTH levels and other indices of mineral metabolism in 13 postmenopausal glucocorticoid-treated women and 16 normal age-matched controls. The glucocorticoid-treated women received a mean prednisone dose of 15.8 ± 3.1 mg/day for 12.9 ± 3.1 years. A linear regression analysis between intact PTH levels and a wide range of prednisone doses in these 13 glucocorticoid-treated women and 26 additional male and female subjects receiving chronic glucocorticoid therapy for a variety of rheumatic and pulmonary disorders (n = 39) was also performed. Intact PTH levels using the sensitive immunoradiometric assay (IRMA, Nichols Institute, San Juan Capistrano, CA) were comparable in the glucocorticoid-treated and normal control women (35.3 ± 4.4 vs 31.3 ± 3.2 ng/l, respectively) as were the total calcium concentrations (9.67 ± 0.12 vs 9.52 ± 0.11 mg/dl). In the glucocorticoid-treated women, the 25-hydroxyvitamin D levels, measured by competitive protein assay were similar to those of the control subjects (29.2 ± 2.8 vs 29.1 ± 2.3 ng/ml), and no patient was treated with vitamin D in excess of 400 IU daily. In the combined 39 male and female patients, there were also no significant regression relationships between daily prednisone dose and intact PTH levels. Thus, secondary hyperparathyroidism does not accompany chronic oral glucocorticoid therapy in women on low to moderate doses of oral glucocorticoids. The lack of an elevation in intact PTH levels in the presence of chronic glucocorticoid therapy may represent an increased sensitivity of bone to PTH, or an alteration in the relationship between calcium and PTH, or both.
Inhibition of antigen-presenting cell function by alendronate in vitro
Authors: Sansoni Paolo , Passeri Giovanni , Fagnoni Francesco , Mohagheghpour Nahid , Snelli Gianluca , Brianti Vincenzo , Dr. Engleman Edgar G.
Published: February 2010

Bisphosphonates are potent inhibitors of bone resorption in vivo and are emerging as important and widely used drugs for the treatment of a variety of abnormal bone resorptive processes. In the current study we investigated the in vitro effects of 4-amino-1-hydroxybutylidene-1,1-bisphosphonate (alendronate), a recently developed, extremely potent bisphosphonate, on the immune functions of human peripheral blood mononuclear cells (PBMCs). PBMC proliferation induced by lectins, alloantigens, and a nominal antigen (tetanus toxoid) was inhibited in a dose-dependent manner by alendronate. Pretreatment of monocytes, but not T cells, with the compound at concentrations ranging from 10−4 to 10−8 M was inhibitory, indicating that alendronate acts selectively on antigen-presenting cells (APCs). Alendronate did not affect the viability of monocytes or T cells or the expression of cell surface molecules known to play critical roles in antigen presentation. Alendronate exhibited dose-dependent inhibition of the production of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) by activated monocytes. The inhibitory effect of 10−6 M alendronate on PBMC proliferation was reversed by 10 U/ml recombinant rIL-1β, whereas other cytokines such as IL-6, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) had no effect. Thus, alendronate acts on monocytes to inhibit their antigen-presenting/accessory cell functions through a mechanism that can be overcome by exogenous IL-1. The inhibitory effect of this agent on cytokine production may contribute to its inhibitory effect on bone resorption.
Effects of physical training on bone adaptation in three zones of the rat tibia
Authors: Dr. Bourrin Sandrine , Palle Sabine , Pupier René , Vico Laurence , Alexandre Christian
Published: February 2010

This study has been conducted to examine the effects of physical exercise on the bone trabecular network and the cellular adaptations in three different areas of a single bone, the tibia. Male Wistar rats (9 weeks old) were treadmill-trained for 0, 3, 4, or 5 weeks at 60% of their measured maximal O2 consumption (VO2max). Histomorphometric analysis of the proximal tibia of running and age-matched control groups was performed in the epiphyseal trabecular bone, in the primary spongiosa and in the secondary spongiosa. Dynamic and static bone cell activities and serum calcium and phosphorus levels were measured. VO2max increased significantly by 18.4% after 5 weeks of training. In the epiphysis, a 9% increase in bone volume, associated with more numerous trabeculae (8%) was detected the third week of training. In primary spongiosa a significant increase (6.7%) in newly formed trabeculae was found. In secondary spongiosa bone volume increased significantly by 26.2% the fifth week of exercise and was associated with thicker trabeculae. The number of osteoclast profiles was significantly depressed. Osteoid surfaces and bone formation rate increased significantly in weeks 3 to 5. Serum calcium levels were found to be significantly decreased in weeks 3 and 4. There was no change in osteoid thickness or mineral apposition rate. These results suggest 1) a rapid increase in osteoblastic recruitment without change of the cell activity in response to moderate exercise; 2) a decreased bone resorption associated with a marked increase in bone formation from the third week of training; 3) adaptation of the trabecular network to exercise that seems to be bone-site-dependent, suggesting a cell sensitivity to training-engendered strain distribution within the bone or to strain-related local factors.
Markers of bone remodeling in the elderly subject: Effects of vitamin D insufficiency and its correction
Authors: Brazier M. , Kamel S. , Maamer M. , Agbomson F. , Elesper I. , Garabedian M. , Desmet G. , Sebert J.L.
Published: February 2010

The elderly subject is prone to both vitamin D insufficiency and calcium insufficiency due to a low calcium intake and calcium malabsorption. These two alterations may lead to secondary hyperparathyroidism, and thus to increased bone loss. We investigated 72 elderly subjects (16 men and 56 women) with vitamin D insufficiency and 25 healthy elderly women with normal vitamin D status, with respect to their indices of calcium metabolism and of bone remodeling: serum total alkaline phosphatases (AP), bone AP (BAP), osteocalcin (BGP), tartrate-resistant acid phosphatase (TRAP), urine hydroxyproline (HYP), and the 3-OH-pyridinium derivatives pyridinoline (PYD) and deoxypyridinoline (DPD), which are new markers of bone resorption. We then studied the modifications of these markers in the patients with vitamin D insufficiency at 3 months and 6 months after onset of a daily vitamin D and calcium supplementation. When compared with elderly subjects with normal vitamin D status, patients with vitamin D insufficiency had increased intact parathyroid hormone (iPTH) levels (60.1 ± 10.2 vs 30.2 ± 4.5, p < 0.001) and a high bone turnover as reflected by increased values of most serum and urine markers of bone remodeling. PYD and DPD levels were significantly correlated with all indices of bone turnover, unlike HYP, which showed no correlation with bone formation markers (AP, BAP, and BGP). A daily supplement of 800 IU vitamin D3 and 1 g of elemental calcium increased 25(OH)D levels and induced a dramatic decrease of iPTH levels; at 3 and 6 months, the mean iPTH level decreased by 50% (p < 0.0001), reaching the mean value of healthy vitamin D sufficient elderly women. All markers of bone turnover, except TRAP, decreased significantly at 3 and 6 months. The PYD/DPD ratio increased significantly at 3 and 6 months. The decrease of bone markers was more marked in patients with more severe hyperparathyroidism, the greatest variations being obtained with BAP (45%, p = 0.006) and DPD (43%, p = 0.036) levels. Most markers of bone remodeling are increased in elderly subjects with vitamin D insufficiency and vary with its correction. However, BAP and DPD are the most sensitive indicators of increased bone turnover due to secondary hyperparathyroidism.
Determinants of bone mineral density in older men
Authors: Glynn Warhaftig Nancy , Meilahn Elaine N. , Charron Martin , Anderson Stewart J. , Kuller Lewis H. , Dr. Cauley Jane A.
Published: February 2010

Osteoporosis is a significant health problem and contributor to disability and premature mortality among older men. Incidence rates for hip fracture have stabilized in women, but continue to increase in men. A major risk factor for hip fracture is bone mineral density level. The determinants of bone mineral density (BMD) are well defined in women, but not in men. The primary goal of the current research was to increase our understanding of the determinants of BMD of the proximal femur in a large community-based sample of older male volunteers. Eligibility requirements included age of 50 years or older, ambulatory, and not having undergone a bilateral hip replacement. Five hundred twenty-three men, mean age 66.6 years, met all eligibility requirements and participated in the Study of Osteoporotic Risk in Men or STORM. Information on demographics, medical history, anthropometry, leisure-time and occupational physical activity, muscular strength, cigarette smoking, alcohol consumption, dietary calcium intake, and medication use (thiazide diuretics and glucocorticoids) were obtained by questionnaire, interview, and examination. BMD of the proximal femur (femoral neck, greater trochanter, and Ward's triangle) was measured by dual-energy X-ray absorptiometry using the Hologic QDR-1000 and QDR-2000. The cross-sectional determinants of BMD included age, blond hair color, current body weight, thiazide diuretic use, historical physical activity, and quadriceps strength. Several variables commonly thought to be determinants of BMD were not related to BMD in this population of older men, including current cigarette smoking, alcohol consumption, current leisure-time physical activity, dietary calcium intake, vitamin D use, and caffeine intake. However, failure to find associations among BMD and some of the potential determinants may be due to lack of statistical power. Identification of the determinants of BMD could lead to the development of interventions aimed at maximizing BMD in men and could potentially decrease the risk of hip fractures.