Different Effects of Fluoride Exposure on the Three Major Bone Cell Types

  • Ningning Jiang
  • Fengyang Guo
  • Boyao Sun
  • Xiuyun Zhang
  • Hui XuEmail author


Fluoride accumulates and is toxic to bones. Clinical bone lesions occur in a phased manner, being less severe early in the natural course of skeletal fluorosis. Previous research rarely focused on osteocyte, osteoclast, and osteoblast at the same time, although these three types of cells are involved in the process of fluorosis. In this study, commitment of bone cells was performed according to their respective characteristics. Osteocyte-like cells were verified by protein expression of sclerostin (SOST) in IDG-SW3 cell culture with mineral medium. Positive tartrate-resistant acid phosphatase (TRACP) staining, characteristic of osteoclasts, is observed in RAW264.7 cells after administration of RANKL. We successfully purified a high percentage (94%) of bone mesenchymal stem cells (BMSCs) co-expressing CD34 and CD44. Parallel studies were performed to observe cell viability and apoptosis rates in osteocyte, osteoclast, and osteoblast like cells by using MTT and Annexin V FITC assays. Our results demonstrated that osteocytes have a strong tolerance to high fluoride concentrations, while osteoclasts are more sensitive to changes of fluoride dose. The range of anabolic action of fluoride concentration on osteoblast was narrow. Notably, fluoride exposure aggravated apoptosis of osteocyte and osteoclast induced by administration of PTH and TGF-β, respectively. In short, three types of bone cells display disparate responses to fluoride exposure and to PTH- and TGF-β-induced apoptosis.


Sodium fluoride Osteoblast Osteoclast Osteocyte Apoptosis 



This study was supported by the National Natural Science Foundation of China (grant number 81673111, Project “Study on role of osteocyte and PTH/TGF-beta signaling pathway in the mechanism of bone turnover occurred in skeletal fluorosis”), the Natural Science Foundation of Jilin Province of China (20180101151JC).


This study was funded by Project [Study on role of osteocyte and PTH/TGF-beta signaling pathway in the mechanism of bone turnover occurred in skeletal fluorosis] supported by National Natural Science Foundation of China (grant number 81673111) and by the Natural Science Foundation of Jilin Province of China (20180101151JC).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Brothwell D, Limeback H (2003) Breastfeeding is protective against dental fluorosis in a nonfluoridated rural area of Ontario, Canada. J Hum Lact 19:386–390CrossRefPubMedGoogle Scholar
  2. 2.
    Hellwig E, Lennon AM (2004) Systemic versus topical fluoride. Caries Res 38:258–262CrossRefPubMedGoogle Scholar
  3. 3.
    Bhawal UK, Lee HJ, Arikawa K, Shimosaka M, Suzuki M, Toyama T, Sato T, Kawamata R, Taguchi C, Hamada N, Nasu I, Arakawa H, Shibutani K (2015) Micromolar sodium fluoride mediates anti-osteoclastogenesis in Porphyromonas gingivalis-induced alveolar bone loss. Int J Oral Sci 7(4):242–249CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Fina BL, Lombarte M, Rigalli JP, Rigalli A (2014) Fluoride increases superoxide production and impairs the respiratory chain in ROS 17/2.8 osteoblastic cells. PLoS One 9(6):e100768CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Woo SM, Rosser J, Dusevich V, Kalajzic I, Bonewald LF (2011) Cell line IDG-SW3 replicates osteoblast-to-late-osteocyte differentiation in vitro and accelerates bone formation in vivo. J Bone Miner Res 26(11):2634–2646CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Cuetara BLV, Crotti TN, O'Donoghue AJ et al (2006) Cloning and characterization of osteoclast precursors from the RAW264.7 cell line. In Vitro Cell Dev Biol Anim 42(7):182–188CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Nadri S, Soleimani M (2007) Isolation murine mesenchymal stem cells by positive selection. Vitro Cell Dev Biol Anim 43(8–9):276–282CrossRefGoogle Scholar
  8. 8.
    Bonewald LF, Johnson ML (2008) Osteocytes, mechanosensing and Wnt signaling. Bone 42(4):606–615CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Xiong J, Onal M, Jilka RL, Weinstein RS, Manolagas SC, O'Brien CA (2011) Matrix-embedded cells control osteoclast formation. Nat Med 17:1235–1241CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Lundy MW, Stauffer M, Wergedal JE, Baylink DJ, Featherstone JDB, Hodgson SF, Riggs BL (1995) Histomophometric analysis of iliac crest bone biopsies in placebo-treated versus fluoride-treated subjects. Osteoporosis Int 5:115–129CrossRefGoogle Scholar
  11. 11.
    Chachra D, Turner CH, Dunipace AJ, Grynpas MD (1999) The effect of fluoride treatment on bone mineral in rabbits. Calcif Tissue Int 64(4):345–351CrossRefPubMedGoogle Scholar
  12. 12.
    Mousny M, Omelon S, Wise L, Everett ET, Dumitriu M, Holmyard DP, Banse X, Devogelaer JP, Grynpas MD (2008) Fluoride effects on bone formation and mineralization are influenced by genetics. Bone 43(6):1067–1074CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Matsuda SS, Silva TL, Buzalaf MA, Rodrigues AC, de Oliveira RC (2014) Differential effects of fluoride during osteoblasts mineralization in C57BL/6J and C3H/HeJ inbred strains of mice. Biol Trace Elem Res 161(1):123–129CrossRefPubMedGoogle Scholar
  14. 14.
    Everett ET (2011) Fluoride's effects on the formation of teeth and bones, and the influence of genetics. J Dent Res 90(5):552–560CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Yu H, Jiang N, Yu X, Zhao Z, Zhang X, Xu H (2018) The role of TGF-β receptor 1-smad3 signaling in regulating the osteoclastic mode affected by fluoride. Toxicology 393:73–82CrossRefPubMedGoogle Scholar
  16. 16.
    Kaminsky LS, Mahoney MC, Leach J (1990) Fluoride: benefits and risks of exposure. Crit Rev Oral Biol Med 1:261–281CrossRefPubMedGoogle Scholar
  17. 17.
    Koide M, Kobayashi Y (2018) Regulatory mechanisms of sclerostin expression during bone remodeling. J Bone Miner Metab.
  18. 18.
    Fulzele K, Dedic C, Lai F, Bouxsein M, Lotinun S, Baron R, Divieti Pajevic P (2018) Loss of Gsα in osteocytes leads to osteopenia due to sclerostin induced suppression of osteoblast activity. Bone 117:138–148CrossRefPubMedGoogle Scholar
  19. 19.
    Hayashi M, Nakashima T, Yoshimura N et al (2019) Autoregulation of osteocyte Sema3A orchestrates estrogen action and counteracts bone aging. Cell Metab S1550-4131(18):30758–30757Google Scholar
  20. 20.
    Bellido T, Saini V, Pajevic PD (2013) Effects of PTH on osteocyte function. Bone 54(2):250–257CrossRefPubMedGoogle Scholar
  21. 21.
    Houde N, Chamoux E, Bisson M, Roux S (2009) Transforming growth factor-beta1 (TGF-beta1) induces human osteoclast apoptosis by up-regulating Bim. J Biol Chem 284(35):23397–23404CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Pharmaceutical SciencesJilin UniversityChangchunPeople’s Republic of China
  2. 2.China-Japan Union HospitalJilin UniversityChangchunPeople’s Republic of China

Personalised recommendations