Cell Biology and Toxicology

, Volume 24, Issue 3, pp 253–263 | Cite as

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

  • Daniel M. Tiago
  • Vincent Laizé
  • M. Leonor Cancela
  • Manuel Aureliano


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.


Bone-derived cell line Vanadate Decavanadate Insulin-mimetic properties Vertebrate bone formation Teleost fish Sparus aurata 



Dulbecco’s modified Eagle medium


Extracellular matrix


Extracellular signal-regulated kinase


Fetal bovine serum


Mitogen-activated protein kinase




Phosphate-buffered saline


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.


  1. Amado AM, Aureliano M, Ribeiro-Claro PJA, Teixeira-Dias JJ. Combined Raman and V-51 NMR spectroscopic study of vanadium (V) oligomerization in aqueous alkaline-solutions. J Raman Spectrosc 1993;24:699–703.CrossRefGoogle Scholar
  2. Anke M. Vanadium – An element both essential and toxic to plants, animals and humans? Anal Real Acad Nac Farm 2004;70:961–99.Google Scholar
  3. Aureliano M, Madeira VM. Interactions of vanadate oligomers with sarcoplasmic reticulum Ca(2+)-ATPase. Biochim Biophys Acta 1994;1221:259–71.PubMedCrossRefGoogle Scholar
  4. Aureliano M, Gandara RM. Decavanadate effects in biological systems. J Inorg Biochem 2005;99:979–85.PubMedCrossRefGoogle Scholar
  5. Aureliano M, Joaquim N, Sousa A, Martins H, Coucelo JM. Oxidative stress in toadfish (Halobactrachus didactylus) cardiac muscle. Acute exposure to vanadate oligomers. J Inorg Biochem 2002;90:159–65.PubMedCrossRefGoogle Scholar
  6. Barrio DA, Etcheverry SB. Vanadium and bone development: putative signaling pathways. Can J Physiol Pharmacol 2006;84:677–86.PubMedCrossRefGoogle Scholar
  7. Barrio DA, Braziunas MD, Etcheverry SB, Cortizo AM. Maltol complexes of vanadium (IV) and (V) regulate in vitro alkaline phosphatase activity and osteoblast-like cell growth. J Trace Elem Med Biol 1997;11:110–5.PubMedGoogle Scholar
  8. Bracken WM, Sharma RP, Elsner YY. Vanadium accumulation and subcellular distribution in relation to vanadate induced cytotoxicity in vitro. Cell Biol Toxicol 1985;1:259–68.PubMedCrossRefGoogle Scholar
  9. Canalis E. Effect of sodium vanadate on deoxyribonucleic acid and protein synthesis in culture rat calvariae. Endocrinology 1985a;116:855–62.PubMedGoogle Scholar
  10. Canalis E. Effect of sodium vanadate on deoxyribonucleic acid and protein syntheses in cultured rat calvariae. Endocrinology 1985b;116:855–62.PubMedCrossRefGoogle Scholar
  11. Cheatham B, Kahn CR. Insulin action and the insulin signaling network. Endocr Rev 1995;16:117–42.PubMedCrossRefGoogle Scholar
  12. Cohick WS, Clemmons DR. The insulin-like growth factors. Annu Rev Physiol 1993;55:131–53.PubMedCrossRefGoogle Scholar
  13. Cortizo AM, Etcheverry SB. Vanadium derivatives act as growth factor-mimetic compounds upon differentiation and proliferation of osteoblast-like UMR106 cells. Mol Cell Biochem 1995;145:97–102.PubMedCrossRefGoogle Scholar
  14. Cortizo AM, Bruzzone L, Molinuevo S, Etcheverry SB. A possible role of oxidative stress in the vanadium-induced cytotoxicity in the MC3T3E1 osteoblast and UMR106 osteosarcoma cell lines. Toxicology 2000;147:89–99.PubMedCrossRefGoogle Scholar
  15. Cortizo AM, Molinuevo MS, Barrio DA, Bruzzone L. Osteogenic activity of vanadyl(IV)-ascorbate complex: evaluation of its mechanism of action. Int J Biochem Cell Biol 2006;38:1171–80.PubMedCrossRefGoogle Scholar
  16. Crans DC, Smee JJ, Gaidamauskas E, Yang L. The chemistry and biochemistry of vanadium and the biological activities exerted by vanadium compounds. Chem Rev 2004;104:849–902.PubMedCrossRefGoogle Scholar
  17. Dubyak GR, Kleinzeller A. The insulin-mimetic effects of vanadate in isolated rat adipocytes. Dissociation from effects of vanadate as a (Na+-K+) ATPase inhibitor. J Biol Chem 1980;255:5306–12.PubMedGoogle Scholar
  18. Etcheverry SB, Cortizo AM. Bioactivity of vanadium compounds on cells in culture. In: Nriagu JO, editor. Vanadium in the environment. New York: John Wiley & Sons; 1998.Google Scholar
  19. Etcheverry SB, Apella MC, Baran EJ. A model study of the incorporation of vanadium in bone. J Inorg Biochem 1984;20:269–74.PubMedCrossRefGoogle Scholar
  20. Etcheverry SB, Crans DC, Keramidas AD, Cortizo AM. Insulin-mimetic action of vanadium compounds on osteoblast-like cells in culture. Arch Biochem Biophys 1997;338:7–14.PubMedCrossRefGoogle Scholar
  21. Goldwaser I, Gefel D, Gershonov E, Fridkin M, Shechter Y. Insulin-like effects of vanadium: basic and clinical implications. J Inorg Biochem 2000;80:21–5.PubMedCrossRefGoogle Scholar
  22. Heyliger CE, Tahiliani AG, McNeill JH. Effect of vanadate on elevated blood glucose and depressed cardiac performance of diabetic rats. Science 1985;227:1474–7.PubMedCrossRefGoogle Scholar
  23. Kato Y, Iwamoto M, Koike T, Suzuki F. Effect of vanadate on cartilagematrix proteoglycan synthesis in rabbit costal chondrocyte cultures. J Cell Biol 1987;104:311–9.PubMedCrossRefGoogle Scholar
  24. Kono SJ, Oshima Y, Hoshi K, Bonewald LF, Oda H, Nakamura K, et al. Erk pathways negatively regulate matrix mineralization. Bone 2007;40:68–74.PubMedCrossRefGoogle Scholar
  25. Lau KH, Tanimoto H, Baylink DJ. Vanadate stimulates bone cell proliferation and bone collagen synthesis in vitro. Endocrinology 1988;123:2858–67.PubMedGoogle Scholar
  26. LeRoith D. Insulin-like growth factor I receptor signaling-overlapping or redundant pathways? Endocrinology 2000;141:1287–8.PubMedCrossRefGoogle Scholar
  27. Malich G, Markovic B, Winder C. The sensitivity and specificity of the MTS tetrazolium assay for detecting the in vitro cytotoxicity of 20 chemicals using human cell lines. Toxicology 1997;124:179–92.PubMedCrossRefGoogle Scholar
  28. McCarthy TL, Ji C, Centrella M. Links among growth factors, hormones, and nuclear factors with essential roles in bone formation. Crit Rev Oral Biol Med 2000;11:409–22.PubMedCrossRefGoogle Scholar
  29. Meyerovitch J, Farfel Z, Sack J, Shechter Y. Oral administration of vanadate normalizes blood glucose levels in streptozotocin-treated rats. Characterization and mode of action. J Biol Chem 1987;262:6658–62.PubMedGoogle Scholar
  30. Moriyama S, Ayson FG, Kawauchi H. Growth regulation by insulin-like growth factor-I in fish. Biosci Biotechnol Biochem 2000;64:1553–62.PubMedCrossRefGoogle Scholar
  31. Murshed M, Harmey D, Millan JL, McKee MD, Karsenty G. Unique coexpression in osteoblasts of broadly expressed genes accounts for the spatial restriction of ECM mineralization to bone. Genes Dev 2005;19:1093–104.PubMedCrossRefGoogle Scholar
  32. Nielsen FH, Uthus EO. The essentiality and metabolism of vanadium. In: Chasteen ND, editor. Vanadium in biological systems. Boston: Kluwer Academic Publishers; 1990.Google Scholar
  33. Pombinho AR, Laize V, Molha DM, Marques SM, Cancela ML. Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res 2004;315:393–406.PubMedCrossRefGoogle Scholar
  34. Quarto R, Campanile G, Cancedda R, Dozin B. Thyroid hormone, insulin and glucocorticoids are sufficient to support chondrocyte differentiation to hypertrophy: a serum-free analysis. J Cell Biol 1992;119:989–95.PubMedCrossRefGoogle Scholar
  35. Rehder D. The coordination chemistry of vanadium as related to its biological functions. Coord Chem Rev 1999;182:297–322.CrossRefGoogle Scholar
  36. Rehder D. Biological and medicinal aspects of vanadium. Inorg Chem Commun 2003;6:604–17.CrossRefGoogle Scholar
  37. Salice VC, Cortizo AM, Gomez Dumm CL, Etcheverry SB. Tyrosine phosphorylation and morphological transformation induced by four vanadium compounds on MC3T3E1 cells. Mol Cell Biochem 1999;198:119–28.PubMedCrossRefGoogle Scholar
  38. Shechter Y. Perspective in diabetes: insulin-mimetic effects of vanadate. Possible implications for future treatment of diabetes. Diabetes 1990;39:1–5.PubMedCrossRefGoogle Scholar
  39. Shechter Y, Karlish SJ. Insulin-like stimulation of glucose oxidation in rat adipocytes by vanadyl (IV) ions. Nature 1980;284:556–8.PubMedCrossRefGoogle Scholar
  40. Shechter Y, Li J, Meyerovitch J, Gefel D, Bruck R, Elberg G, et al. Insulin-like actions of vanadate are mediated in an insulin-receptor-independent manner via non-receptor protein tyrosine kinases and protein phosphotyrosine phosphatases. Mol Cell Biochem 1995;153:39–47.PubMedCrossRefGoogle Scholar
  41. Shisheva A, Shechter Y. A cytosolic protein tyrosine kinase in rat adipocytes. FEBS Lett 1992;300:93–6.PubMedCrossRefGoogle Scholar
  42. Shisheva A, Shechter Y. Role of cytosolic tyrosine kinase in mediating insulin-like actions of vanadate in rat adipocytes. J Biol Chem 1993;268:6463–9.PubMedGoogle Scholar
  43. Soares SS, Martins H, Aureliano M. Vanadium distribution following decavanadate administration. Arch Environ Contam Toxicol 2006;50:60–4.PubMedCrossRefGoogle Scholar
  44. Soares SS, Martins H, Duarte RO, Moura JJG, Coucelo J, Gutiérrez-Merino C, et al. Vanadium distribution, lipid peroxidation and oxidative stress markers upon decavanadate in vivo administration. J Inorg Biochem 2007;101:80–8.PubMedCrossRefGoogle Scholar
  45. Tiago T, Aureliano M, Gutierrez-Merino C. Decavanadate binding to a high affinity site near the myosin catalytic centre inhibits F-actin-stimulated myosin ATPase activity. Biochemistry 2004;43:5551–61.PubMedCrossRefGoogle Scholar
  46. Yang XG, Yang XD, Yuan L, Wang K, Crans DC. The permeability and cytotoxicity of insulin-mimetic vanadium compounds. Pharm Res 2004;21:1026–33.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Daniel M. Tiago
    • 1
  • Vincent Laizé
    • 1
  • M. Leonor Cancela
    • 1
  • Manuel Aureliano
    • 1
    • 2
  1. 1.Centre of Marine Sciences (CCMAR)University of Algarve, Campus GambelasFaroPortugal
  2. 2.Faculdade de Ciências e Tecnologia (FCT)Universidade do Algarve, Campus de GambelasFaroPortugal

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