Protective Effect of 4% Titanium Tetrafluoride Varnish on Dentin Demineralization Using a Microcosm Biofilm Model.

This study evaluated the effect of titanium tetrafluoride (TiF4) varnish on the development of dentin carious lesions. Bovine root dentin samples were treated for 6 h with: (A) 4% TiF4 varnish (2.45% F); (B) 5.42% sodium fluoride (NaF) varnish (2.45% F); (C) 2% chlorhexidine (CHX) gel - positive control; (D) placebo varnish; or (E) untreated - negative control (n = 4 × biological triplicate, n = 12). Treated dentin samples were exposed to human saliva mixed with McBain saliva (1:50) for the first 8 h in 24-well plates. Thereafter, the medium was removed, and McBain saliva containing 0.2% sucrose was applied for 16 h. From days 2 to 5, McBain saliva with sucrose was replaced daily (37°C, 5% CO2). The demineralization was measured using transverse microradiography, while the effect on biofilm was analyzed using viability, extracellular polysaccharide (EPS), and lactic acid production assays. The data were statistically analyzed (p < 0.05). All treatments (fluorides and CHX) significantly reduced the biofilm viability compared to placebo varnish and negative control. However, none of them was able to reduce the colony-forming unit counting for total microorganism, total streptococci, and Streptococcus mutans. NaF significantly reduced the number of Lactobacillus sp. compared to negative control. No effect was seen on lactic acid production neither on EPS synthesis, except that CHX significantly reduced the amount of insoluble EPS. Both fluorides were able to reduce dentin demineralization compared to placebo varnish and negative control; TiF4 had a better effect in reducing mineral loss and lesion depth than NaF. Therefore, TiF4 varnish has the best protective effect on dentin carious lesion formation using this model.

TiF4 and NaF varnishes induce low levels of apoptosis in murine and human fibroblasts through mitochondrial Bcl-2 family and death receptor signalling.

OBJECTIVES: This study evaluated the level and mechanism of apoptosis in human gingival fibroblasts (HGF) and murine fibroblasts (NIH/3T3) treated with a titanium tetrafluoride (TiF4) varnish compared those treated with a sodium fluoride (NaF) varnish. METHODS: Cells were treated with a TiF4, NaF (both 2.45%F) or placebo varnish for 6 h and were then examined using the TUNEL method. The activities of caspase-3, -8 and -9 were assessed. cDNA for Bax, Bad, Bcl-2 and Fas-L was amplified by quantitative PCR. Bax, Bcl-2 and Fas-L were further detected by western blot analysis. RESULTS: Both fluorides similarly increased the percentage of apoptosis, while they failed to activate caspases. The Bax/Bcl-2 gene expression ratio was not altered by either fluoride treatment regardless of the type of cell. NaF varnish increased the amplification of the Fas-L gene in NIH/3T3 and HGF cells, while treatment with the TiF4 varnish resulted in a lower Bad/Bcl-2 expression ratio compared to that of the control for NIH/3T3 cells, but not for HGF cells. No effect of the fluorides was detected in the protein analysis. CONCLUSIONS: NaF and TiF4, at the studied conditions, similarly induce a low level of apoptosis, with consequent modest activation of the Bcl-2 and Fas-l-dependent signalling pathways. Generally, HGF cells are more susceptible to the fluoride effect than NIH/3T3 cells.