Skip to main content

Methodology developed for the simultaneous measurement of bone formation and bone resorption in rats based on the pharmacokinetics of fluoride

Abstract

This paper describes a novel methodology for the simultaneous estimation of bone formation (BF) and resorption (BR) in rats using fluoride as a nonradioactive bone-seeker ion. The pharmacokinetics of flouride have been extensively studied in rats; its constants have all been characterized. This knowledge was the cornerstone for the underlying mathematical model that we used to measure bone fluoride uptake and elimination rate after a dose of fluoride. Bone resorption and formation were estimated by bone fluoride uptake and elimination rate, respectively. ROC analysis showed that sensitivity, specificity and area under the ROC curve were not different from deoxypiridinoline and bone alkaline phosphatase, well-known bone markers. Sprague–Dawley rats with modified bone remodelling (ovariectomy, hyper, and hypocalcic diet, antiresorptive treatment) were used to validate the values obtained with this methodology. The results of BF and BR obtained with this technique were as expected for each biological model. Although the method should be performed under general anesthesia, it has several advantages: simultaneous measurement of BR and BF, low cost, and the use of compounds with no expiration date.

This is a preview of subscription content, access via your institution.

Fig. 1

References

  1. Puche RC, Rigalli A, Bocanera R, Tozzini R, Poudes G, Roberti A (1990) A Clinical test for calcium absorption in climacteric women. A two traces, single blood sample procedure with corrections for body size and skeletal turnover. Medicina 50:16–20

    CAS  PubMed  Google Scholar 

  2. Puche RC, Rigalli A, Trumper L, Dip O, Pereyra JL, Poudes G, Roberti A, Bocanera R, Tozzini R (1991) Estimation of bone turnover in climacteric women by the whole body retention of fluoride. Maturitas 14:57–64

    CAS  PubMed  Article  Google Scholar 

  3. Rigalli A, Ballina JC, Beinlich A, Alloatti R, Puche RC (1994) Pharmacokinetics differences between sodium fluoride and sodium monofluorphosphate and comparative bone mass increasing activity of both compounds, in the rat. Arzneimittelforschung 44:762–766

    CAS  PubMed  Google Scholar 

  4. Lupo M, Buzalaf MAR, Rigalli A (2011) Effect of fluoridated water on plasma insulin levels and glucose homeostasis in rats with renal deficiency. Biol Trace Elem Res 140:198–207

    CAS  PubMed  Article  Google Scholar 

  5. Olfert ED, Cross BM, McWilliam AA (1993) Canadian Council on animal care Guidelines. Guide to the care and use of experimental animal. http://www.ccac.ca

  6. De Candia F, Rigalli A, Di Loreto V (2009) Anesthesia and analgesia. In: Rigalli A, Di Loreto V (eds) Experimental surgical models in the laboratory rat. Taylor and Francis Group, CRC Press, Boca Ratón, pp 21–30

    Google Scholar 

  7. Lupo M, Di Loreto V, Rigalli A (2009) Substances administration. In: Rigalli A, Di Loreto V (eds) Experimental surgical models in the laboratory rat. Taylor and Francis Group, CRC Press, Boca Ratón, pp 55–61

    Google Scholar 

  8. Rigalli A, Alloatti R, Puche RC (1999) Measurement of total and diffusible serum fluoride. J Clin Lab Anal 13:151–157

    CAS  PubMed  Article  Google Scholar 

  9. Farley JR, Chesnut CJ, Baylink DJ (1981) Improved method for quantitative determination in serum alkaline phosphatase of skeletal origin. Clin Chem 27:2002–2007

    CAS  PubMed  Google Scholar 

  10. Di Loreto V, Pera L, Rigalli A (2009) Ovariectomy. In: Rigalli A, Di Loreto V (eds) Experimental Surgical Models in the Laboratory rat. Taylor and Francis Group, CRC Press, Boca Ratón, pp 149–153

    Google Scholar 

  11. Brance ML, Brun LR (2009) Histological procedures. In: Rigalli A, Di Loreto V (eds) Experimental surgical models in the laboratory rat. Taylor and Francis Group, CRC Press, Boca Ratón, pp 221–227

    Google Scholar 

  12. Robin X, Turck N, Hainard A, Tiberti N, Lisacek F, Sanchez J-C, Müller M (2011) pROC: an open-source package for R and S + to analyze and compare ROC curves. BMC Bioinformatics 12:77. doi:10.1186/1471-2105-12-77

    PubMed Central  PubMed  Article  Google Scholar 

  13. Brun L, Pera L, Rigalli A (2010) Bone morphometry and differences in bone fluorine containing compounds in rats treated with NaF and MFP. Biomed Pharmacother 64:1–6

    CAS  PubMed  Article  Google Scholar 

  14. Fawell J, Bailey K, Chilton J, Dahi E, Fewtrell L, Magara Y (2006) Fluoride in drinking-water. World Health Organization, IWA Publishing, London

    Google Scholar 

Download references

Acknowledgments

The authors thank Barbara Garcia and Hilda Moreno for technical assistance. This work was founded by Consejo Nacional de Investigaciones Científicas y Tecnologicas (PIP 112-200801-00462).

Conflict of interest

All authors declare that they have no conflicts of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Maela Lupo.

About this article

Cite this article

Lupo, M., Brance, M.L., Fina, B.L. et al. Methodology developed for the simultaneous measurement of bone formation and bone resorption in rats based on the pharmacokinetics of fluoride. J Bone Miner Metab 33, 16–22 (2015). https://doi.org/10.1007/s00774-013-0557-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00774-013-0557-3

Keywords

  • Bone remodeling
  • Mathematical model
  • Fluoride pharmacokinetics
  • Bone marker
  • Rat