Comparison of three methods for monitoring dentin erosion progression

doi:10.21203/rs.2.22751/v1

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Comparison of three methods for monitoring dentin erosion progression

https://doi.org/10.21203/rs.2.22751/v1

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Background: Clinical assessment and monitoring of dental erosion in vivo is challenging, although a number of clinical indices have been proposed but these are unlikely to be sensitive enough to quantify erosion with sufficient sensitivity to allow comparison of different therapies. In this respect, some of the techniques predominantly employed for the assessment of dental caries such as Quantitative light- induced fluorescence (QLF) and Optical coherence tomography (OCT) may also have utility for the assessment for erosion both in vivo and in vitro. The aim of the present study was to test three techniques (QLF, OCT and SMH) for the assessment of dentine erosion in a product testing model, using dentifrices to reduce erosion susceptibility. Dentine erosion was evaluated comparing the methods. Methods: Human dentine specimens were treated with one of two dentifrice slurries or mineral water (control group), followed by an erosive challenge and storage overnight in artificial saliva. These procedures were performed over 5 days. The following two dentifrices were tested: Duraphat™ 5000 (5000ppm Sodium Fluoride) and Colgate Sensitive Pro-Relief™ (8% arginine plus calcium carbonate and 1450ppm Fluoride). All groups showed a progressivedecrease in SMH and increase in scattering for OCT (P<0.05) indicating mineral loss. Results: The two dentifrice groups demonstrated significantly less softening than the control group (P<0.05). No statistically significantdifferences were detected for dentine erosion using QLF (P>0.05). Conclusion: SMH and OCT techniques were able to detect a significant protective effect against in vitro erosion whensamples were pre-treated with either dentifrice formulation. Dentifrices containing high concentrations of fluoride andarginine associated with calcium carbonate and fluoride have a protective effect against dentinal erosion.

Head & Neck Surgery

Dentistry

Tooth erosion

Dentifrices

Diagnostic Imaging

Epidemiological studies suggest that the prevalence and incidence of pathological tooth wear is increasing [1]. The aetiology of the spectrum of tooth wear presentations is complex but early detection, therapeutic interventions to prevent further tooth wear, and education to minimize causative factors are key to preventing the condition becoming a significant problem later in life [2, 3]. The restoration of worn dentitions, especially those where erosion has occurred, in addition to either attrition or abrasion, is complex often requiring secondary specialist care [3].

Clinical assessment and monitoring of dental erosion in vivo is challenging [3]. A number of clinical indices have been proposed but these are unlikely to be sensitive enough to quantify erosion with sufficient sensitivity to allow comparison of different therapies – certainly within the time frame of most clinical trial study designs [4]. It is possible that some of the techniques predominantly employed for the assessment of dental caries such as Quantitative light-induced fluorescence (QLF) and Optical coherence tomography (OCT) may also have utility for the assessment for erosion both in vivo and in vitro [5].

Quantitative light-induced fluorescence (QLF) is a visible light system that detects early enamel demineralisation and enables, through repeated imaging, longitudinal monitoring of changes over time [5]. Previous studies have reported a direct relationship between the mineral contente of the enamel and its optical properties, particularly fluorescence loss measured by QLF [6]. Although QLF has been used in the past for the assessment of enamel erosion and caries by looking at the loss of fluorescence due to increased porosity and light scattering, [7] this mechanism might be hampered when directly assessing dentine as the organic material responsible for the source of fluorescence could potentially be overexposed. Based on the physical principle, this method may be able to detect an increase of fluorescence of dentine eroded. To date, there is little evidence to support the use of QLF to assess mineral loss from dentine – either due to caries or erosion [8].

Optical coherence tomography is a high-resolution non-destructive optical technique. It produces cross-sectional images of internal biological structures, employing near-infra red low coherence light, to obtain images at a depth of 1–2 mm in scattering tissues such as human dentine.

Therapeutic approaches for the control of enamel and dentine erosion are also clearly important. Studies have evaluated the use of fluoride to protect dental tissues from an acid challenge and precipitation of calcium fluoride may reduce enamel and dentine solubility [9]. It has also been suggested that the use of dentifrice with high fluoride concentration may reduce tooth erosion [9]. Although conflicting results of the efficacy of fluoride has been reported due to different experimental study designs, there are some evidence that fluoride may be effective in the reduction of dental erosion [10].

A new dentifrice formulation based upon 8% arginine, calcium carbonate, and 1450 ppm fluoride as sodium monofluorophophate has shown to be an effective treatment for dentine hypersensitivity [11–13]. The mode of action relies on the precipitation of calcium- and phosphate-containing material on the tooth surface and it is possible that this layer may also afford some protection against acid erosion as well as providing the aforementioned fluoride protection [11, 12, 14].

The aims of the present in vitro study were to compare detection methods (OCT and QLF) against SMH, as well as to evaluate a high fluoride dentifrice (Duraphat^TM5000 ppm) and 8% arginine, calcium carbonate, sodium monofluorophosphate dentifrice (Colgate Sensitive Pro-Relief™) as a possible preventive treatment for dentine erosion.

Preparation of Dentine Specimens

Twenty-four extracted human third molars were selected for not being in contact with the oral cavity [15]. These teeth were previously stored in normal saline and obtained from Sao Paulo State University, Araraquara Dental School, under the ethical approval number #34/10.

A high-speed cylindrical diamond burr (KG Sorensen, Barueri, SP, Brazil) was used under copious irrigation to make two parallel grooves 0.5mm deep on the root surfaces of each tooth (buccal and lingual): one at the cementoenamel junction and the other 4 mm apical to the first. The área between the two grooves was flattened with the same bur, obtaining root dentin exposition [16]. The roots were then cut in the first groove in order to remove the crown. Two root dentine blocks, approximately 3x5 mm, were obtained from each tooth, one from the buccal and one from the lingual surface, obtaining a total of 48 specimens randomly allocated in 3 Groups.

One half of the exposed surface was protected using masking tape while the other half was coated with a thin layer of acid resistant nail varnish that served to protect the sound root as the control area before exposure to the demineralization solution [15]. After the application of the varnish was completed, the masking tape was removed leaving an area exposed to the acidic challenge.

During the evaluations, specimens were kept in 1 ml of distilled water. The specimens were air dried for 10 seconds with compressed air fixed at 10 cm from the teeth before QLF and OCT assessment.

Baseline: Specimens were stored in artificial saliva for 1 day before evaluating with QLF, OCT and surface micro hardness (Figure 1).

Treatment

Each dentine specimen was immersed in dentifrice (3 g dentifrice / 10 ml distilled water) for 3 min at room temperature and pH controlled under constant stirring on a magnetic stirrer (IKA^TM C-MAG MS10, IKA^TM Werke GmbH & Co.KG/ Germany) [17]. The following groups were tested

- Group 1 – Control: Negative Control (Volvic^TM, Mineral water, pH 7.8)

- Group 2 – 5000F: Duraphat^TM 5000 Toothpaste (Colgate Palmolive – 5000ppm Fluoride, pH 8.3).

- Group 3 – Arginine: Colgate Sensitive Pro-Relief^TM (Colgate Palmolive – 8% arginine, calcium carbonate, pH 8.8).

For the negative control group, specimens were kept in 600 ml of non-carbonated mineral water (Volvic^TM, Danone Ltd, Manchester, UK), since it had been shown not to induce any de- or remineralizing effects, [18] under 3 min of constant stirring.

After treatment, the specimens were carefully rinsed with distilled water using a syringe (15 ml). The OCT, QLF and SMH evaluations were repeated.

Erosion-cycle (Acid-challenge intervals)

Specimens were suspended with plastic rods in orange juice (Sainsbury’s^TM Orange Juice, Manchester, UK) (pH 3.72±0.03, temperature: 25^oC). Two 1-litre beakers were used with eight specimens suspended in 600 ml of orange juice in each beaker, and the orange juice was gently agitated with a magnetic stirrer (IKA^TM C-MAG MS10, IKA^TM Werke GmbH & Co.KG/ Germany) for 15 minutes [18] . The specimens were removed from the orange juice and carefully rinsed with 15 ml of distilled water using a syringe to remove acid excess from the surface.

The measurements were performed over 5 days (day 1- baseline; after treatment and after erosion; day 2-5- after treatment and after erosion). The orange juice was changed after every 15 minutes of erosion (Figure 1).

Storage overnight

After every cycle, each specimen was incubated overnight (10 hours) in 5 ml artificial saliva under a controlled temperature of 25^o±2 Celcius and pH of 7.15±0.3. The composition of the artificial saliva prepared was: 1.5 mmol/l Ca(NO₃)2 4H₂O; 0.9 mmol/l Na₂HPO4 2H₂O; 150 mmol/l KCl; 0.02 mol/l H₂NC(CH₂OH)₃ (TRIS); 0.05 μg/ml NaF, pH 7.0 (10).

Surface Microhardness

Surface Microhardness measures softening of the dentine surface following an erosion challenge. The measurements were performed with a Knoop diamond under a force of 10 g applied for 5 seconds (Micro Hardness Tester FM-700; Future-Tech Corporation, Japan) [19]. The teeth were placed flat on the translation stage and fixed at a reproducible position within the micro-indentor. A surface area of approximately 1 x 1 mm2 that was perpendicular to the direction of the load of the indenter was identified on the uncoated half of the dentine window for indentation. Three indentations were made on each specimen during each measurement time point. The Knoop numbers (K) were calculated and averaged. The outcome of surface micro hardness change (SMC) was calculated based upon the differences between the surface micro hardness at baseline and the subsequent time intervals (normalized data). The SMC was calculated as:

SMC=(K_b –K_d)
K_b

with K_b as the mean Knoop micro hardness number at baseline and K_d as the mean Knoop number after demineralization and treatment.

Specimens were stored in distilled water among the assessments to keep them constantly hydrated, as drying of the dentine would cause changes in hardness and contraction (18).

Optical Coherence Tomography (OCT)

A commercially available OCT system (OCS1300SS, Thorlabs Ltd, UK) was used to capture cross sectional images of the eroded dentine area. The OCS1300SS uses the Fourier Domain technology and incorporates a broadband, frequency swept laser and the output wavelength centred at 1325 nm. It has an axial resolution of 9 μm and transverse resolution of 15 μm in air. The hand probe was mounted on a ‘stand’ with the beam facing downwards. The specimens were placed on a translational stage perpendicular to the hand probe. The stage was fixed with a repositioning jig that enabled the samples to be repositioned to the same position and alignment during each examination.

The OCT light beam was configured to scan a length of 5 mm at x-axis direction and an axial depth of 3 mm. The y-axis position of the light beam on the tooth surface was located at a cross section with the least observed specular reflection. The X and Y coordinate of the light beam for each specimen was recorded for replication at each examination. The distance of the specimen surface to the probe was determined using the most convex area at 3.0 ± 0.1 mm from the top margin in the display window of the image capture software. The dynamic range of the OCT light was maintained around 20 – 30 decibels (dB). Background noise was removed before the acquisition of each image.

The OCT measures the increase of surface light scattering due to an increase in porosity of the dentine surface following an erosion challenge. As the collagen matrix gets dissolved, gaps are between the tubules left that would further scatter the light.

A customized MATLAB-generated program (MathWorks, California, US) was used to analyse the changes of the intensity for OCT backscattered light in time (Figure 2). The B- scans of each sample from the different measuring time points were aligned and a similar region of interest was selected for all examinations (16 A-scans in each specimen). After normalization of the data, a mean depth-resolved intensity curve was generated. The attenuation or extinction (A_t) of the backscattered light intensity due to demineralisation is estimated by the function below:

At = I_subsurface I _surface

I_subsurface is the backscattered light intensity at a subsurface level, which would not be affected by demineralization (50 pixels below of dentine surface). Isurface is the backscattered light intensity at a superficial level where demineralisation had occurred (10 pixels below of dentine surface).

Quantitative Light-induced Fluorescence (QLF)

A custom QLF set up was employed. The set-up consisted of blue light-emitting diodes in a ring illuminator casing and a 3-charged coupled device (3CCD) colour camera (FV-F31, Hitachi Kukasai Electric U.K.) installed with a long-pass yellow filter. A 50 mm focal length-imaging lens with a 20 mm extension tube was used to capture a field of view of focal length-imaging lens with a 20 mm extension tube was used to capture a field of view of approximately 330 mm by 450 mm. The images were taken in a dark enclosure and the capturing of the images was done via bespoke software.

Optimum lighting properties such as the gain factor and brightness were determined after obtaining histograms of the pixel value of the green channel for all samples before subjected to erosion to ensure that saturation of signal did not occur towards the end of the study.

After the data were normalized by baseline, the loss of green fluorescence, ΔF, at the non-coated area as compared to the coated area (Figure 3) was calculated as follows:

ΔF = F(NC) – FI

With F(NC) as the intensity of green fluorescence at the non-coated area and FI the intensity of green fluorescence at the coated area.

Statistics

The primary outcome variables were the change in Knoop Hardness Number (SMH), Quantitative Light Fluorescence (QLF) and Optical Coherence Tomography (OCT) during the demineralization/remineralization cycles. Comparison among the time periods for each technique was undertaken using ANOVA with Repeated Measures Test, as these values are dependent. The groups were compared in each time period using One-Way ANOVA. These tests were performed using SPSS version 19.1 (SPSS Inc., IBM Company, United States).

The measurements of QLF, OCT and surface microhardness were found to follow a normal distribution.

Microhardness

Mean and standard deviation values of surface micro hardness (SMH) in sound and following pre-softening, as well as during the cycles, are provided in Table 1. It shows that, at the completion of the study (5 days of treatment and erosive challenge – once per day) all groups presented lower values compared to their corresponding pre-softened readings.

As expected, Group 1 – Control presented a higher decrease of SMH values, revealing more dentine mineral loss than Groups 2 – 5000ppm F and 3 – Arginine. A statistically significant difference from baseline was observed for all time intervals between control and Group 3 – Figure 4.

The statistical analysis revealed significant differences (One-way ANOVA, P< 0.05) between groups for each time point. Duraphat 5000ppm and Arginine delivered a broadly similar protective effect with no significant differences found between then after 45 minutes of erosive exposure (Figure 4).

Quantitative Light-induced Fluorescence (QLF)

Table 1 gives an overview of mean and standard deviation of QLF metrics measured at each cycle. No statistically significant differences were found when a comparison among time intervals was evaluated (Repeated Measures ANOVA, P>0.05). Comparison among groups for each time intervals revealed greater erosive effect for Group 1- Control than Group 2- 5000F and 3- Arginine after 15 min stirring in orange juice.

No statistical significant differences were found among groups for the remaining time intervals (Figure 5).

Optical Coherence Tomography (OCT)

Table 1 shows mean values and standard deviations of optical coherence tomography

measurements at each cycle of evaluation. It shows that, at the end of the cycles, Groups 1- Control and 3- Arginine presented lower values compared to their corresponding pre-softened readings. No statistically significant differences were found between baseline and after erosion on the last cycle (cycle 5) for Group 2- 5000F and 3- Arginine (ANOVA with repeated measures, P>0.05).

Comparison among the time intervals showed significant more scattering after 75 min of orange juice exposure for Group 1. Group 1 also presented statistically significant differences comparing baseline with 15 min, 30 min, 60 min and 75 min of acid challenge.

However, no differences have been shown among the acid-challenge intervals (evaluations after orange juice exposure) (Figure 6).

Demineralization was most severe in the Group 1 – Control. As with the SMH findings, both of the dentifrice groups showed a significant protective effect when compared to the control group. Figure 6 presents these data showing the time points at which significance was detected.

The purpose of this study was to compare different methodologies to assess dentine erosion. Considering the possibility of determining whether quantitative light-induced fluorescence (QLF) and optical coherence tomography (OCT) could be used to nondestructively assess dentine erosion on natural root surfaces. A further aim was to demonstrate whether the methodologies are able to detect if the lesions were less susceptible to erosive challenge after treated with two different dentifrices.

SMH is an effective means of obtaining accurate information about early erosion in vitro [20, 21] and the measurements can be performed with either a Knoop or a Vickers diamond indenter. In the present study, a Knoop diamond indenter was selected as it is more sensitive to changes in the most superficial layer of an eroded lesion [7]. Herkstroter et al. [21] had shown that microhardness measurement of dentine surface should be taken 24 hours after indentation to account for elastic retraction of the organic collagen matrix. Although this recommendation was not adhered to in this study, we have ensured that the measurements in this study were taken consistently within a short period of time immediately after indentation. A consistent overestimation in the surface softness of the sample could be expected. Nevertheless this would only affect the absolute microhardness values and result in larger statistical deviations of the mean but not the relative change of the microhardness measurement (SMC).

Natural unpolished samples were used in this study as its objectives were to evaluate the potential of QLF and OCT in detecting and monitoring dentine erosion in vivo. Although surface microhardness had been used predominantly to measure flat polished samples, few studies have explored the possibility of measuring unpolished surfaces. Chu et al. [22] found that after excluding asymmetrical indentations, the variation in hardness of intact surfaces is usually the result of local differences in hardness of the surfaces and not attributable solely to mechanical difficulties in obtaining symmetrical indentations [22].

In this study SMH and OCT data have demonstrated a similar trends over time. Previous studies have shown that OCT may be an effective means of assessing dentine lesion depth and mineral loss [23]. The present study demonstrated good agreement between techniques for the initial measurements; however changes during time intervals for days 2, 3, 4 and 5 were not as evident for OCT suggesting some degree of measurement saturation.

Quantitative light-induced fluorescence metrics did not demonstrate a relationship with SMH and OCT. Although QLF has been used as a successful technique to detect initial caries in enamel, the same has not observed for carious dentine lesions. The measurement of erosion has been demonstrated in enamel – but again not in dentine. The QLF measures the reduction in light fluorescence due to an increased porosity of the enamel surface. However, as the QLF signal originates from the collagen in dentine, an increase in fluorescence can be observed as a result of dentine erosion as more exposed collagen can get exposed in the dissolution process. Hence where one would expect to see fluorescence loss in demineralized enamel, the reverse would be true in dentine – a fluorescence gain. The failure to detect a dose response using QLF in dentine is explained by this optical phenomenon. Given the high degree of fluorescence produced by exposed dentine, small gains caused by the porosity resulting from demineralisation is unlikely to be detected by the current systems as images become rapidly saturated [24].

The QLF results showed higher fluorescence (as predicted) after 75 min stirring in orange juice. These results also seemed to be affected by the reference area, which presented a loss of fluorescence over time. The presence of stains due to repeated exposure to orange juice in association with varnish might be an explanation for the decreasing of fluorescence in the reference area. A further explanation for the fluorescence increase may be related to the high collagen exposure after repeated exposure to acid challenges. Considering the optical properties of dentine, unlike for enamel the current QLF technique may not be a suitable means of measuring dentine erosion [25].

Due to the reported increase in the prevalence of dental erosion both in enamel and dentine [1, 2], approaches to prevent this cause of tooth surface loss will become increasingly important [26]. The study has tested two dentifrices as potential inhibitors of dentine erosion, Duraphat™ 5000 (5000 ppm Fluoride) and Colgate Sensitive ProRelief™ (1450 ppm Fluoride) (Colgate-Palmolive Ltd). The results suggest that both deliver an effective protective effect against erosion in vitro.

Rios et al. [27] have tested two concentrations of fluoride dentifrice (1,100 ppm and 5,000 ppm) in enamel erosion and no statistical significant differences were found between them. However, Ganss et al. [28] found higher preventive effect of dentine erosion when intensive fluoridation was applied. The dissimilarities in experimental parameters, such as different de-remineralization ratios and mineral content, distribution of initial artificial lesions (enamel or dentine), different periods of acids challenge and treatments may explain the different results reported to date in the literature.

In agreement with previous studies, [10] this current study has demonstrated that fluoride application is effective in increasing dentine’s resistance to erosive challenge. Surface adsorption and hetero-ionic exchange with surface hydroxyl ions are important mechanisms to fluoride uptake by apatite crystals [9]. In the present study, high and low fluoride concentrations were applied to partially demineralized dentine over a large surface, which provides a wide reactive area prior to each period of remineralization. It is known that fluoride can cause a marked inhibition of enamel and dentine dissolution by acid [10].

Besides fluoride, other dentifrice components have been considered for preventing tooth surface loss. The presence of calcium and calcium sources are also an important influence in the overall balance of dental re- and demineralization in erosive processes [11, 12].

An innovative technology by combining arginine and calcium carbonate with sodium monofluorophosphate, Colgate Sensitive Pro-Relief™, has been developed for treating dentine hypersensitivity. In this study, ProRelief™ was as effective as Duraphat™ dentifrice in increasing the dentine’s resistance to erosive losses. It has been clearly shown that the arginine associated with calcium carbonate is highly effective in occluding dentinal tubules [11]. Of relevance to the current study is that the authors also demonstrated the protective effect of this toothpaste against acid challenges.

As demonstrated in the present study, dentifrice application prior to an erosive challenge seems to confer a protective effect but did not completely inhibit progressive erosion. It should be noted that the challenge in this in vitro study was substantial and likely to be in excess of that experienced in vivo.

SMH and OCT have demonstrated considerable potential as a device for the monitoring dentine erosion following acid challenges in vitro. The use of non-polished surfaces in the current study lends authenticity to the experimental design. This study suggested that after the erosion challenge interval used (10 minutes), the OCT signal presents the largest change from baseline. Further studies are still required to evaluate the shortest erosion challenge that OCT can detect before exploring possible routes towards in-vivo measurements. QLF does not appear, in its current configuration, to offer the same potential. The results of the current study suggest that both of the dentifrices present a protective effect against acid challenges on exposed dentine surfaces.

SMH and OCT techniques were able to detect a significant protective effect against in vitro erosion when samples were pre-treated with either dentifrice formulation. Dentifrices containing high concentrations of fluoride and arginine associated with calcium carbonate/fluoride have a protective effect against dentinal erosion.

3CCD 3-charged coupled device

dB Decibels

K Knoop numbers

OCT Optical coherence tomography

QLF Quantitative light-induced fluorescence

SMC Micro hardness change

SMH Surface micro hardness

Acknowledgements

Not applicable.

Ethics approval and consent to participate

The study was approved by the ethics local committee of the faculty of dentistry of the UNESP (approval number #34/10) and the teeth were obtained from the tooth bank.

Consent for publication

The authors give their consent for the publication of the article in the journal.

Availability of data and material

No repository.

Competing interests

The authors declare that they have no conflicts of interests and the authors do not have any financial interest in the companies or products used in this study.

Funding

The work was supported by CAPES.

Authors' contributions

Phase of Design and planning of the study: SP, MB, CZ, IP, RE

Experimental phase and data collection: SP, SL, MN, HC, JS

Phase of data analysis: JS, RE, IP

Preparation of the article: SP, SL, CZ, MB

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Table 1. Mean and standard deviation of Surface Microhardness (SMH) values and the corresponding normalized SMH compared by baseline measured at each time point for all groups.

	DELTA VALUES SMH		DELTA VALUES QLF		DELTA VALUES OCT
			BASELINE
Group	Baseline		Baseline		Baseline
Control	0 (±0,0)		0 (±0,0)		0 (±0,0)
Duraphat 5000	0 (±0,0)		0 (±0,0)		0 (±0,0)
Pro-Relief
			CYCLE 1
Group	After Treatment	After erosion	After Treatment	After erosion	After Treatment	After erosion
Control	−0.03 (±0.04)	−0.15 (±0.07)	−0.01 (±0.04)	−0.06 (±0.06)	−0.02 (±0.09)	−0.42 (±0.27)
Duraphat 5000	0.05 (±0.05)	0.01 (±0.04)	0.04 (±0.08)	0.03 (±0.11)	0.008(±0.15)	−0.02 (±0.18)
Pro-Relief	−0.001 (±0.03)	−0.03 (±0.04)	0.02 (±0.11)	0.03 (±0.10)	0.16 (±0.30)	−0.001 (±0.14)
			CYCLE 2
Group	After treatment	After erosion	After treatment	After erosion	After treatment	After erosion
Control	−0.13 (±0.104)	−0.27 (±0.08)	−0.02 (±0.05)	−0.06 (±0.08)	−0.40 (±0.19)	−0.50 (±0.18)
Duraphat 5000	0.04 (±0.04)	0.01 (±0.04)	−0.01 (±0.07)	0.001 (±0.07)	0.07(±0.27)	−0.05(±0.25)
Pro-Relief	−0.02 (±0.04)	−0.04 (±0.04)	0.006 (±0.14)	0.0013(±0.14)	− 0.06(±0.34)	0.008 (±0.23)
			CYCLE 3
Group	After treatment	After erosion	After treatment	After erosion	After treatment	After erosion
Control	−0.23 (±0.07)	−0.39 (±0.07)	−0.05 (±0.07)	−0.06(±0.07)	−0.38 (±0.24)	−0.43 (±0.23)
Duraphat 5000	0.02 (±0.04)	−0.07 (±0.04)	−0.01 (±0.04)	−0.01 (±0.06)	−0.06(±0.20)	−0.013 (±0.29)
Pro-Relief	0.04 (±0.04)	−0.10 (±0.03)	0.005 (±0.12)	−0.001 (±0.10)	0.09 (±0.23)	0.02 (±0.20)
			CYCLE 4
Group	After treatment	After erosion	After treatment	After erosion	After treatment	After erosion
Control	−0.38 (±0.06)	−0.43 (±0.08)	−0.07 (±0.07)	−0.06 (±0.08)	−0.43 (±0.22)	−0.43 (±0.21)
Duraphat 5000	−0.06 (±0.04)	−0.09 (±0.03)	−-0.03 (±0.05)	−0.01(±0.08)	−0.01(±0.24)	0.02 (±0.28)
Pro-Relief	−0.10 (±0.03)	−0.13 (±0.03)	0.002(±0.10)	−0.007 (±0.09)	−0.005(±0.17)	0.06 (±0.20)
			CYCLE 5
Group	After treatment	After erosion	After treatment	After erosion	After treatment	After erosion
Control	−0.40 (±0.09)	−0.46 (±0.08)	−0.05 (±0.07)	−0.04 (±0.10)	−0.40 (±0.22)	−0.42 (±0.23)
Duraphat 5000	−0.09(±0.03)	−0.13 (±0.04)	−0.04(±0.08)	0.01 (±0.09)	0.03(±0.23)	−0.02 (±0.24)
Pro-Relief	−0.13(±0.03)	−0.18 (±0.04)	−0.005(±0.11)	0.002 (±0.08)	0.08(±0.24)	0.11(±0.24)

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Comparison of three methods for monitoring dentin erosion progression

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Abstract

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1. Background

2. Methods

3. Results

4. Discussion

5. Conclusions

Abbreviations

Declarations

References

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