Effect of TiF4, ZrF4, HfF4 and AmF on erosion and erosion/abrasion of enamel and dentin in situ

Objective

This in situ study aimed to analyse the impact of different tetrafluorides (TiF₄, ZrF₄ and HfF₄) and AmF on erosion and erosion plus abrasion of enamel and dentin.

Design

Ten volunteers took part in this crossover and double-blind study performed in 8 phases of each 3 days. In each phase, 2 bovine enamel and 2 dentin specimens were fixed in intraoral appliances. One enamel and one dentin sample were pretreated once with TiF₄, ZrF₄, HfF₄ or AmF (all 0.5 M F) for 60 s, while the other samples remained unfluoridated and served as control. Then, all samples were subjected to either erosion only (4 times/day, 90 s) or to erosion and abrasion (2 times/day, 30 s/sample). Toothbrushing abrasion was performed 90 min after the first and last erosion with an electrical toothbrush and fluoridated toothpaste at 1.2 N. After 3 days, enamel and dentin loss was assessed by profilometry (μm) and analysed by repeated measures ANOVA and paired t-test (p < 0.05).

Results

All fluoride solutions reduced enamel and dentin loss significantly compared to the controls. Generally, eroded samples showed less wear than eroded and abraded samples. The protective potential of the fluorides was not significantly different and was only slightly, but mostly not significantly, decreased by abrasion. The protective effect of the fluoride solutions was similar in enamel and dentin.

Conclusion

Tetrafluorides and AmF are able to reduce erosion and erosion plus abrasion in situ and are almost equally effective.     

Protective effect of experimental mouthrinses containing NaF and TiF4 on dentin erosive loss in vitro

Objective

This in vitro study assessed the anti-erosive effect of experimental mouthrinses containing TiF₄ and NaF on dentin erosive loss.

Material and Methods

Bovine dentin specimens were randomly allocated into the groups (n=15): 1) SnCl₂/NaF/AmF (Erosion Protection^®/GABA, pH 4.5, positive control); 2) experimental solution with 0.0815% TiF₄ (pH 2.5); 3) 0.105% NaF (pH 4.5); 4) 0.042% NaF+0.049% TiF₄ (pH 4.4); 5) 0.063% NaF+0.036% TiF₄ (pH 4.5); 6) no treatment (negative control). Each specimen was cyclically demineralized (Sprite Zero, pH 2.6, 4x90 s/day) and exposed to artificial saliva between the erosive challenges for 7 days. The treatment with the fluoride solutions was done 2x60 s/day, immediately after the first and the last erosive challenges of the day. Dentin erosive loss was measured by profilometry (μm). The data were analyzed using Kruskal Wallis/Dunn tests (p<0.05).

Results

Mouthrinses containing TiF₄ or Sn/F were able to show some protective effect against dentin erosive loss compared to negative control. The best anti-erosive effect was found for experimental solution containing 0.0815% TiF₄ (100% reduction in dentin loss), followed by 0.042% NaF+0.049% TiF⁴ (58.3%), SnCl₂/NaF/AmF (52%) and 0.063% NaF+0.036% TiF₄ (40%). NaF solution (13.3%) did not significantly differ from control.

Conclusion

The daily application of experimental mouthrinse containing TiF₄ and NaF has the ability to reduce dentin erosion, as well as Erosion Protection^® and TiF₄ alone.     

Effect of a single application of TiF4 and NaF varnishes and solutions on dentin erosion in vitro

Objectives

This in vitro study aimed to analyse the effect of a single application of TiF₄ and NaF varnishes and solutions to protect against dentin erosion.

Methods

Bovine root dentin samples were pre-treated with NaF-Duraphat varnish (2.26%F, pH 4.5), NaF/CaF₂-Duofluorid varnish (5.63%F, pH 8.0), NaF-experimental varnish (2.45%F, pH 4.5), TiF₄-experimental varnish (2.45%F, pH 1.2), NaF solution (2.26%F, pH 4.5), TiF₄ solution (2.45%F, pH 1.2) and placebo varnish (pH 5.0, no-F varnish control). Controls remained untreated. Ten samples in each group were then subjected to an erosive demineralisation (Sprite Zero, 4× 90 s/day) and remineralisation (artificial saliva, between the erosive cycles) cycling for 5 days. Dentin loss was measured profilometrically after pre-treatment and after 1, 3 and 5 days of de-remineralisation cycling. The data were statistically analysed by two-way ANOVA and Bonferroni's post hoc test (p < 0.05).

Results

After pre-treatment, TiF₄ solution significantly induced surface loss (1.08 ± 0.53 μm). Only Duraphat reduced the dentin loss overtime, but it did not significantly differ from placebo varnish (at 3rd and 5th days) and TiF₄ varnish (at 3rd day).

Conclusions

Duraphat varnish seems to be the best option to partially reduce dentin erosion. However, the maintenance of the effects of this treatment after successive erosive challenges is limited.     

Etching effect of acidic fluorides on human tooth enamel in vitro

ObjectiveThis in vitro study aimed to examine the etching effect of acidic fluoride solutions on enamel.Materials and methods24 human teeth divided into 48 enamel-specimens were partly isolated with impression material. Specimens were exposed for 10 min to 20 ml of the following solutions: 1.6% TiF₄, 3.9% SnF₂, 0.2% HF and 1.8% citric acid (CA). The isolation was removed and 24 specimens analysed by profilometry (Δheight: exposed/isolated enamel surfaces, surface roughness parameters). For the remaining 24 specimens [Ca²⁺] in the test solutions was analysed by atomic absorption spectroscopy.ResultsMedian Δheights (μm) after exposure were: TiF₄ 0.07, SnF₂ −0.03, HF −0.14 and CA −5.92. TiF₄-exposed surfaces showed both deposits and etched areas and exhibited statistically significant different surface roughness parameters compared to the HF- and SnF₂-exposed surfaces. Median [Ca²⁺] values (ppm): TiF₄ 1.88, SnF₂ 0.11, HF 0.10 and CA 2.17.ConclusionAt the [F] tested in this study it can be concluded that SnF₂- and HF solutions had negligible erosive effects on enamel. TiF₄ solution resulted in an incomplete surface deposition associated with calcium dissolution suggesting that TiF₄ applied as solution may not be advisable.     

Evaluation of fluoride release from experimental TiF4 and NaF varnishes in vitro

Fluoride varnishes play an important role in the prevention of dental caries, promoting the inhibition of demineralization and the increase of remineralization.

Objective

This study aimed to analyze the amount of fluoride released into water and artificial saliva from experimental TiF₄ and NaF varnishes, with different concentrations, for 12 h.

Material and Methods

Fluoride varnishes were applied on acrylic blocks and then immersed in 10 ml of deionized water and artificial saliva in polystyrene bottles. The acrylic blocks were divided in seven groups (n=10): 1.55% TiF₄ varnish (0.95% F, pH 1.0); 3.10% TiF₄ varnish (1.90% F, pH 1.0); 3.10% and 4% TiF₄ varnish (2.45% F, pH 1.0); 2.10% NaF varnish (0.95% F, pH 5.0); 4.20% NaF varnish (1.90% F, pH 5.0); 5.42% NaF varnish (2.45% F, pH 5.0) and control (no treatment, n=5). The fluoride release was analyzed after 1/2, 1, 3, 6, 9 and 12 h of exposure. The analysis was performed using an ion-specific electrode coupled to a potentiometer. Two-way ANOVA and Bonferroni''s test were applied for the statistical analysis (p<0.05).

Results

TiF₄ varnishes released larger amounts of fluoride than NaF varnishes during the first 1/2 h, regardless of their concentration; 4% TiF₄ varnish released more fluoride than NaF varnishes for the first 6 h. The peak of fluoride release occurred at 3 h. There was a better dose-response relationship among the varnishes exposed to water than to artificial saliva.

Conclusions

The 3.10% and 4% TiF₄ -based varnishes have greater ability to release fluoride into water and artificial saliva compared to NaF varnish; however, more studies must be conducted to elucidate the mechanism of action of TiF₄ varnish on tooth surface.     

TiF4 and NaF varnishes as anti-erosive agents on enamel and dentin erosion progression in vitro

Objective

This study assessed the effect of fluoride varnishes on the progression of tooth erosion in vitro. Material and Methods: Forty-eight enamel and 60 root dentin samples were previously demineralized (0.1% citric acid, pH 2.5, 30 min), leading to a baseline and erosive wear of 12.9 and 11.4 µm, respectively. The samples were randomly treated (6 h) with a 4% TiF₄ varnish (2.45%F-, pH 1.0), a 5.42% NaF varnish (2.45%F-, pH 5.0), a placebo varnish and no varnish (control). The samples were then subjected to erosive pH cycles (4x90 s/day in 0.1% citric acid, intercalated with artificial saliva) for 5 days. The increment of the erosive tooth wear was calculated. In the case of dentin, this final measurement was done with and without the demineralized organic matrix (DOM). Enamel and dentin data were analyzed using ANOVA/Tukey’s and Kruskal-Wallis/Dunn tests, respectively (p<0.05).

Results

The TiF₄ (mean±s.d: 1.5±1.1 µm) and NaF (2.1±1.7 µm) varnishes significantly reduced enamel wear progression compared to the placebo varnish (3.9±1.1 µm) and control (4.5±0.9 µm). The same differences were found for dentin in the presence and absence of the DOM, respectively: TiF₄ (average: 0.97/1.87 µm), NaF (1.03/2.13 µm), placebo varnish (3.53/4.47 µm) and control (3.53/4.36 µm).

Conclusion

The TiF₄ and NaF varnishes were equally effective in reducing the progression of tooth erosion in vitro.     

Combined effect of amoxicillin and sodium fluoride on the structure of developing mouse enamel in vitro

Objective

Excess fluoride intake during tooth development is known to cause dental fluorosis. It has also been suggested that amoxicillin use in early childhood is associated with enamel hypomineralization. The aim was to investigate separate and combined effects of sodium fluoride (NaF) and amoxicillin on enamel formation in vitro.

Design

Mandibular molar tooth germs of E18 mouse embryos were cultured for 10 days in a medium containing NaF (10, 12 or 15 μM) and/or amoxicillin (0.5, 1, 2 or 3.6 mg/mL) or sodium clavulanate (0.07 mg/mL) alone or in combination with 0.5 mg/mL of amoxicillin. Morphological changes were studied from the whole tooth photographs and histological tissue sections with light microscope.

Results

Only with the highest concentrations of NaF or amoxicillin alone the extent of enamel in the first molars measured as the vertical enamel height/crown height ratio was reduced (p < 0.01, p < 0.001, respectively). At lower concentrations, combination of NaF (12 μM) and amoxicillin (2 mg/mL) significantly reduced enamel extent compared with the controls (p < 0.001). Histologically, the ameloblasts were still columnar but poorly organized and the nascent enamel was often non-homogeneous. Enamel formation was not seen in any second molars exposed to 12 μM NaF and 2 mg/mL of amoxicillin (or higher concentrations) compared with the presence of enamel in half of the controls (p < 0.001).

Conclusions

Amoxicillin and NaF dose dependently affect developing enamel of mouse molars in vitro and the effects are potentiative. The clinical significance of the results remains to be studied.     

Validation of enamel erosion in vitro

Objectives

Many tools are available to quantify dental erosion, but each technique has its own inherent disadvantages. This study aims to validate the use of quantitative light-induced fluorescence (QLF) and non-contacting surface profilometry compared to the gold standard transverse microradiography (TMR) for the quantification of enamel erosion in vitro.

Methods

This was an in vitro laboratory based study. 60 bovine incisors were divided into 6 groups of 10. Each tooth's labial surface was completely varnished except for a window of enamel approximately 3 mm × 5 mm. Each was baseline imaged with QLF and non-contacting surface profilometry before being subjected to an erosive solution (pH 3.4) for up to 36 h. The lesions were imaged using non-contacting surface profilometry and QLF, sectioned and analysed with TMR. Correlation coefficients were calculated to assess the validity of the methods of measurement as compared to TMR.

Results

A range of lesion severities resulted. Mineral loss measured as ΔQ (QLF) and step height (profilometry), was recorded and confirmed by TMR. A correlation was found between ΔZ (TMR) and profilometry lesion depth of r = 0.648 (p < 0.001). A poorer correlation was found between ΔZ and ΔQ: r = 0.217 (p = 0.096).

Conclusions

Profilometry lesion depth and ΔZ correlated significantly. Both methods allow for quantification of erosive crater depth. QLF correlated poorly with ΔZ, but is useful for measuring subsurface loss of mineralisation. TMR is valuable but is destructive and can only be used in vitro. Currently only QLF can be used in vivo. Advances in these technologies may allow the development of non-destructive in vivo measurements of mineral loss, combining the positive features of each measurement method.     

Effect of tricalcium silicate (Ca3SiO5) bioactive material on reducing enamel demineralization: An in vitro pH-cycling study

Objectives

The aim of this study was to investigate the effect of Ca₃SiO₅ on reducing enamel demineralization under pH-cycling conditions.

Methods

Forty bovine enamel samples were treated under four conditions: group 1, double distilled water (negative control); group 2, 1000 ppm F (as NaF, positive control); group 3, Ca₃SiO₅ slurry; and group 4, Ca₃SiO₅–F slurry (Ca₃SiO₅ with 1000 ppm F aq.). All the specimens were treated with treatment materials 4 times each day. Samples in groups 1 and 2 were soaked in test solutions for 2 min and samples in groups 3 and 4 were painted in treatment slurry for 2 min. At times between treatments, they were immersed in citric acid solution 3 times a day and 15 s each time. All the procedures were repeated for 7 days. Knoop microhardness, scanning electron microscopy (SEM), X-ray diffraction (XRD) and atom force microscopy (AFM) were used to examine samples.

Results

After treatment for 7 days, enamels in all the groups were significantly softened. The extents of microhardness reduction were 52.3%, 28.5%, 28.5% and 20.2% for groups 1, 2, 3 and 4, respectively. Samples in the negative control group showed a typical acid etched pattern while enamels in other groups were relatively compact. There was no significant difference between samples treated with Ca₃SiO₅ and F. The combination of Ca₃SiO₅ with F showed the best effect on reducing enamel demineralization.

Conclusions

Ca₃SiO₅ is an effective material against enamel demineralization alone but in combination with F a better anti-demineralization effect may be obtained.     

Physical chemical effects of zinc on in vitro enamel demineralization

Objectives

Zinc salts are formulated into oral health products as antibacterial agents, yet their interaction with enamel is not clearly understood. The aim was to investigate the effect of zinc concentration [Zn²⁺] on the in vitro demineralization of enamel during exposure to caries-simulating conditions. Furthermore, the possible mechanism of zinc's action for reducing demineralization was determined.

Methods

Enamel blocks and synthetic hydroxyapatite (HAp) were demineralized in a range of zinc-containing acidic solutions (0–3565 ppm [Zn²⁺]) at pH 4.0 and 37 °C. Inductively coupled-plasma optical emission spectroscopy (ICP-OES) was used to measure ion release into solution. Enamel blocks were analysed by Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR), and HAp by X-ray diffraction (XRD) and neutron diffraction (ND).

Results

ICP-OES analysis of the acidic solutions showed a decrease in [Ca²⁺] and [PO₄³⁻] release with increasing [Zn²⁺]. FTIR revealed a α-hopeite (α-Zn₃(PO₄)₂.4H₂O)-like phase on the enamel surfaces at >107 ppm [Zn²⁺]. XRD and ND analysis confirmed a zinc-phosphate phase present alongside the HAp.

Conclusions

This study confirms that zinc reduces enamel demineralization. Under the conditions studied, zinc acts predominantly on enamel surfaces at PO₄³⁻ sites in the HAp lattice to possibly form an α-hopeite-like phase.

Clinical significance

These results have a significant implication on the understanding of the fundamental chemistry of zinc in toothpastes and demonstrate its therapeutic potential in preventing tooth mineral loss.     

Effect of air-abrasion pre-treatment with bioactive glass 45S5 on enamel surface loss after erosion/abrasion challenge

ObjectivesThe aim of this in vitro study was to evaluate the effect of air-abrasion surface pre-treatment with bioactive glass (BAG) 45S5 on enamel surface loss after erosion/abrasion challenge.MethodsTwenty-four sound bovine incisors were used for this study. Four experimental groups (n = 12) were assigned as follows: Group 1 was the negative control group, Group 2 specimens were treated with a SnF₂ gel (positive control group), Group 3 specimens were air-abraded with BAG 45S5 (ProSylc) and Group 4 received both treatments. The specimens were submitted to erosion/abrasion challenge using a common soft drink. Enamel surface loss was evaluated using an optical profilometer. Additionally, surface roughness (VSI method) and surface microhardness (Vickers method) changes were evaluated, as well as SEM and EDS analyses were performed on enamel surface. The data were statistically analyzed using one-way ANOVA and Tukey’s post-hoc test at a level of significance a = 0.05.ResultsSurface pre-treatment with BAG 45S5 reduced surface loss in comparison with negative control group (p < 0.001), which exhibited the highest surface loss of the experimental groups (p < 0.05). The positive control group (SnF₂ treatment) and Group 4 specimens, which received both air-abrasion pre-treatment and SnF₂ treatment, presented the lowest surface loss (p < 0.05), but did not show significant differences to each other (p = 0.65).SignificanceAir-abrasion pre-treatment with BAG 45S5 may be beneficial as an in-office preventive method for the limitation of enamel erosive wear induced by excessive consumption of soft drinks. The clinical significance of the results regarding this preventive method should be confirmed by clinical studies.     

Effect of fluoride varnish supplemented with sodium trimetaphosphate on enamel erosion and abrasion: An in situ/ex vivo study

Objective

To evaluate the effect of fluoride (F) varnishes supplemented or not with sodium trimetaphosphate (TMP) on enamel erosive wear followed or not by abrasion in situ.

Methods

Ten volunteers were selected and randomly divided into four groups, according to the varnishes tested: placebo (no F or TMP), 5% NaF (positive control), 2.5% NaF and 2.5% NaF/5% TMP. Enamel blocks (n = 4) were mounted in palatal devices and received an application of each test varnish, following a double-blind, crossover protocol. After 6 h, varnishes were completely removed and the blocks were subjected to erosive challenges by ex vivo immersion in citric acid (5 min, 4×/dia, 5 days). Following, half of the blocks were subjected to abrasion by brushing with a placebo dentifrice slurry for 15 s. Enamel wear (μm), surface hardness (SH_f) and cross-sectional hardness (ΔKHN) were assessed after each experimental period. Results were analyzed by ANOVA, Student–Newman–Keuls's test and Pearson correlation coefficient (p < 0.05).

Results

The fluoride varnish supplemented with TMP promoted significantly lower wear and ΔKHN when compared to the other groups after erosive challenges, followed or not by abrasion (p < 0.05). As for (SH_f) the fluoride varnish supplemented with TMP promoted similar results to the 5% NaF product, being significantly higher than the remaining groups after erosive and erosive + abrasive challenges (p < 0.05).

Conclusion

TMP significantly enhanced the effects of F on enamel wear after erosive challenges, followed or not by abrasion.     

Efficacy of the stannous ion and a biopolymer in toothpastes on enamel erosion/abrasion

Objectives

Sn²⁺ has promising erosion-inhibiting properties in solutions, but little is known about respective effects in toothpastes. In addition, biopolymers might have protecting potential. Aim of this study was to investigate the effects of Sn²⁺ in toothpastes and of a biopolymer (chitosan) added to a Sn²⁺ formulation on erosion/abrasion.

Methods

Enamel samples were subjected to cyclic erosion procedures (10 days; 0.50% citric acid, pH 2.5; 6× 2 min/day), and brushing (2× 15 s/day, load 200 g) during immersion in slurries (2 min). The toothpastes were NaF formulations (NaF/1, NaF/2, NaF/3) and Sn²⁺ formulations (NaF/SnCl₂, AmF/SnF₂, AmF/NaF/SnCl₂) and AmF/NaF/SnCl₂ + 0.5% chitosan.Declared concentrations of active ingredients in toothpastes were 1400–1450 μg/g F⁻ and 3280–3500 μg/g Sn²⁺. Negative controls were erosion only and placebo, positive control was a SnF₂ gel. Tissue loss was quantified profilometrically, Sn on enamel surfaces was measured by energy dispersive X-ray spectroscopy.

Results

Loss values (μm) for erosion only and placebo were 14.4 ± 4.5 and 20.2 ± 3.8, respectively, and 4.6 ± 1.9 for the positive control (p ≤ 0.001 each compared to erosion only). The other loss values were: NaF/1 16.5 ± 3.0, NaF/2 14.0 ± 2.7, NaF/3 12.6 ± 3.9, NaF/SnCl₂ 14.7 ± 5.1, AmF/SnF₂ 13.5 ± 4.8, AmF/NaF/SnCl₂ 12.4 ± 4.2, AmF/NaF/SnCl₂ + chitosan 6.6 ± 3.5 (except NaF/1 all p ≤ 0.01 compared to placebo). AmF/NaF/SnCl₂/chitosan was more effective than all other toothpastes (p ≤ 0.01 each). Sn on the enamel surface ranged between 1.3 ± 0.3 and 2.8 ± 0.04 wt.% with no obvious relationship with efficacy.

Conclusions

The NaF and Sn²⁺ toothpastes without chitosan exhibited similar anti-erosion and abrasion-prevention effects. The experimental Sn²⁺ formulation with chitosan revealed promising results similar to those of the positive control.

Clinical significance

NaF toothpastes offer a degree of protection against erosion/abrasion, which is likely sufficient for most subjects with average acid exposures. For patients with initial erosive lesions, however, more effective toothpaste is desirable. The combination of Sn²⁺ and a biopolymer appears promising in this context.     

Effect of rinsing with water immediately after neutral gel and foam fluoride topical application on enamel remineralization: An in situ study

Objective

The aim of this study was to evaluate, in situ, the effect of rinsing with water immediately after neutral fluoride foam application (Foam) or fluoride gel application (Gel).

Design

Ten volunteers wore acrylic palatal appliances containing 4 enamel blocks selected by surface hardness with artificial caries. Five experimental regimes of 3 days each were set according to treatment: placebo; Gel and Foam followed by no rinsing or consuming of liquids or solids for the next 30 min; Gel and Foam followed by immediately washing with water jet. After each phase, surface hardness was again measured for analysis of mineral gain, evaluated through percentage of surface hardness recovery (%SHR) and integrated loss of subsurface hardness (ΔKHN). The concentration of loosely bound fluoride (CaF2) and firmly bound fluoride (FA-like) formed and retained were also determined.

Results

Fluoride treatments produced greater remineralization (%SHR and ΔKHN) compared to placebo group (p < 0.05). There was no difference in the ability to promote remineralization and in the concentration of fluoride formed and retained, in each analysis, between Gel and Foam (p > 0.05).

Conclusion

The data suggest that rinsing with water immediately after topical application does not seem to have an influence on the ability of fluoride to promote remineralization.     

Light cola drink is less erosive than the regular one: An in situ/ex vivo study

Objective

This in situ/ex vivo study assessed the erosive potential of a light cola drink when compared to a regular one.

Methods

During 2 experimental 14-days crossover phases, eight volunteers wore palatal devices with 2 human enamel blocks. The groups under study were: group light, erosive challenge with light cola drink and group regular, erosive challenge with regular cola drink. During 14 days, erosive challenges were performed extraorally 3X/day. In each challenge, the device was immersed in 150 ml of light cola (group light) or regular cola (group regular) for 5 min. Erosion was analysed by surface profilometry (μm) and surface microhardness change (%SMH). The data were statistically analyzed using paired t test (p < 0.05).

Results

Group light (0.6 ± 0.2 μm) showed significantly lesser wear than group regular (3.1 ± 1.0 μm). There was no significant difference between the groups for the %SMH (group light −63.9 ± 13.9 and group regular −78.5 ± 12.7).

Conclusions

The data suggest that the light cola drink is less erosive than the regular one.     

Effect of stannous and fluoride concentration in a mouth rinse on erosive tissue loss in enamel in vitro

The aim of this in vitro study was to investigate the influence of stannous and fluoride ion concentrations in various experimental solutions on erosion progression in enamel. Human enamel specimens were subjected to a cyclic de- and remineralisation procedure for 10 days, with six demineralisation periods per day, of 5 min each. Erosive demineralisation was performed with 0.05 M citric acid (pH 2.3). Except in the control group, specimens were treated for 2 min with test solutions after the first and the sixth demineralisation. Test solutions were: 1500 mg/L F⁻ groups: group 1: 2800 mg/L Sn²⁺; group 2: 2100 mg/L Sn²⁺; group 3: 1400 mg/L Sn²⁺; group 4: 700 mg/L Sn²⁺; 1000 mg/L F⁻ groups: group 5: 2100 mg/L Sn²⁺; group 6: 1400 mg/L Sn²⁺. All preparations were adjusted to pH 4.5. Tissue loss was determined profilometrically after the last experimental day. As expected, the greatest tissue loss (μm, mean ± S.D.) was found in the control group (72.6 ± 11.5). All test solutions were able to reduce tissue loss significantly (p ≤ 0.001). The reduction of tissue loss by test solutions depended on the ratio of the tin concentration to the fluoride concentration. Lowest values were obtained by the application of the solutions of group 1 (7.8 ± 2.5) and group 5 (7.6 ± 5.2). Solutions with high concentrations of tin and fluoride are very effective in reducing erosive tissue loss, and their efficacy increased with increasing ratios of tin to fluoride concentrations.     

TiF4 and NaF at pH 1.2 but not at pH 3.5 are able to reduce dentin erosion

Objective

This study aimed to analyse and compare the protective effect of buffered (pH 3.5) and native (pH 1.2) TiF₄ in comparison to NaF solutions of same pH on dentin erosion.

Design

Bovine samples were pretreated with 1.50% TiF₄ or 2.02% NaF (both 0.48 M F) solutions, each with a pH of 1.2 and 3.5. The control group received no fluoride pretreatment. Ten samples in each group were eroded with HCl (pH 2.6) for 10 × 60 s. Erosion was analysed by determination of calcium release into the acid. Additionally, the surface and the elemental surface composition were examined by scanning electron microscopy (two samples in each group) and X-ray energy-dispersive spectroscopy in fluoridated but not eroded samples (six samples in each group). Cumulative calcium release (nmol/mm²) was statistically analysed by repeated measures ANOVA and one-way ANOVA at t = 10 min.

Results

TiF₄ and NaF at pH 1.2 decreased calcium release significantly, while TiF₄ and NaF at pH 3.5 were not effective. Samples treated with TiF₄ at pH 1.2 showed a significant increase of Ti, while NaF pretreatment increased F concentration significantly. TiF₄ at pH 1.2 led to the formation of globular precipitates occluding dentinal tubules, which could not be observed on samples treated with TiF₄ at pH 3.5. NaF at pH 1.2 but not at pH 3.5 induced the formation of surface precipitates covering dentinal tubules.

Conclusion

Dentin erosion can be significantly reduced by TiF₄ and NaF at pH 1.2, but not at pH 3.5.     

In vitro evaluation of the early erosive lesion in polished and natural human enamel

Objective

This study evaluated the capability of profilometry, microhardness, Optical Coherence Tomography (OCT) and Tandem Scanning Confocal Microscopy (TSM) in characterising the early erosive lesion in polished and natural human enamel in vitro.

Effect of nano-carbonate apatite to prevent re-stain after dental bleaching in vitro

Objectives

This study examined the effect of nano-carbonate apatite (n-CAP) to prevent re-staining and the change of enamel surface after dental bleaching in vitro.

Methods

Twenty-four bovine specimens were bleached for 2 weeks with 10% carbamide peroxide (CP). After bleaching, the specimens were divided into the following four groups: distilled and deionized water (DDW, negative control), 10% n-CAP, NaF (positive control) and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP, positive control). Each group was subjected to pH cycling for 7 days. The specimens were treated for 4 min 3 times per day and re-staining was induced naturally by artificial saliva in the remineralization process. After pH cycling, the changes in colour were evaluated with spectrophotometry and scanning electron microscopy (SEM). The difference in colour between before and after pH cycling was evaluated using an ANOVA and Tukey test.

Results

After pH cycling, the colour difference of n-CAP group was significantly lower than that of the DDW and CPP-ACP groups (p < 0.05). SEM showed that n-CAP particles were deposited regularly on the damaged surface compared to the other groups.

Conclusion

10% n-CAP could significantly maintain the initial colour and protect the damaged enamel structure after bleaching.     

MMP20 and KLK4 activation and inactivation interactions in vitro

Objective

Enamelysin (MMP20) and kallikrein 4 (KLK4) are believed to be necessary to clear proteins from the enamel matrix of developing teeth. MMP20 is expressed by secretory stage ameloblasts, while KLK4 is expressed from the transition stage throughout the maturation stage. The aim of this study is to investigate the activation of KLK4 by MMP20 and the inactivation of MMP20 by KLK4.

Design

Native pig MMP20 (pMMP20) and KLK4 (pKLK4) were isolated directly from enamel scrapings from developing molars. Recombinant human proKLK4 (rh-proKLK4) was activated by incubation with pMMP20 or recombinant human MMP20 (rhMMP20), and the resulting KLK4 activity was detected by zymography. Reaction products were isolated by reverse-phase high performance liquid chromatography (RP-HPLC), and their N-termini characterized by Edman degradation. The pMMP20 was incubated with pKLK4 under mildly acidic or under physiologic conditions, and enzyme activity was analyzed by zymography. The catalytic domain of rhMMP20 was incubated with pKLK4 or recombinant human KLK4 (rhKLK4) and the digestion products were characterized by zymography and Edman degradation.

Results

Both pMMP20 and rhMMP20 activated rh-proKLK4 by cleaving at the propeptide-enzyme junction used in vivo. The pMMP20 was inactivated by pKLK4 under physiologic conditions, but not under mildly acidic conditions. Both pKLK4 and rhKLK4 cleaved MMP20 principally at two sites in the catalytic domain of MMP20.

Conclusions

MMP20 activates proKLK4 and KLK4 inactivates MMP20 in vitro, and these actions are likely to occur during enamel formation in vivo.