Abstract:The effect of different vanadium compounds on proliferation and differentiation was examined in osteoblast-like UMR106 cells. Vanadate increased the cell growth in a biphasic manner, the higher doses inhibiting cell progression. Vanadyl stimulated cell proliferation in a dose-responsive manner. Similar to vanadate, pervanadate increased osteoblast-like cell proliferation in a biphasic manner but no inhibition of growth was observed. Vanadyl and pervanadate were stronger stimulators of cell growth than vanadate… Show more
“…UMR106 rat osteosarcoma cells were grown in DMEM containing 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin at 37°C in 5% CO 2 atmosphere (Cortizo and Etcheverry, 1995). Cells were seeded on 75 cm 2 flasks and subcultured using trypsin-EDTA.…”
Section: Cell Culture and Spheroid Preparationmentioning
Bisphosphonates are nonhydrolysable pyrophosphate analogues that prevent bone loss in several types of cancer. However, the mechanisms of anticancer action of bisphosphonates are not completely known. We have previously shown that nitrogen-containing bisphosphonates directly inhibit alkaline phosphatase of UMR106 rat osteosarcoma cells. In this study, we evaluated the effects of alendronate on the migration of UMR106 osteosarcoma using a model of multicellular cell spheroids, as well as the alendronate effect on neutral phosphatases. Alendronate significantly inhibited the migration of osteoblasts in a dose-dependent manner (10 − 6 -10 − 4 M). This effect was also dependent on calcium availability. The spheroid morphology and distribution of actin fibers were also affected by alendronate treatment. Alendronate dose-dependently inhibited neutral phosphatase activity in cell-free osteoblastic extracts as well as in osteoblasts in culture. Our results show that alendronate inhibits cell migration through mechanisms dependent on calcium, and that seem to involve inhibition of phosphotyrosine-neutral-phosphatases and disassembly of actin stress fibers.
“…UMR106 rat osteosarcoma cells were grown in DMEM containing 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin at 37°C in 5% CO 2 atmosphere (Cortizo and Etcheverry, 1995). Cells were seeded on 75 cm 2 flasks and subcultured using trypsin-EDTA.…”
Section: Cell Culture and Spheroid Preparationmentioning
Bisphosphonates are nonhydrolysable pyrophosphate analogues that prevent bone loss in several types of cancer. However, the mechanisms of anticancer action of bisphosphonates are not completely known. We have previously shown that nitrogen-containing bisphosphonates directly inhibit alkaline phosphatase of UMR106 rat osteosarcoma cells. In this study, we evaluated the effects of alendronate on the migration of UMR106 osteosarcoma using a model of multicellular cell spheroids, as well as the alendronate effect on neutral phosphatases. Alendronate significantly inhibited the migration of osteoblasts in a dose-dependent manner (10 − 6 -10 − 4 M). This effect was also dependent on calcium availability. The spheroid morphology and distribution of actin fibers were also affected by alendronate treatment. Alendronate dose-dependently inhibited neutral phosphatase activity in cell-free osteoblastic extracts as well as in osteoblasts in culture. Our results show that alendronate inhibits cell migration through mechanisms dependent on calcium, and that seem to involve inhibition of phosphotyrosine-neutral-phosphatases and disassembly of actin stress fibers.
“…Vanadate was also shown to exhibit strong metabolic and mitogenic effects, as demonstrated by in vitro studies using mammalian bone-derived cell lines (and also using rat calvaria and chondrocyte primary cultures) (Canalis 1985a;Kato et al 1987;Lau et al 1988). These effects have been extensively studied using two mammalian cell lines, MC3T3-E1 and UMR106, and important features, such as tyrosine phosphorylation and activation of signalling mechanisms, have been characterized (Barrio and Etcheverry 2006;Cortizo and Etcheverry 1995;Salice et al 1999). Although repetitively studied in mammals, vanadate effects have not been investigated in marine vertebrates (e.g.…”
Vanadium, a trace metal known to accumulate in bone and to mimic insulin, has been shown to regulate mammalian bone formation using in vitro and in vivo systems. In the present work, short-and long-term effects of metavanadate (containing monomeric, dimeric, tetrameric and pentameric vanadate species) and decavanadate (containing decameric vanadate species) solutions on the mineralization of a fish bone-derived cell line (VSa13) were studied and compared to that of insulin. After 2 h of incubation with vanadate (10 μM in monomeric vanadate), metavanadate exhibited higher accumulation rates than decavanadate (6.85±0.40 versus 3.95± 0.10 μg V/g of protein, respectively) in fish VSa13 cells and was also shown to be less toxic when applied for short periods. In longer treatments with both metavanadate and decavanadate solutions, similar effects were promoted: stimulation of cell proliferation and strong impairment (75%) of extracellular matrix (ECM) mineralization. The effect of both vanadate solutions (5 μM in monomeric vanadate), on ECM mineralization was increased in the presence of insulin (10 nM). It is concluded that chronic treatment with both vanadate solutions stimulated fish VSa13 cells proliferation and prevented ECM mineralization. Newly developed VSa13 fish cells appeared to be appropriate in the characterization of vanadate effects on vertebrate bone formation, representing a good alternative to mammalian systems.
“…The results have shown that due to the competition of the SO 4 2-ion in the equilibrium, only the first complexed species (ML) could be detected, before the whole system precipitated. These solutions however were investigated by EPR spectrometer in order to measure the A o and g o parameters.…”
Section: Resultsmentioning
confidence: 99%
“…The vanadyl solutions were prepared in two different ways as follows. In the first one, a proper mass of the VOSO 4 …”
Section: Methodsmentioning
confidence: 99%
“…As a micronutrient in living organisms, vanadium is important to prevent growth retardation and skeletal deformations in animals, and possibly participates in regulation of enzymatic processes. Further some complexes of V(IV) and V(V) have shown insulin-mimetic actions [2][3][4] . Humic acids present heterogeneous compositions that depend on the chemistry of how they were generated.…”
As constantes de estabilidade e parâmetros isotrópicos de EPR (Ao e go) foram obtidos por titulações potenciométricas e espectroscopia de EPR, empregando-se soluções aquosas, 15% v/v etanol, de compostos modelo de ácido húmico -ácido salicílico (SALA) -e do artefato de laboratório, ácido nitrohúmico -ácidos 3-nitrossalicílico (3-NSA), 5-nitrossalicílico (5-NSA) e 3,5-dinitrossalicílico -e cátions oxovanádio.Espectros The stability constants and the isotropic EPR parameters Ao (hyperfine splitting constant) and go (g value) were obtained by potentiometric titrations and EPR spectroscopy, respectively, of 85%v/v aqueous solutions of model compounds of humic acids -salicylic acid (SALA) -and both nitrohumic acids, a laboratory artifact -nitrosalicylic acids, 3-nitrosalicylic acid (3-NSA), 5-nitrosalicylic acid (5-NSA) and 3,5-dinitrosalicylic acid (3,5-DNSA) and oxovanadium cations.It was possible to record EPR spectra of those model compounds and the ion VO 2+ (V(IV)), and the stability constants were obtained from a solution of VO 3+ (V(V)), the values for the logarithms of the stability constants ranging from 12.77 ± 0.04 to 7.06 ± 0.05 for the species ML, and from 9.90 ± 0.04 to 4.06 ± 0.05 for the species ML2 according to the decrease in the acidity of the carboxylic and the hydroxyl groups in the aromatic ring of the model compounds studied as the -NO2 substituents were added. Species distribution diagrams were also obtained for the equilibria studied.The EPR parameters showed that as the logarithm of the overall stability constants increase, go values also increase, while Ao values show a tendency to decrease.
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