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  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochemical and Biophysical Research Communications 65 (1975), S. 1343-1349 
    ISSN: 0006-291X
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochemical and Biophysical Research Communications 162 (1989), S. 435-440 
    ISSN: 0006-291X
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochemical and Biophysical Research Communications 65 (1975), S. 1343-1349 
    ISSN: 0006-291X
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochemical and Biophysical Research Communications 174 (1991), S. 181-188 
    ISSN: 0006-291X
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
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  • 5
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochemical and Biophysical Research Communications 164 (1989), S. 119-127 
    ISSN: 0006-291X
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 22 (1977), S. 137-147 
    ISSN: 1432-0827
    Keywords: Bone cells ; Bone culture ; Calcium ionophore ; Cyclic AMP ; Calcium transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The ionophore A23187 produced a rapid transient increase in the rate of calcium uptake by isolated fetal rat bone cells. There was no effect on calcium efflux or total cellular calcium. The magnitude of the effect on influx was amplified when the cell were incubated at 4°C. Cellular metabolic functions and resorption of cultured fetal rat bones (release of45Ca from pre-labeled long bone) were affected by A23187 in a biphasic manner: cell cyclic AMP (cAMP) was increased by 0.1 and 0.3 μg/ml of the ionophore, whereas 10 μg/ml was either ineffective or lowered the cAMP levels. The high A23187 concentration abolished the stimulatory effects of parathyroid hormone and methylisobutylxanthine. Concentrations of 0.1 and 0.3 μg/ml A23187 stimulated bone resorption. The effect was abolished by calcitonin. Ionophore concentrations above 1 μg/ml produced less bone resorption. These higher concentrations antagonized the bone-resorbing effect of parathyroid hormone and 1,25-dihydroxyvitamin D3. A23187 at 5 and 10 μg/ml decreased bone cell lactate and ATP. Thus at low concentrations, A23187 produced effects on bone similar to those of parathyroid hormone, suggesting that calcium is the primary initiator of PTH-induced bone resorption. At the higher concentrations A23187 may have a general inhibitory effect on cell metabolism.
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  • 7
    ISSN: 1432-0827
    Keywords: Tissue plasminogen activator ; Osteoblasts ; Interleukin-1 ; Tumor necrosis factor β ; Forskolin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The effects of interleukin-1 (IL-1), forskolin, and tumor necrosis factor β (TNF-β) on tissue plasminogen activator (t-PA) activity were studied in the human osteoblastic osteosarcoma cell line, G292. t-PA activity was measured in the cell media using the chromogenic substrate, S-2251. After a 24 hour incubation period, IL-1 increased t-PA in a dose-dependent manner. The effect of IL-1 at 10.0 U/ml was partially inhibited in the presence of indomethacin. Forskolin (1.0 μM) increased t-PA activity after 24 hours with the effects of combined treatment of IL-1 (1.0 U/ml, 10.0 U/ml) and forskolin being apparently additive in nature. TNF-β (10-8–10-7 M) also produced increased t-PA activity in the cell medial after a 24 hour incubation period. These results suggest that the cytokines, IL-1 and TNF-β, can increase t-PA activity in G292 cells and that there is both a cAMP-dependent as well as a cAMP-independent pathway involved in the regulation of this osteoblastic cell function.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 55 (1994), S. 128-133 
    ISSN: 1432-0827
    Keywords: Osteoblast ; Plasma membrane ; Calcium ; Ion channel ; Cell proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Patch clamp physiological techniques were used to characterize the voltage-activated calcium currents (VACC) expressed in the plasma membrane of osteoblastic cells as a function of time in culture and proliferative state of the cell. Osteoblast-enriched preparations were isolated by collagenase digestions of newborn rat calvaria and cultured under different conditions which affected cell proliferation (i.e., low serum in the media to arrest proliferation). VACC were isolated by replacing the intracellular potassium with cesium, and adding 1 μM tetrodotoxin to the bath. Under conditions that favored cell proliferation, low cell density, and media supplemented with 10% fetal calf serum (FCS), a transient calcium current was not expressed until day 3 in culture. There was a statistically significant relationship between the precentage of cells expressing this current and the time in culture. The magnitude of the current significantly increased as days in culture increased. Under the same conditions, the sustained VACC was detected after 7 or 8 days in culture. However, arresting cell proliferation after 2 days in culture by reducing the FCS concentration to 0.01% induced the expression of the sustained VACC the next day. The data suggest that the expression of VACC in the plasma membrane of rat calvarial osteoblasts depends on the time in culture and the state of proliferation of the cells. These results should prove to be valuable in studying the functional significance of VACC in osteoblastic cells and their regulation by various bone regulatory agents.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 35 (1983), S. 243-249 
    ISSN: 1432-0827
    Keywords: Prostaglandin E2 ; Bone cells ; Cyclic AMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The binding of prostaglandin E2 (PGE2) to bone cells was studied to provide direct evidence for the existence of specific receptors in bone. Bone cells were isolated by collagenase digestion of fetal and newborn rat calvaria. Isolated cells were incubated with3H-PGE2 and collected on Millipore filters. Specific binding was determined by subtracting the binding that occurred with 10−6 M non-radioactive PGE2 and3H-PGE2 from that with3H-PGE2 alone. With heterogeneous cell preparations and at PGE2 concentrations from 10−9 − 1.7 × 10−8 M at 37°C, specific binding reached steady state within 10 min. Bound3H-PGE2 was displaced by the addition of increasing amounts of unlabeled PGE2. Inhibition of PGE2 binding was observed with PGE1 and the endoperoxide analog, U44069, but not with PGE2α, a lipopolysaccharide, or 13,14-dihydro 15-keto PGE2. Studies with bone cell populations, obtained by sequential digestions, indicated that an osteoclastic population binds 30-fold more PGE2 than osteoblastic cells. Scatchard analyses revealed that the osteoclastic cells have an affinity constant for PGE2 binding similar to that obtained with heterogeneous populations. However, the PGE2 binding capacity in this osteoclastic population was fivefold greater than in the heterogeneous population. The osteoclastic population responded with an increase in cyclic AMP to lower concentrations of PGE2 than the osteoblastic populations. These studies suggest that differences in the binding capacity of PGE2 receptors exist among bone celltypes and that these differences are reflected in the cellular cyclic AMP response.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 54 (1994), S. 409-413 
    ISSN: 1432-0827
    Keywords: Osteoblasts ; Epidermal growth factor ; Tyrosine kinase ; G proteins ; Mitogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Epidermal growth factor (EGF) has been found to stimulate proliferation in a variety of cell types. The EGF receptor is known to have tyrosine kinase activity [1], however, the role of this signal mechanism has not been established in bone cells. The aim of this study was to determine whether tyrosine kinase activity and G inhibitory (Gi) proteins are involved in EGF-stimulated proliferation in the osteoblastic cell line G292 and in primary culture osteoblasts isolated from neonatal rat calvaria. Cell proliferation was measured by 3H-thymidine incorporation using liquid scintillation spectrometry. EGF stimulates a dose-dependent increase in proliferation of G292 and primary culture cells above control. Genistein was able to inhibit the effects of EGF in the G292 cells. In the primary culture cells, genistein with EGF appeared to enhance proliferation compared with EGF alone or genistein alone. Tyrphostin 25, on the other hand, inhibited the EGF response in both of these cell types. Inactivation of Gi proteins with pertussis toxin was able to inhibit EGF-induced mitogenesis in the neonatal rat osteoblasts but did not appear to specifically inhibit this response in the G292 cells. These results suggest that although both of these osteoblastic cell types increase proliferation in response to EGF, their signal pathways are different.
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