bluuebunny/crossref_metadata_2025_split huggingface dataset:
The expression of cytokine activity by fracture callus
Authors: Einhorn Thomas A. , Majeska Robert J. , Rush Elisa B. , Levine Pamela M. , Horowitz Mark C. 
Published: February 2010

Abstract
Cytokines, a group of proteins known to regulate hemopoietic and immune functions, are also involved in inflammation, angiogenesis, and bone and cartilage metabolism. Since all of these processes occur following bone injury, or are known to contribute to wound repair mechanisms, this investigation sought to test the hypothesis that cytokines are involved in fracture healing. Two sets of 60 male Sprague-Dawley rats underwent the production of standard closed femoral fractures. The animals were then euthanized in groups of 15 on days 3, 7, 14, and 21 postfracture. A separate control group was also used for the harvesting of intact unfractured bone. At the time of euthanasia, calluses or bone specimens were explanted to organ culture and treated with either media alone or media containing the inducing agents lipopolysaccharide or concanavalin A. A titration of conditioned medium from these cultures was then added to factor-dependent clonal cell lines that are known to be specifically responsive to interleukin-1, interleukin-6, granulocyte-macrophage colony stimulating factor or macrophage-colony stimulating factor. To confirm the identities of each of these cytokines, neutralizing antibody studies were performed. The results showed that interleukin-1 is expressed at very low constitutive levels throughout the period of fracture healing but can be induced to high activities in the early inflammatory phase (day 3). Granulocyte-macrophage colony stimulating factor showed no constitutive activity but could also be induced to high activities with lipopolysaccharide. The ability of these two cytokines to be induced declined progressively as fracture healing proceeded. Interleukin-6 showed high constitutive activity early in the healing process (day 3), and treatment with inducing agent did not increase the activity of this cytokine at this timepoint. Lipopolysaccharide did increase interleukin-6 activity in day 7 and 14 fracture calluses. Although macrophage-colony stimulating factor is thought to be involved in a variety of metabolic bone conditions, it could not be detected or induced from any of the callus samples. Moreover, none of the samples of unfractured bone showed constitutive or inducible activities for any of these cytokines. A separate experiment in which calluses and samples of unfractured bone from similar cultures were examined histologically and tested for DNA or protein synthesis at two timepoints in the culture period (days 1 and 4) showed that tissue viability was maintained. Thus the inability to detect macrophage colony-stimulating factor in fracture callus or any cytokine activity in unfractured bones was not due to cell death. These findings suggest that the occurrence of a fracture results in a cascade of events that includes the production of cytokines. The time-dependent expression of specific cytokine activities, which are differentially regulated, supports the concept that fracture healing may be partially mediated by these factors.
Additive effects of weight-bearing exercise and estrogen on bone mineral density in older women
Authors: Kohrt Wendy M. , Snead David B. , Slatopolsky Eduardo , Birge Stanley J. 
Published: February 2010

Abstract
The separate and combined effects of weight-bearing exercise and hormone replacement therapy (HRT) on bone mineral density (BMD) were studied in 32 women, 60 to 72 years of age. HRT consisted of continuous conjugated estrogens 0.625 mg/day and trimonthly medroxyprogesterone acetate 5 mg/day for 13 days. Exercise consisted of 2 months of low-intensity exercise followed by 9 months of more vigorous weight-bearing exercise ˜45 minutes/day, ≥3 days/week, at 65–85% of maximal heart rate. Lumbar spine and proximal femur BMD were significantly increased in response to exercise and to HRT, and total body BMD was significantly increased in response to HRT; neither exercise nor HRT had an effect on wrist BMD. The combination of exercise + HRT resulted in increased BMD at all sites except the wrist, with effects being additive for the lumbar spine and Ward's triangle and synergistic for the total body. Based on reductions in serum osteocalcin levels, it appears that increases in BMD in response to HRT and exercise + HRT were due to decreased bone turnover. The lack of change in serum osteocalcin and IGF-I in response to exercise alone suggests that increases in BMD were due to decreased bone resorption and not increased formation. Results indicate that weight-bearing exercise + HRT may be effective in preventing and/or treating osteoporosis. It is likely that the additive effects of weight-bearing exercise and HRT on bone mineral accretion, coupled with other adaptations to the exercise (i.e., increased strength and functional capacity), could effectively reduce the incidence of falls and osteoporotic fractures.
Treatment with growth hormone and IGF-I in growing rats increases bone mineral content but not bone mineral density
Authors: Rosen Harold N. , Chen Vicki , Cittadini Antonio , Greenspan Susan L. , Douglas Pamela S. , Moses Alan C. , Beamer Wesley G. 
Published: February 2010

Abstract
Human growth hormone (hGH) and insulin-like growth factor I (IGF-I) both stimulate bone formation and have been proposed as therapeutic agents for osteoporosis. We examined the effect of hGH and IGF-I alone and in combination on bone size, bone mineral content (BMC), and bone mineral density (BMD) in 10- to 12-week-old growing female Sprague-Dawley rats. Sixty rats were assigned to treatment with either placebo, hGH, IGF-I, or both for 4 weeks. After 4 weeks, the right femurs and tibias were excised, and ex vivo BMC and the area of the tibia and femur were measured by dual-energy X-ray absorptiometry (DXA); volume of these bones was measured by Archimedes' principle. In addition, proximal tibial bone density was measured directly by peripheral quantitative computerized tomography (pQCT). Bone length, area, and volume in all treated groups was greater than controls. Areal bone density by DXA (BMC/area) was higher in IGF-treated rats and lower in GH-treated rats than in controls. Volumetric bone density (BMC/volume) was lower in treated groups than in controls. Measurements by pQCT confirmed that true bone density was lower in all treated groups than in controls. We conclude that treatment with hGH or IGF-I increased bone size and mineral content but decreased bone density in growing rats. Because areal correction of BMC did not adequately correct for the increased bone volume in IGF-treated rats, results of areal bone density by DXA should be interpreted with caution when treatment causes a disparity in bone size between groups.
Chondrocytes isolated from mature articular cartilage retain the capacity to form functional gap junctions
Authors: Donahue Henry J. , Guilak Farshid , Vander Molen Monique A. , Mcleod Kenneth J. , Rubin Clinton T. , Grande Daniel A. , Brink Peter R. 
Published: February 2010

Abstract
The distribution, expression, and functionality of gap junctions was examined in bovine chondrocytes (BCs) isolated from mature articular cartilage. BC cells displayed immunoreactivity for connexin 43 (Cx43), a specific gap junction protein. Cx43 protein expression was confirmed by Western blot analysis, and Cx43 mRNA was detected by nuclease protection assay. Additionally, BCs were shown to be functionally coupled, as revealed by dye transfer studies, and octanol, a gap junction uncoupler, greatly attenuated coupling. Furthermore, confocal microscopy of fluo-3 loaded BC cells revealed that deformation-induced cytosolic Ca2+ ion (Ca2+) signals propagated from cell-to-cell via gap junctions. To our knowledge, this is the first evidence suggesting that chondrocytes isolated from adult articular cartilage express functional gap junctions.
Energy-shunting external hip protector attenuates the peak femoral impact force below the theoretical fracture threshold: An in vitro biomechanical study under falling conditions of thel elderly
Authors: Parkkari Jari , Kannus Pekka , Heikkilä Jussi , Poutala Jarmo , Sievanen Harri , Vuori Ilkka 
Published: February 2010

Abstract
The first objective of this study was to design a hip protector that would effectively attenuate and shunt away from the greater trochanter the impact energies created in typical falls of the elderly. As the shock absorption material, the protector included the 12 mm-thick Plastazote, which was found to be the most efficient energy-absorbing material in our previous in vitro biomechanical tests. With an anatomically designed semiflexible outer shield of the protector (high density polyethylene), the impact surface was increased and the impact energy shunted away from the greater trochanter. In the second phase of the study, we determined the force attenuation capacity of this device in realistic (in vitro) falling conditions of the elderly. With the impact force of 6940 N used (a typical hip impact force measured in in vitro falling tests), the trochanteric soft tissue (25 mm-thick polyethylene foam) attenuated the peak femoral impact force to 5590 N and the tested protector to 1040 N. In the second series of this experiment, the peak femoral impact force was set to be so high (13,130 N) that the protector, if effective, should prevent the hip fracture in almost all cases. The trochanteric soft tissue attenuated this peak impact force to 10,400 N and the tested protector to 1810 N. Thus, the force received by the proximal femur still remained clearly below 4170 N, the average force required to fracture in vitro the proximal femur of the elderly in a fall loading configuration. In conclusion, our test results suggest that an anatomically designed energy-shunting and energy-absorbing hip protector can provide an effective impact force attenuation in typical falling conditions of the elderly. However, the efficacy of the protector in the prevention of hip fractures can only be evaluated in randomized clinical trials.
Monoclonal antibodies recognize antigen expressed by osteoblasts
Authors: Dr. Benayahu Dafna , Efrati M. , Wientroub S. 
Published: February 2010

Abstract
A marrow stromal osteogenic cell line (MBA-15) was used to create monoclonal antibodies (MoAbs). In this study, we describe a series of MoAbs for mouse marrow stroma (MMS) (MMS-25/17, MMS-85/12, MMS-302/40, and MMS-319/4) that recognized antigens expressed by stromal cells including osteoblastic cells. The MoAbs were screened against various cell and tissue types. MMS-85/12 was positive in detecting an antigen that was highly abundant in osteoblastic cells and primary adherent bone marrow cultures (BMC) but was negative for the marrow adipocytes copartner. The MMS-85/12 MoAb is an IgGl immunoglobulin. The immunohistochemical staining pattern is suggestive of the antigen being associated with the osteoblasts' plasma membrane and with the extracellular matrix constituent secreted by these cells. Western blotting and immunoprecipitation indicated that the antigen that was recognized by MMS-85/12 apparently had a molecular weight of 84 kD.
In vitro and Ex vivo evidence that estrogens suppress increased bone resorption induced by ovariectomy or PTH stimulation through an effect on osteoclastogenesis
Authors: Most Werner , Schot Lodewijk , Ederveen Antwan , van der Wee-Pals Lianne , Papapoulos Socrates , Dr. Löwik Clemens 
Published: February 2010

AbstractThe mechanism of bone loss following cessation of ovarian function is still unclear. Several studies have shown an increase in bone turnover following natural or surgical menopause which could be prevented by estrogen administration. However, a direct effect of estrogen on osteoclast-mediated bone resorption has been difficult to demonstrate in vitro. Recent evidence suggested that estrogen withdrawal stimulates the production of bone resorbing cytokines, (e.g., interleukin-6, IL-6), which regulate osteoclast formation in the bone marrow microenvironment. We studied the effects of 17β-estradiol on osteoclastic resorption, measured as 45calcium release, in vitro using cultures of fetal mouse long bone explants in which different stages of osteoclast development and activity are represented. 17β-estradiol (10−12-10−8 M) had no effect on basal or parathyroid hormone (PTH)-stimulated resorption of bone explants in which mature osteoclasts (radii/ulnae) or osteoclast precursors/progenitors (metacarpals) are present. 17β-estradiol, however, inhibited significantly the PTH-stimulated resorption of osteoclast-free metacarpals cultured together with mouse fetal liver as a source of early osteoclast progenitors; basal resorption was also not inhibited in this system. In ex vivo studies we further examined the effects of culturing bone marrow cells from ovariectomized (OVX) or sham-operated mice as an osteoclastic source together with osteoclast-free metacarpals on 45calcium release and bone histology. Cocultures of the bone marrow cells from OVX mice with osteoclast-free metacarpals increased significantly the osteoclast formation and subsequent osteoclastic resorption compared with control cocultures. This increase in resorption was prevented by either treatment of the OVX animals with estrogen for 1 week starting immediately after OVX or injection of the OVX animals with an IL-6 neutralizing antibody. We conclude that estrogens suppress the increased osteoclastic resorption induced by PTH or OVX through an effect on hematopoietic progenitor cells of the osteoclast lineage. Furthermore our data suggest that IL-6 is involved in the increase in osteoclastic resorption following OVX.
Aging changes mechanical loading thresholds for bone formation in rats
Authors: Turner Charles H. , Takano Y. , Owan I. 
Published: February 2010

AbstractThe effect of aging on the mechanical loading thresholds for osteogenesis was investigated in rats. We applied mechanical loads varying from 30 to 64 N to the tibiae of 43 19-month-old rats using a four-point bending apparatus. Bone formation rates were measured on the periosteal and endocortical surfaces of the tibial midshaft using double-label histomorphometry. Bone formation rates from the old rats were compared with results from adult (9-month-old) rats that we reported earlier.(4) Bone formation on the periosteal surface of the old rats was predominantly woven-fibered. Periosteal bone formation was observed in a lower percentage of the old rats compared with the younger adult rats for applied loads of 40 N and greater (59% old, 100% adult). However, in the old rats that formed woven bone there were no significant differences in woven bone area (p = 0.1) or surface (p = 0.24) compared with younger adult rats. Therefore, the periosteum of old rats had a higher threshold for activation by mechanical loading, but after activation occurred, the cells had the same capacity to form woven bone as younger adult animals. On the endocortical surface, relative bone formation rates in old rats showed a marginal (p = 0.06) increase in response to an applied load of 64 N but was not increased at lower loads. The relative bone formation rate in the old rats was over 16-fold less than that reported for the younger adult rats at an applied load of 64 N and the relative bone forming surface in old rats in this study was 5-fold less than it was in younger rats under similar loading conditions. In the younger adult rats, a mechanical threshold for lamellar bone formation of 1050 μstrain was calculated for the endocortical bone surface. The old rats required over 1700 μstrain on the endocortical surface before bone formation was increased. The data suggest that both the periosteal and endocortical surfaces of the tibiae of older rats are less responsive to mechanical stimuli.
Bone mineral metabolism in T lymphocyte–deficient and –replete strains of rat
Authors: Buchinsky Farrel J. , Ma Yanfei , Mann Gary N. , Rucinski Boguslaw , Bryer Haldon P. , Paynton Barbara V. , Jee Webster S.S. , Hendy Geoffrey N. , Dr. Epstein Solomon 
Published: February 2010

AbstractThe immune and skeletal systems are known to interact. We have repeatedly shown that in contrast to in vitro data, the administration of T lymphocyte immunosuppressants, such as cyclosporin A, leads to an increase in bone resorption and a high turnover osteopenia. The purpose of this study was to characterize the bone metabolism of the T lymphocyte deficient Rowett athymic homozygous (rnu/rnu) nude rat. We wished to determine whether these rats share the bone abnormalities of cyclosporin A–treated rats. Eleven 10-week-old Sprague–Dawley rats and 12 similarly aged nude rats were studied over a 4-week period. Metaphyseal cancellous bone histomorphometry was similar in the two groups of rats and only differed with regard to percentage eroded perimeter (lower in nude rats, p = 0.008) and longitudinal growth rate (49% lower in nude rats, p &lt; 0.001). The nude rats had less body mass (p &lt; 0.001) but nevertheless gained the same percentage of their body weight over the study period. The athymic rats had lower levels of serum 1,25-dihydroxyvitamin D (p &lt; 0.014) and serum osteocalcin (p &lt; 0.009), and at the age of 14 weeks the nude rats had lower concentrations of serum creatinine (p = 0.001) and blood ionized calcium (p = 0.0002), yet serum PTH was similar throughout. RNA isolated from the contralateral tibias revealed that the nude group had lower steady-state levels of osteocalcin mRNA despite similar rates of bone formation. In its entirety, the data suggest that T cell deficiency per se is not necessarily associated with high turnover osteopenia.
Isolation of calcium-phosphate crystals of bone by non-aqueous methods at low temperature
Authors: Kim Hyun-Man , Rey Christian , Dr. Glimcher Melvin J. 
Published: February 2010

Abstract
We have developed low temperature nonaqueous solution methods as well as low power plasma ashing for the degradation of the organic matrix of bone powder which have permitted us to obtain bone crystals essentially free of organic matrix constituents without any significant change in their composition, overall structure, or internal short-range order. We have also been able to disperse the crystals, which has made it possible to examine the isolated crystals by X-ray diffraction and resolution-enhanced Fourier transform infrared (FTIR) spectroscopy and isolated single crystals by high resolution transmission electron microscopy (TEM) and electron diffraction. TEM of isolated single crystals of fish, chicken, mouse, and bovine bone have clearly demonstrated that the crystals are very thin plates. No rod or needle-like crystals were observed in any of the bone samples in the four species studied including the earliest crystals deposited. Although there were variations in the size distribution of the crystals in the different species studied, in general the average crystal dimensions were very similar.
Mineralization of bone-like extracellular matrix in the absence of functional osteoblasts
Authors: Marsh Mary E. , Munne Anna M. , Vogel James J. , Cui Yingqi , Dr. Franceschi Renny T. 
Published: February 2010

Abstract
When grown in medium containing ascorbic acid and β-glycerol phosphate, mouse MC3T3-E1 cells express an osteoblast phenotype and produce a highly mineralized extracellular matrix. The purpose of this study was to independently examine the role of the collagenous matrix and functional osteoblasts on the mineralization process. Cultures with and without an extensive collagenous matrix were prepared by growing MC3T3-E1 cells in the presence and absence of ascorbic acid. Matrix-rich cultures mineralized at much lower calcium phosphate ion products than nonmatrix cultures. At higher ion products, spontaneous precipitation in the medium and cell layers of nonmatrix cultures were observed. In contrast, mineral in matrix-rich cultures was still exclusively associated with collagen fibrils and not with ectopic sites in the cell layer or medium. To examine the effect of cell viability on matrix mineralization, cells were grown 8 or 16 days in the presence of ascorbic acid, then killed and incubated in a mineralizing medium. Significant mineralization was not observed in the collagenous matrix of 8-day killed cultures or age-matched controls. At 16 days mineral was associated with collagen fibrils at specific foci in the matrix of both viable and killed cultures. This observation is consistent with the concept that collagenous matrices must undergo a maturation process before they can support mineral induction and growth. It further shows that osteoblast-like cells are not required for mineralization of mature matrices, but are required for matrix maturation.
Leukemia inhibitory factor as a mediator of JAK/STAT activation in murine osteoblasts
Authors: Dr. Lowe Carolyn , Gillespie Gerald A.J. , Pike Wesley J. 
Published: February 2010

Abstract
A number of cytokines have been shown to exert their effects via a recently discovered signaling cascade. One step in this pathway is mediated by a family of nonreceptor protein tyrosine kinases, the Janus kinases or JAK kinases, which become phosphorylated upon ligand-receptor binding and receptor phosphorylation. This in turn is followed by phosphorylation of certain members of a family of latent transcription factors, called signal transducers and activators of transcription (STATs), which subsequently enter the nucleus, bind to DNA in a sequence-specific fashion, and modulate transcription. In view of the apparent role of leukemia inhibitory factor (LIF) in bone remodeling, we sought to determine which, if any, of the JAK/STAT family members are involved in mediating the actions of LIF using the MC3T3-E1 cell line (a spontaneously immortalized osteoblast) and normal murine calvarial osteoblasts. We report here rapid and transient phosphorylation of the LIF receptor, and similarly, we detect phosphorylation of predominantly JAK1 and to a minor extent JAK2 in response to LIF treatment in MC3T3-E1 cells. In these experiments we also detect phosphorylation of STAT1 and to a much lesser degree STAT3 upon addition of LIF. Phosphorylation of the STAT proteins correlates directly with their ability to bind DNA in a gel mobility shift assay in MC3T3-E1 cells and in normal calvarial osteoblasts. These studies suggest that LIF action in these cells, as in other cell types, is mediated in part via specific members of the JAK/STAT pathway.