The cytotoxic effect of TiF4 and NaF on fibroblasts is influenced by the experimental model, fluoride concentration and exposure time

PLoS One. 2017 Jun 14;12(6):e0179471. doi: 10.1371/journal.pone.0179471. eCollection 2017.

Abstract

Objective: Titanium tetrafluoride (TiF4) has shown promising effect in preventing tooth lesions. Therefore, we compared the cytotoxicity of TiF4 with sodium fluoride (NaF) (already applied in Dentistry) considering different fluoride concentrations, pH values and experimental models.

Materials and methods: Step 1) NIH/3T3 fibroblasts were exposed to mediums containing NaF or TiF4 (from 0.15 to 2.45% F), both at native and adjusted pH, for 6 h. Step 2) NIH/3T3 were exposed to NaF or TiF4 varnishes with 0.95, 1.95 or 2.45% F (native pH), for 6, 12 or 24 h. We applied MTT (1st and 2nd steps) and Hoescht/PI stain (2nd step) assays. Step 3) NIH/3T3 were exposed to NaF or TiF4 varnish (2.45% F), at native pH, for 6 or 12 h. The cell stiffness was measured by atomic force microscopy (AFM).

Results: Step 1) All cells exposed to NaF or TiF4 mediums died, regardless of the F concentration and pH. Step 2) Both varnishes, at 1.90 and 2.45% F, reduced cell viability by similar extents (33-86% at 6 h, 35-93% at 12 h, and 87-98% at 24 h) compared with control, regardless of the type of fluoride. Varnishes with 0.95% F did not differ from control. Step 3) TiF4 and NaF reduced cell stiffness to a similar extent, but only TiF4 differed from control at 6 h.

Conclusions: Based on the results of the 3 experimental steps, we conclude that TiF4 and NaF have similar cytotoxicity. The cytotoxicity was dependent on F concentration and exposure time. This result gives support for testing the effect of TiF4 varnish in vivo.

MeSH terms

  • Animals
  • Cariostatic Agents / pharmacology
  • Cell Physiological Phenomena / drug effects*
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Fluorides / pharmacology*
  • Hydrogen-Ion Concentration
  • Mice
  • Microscopy, Atomic Force
  • Models, Theoretical
  • NIH 3T3 Cells
  • Sodium Fluoride / pharmacology*
  • Time Factors
  • Titanium / pharmacology*

Substances

  • Cariostatic Agents
  • Sodium Fluoride
  • Titanium
  • Fluorides
  • titanium tetrafluoride

Grants and funding

This work was supported by FAPESP [Grant numbers: 09/17360-4, 10/14984-4, 13/20372-0, 13/13484-6, 14/16415-8 and 15/05009-1]; and Kidney Research UK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.