Impairment of mineralization by metavanadate and decavanadate solutions in a fish bone-derived cell line
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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.
KeywordsBone-derived cell line Vanadate Decavanadate Insulin-mimetic properties Vertebrate bone formation Teleost fish Sparus aurata
Dulbecco’s modified Eagle medium
Extracellular signal-regulated kinase
Fetal bovine serum
Mitogen-activated protein kinase
Phosphatidyl inositol-3 kinase
The authors thank Dr. Hélio Martins from the Faculty of Sciences and Technology (FCT) of the University of Algarve for his technical help in the course of atomic absorption spectrometry experiments. Thanks also to J.J.G. Moura, REQUIMTE, New University of Lisbon, for the esteemed collaboration in the NMR studies. DMT was the recipient of a Ph.D. fellowship (BD/12773/2003) from the Portuguese Science and Technology Foundation.
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