bluuebunny/crossref_metadata_2025_split huggingface dataset:
Elevated progesterone during pseudopregnancy may prevent bone loss associated with low estrogen
Authors: Bowman Beth M. , Miller Scott C. 
Published: February 2010

Abstract
There is increasing evidence for a role of endogenous progesterone in mineral and skeletal metabolism. The purpose of this study was to compare skeletal changes that occur during a condition of high endogenous progesterone but low estrogen (pseudopregnancy) with a condition of low endogenous progesterone and low estrogen (ovariectomy). Pseudopregnancy was selected over pregnancy to eliminate placental factors that may influence mineral metabolism. Rats were ovariectomized (OVX) or pseudopregnancy initiated, and bones were collected 13 days later. In some animals, blood was collected by indwelling catheters for determination of progesterone and estrogen levels. At mid-pseudopregnancy, there were substantial elevations in progesterone but estrogen was below the level of detection. Progesterone and estrogen were below the level of detection in the OVX rats. Longitudinal growth rates were increased compared with the normal cycling rats in both the pseudopregnant and OVX groups, indicative of decreased estrogen levels, but they were greatest in the OVX rats. Cancellous bone mass was maintained in the pseudopregnant rats compared with normal cycling rats but significantly reduced in the OVX animals. As expected, increased bone formation and turnover rates were observed in the OVX animals although some indices of bone formation were also increased in the pseudopregnant animals. Osteoclasts were significantly increased in the OVX but not the pseudopregnant animals compared with normal cycling rats. Increased periosteal bone formation indices are known to occur following OVX, but the greatest periosteal formation rates were observed in the pseudopregnant animals. While possible roles for some other endocrine agents cannot be excluded at this time, the data from the present study suggest that endogenous progesterone may have a role in the maintenance of bone mass perhaps by decreasing bone resorption while maintaining or increasing bone formation during physiological periods of low estrogen such as occurs during early pregnancy.
Ontogeny of the 1,25-dihydroxyvitamin D3 receptor in fetal rat bone
Authors: Johnson Julie A. , Grande Joseph P. , Roche Patrick C. , Dr. Kumar Rajiv 
Published: February 2010

Abstract
To gain insights into 1,25-dihydroxyvitamin D3 receptor (VDR) function during fetal bone development, we examined fetal rat tissues from gestational days 13–21 for the presence and distribution of VDR using immuno-histochemistry. Prior to ossification, VDR epitopes were observed in the mesenchyme condensing to form skeletal tissues, on day 13 in the developing vertebral column and limbs, and on day 17 of gestation in developing calvaria. Immunostaining for VDR was seen in proliferating and hypertrophic chondrocytes and in osteoblasts of limb buds and the vertebral column by day 17 of gestation. In calvaria, VDR epitopes were observed in osteoblasts by gestational day 19. VDR immunostaining was also evident in the skin of fetal limbs at all gestational ages examined. We show for the first time that the VDR appears very early in the developing fetal rat skeleton, suggesting that the VDR, in concert with its ligand, 1,25-dihydroxyvitamin D3, may play a role in the differentiation of mesenchymal precursors into bone tissue.
Expression of amelogenin mRNA sequences during development of rat molars
Authors: Wurtz Tilmann , Lundmark Carin , Christersson Cecilia , Bawden James W. , Slaby Ivan , Hammarström Lars 
Published: February 2010

Abstract
The expression of amelogenin mRNA in growing rat molars was studied. Northern blotting and the analysis of cDNA isolates revealed two predominant variants. One group of cDNA inserts contained sequences of a long mRNA version and the other group contained mRNA sequences of the shorter leucin-rich amelogenin polypeptide (LRAP). The LRAP group was deficient in an internal stretch which coded for a peptide with a high potential for beta turns. Northern blot experiments showed that most amelogenin RNA in rat teeth was represented by two bands of 1.1 and 0.8 kb. Two oligonucleotide probes were designed that were specific for the long version and for the LRAP variant. The probes were used for in situ hybridization experiments on sections of developing maxillar teeth of rats between day 2 and day 15 after birth. Both RNA species were accumulated concomitantly and exclusively in cells of the inner enamel epithelium. Expression was first observed at the mesial cusp sides and finally involved the whole ameloblast layer except for the cells adjacent to the enamel-free region at the tip of the cusps. The early amelogenin RNA expression occurred adjacent to the initial deposition of the dentin matrix. Low amounts of amelogenin RNA persisted after the differentiation of ameloblasts into the maturative stage. The sequence of events was similar in all three molars.
Expression of the nucleoside triphosphate pyrophosphohydrolase PC-1 is induced by basic fibroblast growth factor (bFGF) and modulated by activation of the protein kinase A and C pathways in osteoblast-like osteosarcoma cells
Authors: Solan Joell L. , Deftos Leonard J. , Coding James W. , Terkeltaub Robert A. 
Published: February 2010

Abstract
The closely related cytokines bFGF and aFGF regulate the function of bone cells and mineralization. Osteoblasts express PPi-generating nucleoside triphosphate pyrophosphohydrolase (NTPPPH)/nucleotide phosphodiesterase I activity. bFGF and aFGF (10 ng/ml) up-regulated NTPPPH in human SaOS-2 and U2OS osteosarcoma cells, which express osteoblast-like features in culture. The induction was selective as alkaline phosphatase activity was down-regulated and specific as insulin-like growth factor-1 (IGF-1) and interleukin-1β (IL-1β) were not active. Furthermore, IL-β but not IGF-1 inhibited bFGF-induced up-regulation of NTPPPH. The induced NTPPPH remained predominantly associated with cells. bFGF can induce signaling through pathways including protein kinase A (PKA) and protein kinase C (PKC)-mediated transduction. An activator of the PKA pathway (8-bromo cyclic adenosine monophosphate [cAMP]) induced NTPPPH. Furthermore, pretreatment with the PKC activator phorbol myristate acetate (PMA) (80 nM) markedly increased subsequent NTPPPH induction by both bFGF and cAMP. The PMA effect was associated with morphologic changes characterized by long, thin intercellular extensions. PKC desensitization also potentially contributed to this effect because the PKC inhibitors staurospo-rine and H-7 enhanced bFGF-induced and cAMP-induced NTPPPH expression in the absence of morphologic changes. We observed that bFGF induced expression of PC-1, a member of the NTPPPH gene family. The majority of NTPPPH activity was depleted by immunoadsorption using a monoclonal antibody to native human PC-1. bFGF- and aFGF-induced production of PC-1/NTPPPH in osteoblastoid cells may contribute to the effects of FGFs on bone metabolism.
Quantification of the collagenolytic activity of isolated osteoclasts by enzyme-linked immunosorbent assay
Authors: Foged Niels Taekker , Delaisse Jean-Marie , Hou Peng , Lou Henriette , Sato Takuya , Winding Bent , Bonde Martin 
Published: February 2010

Abstract
Difficulties in the geometrical definition and measurement of resorption pits is a major problem for the quantitative analysis of bone resorption by isolated osteoclasts cultured on bone or dentin substrates. In this study we developed an enzyme-linked immunosorbent assay (ELISA) for quantification of bone resorption in vitro, which specifically quantifies type I collagen fragments released into the culture medium by the resorptive action of bone cells cultured on slices of bone or dentin. A consistently high correlation between the formation of resorption pits and the release of antigenic collagen fragments was observed for isolated rabbit osteoclasts seeded at various densities and cultured for various periods on bovine, elephant, and human substrates. In a further support of the osteoclastic nature of the collagenolytic effects, a high consistency between pit formation and collagenolysis was also observed when the rabbit bone cells were cultured in the presence of very differently acting but typical inhibitors of pit formation, i.e., the carbonic anhydrase inhibitor acetazolamide, the cysteine proteinase inhibitor epoxysuccinyl-L-leucylamido-(4-guanodino)butane (E-64), the phosphatidyl-inositol 3-kinase inhibitor wortmannin, and the bisphosphonate ibandronate (BM 21.0955). In conclusion, the ELISA represents a simple, precise, and objective way to dynamically monitor bone resorption in vitro through quantification of the collagenolytic activity of isolated osteoclasts.
Prediction of perimenopausal fractures by bone mineral density and other risk factors
Authors: Torgerson D.J. , Campbell M.K. , Thomas R.E. , Reid D.M. 
Published: February 2010

Abstract
In a prospective population-based cohort study, we assessed whether bone mineral density (BMD) measurements of perimenopausal women and other risk factors for osteoporosis are predictive of subsequent fracture. Women aged 47-51 years chosen randomly from a population register who underwent a bone density measurement 2 years previously were followed up by questionnaire to assess the 2-year incidence of any self-reported fractures. We found that 44 women, out of 1857 who completed the questionnaire, sustained at least one fracture within a 2-year follow-up period. After adjustment for covariates, the odds ratio of sustaining a fracture was 1.6 (95% confidence interval [CI] 1.16-2.34) for every standard deviation reduction in BMD at the spine, for women with a prior history of fracture the odds ratio of a subsequent fracture was approximately 2 (95% CI 1.31–3.03), a family history of hip fracture (maternal grandmother) carried an odds ratio of 3.7 (95% CI 1.55-8.85), while being postmeno-pausal or having a hysterectomy resulted in an odds ratio of 1.98 (1.02-3.56). This study has shown that BMD measurements at the hip and spine and other risk factors predict any nonhip and nonspine perimenopausal fractures. Further follow-up is required to assess the predictive performance of BMD measurements and other risk factors for hip and spine fractures.
Nitric oxide and bone
Authors: Dr. Evans Deborah M. , Ralston Stuart H. 
Published: February 2010

AbstractNitric oxide (NO), a mediator of cardiovascular homeostasis, neurotransmission, and immune function, has recently been found to have important effects in bone. Both constitutive and inducible forms of NO synthase are expressed by bone-derived cells, and cytokines such as interleukin-1 (IL-1), tumor necrosis factor (TNF), and interferon gamma (IFN-γ), are potent stimulators of NO production. When combined with other cytokines, IFN-γ markedly induces NO production, which suppresses osteoclast formation and activity of mature osteoclasts. This “superinduction” of NO is largely responsible for the selective inhibitory effect of IFN-γ on cytokine-induced bone resorption. High concentrations of NO are also inhibitory for cells of the osteoblast lineage, and NO production appears to be partly responsible for the inhibitory effects of cytokines on osteoblast proliferation. At lower concentrations, however, NO has different effects. Moderate induction of NO potentiates bone resorption, and the constitutive production of NO at low concentrations promotes the proliferation of osteoblast-like cells and modulates osteoblast function. NO therefore appears to be an important regulatory molecule in bone with effects on cells of the osteoblast and osteoclast lineage and represents one of the molecules produced by osteoblasts which directly regulate osteoclastic activity. Stimulation of NO production in bone by proinflammatory cytokines raises the possibility that NO may be involved as a mediator of bone disease in conditions associated with cytokine activation, such as rheumatoid arthritis, tumor associated osteolysis, and postmenopausal osteoporosis.
Isolation and characterization of osteoblast precursor cells from human bone marrow
Authors: Rickard David J. , Kassem Moustapha , Hefferan Theresa E. , Sarkar Gobinda , Spelsberg Thomas C. , Riggs B. Lawrence 
Published: February 2010

Abstract
Osteoblasts are derived from precursor cells present in low frequency in the stromal element of bone marrow. Because of the lack of a practical procedure to isolate osteoblast precursors from early cultures of plastic adherent cells from bone marrow, previous studies of marrow stromal cells have been made in confluent cultures of bone marrow when the osteoblast (OB) precursors are already differentiated. Also these studies utilized cultures containing mixed populations of cells including hematopoietic cells. Thus we have employed a negative immunoselection procedure to remove contaminating hematopoietic cells and to isolate nearly homogeneous populations of early human stromal cells derived from the plastic-adherent mononuclear marrow cells cultured in the presence of serum. By reverse transcriptase polymerase chain reaction (RT-PCR) analysis for mRNA, and by immunocytochemical study for protein, we studied the sequential expression in culture of multiple markers of the osteoblast phenotype-alkaline phosphatase, osteopontin, parathyroid hormone receptor, types I and III procollagen, and osteocalcin-as well as lipoprotein lipase (LPL), a marker of the adipocyte phenotype. At an early stage of culture (7-9 days), human OB precursors formed colonies of variable sizes that expressed low levels of mRNA and protein concentrations of OB markers, and their concentration increased on growth to a confluent monolayer (approximately 14 days). LPL mRNA was expressed at high levels in the colony stage, and its level decreased upon confluency, suggesting a loss of potential for commitment to the adipocyte lineage. Interestingly, treatment with dexamethasone at 10−8 M increased the expression for some of the osteoblast markers and for the LPL gene and was required for the deposition of mineralized matrix and for the formation of adipocytes containing cytoplasmic lipid droplets in confluent cultures. Cloned single early colonies were able to coexpress the osteoblast and adipocyte markers (as assessed by RT-PCR). Thus these immunoselected marrow stromal cells have the characteristics of authentic human osteoblast precursor cells which also are capable of differentiating into adipocytes.
Effects of a synthetic vitamin D analog, ED-71, on bone dynamics and strength in cancellous and cortical bone in prednisolone-treated rats
Authors: Tanaka Yuzo , Dr. Nakamura Toshitaka , Nishida Satoshi , Suzuki Katsumi , Takeda Satoshi , Sato Katsuhiko , Nishii Yasuho 
Published: February 2010

Abstract
To determine the action of corticosteroid on bone metabolism and assess the effects of a synthetic vitamin D analog, ED-71, on them, 56 SD rats, 8 weeks of age, were assigned to seven groups of eight animals each. Group 1 was the basal control. Group 2 was the nontreated control. Groups 3–7 were given prednisolone at 30 mg/kg of body weight (BW) twice a week and concomitantly administered ED-71 with respective doses of 0, 0.0125, 0.025, 0.05, and 0.1 μg/kg of BW for 12 weeks. In group 3, urinary calcium (U-Ca) and deoxypyridinoline (U-Dpy) were significantly increased compared with group 2. In groups 4–7, U-Ca values were increased but U-Dpy values were dose-dependently decreased. Age-dependent increases in the parameter values of BMD, compressive strength, trabecular bone volume (BV/TV), and trabecular thickness (Tb.Th) of the lumbar body were significantly suppressed in group 3 but dose-dependently increased in groups 4–7, and the values of group 7 exceeded those of group 2. The parameters of bone mineral density (BMD) and the bending strength of the femur in groups 4–7 were larger than the values in group 3 but did not reach the levels of group 2. The trabecular bone formation rate (BFR/BS) of the lumbar body measured by calcein labeling in group 3 was reduced when compared with group 2, but the values were not further decreased in groups 4–7. The perimeter ratios of double labels over single labels (dLS/sLS) greatly decreased by prednisolone, were dose-dependently increased to the level of the normal control by ED-71. Double-labeled perimeters and the dLS/sLS ratios were also increased in the periosteal envelope of the midfemur. These findings clearly demonstrate that prednisolone administration affects the age-related changes in bone metabolism, and ED-71 administration counteracts the effects by increasing intestinal calcium absorption, reducing bone resorption, and enhancing mineralization. The action of ED-71, however, seems to be less potent in the cortical bone.
TGF-β and basement membrane matrigel stimulate the chondrogenic phenotype in osteoblastic cells derived from fetal rat calvaria
Authors: Basic Nikolina , Basic Vanja , Bulic Kresimir , Grgic Marko , Kleinman Hynda K. , Luyten Frank P. , Vukicevic Slobodan 
Published: February 2010

Abstract
Primary cultures of fetal rat calvarial cells contain a spectrum of osteogenic phenotypes including undifferentiated mesenchymal cells, osteoprogenitor cells, and osteoblasts. We recently demonstrated that rat calvarial osteoblast-like cells grown on basement membrane undergo profound morphological changes resembling a canalicular network in bone. In the present study, we examined the effect of reconstituted basement membrane Matrigel on chondroblastic versus osteoblastic differentiation of different cell subpopulations obtained by five consecutive enzymatic digestions of rat calvarial cell populations. We found that the appearance of canalicular cell processes decreased with the later digests. When cells from the fourth and fifth digest were grown on top of Matrigel for 7 days, the majority of the cell aggregates displayed chondrocytic characteristics but none of the cells became hypertrophic. When individual chondroblastic cell aggregates were subsequently transferred from Matrigel to plastic, they started expressing types I and X collagens, alkaline phosphatase, and osteocalcin. Within the next 7 days (days 8–14 of the experiment), the majority of cells increased in size, and at day 17 on plastic (day 24 of the experiment) mineralized bone nodules formed. The chondroblastic differentiation of calvarial cells grown on Matrigel could be inhibited by a specific transforming growth factor-β1 (TGF-β1) but not by a TGF-β2 antibody. Addition of recombinant TGF-β1 to similar cultures promoted the appearance of chondroblastic cell aggregates. The cartilage phenotype could not, on the contrary, be promoted by growing the cells on other extracellular matrices such as a collagen I gel. We suggest that TGF-β1 in concert with the basement membrane extracellular matrix induces chondroblastic differentiation of rat calvarial osteoprogenitor cells.
Involvement of prostaglandin endoperoxide H synthase-2 in osteoclast-like cell formation induced by interleukin-1β
Authors: Sato Takahiro , Dr. Morita Ikuo , Sakaguchi Kouji , Nakahama Ken-Ichi , Smith William L. , Dewitt David L. , Murota Sei-Itsu 
Published: February 2010

Abstract
Interleukin-1β (IL-1β) stimulates osteoclast-like cell formation via prostaglandin E2 (PGE2) production. However, the regulatory mechanism for the production of PGE2 in bone cells is still unclear. Recently, it has been shown that two prostaglandin endoperoxide H synthase (PGHS) isozymes exist, termed PGSH-1 and PGHS-2. We report here that IL-1β induces PGE2 production in bone marrow culture induced by a PGHS-2-dependent mechanism. IL-1β stimulated the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNC) and the production of PGE2 in mouse bone marrow cultures. The dose response curves for the indomethacin inhibition of TRAP-positive MNC formation and PGE2 production were nearly identical. Cycloheximide (CHX) suppressed IL-1β-induced PGE2 production, suggesting that the production of PGE2 induced by IL-1β required de novo protein synthesis. Northern blot analysis determined that IL-1β induced PGHS-2 expression by 30 minutes and mRNA levels were maximal by 1–2 h. Cycloheximide potentiated the accumulation of PGHS-2 mRNA linearly up to 8 h. Dexamethasone, an inhibitor of the induction of PGHS-2, inhibited IL-1β-induced PGHS-2 mRNA expression and also suppressed IL-1β-stimulated formation of TRAP-positive MNC. Furthermore NS-398, as a selective PGHS-2 inhibitor, completely inhibited IL-1β-induced TRAP-positive MNC formation. Moreover, IL-1β-induced PGHS-2 mRNA expression and formation of TRAP-positive MNC were inhibited by calphostin C, a selective inhibitor of protein kinase C (PKC). These results indicate that IL-lβ-induced formation of osteoclast-like cells requires PKC activation, induction of PGHS-2, and subsequent prostaglandin synthesis by this enzyme.
Effect of physical activity as a caddie on ultrasound measurements of the os calcis: A cross-sectional comparison
Authors: Dr. Hoshino Hironobu , Kushida Kazuhiro , Yamazaki Kaoru , Takahashi Masaaki , Ogihara Hiroaki , Naitoh Kenichi , Toyoyama Okimitsu , Doi Syun , Tamai Hideki , Inoue Tetsuo 
Published: February 2010

Abstract
This cross-sectional study investigated the effect of long-term activity as a caddie on ultrasonic properties of the os calcis. We measured 74 healthy women, age 20–59 years, who worked at a golf course as caddies. An age-matched control group of 433 healthy women, who were office workers or housewives, also were recruited for comparison. The ultrasound measurements were performed with an Achilles ultrasound densitometer. The quadriceps muscle strength and the hand grip strength were measured in a perimenopausal subgroup (45-59 years) of the caddies and a subgroup of controls matched for age, height, weight, and body mass index. Urinary pyridinoline and deoxypyridinoline were also measured in these perimenopausal subgroups. Caddies had significantly higher ultrasound values than controls in the 40–49 (stiffness index, 101.6 ± 12.9% versus 87.9 ± 11.9%; p &lt; 0.0001) and 50–59 (stiffness index, 90.5 ± 11.6% versus 77.2 ± 11.6%; p &lt; 0.0001) age-stratified groups. Quadriceps strength and grip strength were significantly higher in caddies than those in controls. In postmenopausal caddies, all ultrasound values were significantly higher than for controls. In caddies there were not significant decreases of any ultrasound values with postmenopausal age. Even for the subgroup within 3 years of menopause there were significant differences between caddies and controls (p &lt; 0.01). There were no significant increases of pyridinoline and deoxypyridinoline after menopause in the caddies. We demonstrated that the caddies had higher ultrasound properties of the os calcis and lower bone resorption after menopause compared with controls.