Impairment of mineralization by metavanadate and decavanadate solutions in a fish bone-derived cell line

Abstract

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.

Vanadate proliferative and anti‐mineralogenic effects are mediated by MAPK and PI‐3K/Ras/Erk pathways in a fish chondrocyte cell line

“…ALP activity was then measured in cellular extracts of mineralized cultures (4 weeks) in the presence of increasing concentrations of vanadate: activity was stimulated at low concentration (5 lM) while strongly inhibited at higher concentrations (from 25 to 500 lM) of vanadate. We propose, in agreement with recent result demonstrating the accumulation of vanadium in VSa13 cells [8], that although cultures were treated with non-inhibitory concentrations of vanadate, these may accumulate intracellularly and reach concentrations inhibitory for ALP activity therefore affecting mechanisms of ECM mineralization. On the contrary, the increase of total collagen content in vanadate-treated mineralized cultures (most probably the result of increased type X collagen as observed in differentiating ATDC5 chondrocyte cells [16]) can hardly explain the decrease of ECM mineralization since increased collagen production is rather likely to benefit ECM mineralization.…”

“…Wortmannin and PD98059 were then tested for their effect on VSa13 ECM mineralization and for their capacity of reverting vanadate and insulin inhibitory effects previously demonstrated [8] and further confirmed here. Mineralizing VSa13 Fig.…”

“…, wortmannin, insulin and IGF-1 stock solutions Vanadate solution (50 mM, pH 6-7) was prepared as described previously [8]. PD98059 and wortmannin were solubilised in DMSO at 6 and 10 mg/ml, respectively.…”

“…A role for vanadium in bone formation has been demonstrated [4,5] but mechanisms of action have remained largely unexplored. In vitro studies, using calvaria primary cultures [6,7] and bone-derived cell lines [5,8,9], have evidenced the regulation of bonerelated enzymes activity (e.g. alkaline phosphatase (ALP) and protein tyrosine phosphatases), the stimulation of bonerelated protein synthesis (e.g.…”

Synthesis, Characterization and Crystal Structure of a Novel Decavanadate Salt, [V0.50(H2O)5]2[H2(V10O28)]·4(H2O)

“…22 In aqueous medium, under some conditions, the vanadate oligomers may be in equilibrium with monomeric (V 1 ), dimeric (V 2 ) and tetrameric (V 4 ) vanadate species. 23 In the literature, the crystal structures of the decavanadate anion, [V 10 O 28 ] 6-, 24 27 ), containing the different transition metal complex cations were characterized. The aim of this research was the synthesis, physicochemical characterization both in the aqueous solution and in the solid state, and the determination of molecular structure of a novel vanadium complex in the presence of chinhydrone.…”