bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of Bisphosphonate (Incadronate) on Fracture Healing of Long Bones in Rats
Authors: Li Jiliang , Mori Satoshi , Kaji Yoshio , Mashiba Tasuku , Kawanishi Jun , Norimatsu Hiromichi 
Published: April 2006

Abstract
This study was designed to test whether bisphosphonates disturb the process of fracture healing. Female Sprague-Dawley rats were injected with either two doses of bisphosphonate (incadronate) (10 μg/kg and 100 μg/kg) or vehicle three times a week for 2 weeks. Right femora were then fractured and fixed with intramedullary wires. Incadronate treatment was stopped in pretreatment groups (P-10 and P-100 groups), while the treatment was continued in continuous treatment groups (C-10 and C-100 groups). Animals were sacrificed at 6 and 16 weeks after surgery. Soft X-ray of all fractured femora was taken. After mechanical testing, fractured femora were stained in Villanueva bone stain and embedded in methyl methacrylate. Cross-sections near fracture line were analyzed by microradiography and histomorphometry. Radiographic study showed that bony callus was present in all the fractures and incadronate treatment led to a larger callus, especially in C-100 group at both 6 and 16 weeks. Histologic study showed that the process of fracture healing in pretreatment groups was delayed at 6 weeks, but reached control level thereafter and showed same characteristics as in control at 16 weeks. Woven bony callus could still be seen in continuous treatment groups at 16 weeks. Mechanical study indicated that the ultimate load of C-100 group was slightly higher than the other treatment groups and control. The results suggest that pretreatment with incadronate did not affect fracture healing at 16 weeks after fracture. However, continuous incadronate treatment could lead to larger callus, but it delayed remodeling process during fracture healing, especially with high-dose treatment.
Bone's Early Responses to Mechanical Loading Differ in Distinct Genetic Strains of Chick: Selection for Enhanced Growth Reduces Skeletal Adaptability
Authors: Dr. Pitsillides Andrew A. , Rawlinson Simon C. F. , Mosley John R. , Lanyon Lance E. 
Published: April 2006

Abstract
Bone's functional competence is established and maintained, at least partly, by mechanisms involving appropriate adaptation to mechanical loading. These appear to fail in chickens selectively bred either for maximum egg (Egg-type) or meat (Meat-type) production, which show high rates of fracture and skeletal abnormality, respectively. By measuring several early strain-induced responses in cultured embryonic tibiotarsi from commercially bred (Egg-type and Meat-type) and wild-type (Wild-type) chicks, we have investigated the possibility that these skeletal failures are the product of a compromised ability to respond appropriately to loading-induced mechanical strain. Axial loads engendering peak dynamic (1 Hz) longitudinal strains of between −1300 με and −1500 με (for 10 minutes) in vitro in tibiotarsi from the three types of 18-day-old chicks increased periosteal osteoblast glucose 6-phosphate dehydrogenase (G6PD) activity in both Wild-type (26%, p &lt; 0.01) and Egg-type (49%, p &lt; 0.001) chicks in situ, while Meat-type chicks did not show any significant changes (11%). Load-induced increases in medium nitrite accumulation (stable nitric oxide [NO] metabolite) were produced in Egg-type and Wild-type tibiotarsi (82 ± 12%, p &lt; 0.01; 39 ± 8%, p &lt; 0.01), respectively. In contrast, loading produced no change in NO release from Meat-type chick tibiotarsi. These changes in NO release correlated with load-related increases in G6PD activity (R2 = 0.98, p &lt; 0.05) in the different chick types. Wild-type and Meat-type tibiotarsal periosteal osteoblasts responded in a biphasic manner to exogenous prostacyclin (PGI2), with maximal stimulation of G6PD activity at 10−7 M and 10−6 M PGI2. However, Egg-type chick osteoblasts showed smaller, progressive increases up to 10−5 M PGI2. These results indicate that early phases of the adaptive response to loading differ in different genetic strains of embryonic chick; that skeletal abnormalities which develop in genetically selected, high growth rate chicks may reflect a compromised ability to respond to load; and that load-induced increases in osteoblastic G6PD activity appear to be closely associated with increased rates of NO release. It is probable that similar genetically related differences in bones' responsiveness to mechanical loading occur in other species.
Longitudinal Bone Mineral Density Changes in Female Child Artistic Gymnasts
Authors: Nickols-Richardson S. M. , O'Connor P. J. , Shapses S. A. , Lewis R. D. 
Published: April 2006

Abstract
Changes in bone mineral density (BMD), and related factors, in female child artistic gymnasts (n = 9) and their age- (±0.3 years), height- (±2.8 cm), and weight- (±1.7 kg) matched controls (n = 9) were prospectively examined. It was hypothesized that gymnasts would possess higher BMD at baseline, 6, and 12 months later and have greater gains in BMD over 1 year compared with controls. BMD (g/cm2) of the total proximal femur (TPF), Ward's triangle (WT), trochanter (Troch), femoral neck (FN), lumbar spine (LS, L1–L4), and total body (TB) were measured by dual-energy X-ray absorptiometry. Physical activity was measured by a 7-day recall; daily dietary intakes of energy and nutrients were estimated from 3-day records. Serum osteocalcin and urinary pyridinium cross-links were measured by radioimmunoassay and high performance liquid chromatography, respectively. Gymnasts versus controls possessed significantly higher BMD at all sites measured. Although not significantly different (p &gt; 0.05), gymnasts compared with controls had moderately larger percentage changes in Troch (% Δ = 8.6 ± 3.0 vs. 3.8 ± 5.1%, d = 0.41), FN (% Δ = 6.1 ± 1.2 vs. 3.9 ± 1.6%, d = 0.55), LS (% Δ = 7.8 ± 1.1 vs. 6.8 ± 1.6%, d = 0.26), and TB BMD (% Δ = 5.6 ± 0.8 vs. 3.4 ± 0.7%, d = 0.98) as evidenced by the magnitude of the effect sizes (d). Gymnasts versus controls possessed a lower percentage body fat (p &lt; 0.01) and engaged in more hours of very hard activity (p &lt; 0.0001). Calcium, as a percentage of adequate intake, decreased over 12 months (p &lt; 0.01), and urinary cross-links significantly decreased over 6 months in both groups. Female child gymnasts possess higher BMD at the TPF and related sites, LS, and TB compared with nongymnast controls, and 1 year of gymnastics training moderately increases Troch, FN, LS, and TB BMD for gymnasts compared with controls. These findings lend support to the idea that gymnastics training in childhood helps maximize peak BMD.
Targeted Expression of Calcitonin Gene-Related Peptide to Osteoblasts Increases Bone Density in Mice
Authors: Ballica Rabia , Valentijn Karine , Khachatryan Armen , Guerder Sylvie , Kapadia Shanta , Gundberg Caren , Gilligan James , Flavell Richard A. , Vignery Agnès 
Published: April 2006

Abstract
The neuropeptide calcitonin gene-related peptide (CGRP) is concentrated in fine sensory nerve endings innervating all tissues, including bone. CGRP inhibits osteoclasts, stimulates insulin-like growth factor I and inhibits tumor necrosis factor alpha production by osteoblasts in vitro. To investigate the role of CGRP in bone in vivo, mice were engineered to express CGRP in osteoblasts by placing the human CGRP gene under the control of the rat osteocalcin promoter (Ost-CGRP tg+ mice). Calvaria cultures from transgene positive (tg+), but not tg− mice, produced bioactive CGRP. Trabecular bone density and bone volume, determined by peripheral quantitative computed tomography and bone histomorphometry, respectively, were higher in tg+ than tg− littermates. This increase in bone volume was associated with an increased bone formation rate. Trabecular bone density decreased in tg+ mice as a result of ovariectomy, but remained higher than in sham tg− mice. Targeting CGRP to osteoblasts appears to favor the establishment of a higher trabecular bone mass in mice.
Mechanical Tension-Stress Induces Expression of Bone Morphogenetic Protein (BMP)-2 and BMP-4, but Not BMP-6, BMP-7, and GDF-5 mRNA, During Distraction Osteogenesis
Authors: Sato Motohiko , Ochi Takahiro , Nakase Takanobu , Hirota Seiichi , Kitamura Yukihiko , Nomura Shintaro , Yasui Natsuo 
Published: April 2006

Abstract
Bone lengthening with osteotomy and gradual distraction was achieved in 57 rats, and the effect of mechanical tension-stress on gene expression of bone morphogenetic proteins (BMPs) was investigated by in situ hybridization and Northern blot analysis using probes of BMP-2, BMP-4, BMP-6, BMP-7, and growth/differentiation factor (GDF)-5. There was a lag phase for 7 days after femoral osteotomy until gradual distraction was carried out for 21 days at a rate of 0.25 mm/12 h using a small external fixator. The signals of the above BMPs mRNA were not detected in the intact rat bone but they were induced after osteotomy except those for BMP-7. By 4 days after osteotomy, BMP-2 and BMP-4 mRNAs were detected in chondrogenic precursor cells in the subperiosteal immature callus. BMP-6 and GDF-5 mRNA were detected in more differentiated cells in chondroid bone. By 7 days after osteotomy, cartilaginous external callus and bony endosteal callus were formed. Meanwhile, the signals of BMP-2 and BMP-4 mRNAs declined to preoperative levels, whereas the signals of BMP-6 and GDF-5 mRNAs were rather elevated. As distraction was started, the callus elongated and eventually separated into proximal and distal segments forming a fibrous interzone in the middle. Expression of BMP-2 and BMP-4 mRNAs was markedly induced at this stage. Their signals were detected widely among chondrogenic and osteogenic cells and their precursor cells sustaining mechanical tension-stress at the fibrous interzone. BMP-6 and GDF-5 mRNAs were detected exclusively in chondrogenic cells at both ends of the fibrous interzone, where endochondral ossification occurred. But neither mRNA was detected in terminally differentiated hypertrophic chondrocytes. As distraction advanced, the cartilage was progressively resorbed from both ends and new bone was formed directly by intramembranous ossification. There was no new cartilage formation in the advanced stage of distraction. The signals of BMP-6 and GDF-5 mRNA declined by this stage, while those of BMP-2 and BMP-4 were maintained at high level for as long as distraction was continued. After completion of distraction, the fibrous interzone fused and the lengthened segment was consolidated. BMP-2, BMP-4, BMP-6, nor GDF-5 was expressed at this stage. The signals of BMP-7 were not detected throughout the experiment. The present results suggest that excellent and uninterrupted bone formation during distraction osteogenesis owes to enhanced expression of BMP-2 and BMP-4 genes by mechanical tension-stress. Abundant gene products of BMP-2 and BMP-4 could induce in situ bone formation by paracrine and autocrine mechanisms.
A Novel T Cell Cytokine Stimulates Interleukin-6 in Human Osteoblastic Cells
Authors: Rifas Leonard , Avioli Louis V. 
Published: April 2006

Abstract
Rheumatoid arthritis (RA) is an autoimmune disease characterized by a heavy lymphocytic infiltration into the synovial cavity, resulting in the secretion of a variety of cytokines which ultimately leads to destruction of joint tissue. Among the infiltrating cells are activated T cells which produce specific cytokines capable of osteoclast progenitor cell expansion, fusion, and activation. Cultures of activated human T cells and human osteoblasts (hOBs) were used to study the possibility that lymphokines may act on osteoblasts to produce the osteoclastogenic factor interleukin-6 (IL-6). Purified T cells were activated with a combination of anti-CD3 and anti-CD28 antibodies, cocultured with hOBs in direct physical contact or separated by a transwell system, and conditioned media (CM) were assayed for IL-6 production. After a 72 h incubation period, activated T cell–hOB interaction resulted in a 100-fold increase of IL-6 production over basal levels. The immunosuppressant cyclosporine A (CsA) inhibited T cell tumor necrosis factor alpha and IL-6 production but did not inhibit the T cell induction of IL-6 from hOB. Assay of activated T-cell CM on hOB revealed that a soluble factor, not cell-cell contact, was the major inducer of IL-6. The induction of IL-6 mRNA by both activated T cell CM and CsA-treated activated T cell CM was confirmed by Northern blot analysis. Neutralizing antibodies to IL-13 and IL-17 did not affect IL-6 production. These findings suggest that activated T cells produce a novel, potent, IL-6 inducing factor that may be responsible for the bone loss observed in RA patients.
Glucocorticoids Decrease Interleukin-6 Levels and Induce Mineralization of Cultured Osteogenic Cells from Children with Fibrous Dysplasia
Authors: Stanton Robert P. , Hobson Grace M. , Montgomery Ben E. , Moses Priscilla A. , Smith-Kirwin Susan M. , Funanage Vicky L. 
Published: April 2006

Abstract
Fibrous dysplasia (FD) is a progressive bone disease in which abnormal fibroblast proliferation results in the replacement of normal cancellous bone with an immature fibrous tissue that is poorly mineralized. The disease manifests itself in the monostotic form in which only one bone is involved and the polyostotic form in which multiple bones at different sites are affected. The McCune–Albright syndrome is a variation of the polyostotic form in which patients demonstrate a greater extent of bone involvement and a variety of endocrinopathies. Somatic activating mutations in the GNAS gene have been demonstrated in the fibrotic lesions of patients affected with either monostotic or polyostotic FD. The increased cAMP levels caused by the G-protein mutations lead to increased interleukin-6 (IL-6) levels in the affected tissues, resulting in abnormal osteoblast differentiation and increased osteoclastic activity. Utilizing cell culture techniques that have been developed for mammalian bone marrow stromal cells, we have successfully cultured osteogenic stem cells from the affected stroma of 11 FD patients. Cells cultured from patients with polyostotic FD showed a high frequency of the Gsα mutation, whereas cells from monostotic FD patients showed a low frequency of the mutation. Both the normal and FD cells displayed the osteogenic phenotype when exposed to medium containing glucocorticoids. Glucocorticoids also caused a dramatic inhibition of IL-6 mRNA and protein levels in osteogenic cells cultured from the FD patients. These findings suggest that chemical alteration of cellular function may lead to new treatment options for patients with FD.
Age-Related Osteogenic Potential of Mesenchymal Stromal Stem Cells from Human Vertebral Bone Marrow
Authors: Dr. D'Ippolito Gianluca , Schiller Paul C. , Ricordi Camillo , Roos Bernard A. , Howard Guy A. 
Published: April 2006

Abstract
Mesenchymal stem cells (MSCs) residing in bone marrow (BM) are the progenitors for osteoblasts and for several other cell types. In humans, the age-related decrease in bone mass could reflect decreased osteoblasts secondary to an age-related loss of osteoprogenitors. To test this hypothesis, BM cells were isolated from vertebral bodies of thoracic and lumbar spine (T1–L5) from 41 donors (16 women and 25 men) of various ages (3–70 years old) after death from traumatic injury. Primary cultures were grown in alpha modified essential medium with fetal bovine serum for 13 days until adherent cells formed colonies (CFU-Fs). Colonies that stained positive for alkaline phosphatase activity (CFU-F/ALP+) were considered to have osteogenic potential. BM nucleated cells were plated (0.5, 1, 2.5, 5, or 10 × 106 cells/10-cm dish) and grown in dexamethasone (Dex), which promotes osteoblastic differentiation. The optimal plating efficiency using BM-derived cells from donors of various ages was 5 × 106 cells/10-cm dish. BM-derived cells were also grown in the absence of Dex at this plating density. At the optimal plating density, in the presence of Dex, the number of CFU-F/ALP+ present in the BM of the younger donors (3–36 years old) was 66.2 ± 9.6 per 106 cells (mean ± SEM), but only 14.7 ± 2.6 per 106 cells in the older donors (41–70 years old). With longer-term culture (4–5 weeks) of these BM cells in medium containing 10 mM β-glycerophosphate and 100 μg/ml ascorbic acid, the extracellular matrix mineralized, a result consistent with mature osteoblastic function. These results demonstrate that the number of MSCs with osteogenic potential (CFU-F/ALP+) decreases early during aging in humans and may be responsible for the age-related reduction in osteoblast number. Our results are particularly important in that the vertebrae are a site of high turnover osteoporosis and, possibly, the earliest site of bone loss in age-related osteoporosis.
Localization of Smads, the TGF-β Family Intracellular Signaling Components During Endochondral Ossification
Authors: Prof. Sakou Takashi , Onishi Toshiyuki , Yamamoto Takuya , Nagamine Tomonori , Sampath T. Kuber , ten Dijke Peter 
Published: April 2006

Abstract
Members of the transforming growth factor-β (TGF-β) family transduce signals from the cell membrane to the nucleus via specific type I and type II receptors and Smad proteins. Smad1 and Smad5 mediate intracellular signaling of bone morphogenetic protein (BMP), whereas Smad2 and Smad3 transduce TGF-β signaling. Smad4 is a common mediator required for both pathways. Smad6 and Smad7 inhibit signaling by members of the TGF-β superfamily. Here, we examined the expression of Smad1 to Smad7 proteins during endochondral ossification of epiphyseal plate of growing rats using immunohistochemical techniques. The expression of Smad proteins was correlated with the expression of TGF-β1 and its receptors, and BMP-2/4 and BMP receptors. The results show that TGF-β1 and BMP-2/4 were actively expressed in chondrocytes that are undergoing proliferation and maturation, which overlaps with expression of their corresponding type I and type II receptors. The Smads, however, exhibited a distinct expression pattern, respectively. For example, Smad1 and Smad5 were highly expressed in proliferating chondrocytes and in those chondrocytes that are undergoing maturation. The TGF-β/activin-restricted Smads were also expressed in a nearly complementary fashion; Smad2 was strongly expressed in proliferating chondrocytes, whereas Smad3 was strongly observed in maturing chondrocytes. Smad4 was broadly expressed in all zones of epiphyseal plate. Inhibitory Smads, Smad6 and Smad7, were strongly expressed in the zone of cartilage that contained mature chondrocytes. Our findings show a colocalization of the pathway-restricted and inhibitory Smads with activating ligands or ligands whose action they antagonize and their receptors in various zones of epiphyseal growth plate, suggesting that TGF-β superfamily Smad signaling pathways plays a morphogenic role during endochondral bone formation.
Hormone Replacement Therapy Prevents Osteoclastic Hyperactivity: A Histomorphometric Study in Early Postmenopausal Women
Authors: Dr. Eriksen Erik Fink , Langdahl Bente , Vesterby Annie , Rungby Jørgen , Kassem Mustapha 
Published: April 2006

Abstract
In a randomized, double blind, clinical prospective trial comprising 35 women treated with either hormone replacement therapy (HRT) (cyclic estradiol/norethisterone acetate) or placebo we performed histomorphometric studies on paired bone biopsies obtained before and after 2 years of treatment. Untreated women developed a progressively more negative balance at individual bone multicellular units (BMUs) (i.e., wall thickness-erosion depth) (2.2 ± 1.7 μm vs. −5.7 ± 1.4 μm; p &lt; 0.01), while women on HRT displayed preservation of bone balance (2.4 ± 2.4 μm vs. 2.5 ± 2.5 μm; NS). No significant differences in wall thickness between the two groups were demonstrable, but the untreated women developed a pronounced increase in erosion depth over 2 years (46.9 ± 1.8 μm vs. 52.0 ± 1.9 μm; p &lt; 0.05), while the HRT group revealed no change (47.8 ± 2.7 μm vs. 44.6 ± 1.7 μm; NS). Furthermore, the placebo group displayed an increased osteoclastic erosion depth (17.8 ± 1.6 μm vs. 25.0 ± 1.7 μm; p &lt; 0.001), compared with unchanged values in the HRT group (20.0 ± 1.6 μm vs. 16.9 ± 1.4 μm/day; NS). While the placebo group revealed a slight increase in volume referent resorption rate (35 ± 8% vs. 38 ± 8%; NS) the HRT group revealed a pronounced decrease (46 ± 8% vs. 28 ± 5%; p &lt; 0.05). No significant changes in marrow star volume (an index of trabecular perforations) were demonstrable in either group. Our results demonstrate that bone remodeling in early postmenopausal women is characterized by progressive osteoclastic hyperactivity, which is reduced by cyclic HRT. This reduction of resorptive activity at the BMU level after HRT seems to precede the reduction in activation frequency demonstrated in previous studies on older postmenopausal women.
Identification and Characterization of a Novel Protein, Periostin, with Restricted Expression to Periosteum and Periodontal Ligament and Increased Expression by Transforming Growth Factor β
Authors: Horiuchi Keisuke , Amizuka Norio , Takeshita Sunao , Takamatsu Hiroyuki , Katsuura Mieko , Ozawa Hidehiro , Toyama Yoshiaki , Bonewald Lynda F. , Kudo Akira 
Published: April 2006

Abstract
We had previously identified the cDNA for a novel protein called osteoblast-specific factor 2 (OSF-2) from an MC3T3-E1 cDNA library using subtraction hybridization and differential screening techniques. Here we describe the localization, regulation, and potential function of this protein. Immunohistochemistry using specific antiserum revealed that in adult mice, the protein is preferentially expressed in periosteum and periodontal ligament, indicating its tissue specificity and a potential role in bone and tooth formation and maintenance of structure. Based on this observation and the fact that other proteins have been called OSF-2, the protein was renamed “periostin.” Western blot analysis showed that periostin is a disulfide linked 90 kDa protein secreted by osteoblasts and osteoblast-like cell lines. Nucleotide sequence revealed four periostin transcripts that differ in the length of the C-terminal domain, possibly caused by alternative splicing events. Reverse transcription- polymerase chain reaction analysis revealed that these isoforms are not expressed uniformly but are differentially expressed in various cell lines. Both purified periostin protein and the periostin-Fc recombinant protein supported attachment and spreading of MC3T3-E1 cells, and this effect was impaired by antiperiostin antiserum, suggesting that periostin is involved in cell adhesion. The protein is highly homologous to βig-h3, a molecule induced by transforming growth factor β (TGF-β) that promotes the adhesion and spreading of fibroblasts. Because TGF-β has dramatic effects on periosteal expansion and the recruitment of osteoblast precursors, this factor was tested for its effects on periostin expression. By Western blot analysis, TGF-β increased periostin expression in primary osteoblast cells. Together, these data suggest that periostin may play a role in the recruitment and attachment of osteoblast precursors in the periosteum.