Effect of Biomin F Toothpaste and Diode Laser on Remineralization of White Spot Lesions (In Vitro Study)

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Tamish, Ahmed M. Madian This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3990752/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Jul, 2024 Read the published version in BMC Oral Health → Version 1 posted 10 You are reading this latest preprint version Abstract Background : White-spot lesions are considered an initial carious stage characterized by an outer enamel layer with significantly reduced mineralization. This study was conducted to assess the combined effect of Biomin F toothpaste and Diode laser on remineralization of white spot lesions . Materials and methods : An invitro study conducted on a total of 30 premolars divided into three groups; Group A (Biomin F Tooth paste), Group B (Biomin F with laser application for 30 sec), Group C (Negative control).The three groups were submitted to three stages; stage 1:Baseline,stage 2:After demineralization ,and stage 3:After remineralization .In each stage, elemental analysis(calcium, phosphorus, and fluoride)was measured quantitatively using Energy Dispersive X-ray (EDX) analysis and qualitatively by micrographs using scanning electron microscope. The data were tested to find significant difference between mineral changes during stages by using (ANOVA) test and Bonferroni test. Results : Calcium, phosphorus and fluoride ions decreased in all groups after demineralization. In stage 3, after application of remineralizing agents, Calcium ions increased significantly in groups A and B where P˂.05. As regards to the phosphorus ions, a significant increase was observed in all groups with group A showed the highest gain as phosphorus level percentage change (%mass) was 56.52±18.02 . Fluoride ions increased significantly in groups A and B (p˂0.05) but decreased significantly in group C. There was no statistical significant difference between group A and B (p≥.05) in calcium, phosphorus, and fluoride level after remineralization . Conclusion : Within the limitation of the present study, we concluded that Biomin F toothpaste is promising in the repairing of white spot lesions on the surface of the demineralized enamel. Diode laser didn’t affect the remineralizing ability of Biomin F toothpaste which add hazard without benefit . Biomin laser therapy WSL tooth remineralization Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 INTRODUCTION Typically, patients seek orthodontic intervention to enhance their aesthetic appeal. Nonetheless, the use of fixed orthodontic devices could result in the development of white spot lesions (WSLs), posing an additional aesthetic challenge for the patient. Consequently, both the patient and orthodontist may experience disappointment upon the removal of the appliances [ 1 ]. White spot lesions are recognized as the initial stage of tooth decay, characterized by a surface enamel layer showing a significant reduction in mineral content [ 2 ]. This condition is prone to deteriorate, potentially necessitating invasive treatment [ 3 ]. Enamel demineralization and the development of WSLs advance swiftly [ 4 ], often developing within a few weeks [ 5 ]. Preventive approaches for WSLs during orthodontic treatment primarily involve the utilization of fluoride-releasing varnishes [ 6 , 7 ], bonding materials, and cements [ 8 , 9 ]. Additionally, the application of concentrated fluoride gels [ 10 ] and daily rinsing with mouthwash [ 11 – 14 ] are employed to reduce enamel demineralization. While fluoride application is effective in halting WSLs, it comes with certain constraints. The consistent use of toothpaste containing fluoride necessitates a significant presence of bioavailable calcium and phosphate ions alongside fluoride ions. [15] Furthermore, fluoride exhibits diminished effectiveness when pH level drops below 4.5, a situation often instigated by bacterial actions [ 16 ]. Although the impact of fluoride relies on the dosage and improves with higher doses, there exists a threshold beyond which increasing the fluoride dose may lead to unwanted side effects such as dental fluorosis and toxicity [ 16 ]. In recent times, bioactive glasses have emerged as a notable advancement in dental applications, extensively researched in various studies targeting the treatment of white spot lesions through remineralization. These materials have the potential to rejuvenate and regenerate dental tissues by triggering apatite formation upon exposure to saliva or any other physiological fluid [ 16 , 17 ]. These apatites can either be hydroxyapatites or fluorapatites, depending on whether fluoride is integrated into the glass structure chemical composition. Glasses contain fluoride exhibit "smart" properties, demonstrating remineralization activity enhancement in low pH environments. Biomin F is recognized as a bioactive glass-based toothpaste that incorporates fluoride to aid in the remineralization of enamel. Laser technology has been utilized to decrease the rate of subsurface demineralization of enamel by modifying its crystalline structure, acid solubility, and permeability. However, it is crucial to apply lasers at a low energy level to maintain enamel integrity [ 18 ]. Following laser irradiation, enamel undergoes chemical and structural changes, including reduction in carbonates fusion and re-crystallization of hydroxyapatite crystals. These alterations enhance enamel's resistance against the acid attacks. Moreover, studies have demonstrated synergistic effects between laser treatment and topical fluoride application, leading to a significant reduction in the rate of enamel decalcification [ 19 , 20 ]. Hence, this study was conducted for the evaluation of the combined impact of Biomin F toothpaste and diode laser on the remineralization of WSLs. The null hypothesis of this study assumed that no significant difference is expected between the effect of Biomin F toothpaste coupled with diode laser and Biomin F alone on remineralization of white spot lesions. MATERIALS AND METHODS This in-vitro study was conducted at the Department of Orthodontics, Faculty of Dentistry, Alexandria University and Scanning Electron Microscope unit, Faculty of Sciences, Alexandria University. The present study was approved by the Research Ethics Committee of the Faculty of Dentistry, Alexandria University (IRB:00010556– IORG:0008839). Sample size calculation The sample size estimate was calculated based on an invitro study by Aidaros et al. (2022) [ 15 ] that aimed to evaluate and compare the remineralizing potential of dentifrices containing fluoride and bioactive glass on enamel by assessing the enamel structure and elemental analysis through Energy Dispersive X-ray Analysis (EDX). During sample size calculation, a beta error of up to 20% is accepted, with a study power of 80%. The alpha level was established at 5%, corresponding to a significance level of 95%. Statistical significance was assessed at a p -value < 0.05 [ 21 ]. The minimum required sample size was determined to be 9 teeth per group (number of groups = 3) (Total sample size = 27 teeth). Any withdrawal for any reason will be compensated by replacement to control for attrition (loss of specimen) bias. Therefore sample size will be increased to 10 teeth per group (number of groups = 3) (Total sample size = 30 teeth). A total of 30 human premolars were collected from patients requiring premolars extraction during their orthodontic treatment in the Orthodontic Department, Faculty of Dentistry, Alexandria University, Egypt. Informed consent was obtained from all subjects and/or their legal guardian(s). All patients were born and lived in areas where the typical concentration of fluoride in the drinking water was 0.36 mg/L [ 22 ]. Any calculus or tissue remnants were removed from the teeth using a scaler. Subsequently, the teeth were stored in saline until the commencement of the study. Procedures for each group are shown in the flowchart (Fig. 1). 1. Collection of teeth Thirty human premolars that were extracted for orthodontic purposes were chosen for inclusion in this study. All teeth were examined macroscopically using a magnifying loupe and fulfilled the following selection criteria: Intact buccal enamel surface, with no decalcifications, cracks, or stains.Teeth previously bonded or received any chemical treatment or decayed premolars were excluded from this study. The teeth were preserved in saline until the start of the experiment. 2. Teeth preparation After recruiting the appropriate teeth, all remnants were removed and teeth were cleaned with fluoride free pumice and running water. The roots of the teeth were cut 2mm under cemento-enamel junction using a diamond disk under water cooling. The crowns were embedded in self cured acrylic resin and the buccal surfaces were directed upward for easy manipulation [ 15 ]. (Fig. 2a) Each sample was covered with acid resistant varnish (nail polish) at all tooth crown surfaces, leaving a window of 4mm X4mm in the middle third of the buccal surface of the premolar. (Fig. 2b) Randomization Each tooth got a number from 1 to 30 typed at the base of the acrylic block using waterproof permanent marker. These numbers were used to randomly and equally assign the samples into 3 groups using computer generated random list. 3. Grouping of the teeth Teeth were divided randomly into 3 groups Group A: Biomin F tooth paste. Group B: Biomin F with laser application for 30 sec . Group C: Negative control group (no treatment(. Each group was placed in its own labeled beaker which contained 150 mL of artificial saliva solution at room temperature of 37°C and neutral PH to replicate the oral environment. (Fig. 2c) The materials used, their specifications, compositions and manufactures are present in (Table 1). 4. Intervention The study was divided into a number of stages with various procedures as the following: First stage (Baseline) assessment The assessment was conducted utilizing an environmental scanning electron microscope (JSM-IT 200-Japan) at Faculty of Sciences, Alexandria University. SEM attached with energy-dispersive X-ray (EDX) unit (Fig. 3). The amount of remineralization was measured qualitatively by comparing the scanning electron microscope pictures and quantitatively by the Energy dispersive X-ray analysis that measured the average of three points selected in the area of concern. These values were taken at different stages of the study (before, after demineralization and after remineralization). Second stage (Demineralization process) The teeth were immersed in the demineralizing solution for 96 hours at 37ᵒC until white spot lesions were obtained (Fig. 4) (Table 1). The samples were removed from the solution and rinsed with distilled water to stop the demineralization process and to remove any residuals of the solution [ 20 , 23 ]. At this stage, evaluations were carried out using Scanning Electron Microscope (SEM) analysis alongside Energy Dispersive X-ray Analysis (EDX). Third stage)Remineralization process) In group A: Biomin F toothpaste was applied in circular motion on the demineralized region using microbrush twice daily, each lasted for two minutes and left undisturbed for 30 seconds (Fig. 5). Then the samples were rinsed carefully with distilled water to remove any excess paste. Then it was stored in artificial saliva to mimic oral environment. This procedure was repeated for two weeks (Table 1) [ 15 , 20 ]. In group B: The toothpaste was applied twice daily each for two minutes in a circular motion using a microbrush and left undisturbed for 30 seconds which lasted for two weeks, then laser was applied at the 14th day (Fig. 6) (Table 2). In group C: No treatment was received. Table (1): The materials used and their specifications Solution Composition Usage Artificial saliva [20] • 2.20 g/L gastric mucin • 1.45 mmol/L CaCl2 2H2O • 5.42 mmol/L KH2PO4 • 6.50 mmol/L NaCl • 14.94 mmol/L KCl. PH was adjusted to 7.0 using KOH. Storage medium for the samples. Demineralizing solution[ 20 ] • 2.2 mM CaCl2 • 2.2 mM NaH2PO4 • 0.05 M lactic acid • 0.2 ppm fluoride. The pH was adjusted to 4.5 with 50% NaOH. Artificial sub-surface lesion formation. BioMinF Armour for teeth toothpaste (BioMin Technologies Ltd., London, UK) • Fluoro Calcium-PhosphoSilicate (Biomin) • Glycerin • Silica • PEG 400 • Sodium-Lauryl Sulphate • Titanium Dioxide • Aroma • Carbomer • Potassium Acesulfame • Contains maximum 530ppm of available fluoride when packed. Remineralizing agent Laser application Laser irradiation (Lastronix-Boland) has been done on the 14th day for 30 seconds after Laser beam activation with carbon particles and laser setting was done as shown in table (2). The practitioner wore eye goggles for self protection from laser irradiation. The distance between the buccal surface of the tooth and laser fiber was kept 5mm using custom- made holder (Fig. 6) (Table 2).[ 15 , 24 ] Table (2): Parameters of laser irradiation Center wave lenghth(nm) 980 Operating mode Continues wave(CW) Power output(W) 0.5 Exposure time(s) 30 Radiant energy(J) 15 Optic conductor fiber diameter(µm) 320 Mode Non contact mode The Scanning Electron Microscope assessment and Energy Dispersive X-ray Analysis (EDX) were repeated after remineralization process. Blinding Technician of Scanning Electron Microscope attached with EDX unit and the statistician were blinded. Statistical analysis The data were gathered and inputted into the computer using the Statistical Package for the Social Sciences (SPSS) software program for statistical analysis (ver 25).[ 25 ] Kolmogorov-Smirnov test of normality revealed no significance in the distribution of the variables, so the parametric statistics were adopted.[ 26 ] Data were described using minimum, maximum, mean, standard deviation, standard error of the mean, and 95% CI of the mean, 25th to 75th percentile.[ 27 ] Comparisons were conducted among more than two independent normally distributed subgroups utilizing the one-way Analysis of Variance (ANOVA) test [ 28 ]. Post-hoc multiple comparisons [ 29 ] were performed using the Bonferroni method [ 30 ]. Repeated measures analysis of variance was used [ 31 ]. Model assumptions were tested and found to be satisfactory with the exception of Mauchly’s test of sphericity [ 32 ], and when it was statistically significant denoting the violation of the assumption of sphericity, Greenhouse-Geisser correction was used [ 33 ]. Pair-wise comparison was done with Bonferroni correction. Linear trend analysis was used to test for within-subjects contrast [ 34 ]. Percentage change was calculated as follows: $$\varvec{P}\varvec{e}\varvec{r}\varvec{c}\varvec{e}\varvec{n}\varvec{t}\varvec{a}\varvec{g}\varvec{e} \varvec{c}\varvec{h}\varvec{a}\varvec{n}\varvec{g}\varvec{e} \left(\varvec{\%}\right)=\frac{Measurement \left(after\right)-Measurement \left(before\right)}{Measurement \left(before\right)}x 100$$ RESULTS This study included 30 premolars extracted for orthodontic purposes, all of which had an intact buccal enamel surface without any apparent decalcifications, cracks, or stains. These premolars were randomly assigned to three groups, with each group consisting of ten premolars: Group a : Biomin F Tooth paste Group b : Biomin F with laser application 30 sec Group c : Negative control Tables (3,4,5) shows the mean value (%mass) of mineral content (calcium –phosphate – fluoride) obtained from elemental analysis by using EDX for each tested group. Table (3): Comparison of calcium level (% mass) at different repeated points of the measurement interval in the three studied groups Ca (% mass) Progression Test of significance p value Biomin (n = 10) Biomin and Laser (n = 10) Control (n = 10) Baseline - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 24.85–34.42 31.34 X,Z ±2.74 0.87 29.38–33.30 30.35–33.17 27.81–32.07 29.98 X,Z ±1.40 0.44 28.98–30.98 29.13–31.51 25.68–37.98 30.91 X ± 3.58 1.13 28.35–33.47 28.99–33.42 F (df=2) = 0.651 p = .529 NS After Demineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 18.95–32.08 22.64 Y ± 4.02 1.27 19.76–25.51 19.52–23.22 16.38–24.42 20.86 Y ± 2.69 0.85 18.93–22.78 20.27–22.52 16.07–23.84 20.16 Y,Z ±2.59 0.82 18.31–22.01 18.70-22.04 F (df=2) = 1.625 p = .216 NS After Remineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 21.68–36.41 29.49 a,b,X,Z ±3.99 1.26 26.64–32.35 28.18–31.48 25.45–31.77 29.14 a,b,X,Z ±2.39 0.76 27.43–30.85 26.51–30.97 19.58–25.85 22.66 c,Y,Z ±2.14 0.68 21.13–24.19 21.18–25.07 F (df=2) = 16.936 p < .001* One-way repeated measures analysis p value F (GG)(df=1.260) = 16.179 p = .001* F (df=2) = 71.922 p < .001* F (GG)(df=1.186) = 44.448 p < .001* Percentage Change (%) (Baseline vs After Demineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -41.15–3.45 -27.56 ± 12.49 3.95 -36.50 - -18.63 -34.88 - -23.74 -44.43 - -13.81 -30.26 ± 10.14 3.21 -37.51 - -23.01 -36.79 - -21.20 -57.69 - -19.24 -33.77 ± 12.26 3.88 -42.54 - -25.00 -44.48 - -23.62 F (df=2) = 0.710 p = .501 NS Percentage Change (%) (Baseline vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -30.09–19.97 -5.61 a,b ±12.43 3.93 -14.50–3.29 -10.20 - -1.84 -17.20–14.24 -2.56 a,b ±9.83 3.11 -9.59–4.47 -8.99–3.16 -37.01 - -16.49 -26.24 c ± 7.21 2.28 -31.39 - -21.08 -31.94 - -17.56 F (df=2) = 16.412 p < .001* Percentage Change (%) (After Demineralization vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -32.42–66.41 34.20 a,b,c ±27.93 8.83 14.21–54.18 27.05–49.95 23.36–61.84 40.88 a,b ±12.23 3.87 32.13–49.63 32.54–51.11 1.73–60.86 13.94 a,c ±18.18 5.75 0.93–26.94 4.71–14.77 F (df=2) = 4.686 p = .018* Min. – Max. : Minimum – Maximum S.D.: Standard Deviation SE: Standard error CI: Confidence interval df: degree of freedom *: Statistically significant ( p < .05) NS: Statistically not significant ( p ≥ .05) Superscript letters for intergroup comparison Biomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c) Bonferroni Pairwise multiple comparison mean of groups that is labeled with similar superscript letter are statistically not significantly different GG: Greenhouse-Geisser correction Superscript letters for intragroup comparison Baseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z) Table (4): Comparison of Phosphorus level (% mass) at different repeated points of the measurement interval in the three studied groups P (% mass) Progression Test of significance p value Biomin (n = 10) Biomin and Laser (n = 10) Control (n = 10) Baseline - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 14.93–17.32 16.38 X,Z ±0.84 0.27 15.78–16.98 16.04–17.01 14.29–17.23 15.90 X,Z ±0.88 0.28 15.27–16.52 15.75–16.39 14.00-17.62 16.21 X ± 0.96 0.30 15.52–16.90 16.10-16.59 F (df=2) = 0.752 p = .529 NS After Demineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 9.40-12.78 10.77 Y ± 0.96 0.30 10.09–11.46 10.35–10.96 8.14–13.48 11.34 Y ± 1.86 0.59 10.02–12.67 9.23–12.50 8.28–12.56 10.76 Y ± 1.27 0.40 9.85–11.67 10.29–11.76 F (df=2) = 0.555 p = .581 NS After Remineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 15.47–18.38 16.72 a,b,X,Z ±0.82 0.26 16.13–17.31 16.29–16.94 13.06–17.11 16.01 a,b,X,Z ±1.23 0.39 15.13–16.90 15.44–16.89 12.30-15.68 14.04 c,Z ±1.18 0.37 13.20-14.89 13.44–15.01 F (df=2) = 16.056 p < .001* One-way repeated measures analysis p value F (df=2) = 140.679 p < .001* F (df=2) = 39.902 p < .001* F (df=2) = 50.919 p < .001* Percentage Change (%) (Baseline vs After Demineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -45.73 - -23.06 -34.07 ± 6.69 2.12 -38.86 - -29.28 -38.86 - -28.86 -49.19 - -15.06 -28.27 ± 13.24 4.19 -37.74 - -18.80 -43.29 - -17.96 -49.05 - -18.79 -33.23 ± 10.02 3.17 -40.39 - -26.07 -37.67 - -24.42 F (df=2) = 0.919 p = .411 NS Percentage Change (%) (Baseline vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -4.25–14.59 2.24 a,b ±5.81 1.84 -1.91-6.39 -1.99-3.34 -24.20–18.33 1.26 a,b ±11.96 3.78 -7.30–9.82 -3.62–7.81 -24.47 - -2.93 -13.15 c ± 8.21 2.60 -19.02 - -7.27 -20.89 - -4.89 F (df=2) = 9.111 p = .001* Percentage Change (%) (After Demineralization vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 28.72–88.72 56.52 ± 18.02 5.70 43.63–69.41 45.26–62.88 24.18–89.68 43.88 ± 21.26 6.72 28.67–59.10 31.41–44.90 11.62–81.28 32.60 ± 23.26 7.35 15.96–49.24 14.29–47.18 F (df=2) = 3.259 p = .054 NS Min. – Max. : Minimum – Maximum S.D.: Standard Deviation SE: Standard error CI: Confidence interval df: degree of freedom *: Statistically significant ( p < .05) NS: Statistically not significant ( p ≥ .05) Superscript letters Biomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c) Bonferroni Pairwise multiple comparison mean of groups that is labeled with similar superscript letter are statistically not significantly different Superscript letters for intragroup comparison Baseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z) Table (5): Comparison of Fluoride level (% mass) at different repeated points of the measurement interval in the three studied groups F (% mass) Progression Test of significance p value Biomin (n = 10) Biomin and Laser (n = 10) Control (n = 10) Baseline - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 0.63–1.44 0.94 X,Z ± 0.27 0.08 0.75–1.13 0.76–1.18 0.52–1.38 0.87 X,Z ± 0.28 0.09 0.67–1.07 0.61–1.04 0.53–1.17 0.78 X ± .20 0.06 0.63–0.92 0.61–0.90 F (df=2) = 1.098 p = .348 NS After Demineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 0.29–1.17 0.62 Y ± .29 0.09 0.42–0.83 0.37–0.83 0.20–0.98 0.61 Y ± 0.29 0.09 0.41–0.82 0.35–0.82 0.50–0.80 0.64 Y ± 0.09 0.03 0.58–0.70 0.61–0.68 F (df=2) = 0.037 p = .967 NS After Remineralization - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile 0.39–1.40 0.97 a,b,X,Z ± 0.29 0.09 0.76–1.17 0.77–1.16 0.45–1.50 0.98 a,b,X,Z ± 0.30 0.09 0.77–1.19 0.72–1.16 0.35–0.84 0.63 c,Z ± 0.14 0.04 0.53–0.73 0.56–0.74 F (df=2) = 6.241 p = .006* One-way repeated measures analysis p value F (df=2) = 9.465 p = .002* F (df=2) = 17.450 p < .001* F (df=2) = 4.719 p = .023* Percentage Change (%) (Baseline vs After Demineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -61.84–31.75 -33.32 ± 27.54 8.71 -53.02 – -13.62 -54.32 – -19.49 -65.57–27.63 -31.71 ± 25.42 8.04 -49.89 – -13.53 -41.57 – -28.70 -32.43–28.30 -13.59 ± 20.26 6.41 -28.08–0.90 -30.00–1.64 F (df=2) = 1.985 p = .157 NS Percentage Change (%) (Baseline vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -58.51–43.21 5.53 ± 28.57 9.04 -14.91–25.97 -7.32–22.22 -26.23–97.37 16.59 ± 35.21 11.13 -8.60–41.77 -6.96–32.69 -61.11–21.31 -14.99 ± 24.63 7.79 -32.61–2.63 -29.67–5.66 F (df=2) = 2.892 p = .073 NS Percentage Change (%) (After Demineralization vs After Remineralization) - Min-Max - Mean ± SD - SE of Mean - 95.0% CI for Mean - 25th Percentile – 75th Percentile -39.06–213.51 83.08 a,b ± 90.41 28.59 18.41–147.75 9.47–185.37 19.39–260.00 83.91 a,b ± 69.61 22.01 34.11–133.71 35.80–97.14 -44.44–19.35 -1.79 c ± 18.65 5.90 -15.13–11.54 -12.33–5.00 F (df=2) = 5.442 p = .010* Min. – Max. : Minimum – Maximum S.D.: Standard Deviation SE: Standard error CI: Confidence interval df: degree of freedom *: Statistically significant ( p < .05) NS: Statistically not significant ( p ≥ .05) Superscript letters for intergroup comparison Biomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c) Bonferroni Pairwise multiple comparison mean of groups that is labeled with similar superscript letter are statistically not significantly different Superscript letters for intragroup comparison Baseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z) Mineral content (Ca,P,F) using EDX : a. Comparison between baseline and (After demineralization ) : At baseline, there was no statistical significant difference between groups A,B and C regarding to mass% value of Ca, P and F. After demineralization, it was shown that calcium level (% mass) significantly decreased in the three studied groups .Calcium level decreased from a mean of 31.34 ,29.98 and 30.91 (%mass) in groups A,B and C respectively at baseline to a mean of 22.64 ,20.86 and 20.16% mass after demineralization (Fig. 7) (table 3). Phosphorus level % mass also significantly decreased from a mean of 16.38,15.90 and 16.21 to 10.77,11.34 and 10.76%mass in groups A,B and C respectively (Fig. 8) (table4). Regarding to fluoride level (%mass), there was significant decrease from 0.94,0.87 and 0.78 to 0.62,0.61and 0.64 in groups A,B and C respectively (p<0.05) (Fig. 9) (table 5). There was no statistical significant difference among the three studied groups in Ca,P,F level after demineralization (p ≥ 0.05). b. Comparison between (after demineralization) and (after remineralization) : After the application of the remineralizing agents, calcium ions gained a significant increase in groups A and B. Group B showed the highest gain of ions as percentage change (%) was 40.88 ± 12.23. There was no statistical difference between group A and B after remineralization (p ≥ 0.05) and both were significantly higher than the control group (p˂0.05) (Table 3) (Fig. 7). As regards the phosphorus ions, a significant increase was observed in all groups. Group A showed the highest gain as P level percentage change(%mass) was 56.52 ± 18.02 and group C showed the least gain as P level percentage change was 32.60 ± 23.26 .There was no significant difference between group A and group B but both were significantly higher than group C after remineralization (p < 0.001) (Fig. 8) (Table 4). Fluoride ions increased significantly in groups A and B (p˂0.05) but decreased significantly in group C as shown in percentage change formula (-1.78 ± 18.65). There was no statistical difference between group A and B (p ≥ .05) in fluoride level after remineralization (Fig. 9) (Table 5). Both groups A and B were significantly higher than group C (p = .006). C. Comparison between baseline and after remineralization When comparing the mean values of minerals % mass between the baseline and the last stage after application of the remineralizing agents, it was obvious that there was no significant difference in minerals percentage (Ca,P,F) between baseline and after remineralization in groups A and B (p ≥ 0.05). While in the control group, there was significant decrease in the Ca,P and F %mass between baseline and after remineralization process. 2. Environmental scanning electron microscope (ESEM) analysis The sample surface characteristics at each stage were described using ESEM at a magnification of x2000. At base line, micrographs showed smooth enamel surface. After demineralization, samples showed honey comb appearance which represent areas of minerals dissolution. After application of the remineralizing agent, samples showed partial restoration of enamel surface structure. (Figs. 10–14) DISCUSSION This study was conducted to evaluate the impact of diode laser and Biomin F toothpaste on the remineralization of WSLs. The null hypothesis was accepted as there was no significant difference between the effect of Biomin F toothpaste coupled with diode laser and Biomin F alone on the remineralization of white spot lesions. The natural physiological processes of demineralization and remineralization in tooth structure may be disrupted when there is an imbalance between pathogenic and protective factors [ 16 , 19 ]. From a scientific standpoint, it has been recognized that salivary dysfunction, fermentable carbohydrates, and cariogenic bacteria play significant roles as pathogenic factors [ 23 ]. Calcium and phosphate ions presence in a supersaturated state within human saliva enables it to potentially facilitate the remineralization of enamel [ 35 ]. However, if acid challenges exceed this physiological remineralization process, alternative therapeutic interventions are needed to strengthen remineralization [ 23 , 36 ]. Throughout orthodontic treatment, tooth enamel faces an elevated risk of developing white spot lesions due to food accumulation triggered by appliances and insufficient oral hygiene practices. When treating visible white spot lesions using concentrated fluoride agents (hypermineralization), the lesion is arrested at the surface instead of allowing saliva to promote remineralization in the deeper areas [ 16 ]. Presently, there are numerous methods available to halt or reverse the advancement of WSLs utilizing low levels of fluoride, such as casein phosphopeptides–amorphous calcium phosphate (CPP-ACP) [ 36 ], Nano-hydroxyapatite, Trimetaphosphate ion and Bioactive glasses [ 16 , 19 ]. Therefore, Biomin F was utilized in this study as it is a bioactive glass material containing a low level of fluoride. Ali et al [ 37 ] noted in their study that Biomin F toothpastes exhibited lower total fluoride content compared to the values asserted by their respective companies (approximately 400 ppm). Biomin F possesses the characteristic of prolonged fluoride delivery over a 12-hour period through gradual dissolution of the glass [ 16 ]. This characteristic is due to the polymer that enhances the bond between the calcium in the bioglass material and the calcium on the enamel. This bonding reduces the leaching of bioactive glass material [ 38 ]. Biomin F contains small bioglass particles that aid in the infiltration of remineralizing agents into subsurface lesions [ 20 , 39 ]. With its high phosphate content, Biomin F facilitates rapid apatite formation (within 6 hours) and contains fewer carbon impurities, thereby rendering enamel less soluble in acid [ 20 , 39 ]. The majority of studies conducted on Biomin F have explored its efficacy in occluding dentinal tubules and addressing hypersensitivity [ 40 , 41 ]. A systematic review focused on investigating the role of bioactive glass in enamel remineralization highlighted the significant contribution of fluoride-containing bioactive glass dentifrice, specifically Biomin F, in enamel regeneration [ 19 ]. Aidaros et al [ 15 ] carried out an in-vitro investigation utilizing SEM and elemental analysis to assess the mineral composition of extracted permanent third molars prior to and following the application of remineralizing agents, which included Biomin F. Their study involved comparing these agents, employing a similar application regimen to that of the current study (two minutes, twice daily for two weeks). They utilized the materials in the form of toothpaste and concluded that the combination of fluoride with bioactive glass technology, as seen in Biomin F toothpaste, had the most significant impact on the demineralized enamel surface. This finding aligned with our study, which demonstrated that Biomin F possessed the ability to remineralize white spot lesions (WSLs) to restore them to their baseline mineral content. In this current study, laser was examined in conjunction with Biomin F, given that numerous studies have evaluated various types of lasers, including CO2, Nd:YAG, Er:YAG, and diode lasers, utilizing different parameters for caries prevention and enamel remineralization, either with or without fluoride-containing agents [ 42 , 43 ]. The present study revealed that the mineral content following remineralization reached the baseline mineral level in both test groups. This finding contradicted the results reported by Omran et al. [ 20 ] as the mean calcium mass percentage after remineralization was significantly lower compared to the baseline calcium mass percentage in the Biomin group. Notably, both studies employed the same Biomin F application protocol (two minutes twice daily for two weeks). This variation might be explained by the fact that Omran-T applied Biomin F as toothpaste slurries and subjected the samples to a shorter demineralization period (72 hours), while the present study utilized it in the form of toothpaste, as commonly used by orthodontic patients. Nonetheless, both studies concurred on the high phosphate content of Biomin F. An intriguing observation emerged from the study comparing laser treatment with a bioactive glass material (Novamin) using SEM: laser therapy did not provide additional benefits to Novamin in the process of remineralizing the enamel surface [ 23 ]. In the current study, where Biomin F demonstrated the capability to remineralize the enamel surface, the diode laser did not exhibit a synergistic effect in enamel remineralization, as indicated by the insignificant difference between group A and B. Similarly, comparable outcomes were observed when Novamin was utilized alongside laser therapy. The potential rationale for the findings of this study could be that the efficacy of the bioactive glass material (Biomin F) relies on its interaction with physiological aqueous solutions, leading to the release of calcium, phosphorus, and fluoride. However, when a diode laser is employed, a certain degree of heat is generated within the treated surface, typically ranging from 1 to 6 degrees Celsius [ 44 ]. As a consequence, this leads to some dryness and removal of moisture from the paste, which is essential for mineral release. The analysis of SEM micrographs enabled us to observe the notable regeneration of the enamel structure and the deposition of mineral crystals, a result that aligns with findings reported by Bakrey et al., who observed the deposition of mineral crystals blocking the dentinal tubules [ 40 ] after using Biomin F. This study has a limitation that we must consider the dynamic complex system in oral environment which may differ from the in-vitro study employed in the present work. CONCLUSION Within the limitation of the present study, we concluded that Biomin F toothpaste is promising in repairing the white spot lesions on the surface of the demineralized enamel. Diode laser didn’t affect the remineralizing ability of Biomin F toothpaste which add hazard without benefit. Recommendation Clinical studies are needed for more evaluation of the benefits of these approaches. It is recommended to evaluate the synergistic effect between laser and biomin F in decreasing the white spot lesions depth with a variation of laser protocols and exposure time. Declarations Ethics approval and consent to participate The present study was approved by the Research Ethics Committee of the Faculty of Dentistry, Alexandria University (IRB:00010556– IORG:0008839). Consent for publication Not applicable Availability of data and materials The data and materials used to support the findings of this study are available from the corresponding authors upon request. Competing interests The authors declare that there is no conflict of interest. Funding This clinical research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Authors' contributions NT, AM Guide the correct methodology for the trial and supervise the work . Help the student in interpreting results and revising the manuscript. AIE contributed to the concept, methodology and the design of the study. Acknowledgements Not applicable References Ogaard B. Prevalence of white spot lesions in 19-year-olds: a study on untreated and orthodontically treated persons 5 years after treatment. Am J Orthod Dentofacial Orthop. 1989;96:423-7. Palamara J, Phakey PP, Rachinger WA, Orams HJ. Ultrastructure of the intact surface zone of white spot and brown spot carious lesions in human enamel. J Oral Pathol. 1986;15:28-35. Featherstone JD. The science and practice of caries prevention. J Am Dent Assoc. 2000;131:887-99. Ogaard B, Rølla G, Arends J, ten Cate JM. Orthodontic appliances and enamel demineralization. Part 2. 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Aoki A, Sasaki KM, Watanabe H, Ishikawa I. Lasers in nonsurgical periodontal therapy. Periodontol 2000. 2004;36:59-97. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 30 Jul, 2024 Read the published version in BMC Oral Health → Version 1 posted Editorial decision: Revision requested 11 Apr, 2024 Reviews received at journal 09 Apr, 2024 Reviews received at journal 07 Apr, 2024 Reviewers agreed at journal 30 Mar, 2024 Reviewers agreed at journal 28 Mar, 2024 Reviewers invited by journal 10 Mar, 2024 Editor assigned by journal 10 Mar, 2024 Editor invited by journal 04 Mar, 2024 Submission checks completed at journal 04 Mar, 2024 First submitted to journal 26 Feb, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3990752","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":276359782,"identity":"eb1593d2-86aa-475d-a609-b77557d08d5f","order_by":0,"name":"Amira Ibrahim Eldeeb","email":"data:image/png;base64,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","orcid":"","institution":"Alexandria University","correspondingAuthor":true,"prefix":"","firstName":"Amira","middleName":"Ibrahim","lastName":"Eldeeb","suffix":""},{"id":276359783,"identity":"6a43f978-f4c1-437f-a73e-39a8c7a22612","order_by":1,"name":"Nazla O. Tamish","email":"","orcid":"","institution":"Alexandria University","correspondingAuthor":false,"prefix":"","firstName":"Nazla","middleName":"O.","lastName":"Tamish","suffix":""},{"id":276359784,"identity":"f025c99f-43bf-4751-ac4a-cfcfa5f8dad8","order_by":2,"name":"Ahmed M. Madian","email":"","orcid":"","institution":"Alexandria University","correspondingAuthor":false,"prefix":"","firstName":"Ahmed","middleName":"M.","lastName":"Madian","suffix":""}],"badges":[],"createdAt":"2024-02-26 11:14:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3990752/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3990752/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12903-024-04589-9","type":"published","date":"2024-07-30T15:57:33+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":52184190,"identity":"00a99e1e-646a-4977-9368-54da44053a95","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":81730,"visible":true,"origin":"","legend":"\u003cp\u003eFlow chart showing procedure that has been done.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/b44f1adb68132f576fb47617.jpg"},{"id":52186404,"identity":"ad64a7d0-241e-442f-b1d4-2e1bb91a2741","added_by":"auto","created_at":"2024-03-07 18:26:04","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":55625,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Each tooth has a number typed at the base of acrylic block.(b) a window of 4x4mm in the middle third of the buccal surface of the tooth. (c) Each group was stored in a separate beaker containing artificial saliva and labeled with the group name.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/19f8d2c479e79f01f4eb5127.jpg"},{"id":52184191,"identity":"fb0c806e-7e17-4731-8190-8d57e382da88","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":92208,"visible":true,"origin":"","legend":"\u003cp\u003eScanning Electron Microscope attached with EDX unit\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/d9205bcfea548fdd0be1387b.jpg"},{"id":52186405,"identity":"caac9dfa-3b93-4741-ae64-182cdd96333e","added_by":"auto","created_at":"2024-03-07 18:26:04","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":60581,"visible":true,"origin":"","legend":"\u003cp\u003eWhite spot lesions obtained after exposure to demineralizing solution\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/6c18332e01600ee818043b83.jpg"},{"id":52184193,"identity":"4c7301cc-4539-4138-ab14-22c2126fdaa5","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":37931,"visible":true,"origin":"","legend":"\u003cp\u003eBiomin F toothpaste was applied in a circular motion\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/bb2371d8e5e7e41b099ab4f4.jpg"},{"id":52186407,"identity":"d6cb694c-0835-4af2-835f-e076044715ce","added_by":"auto","created_at":"2024-03-07 18:26:04","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":81933,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Laser machine (Lasotronix-Boland),(b)and(c) showed The distance between the buccal surface of the tooth and laser fiber was kept at 5mm\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/85421508cbcfefee7865cdc6.jpg"},{"id":52184202,"identity":"24990cb8-b01c-47c5-b3b2-ee4bbad214f6","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":52535,"visible":true,"origin":"","legend":"\u003cp\u003eClustered bar chart of Mean of Ca (% mass) in the studied groups at different times of measurement\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/593f24364ab5a708736a4ec7.jpg"},{"id":52184195,"identity":"bd314cb2-ac61-42d9-a1ef-b9787dd2dffb","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":50004,"visible":true,"origin":"","legend":"\u003cp\u003eClustered bar chart of Mean of P (% mass) in the studied groups at different times of measurement\u003c/p\u003e","description":"","filename":"8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/1028db9074cf7162a6de6bac.jpg"},{"id":52184197,"identity":"87de818d-91a0-4447-8143-3b5d3f0a7593","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":50639,"visible":true,"origin":"","legend":"\u003cp\u003eClustered bar chart of Mean of F (% mass) in the studied groups at different times of measurement\u003c/p\u003e","description":"","filename":"9.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/ebf0ea765e625d943aa9f4f4.jpg"},{"id":52186825,"identity":"e2d49e79-1c21-4eef-806b-da844a10acdb","added_by":"auto","created_at":"2024-03-07 18:34:04","extension":"jpg","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":40324,"visible":true,"origin":"","legend":"\u003cp\u003eAt the base line, micrographs (a)sample 9 in group a (b) sample 6 in group b(c)sample 5 in group c showed sound enamel with smooth surface .The enamel prisms showed minimal visibility, and the presence of scratches suggests potential carbon surface contamination.\u003c/p\u003e","description":"","filename":"10.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/c737290d74281d4dec860386.jpg"},{"id":52184203,"identity":"3ea5e885-043c-4a8f-8dd7-4ef71a2867aa","added_by":"auto","created_at":"2024-03-07 18:18:05","extension":"jpg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":112339,"visible":true,"origin":"","legend":"\u003cp\u003eAfter demineralization ,micrographs (a)sample 3 in group a(b)sample 14 in group b (c)sample 11 in group c showed areas of dissolution and pores characterized by honey comb appearance. Enamel prisms become more visible because of erosion.\u003c/p\u003e","description":"","filename":"11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/bb3ce94b77e26a7243476059.jpg"},{"id":52184198,"identity":"c7eebf42-3ecc-49cd-80ed-763ba7cf5ef6","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":99098,"visible":true,"origin":"","legend":"\u003cp\u003eAfter remineralization ,biomin group showed enamel surface with partial restoration of the surface structure and areas of uniform and smooth enamel suface. Mineral crystals were deposited obliterating prism cores.(a)sample 22(b) sample 27(c) sample 28 in biomin group.\u003c/p\u003e","description":"","filename":"12.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/9220e82e33206bf040168f51.jpg"},{"id":52184201,"identity":"dd541f58-a8c7-4524-b7db-9805f3ca974e","added_by":"auto","created_at":"2024-03-07 18:18:04","extension":"jpg","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":75057,"visible":true,"origin":"","legend":"\u003cp\u003eAfter remineralization in biomin and laser group micrographs showed areas of crystal deposition obliterating prism cores and smooth areas at the surface of enamel(a)sample 23(b) sample 4 (c) sample 14 in biomin and laser group.\u003c/p\u003e","description":"","filename":"13.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/7e745afe82bba0b211de6d32.jpg"},{"id":52186406,"identity":"4318dad5-f556-4a8f-8120-3f0b7250b334","added_by":"auto","created_at":"2024-03-07 18:26:04","extension":"jpg","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":103516,"visible":true,"origin":"","legend":"\u003cp\u003eIn the control goup, micrographs showed areas of erosions with visible prisms\u003cstrong\u003e \u003c/strong\u003eSample 10 (b) sample 17 (c)sample 12 in the control group.\u003c/p\u003e","description":"","filename":"14.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/4f429db2d50683fbc7ff3333.jpg"},{"id":61793484,"identity":"1d86f08c-cea1-4892-8477-e75edf4aae4b","added_by":"auto","created_at":"2024-08-05 16:13:08","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2490544,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3990752/v1/d5572e71-33e4-4eb9-a566-77825f5c2052.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eEffect of Biomin F Toothpaste and Diode Laser on Remineralization of White Spot Lesions (In Vitro Study)\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eTypically, patients seek orthodontic intervention to enhance their aesthetic appeal. Nonetheless, the use of fixed orthodontic devices could result in the development of white spot lesions (WSLs), posing an additional aesthetic challenge for the patient. Consequently, both the patient and orthodontist may experience disappointment upon the removal of the appliances [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWhite spot lesions are recognized as the initial stage of tooth decay, characterized by a surface enamel layer showing a significant reduction in mineral content [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. This condition is prone to deteriorate, potentially necessitating invasive treatment [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Enamel demineralization and the development of WSLs advance swiftly [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], often developing within a few weeks [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePreventive approaches for WSLs during orthodontic treatment primarily involve the utilization of fluoride-releasing varnishes [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], bonding materials, and cements [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Additionally, the application of concentrated fluoride gels [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] and daily rinsing with mouthwash [\u003cspan additionalcitationids=\"CR12 CR13\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] are employed to reduce enamel demineralization.\u003c/p\u003e \u003cp\u003eWhile fluoride application is effective in halting WSLs, it comes with certain constraints. The consistent use of toothpaste containing fluoride necessitates a significant presence of bioavailable calcium and phosphate ions alongside fluoride ions.\u003csup\u003e[15]\u003c/sup\u003e Furthermore, fluoride exhibits diminished effectiveness when pH level drops below 4.5, a situation often instigated by bacterial actions [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAlthough the impact of fluoride relies on the dosage and improves with higher doses, there exists a threshold beyond which increasing the fluoride dose may lead to unwanted side effects such as dental fluorosis and toxicity [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn recent times, bioactive glasses have emerged as a notable advancement in dental applications, extensively researched in various studies targeting the treatment of white spot lesions through remineralization. These materials have the potential to rejuvenate and regenerate dental tissues by triggering apatite formation upon exposure to saliva or any other physiological fluid [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. These apatites can either be hydroxyapatites or fluorapatites, depending on whether fluoride is integrated into the glass structure chemical composition. Glasses contain fluoride exhibit \"smart\" properties, demonstrating remineralization activity enhancement in low pH environments. Biomin F is recognized as a bioactive glass-based toothpaste that incorporates fluoride to aid in the remineralization of enamel.\u003c/p\u003e \u003cp\u003eLaser technology has been utilized to decrease the rate of subsurface demineralization of enamel by modifying its crystalline structure, acid solubility, and permeability. However, it is crucial to apply lasers at a low energy level to maintain enamel integrity [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFollowing laser irradiation, enamel undergoes chemical and structural changes, including reduction in carbonates fusion and re-crystallization of hydroxyapatite crystals. These alterations enhance enamel's resistance against the acid attacks. Moreover, studies have demonstrated synergistic effects between laser treatment and topical fluoride application, leading to a significant reduction in the rate of enamel decalcification [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHence, this study was conducted for the evaluation of the combined impact of Biomin F toothpaste and diode laser on the remineralization of WSLs.\u003c/p\u003e \u003cp\u003eThe null hypothesis of this study assumed that no significant difference is expected between the effect of Biomin F toothpaste coupled with diode laser and Biomin F alone on remineralization of white spot lesions.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eThis in-vitro study was conducted at the Department of Orthodontics, Faculty of Dentistry, Alexandria University and Scanning Electron Microscope unit, Faculty of Sciences, Alexandria University. The present study was approved by the Research Ethics Committee of the Faculty of Dentistry, Alexandria University (IRB:00010556\u0026ndash; IORG:0008839).\u003c/p\u003e \u003cp\u003e \u003cb\u003eSample size calculation\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe sample size estimate was calculated based on an invitro study by Aidaros et al. (2022) [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] that aimed to evaluate and compare the remineralizing potential of dentifrices containing fluoride and bioactive glass on enamel by assessing the enamel structure and elemental analysis through Energy Dispersive X-ray Analysis (EDX). During sample size calculation, a beta error of up to 20% is accepted, with a study power of 80%. The alpha level was established at 5%, corresponding to a significance level of 95%. Statistical significance was assessed at a \u003cem\u003ep\u003c/em\u003e-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The minimum required sample size was determined to be 9 teeth per group (number of groups\u0026thinsp;=\u0026thinsp;3) (Total sample size\u0026thinsp;=\u0026thinsp;27 teeth). Any withdrawal for any reason will be compensated by replacement to control for attrition (loss of specimen) bias. Therefore sample size will be increased to 10 teeth per group (number of groups\u0026thinsp;=\u0026thinsp;3) (Total sample size\u0026thinsp;=\u0026thinsp;30 teeth).\u003c/p\u003e \u003cp\u003eA total of 30 human premolars were collected from patients requiring premolars extraction during their orthodontic treatment in the Orthodontic Department, Faculty of Dentistry, Alexandria University, Egypt. Informed consent was obtained from all subjects and/or their legal guardian(s).\u003c/p\u003e \u003cp\u003eAll patients were born and lived in areas where the typical concentration of fluoride in the drinking water was 0.36 mg/L [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Any calculus or tissue remnants were removed from the teeth using a scaler. Subsequently, the teeth were stored in saline until the commencement of the study.\u003c/p\u003e \u003cp\u003eProcedures for each group are shown in the flowchart (Fig.\u0026nbsp;1).\u003c/p\u003e \u003cp\u003e \u003cb\u003e1. Collection of teeth\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThirty human premolars that were extracted for orthodontic purposes were chosen for inclusion in this study. All teeth were examined macroscopically using a magnifying loupe and fulfilled the following selection criteria: Intact buccal enamel surface, with no decalcifications, cracks, or stains.Teeth previously bonded or received any chemical treatment or decayed premolars were excluded from this study. The teeth were preserved in saline until the start of the experiment.\u003c/p\u003e \u003cp\u003e \u003cb\u003e2. Teeth preparation\u003c/b\u003e \u003c/p\u003e \u003cp\u003eAfter recruiting the appropriate teeth, all remnants were removed and teeth were cleaned with fluoride free pumice and running water. The roots of the teeth were cut 2mm under cemento-enamel junction using a diamond disk under water cooling. The crowns were embedded in self cured acrylic resin and the buccal surfaces were directed upward for easy manipulation [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. (Fig.\u0026nbsp;2a) Each sample was covered with acid resistant varnish (nail polish) at all tooth crown surfaces, leaving a window of 4mm X4mm in the middle third of the buccal surface of the premolar. (Fig.\u0026nbsp;2b)\u003c/p\u003e \u003cp\u003e \u003cb\u003eRandomization\u003c/b\u003e \u003c/p\u003e \u003cp\u003eEach tooth got a number from 1 to 30 typed at the base of the acrylic block using waterproof permanent marker. These numbers were used to randomly and equally assign the samples into 3 groups using computer generated random list.\u003c/p\u003e \u003cp\u003e \u003cb\u003e3. Grouping of the teeth\u003c/b\u003e \u003c/p\u003e \u003cp\u003eTeeth were divided randomly into 3 groups\u003c/p\u003e \u003cp\u003eGroup A: Biomin F tooth paste.\u003c/p\u003e \u003cp\u003eGroup B: Biomin F with laser application for 30 sec .\u003c/p\u003e \u003cp\u003eGroup C: Negative control group (no treatment(.\u003c/p\u003e \u003cp\u003e Each group was placed in its own labeled beaker which contained 150 mL of artificial saliva solution at room temperature of 37\u0026deg;C and neutral PH to replicate the oral environment. (Fig.\u0026nbsp;2c) The materials used, their specifications, compositions and manufactures are present in (Table\u0026nbsp;1).\u003c/p\u003e \u003cp\u003e \u003cb\u003e4. Intervention\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe study was divided into a number of stages with various procedures as the following:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eFirst stage (Baseline) assessment\u003c/b\u003e \u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eThe assessment was conducted utilizing an environmental scanning electron microscope (JSM-IT 200-Japan) at Faculty of Sciences, Alexandria University. SEM attached with energy-dispersive X-ray (EDX) unit (Fig.\u0026nbsp;3).\u003c/p\u003e \u003cp\u003eThe amount of remineralization was measured qualitatively by comparing the scanning electron microscope pictures and quantitatively by the Energy dispersive X-ray analysis that measured the average of three points selected in the area of concern. These values were taken at different stages of the study (before, after demineralization and after remineralization).\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eSecond stage (Demineralization process)\u003c/b\u003e \u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eThe teeth were immersed in the demineralizing solution for 96 hours at 37ᵒC until white spot lesions were obtained (Fig.\u0026nbsp;4) (Table\u0026nbsp;1). The samples were removed from the solution and rinsed with distilled water to stop the demineralization process and to remove any residuals of the solution [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. At this stage, evaluations were carried out using Scanning Electron Microscope (SEM) analysis alongside Energy Dispersive X-ray Analysis (EDX).\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eThird stage)Remineralization process)\u003c/b\u003e \u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eIn group A: Biomin F toothpaste was applied in circular motion on the demineralized region using microbrush twice daily, each lasted for two minutes and left undisturbed for 30 seconds (Fig.\u0026nbsp;5). Then the samples were rinsed carefully with distilled water to remove any excess paste. Then it was stored in artificial saliva to mimic oral environment. This procedure was repeated for two weeks (Table\u0026nbsp;1) [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn group B: The toothpaste was applied twice daily each for two minutes in a circular motion using a microbrush and left undisturbed for 30 seconds which lasted for two weeks, then laser was applied at the 14th day (Fig.\u0026nbsp;6) (Table\u0026nbsp;2).\u003c/p\u003e \u003cp\u003eIn group C: No treatment was received.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(1): The materials used and their specifications\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSolution\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eComposition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eUsage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArtificial saliva\u003csup\u003e[20]\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026bull; 2.20 g/L gastric mucin\u003c/p\u003e \u003cp\u003e\u0026bull; 1.45 mmol/L CaCl2 2H2O\u003c/p\u003e \u003cp\u003e\u0026bull; 5.42 mmol/L KH2PO4\u003c/p\u003e \u003cp\u003e\u0026bull; 6.50 mmol/L NaCl\u003c/p\u003e \u003cp\u003e\u0026bull; 14.94 mmol/L KCl.\u003c/p\u003e \u003cp\u003ePH was adjusted to 7.0 using KOH.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStorage medium for the samples.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDemineralizing solution[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026bull; 2.2 mM CaCl2\u003c/p\u003e \u003cp\u003e\u0026bull; 2.2 mM NaH2PO4\u003c/p\u003e \u003cp\u003e\u0026bull; 0.05 M lactic acid\u003c/p\u003e \u003cp\u003e\u0026bull; 0.2 ppm fluoride.\u003c/p\u003e \u003cp\u003eThe pH was adjusted to 4.5 with 50% NaOH.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eArtificial sub-surface lesion formation.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBioMinF Armour for teeth toothpaste\u003c/p\u003e \u003cp\u003e(BioMin Technologies Ltd.,\u003c/p\u003e \u003cp\u003eLondon, UK)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026bull; Fluoro Calcium-PhosphoSilicate (Biomin)\u003c/p\u003e \u003cp\u003e\u0026bull; Glycerin\u003c/p\u003e \u003cp\u003e\u0026bull; Silica\u003c/p\u003e \u003cp\u003e\u0026bull; PEG 400\u003c/p\u003e \u003cp\u003e\u0026bull; Sodium-Lauryl Sulphate\u003c/p\u003e \u003cp\u003e\u0026bull; Titanium Dioxide\u003c/p\u003e \u003cp\u003e\u0026bull; Aroma\u003c/p\u003e \u003cp\u003e\u0026bull; Carbomer\u003c/p\u003e \u003cp\u003e\u0026bull; Potassium Acesulfame\u003c/p\u003e \u003cp\u003e\u0026bull; Contains maximum\u003c/p\u003e \u003cp\u003e530ppm of available fluoride when packed.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRemineralizing agent\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e\u003cb\u003eLaser application\u003c/b\u003e\u003c/p\u003e\u003cp\u003eLaser irradiation (Lastronix-Boland) has been done on the 14th day for 30 seconds after Laser beam activation with carbon particles and laser setting was done as shown in table (2). The practitioner wore eye goggles for self protection from laser irradiation. The distance between the buccal surface of the tooth and laser fiber was kept 5mm using custom- made holder (Fig.\u0026nbsp;6) (Table\u0026nbsp;2).[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(2): Parameters of laser irradiation\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabb\" border=\"1\"\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCenter wave lenghth(nm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e980\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOperating mode\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eContinues wave(CW)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePower output(W)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExposure time(s)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRadiant energy(J)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOptic conductor fiber diameter(\u0026micro;m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e320\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMode\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNon contact mode\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe Scanning Electron Microscope assessment and Energy Dispersive X-ray Analysis (EDX) were repeated after remineralization process.\u003c/p\u003e \u003cp\u003e \u003cb\u003eBlinding\u003c/b\u003e \u003c/p\u003e \u003cp\u003eTechnician of Scanning Electron Microscope attached with EDX unit and the statistician were blinded.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eThe data were gathered and inputted into the computer using the Statistical Package for the Social Sciences (SPSS) software program for statistical analysis (ver 25).[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eKolmogorov-Smirnov test of normality revealed no significance in the distribution of the variables, so the parametric statistics were adopted.[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eData were described using minimum, maximum, mean, standard deviation, standard error of the mean, and 95% CI of the mean, 25th to 75th percentile.[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eComparisons were conducted among more than two independent normally distributed subgroups utilizing the one-way Analysis of Variance (ANOVA) test [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Post-hoc multiple comparisons [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] were performed using the Bonferroni method [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eRepeated measures analysis of variance was used [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Model assumptions were tested and found to be satisfactory with the exception of Mauchly\u0026rsquo;s test of sphericity [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], and when it was statistically significant denoting the violation of the assumption of sphericity, Greenhouse-Geisser correction was used [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Pair-wise comparison was done with Bonferroni correction.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eLinear trend analysis was used to test for within-subjects contrast [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003ePercentage change was calculated as follows:\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\varvec{P}\\varvec{e}\\varvec{r}\\varvec{c}\\varvec{e}\\varvec{n}\\varvec{t}\\varvec{a}\\varvec{g}\\varvec{e} \\varvec{c}\\varvec{h}\\varvec{a}\\varvec{n}\\varvec{g}\\varvec{e} \\left(\\varvec{\\%}\\right)=\\frac{Measurement \\left(after\\right)-Measurement \\left(before\\right)}{Measurement \\left(before\\right)}x 100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003eThis study included 30 premolars extracted for orthodontic purposes, all of which had an intact buccal enamel surface without any apparent decalcifications, cracks, or stains. These premolars were randomly assigned to three groups, with each group consisting of ten premolars:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eGroup a : Biomin F Tooth paste\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eGroup b : Biomin F with laser application 30 sec\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eGroup c : Negative control\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eTables\u0026nbsp;(3,4,5) shows the mean value (%mass) of mineral content (calcium \u0026ndash;phosphate \u0026ndash; fluoride) obtained from elemental analysis by using EDX for each tested group.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(3): Comparison of calcium level (% mass) at different repeated points of the measurement interval in the three studied groups\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabc\" border=\"1\"\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e \u003cp\u003eCa (% mass)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c7\" namest=\"c3\"\u003e \u003cp\u003eProgression\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003cp\u003eof significance\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eBiomin\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eBiomin and Laser\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eBaseline\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e24.85\u0026ndash;34.42\u003c/p\u003e \u003cp\u003e31.34\u003csup\u003eX,Z\u003c/sup\u003e\u0026plusmn;2.74\u003c/p\u003e \u003cp\u003e0.87\u003c/p\u003e \u003cp\u003e29.38\u0026ndash;33.30\u003c/p\u003e \u003cp\u003e30.35\u0026ndash;33.17\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e27.81\u0026ndash;32.07\u003c/p\u003e \u003cp\u003e29.98\u003csup\u003eX,Z\u003c/sup\u003e\u0026plusmn;1.40\u003c/p\u003e \u003cp\u003e0.44\u003c/p\u003e \u003cp\u003e28.98\u0026ndash;30.98\u003c/p\u003e \u003cp\u003e29.13\u0026ndash;31.51\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25.68\u0026ndash;37.98\u003c/p\u003e \u003cp\u003e30.91\u003csup\u003eX\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;3.58\u003c/p\u003e \u003cp\u003e1.13\u003c/p\u003e \u003cp\u003e28.35\u0026ndash;33.47\u003c/p\u003e \u003cp\u003e28.99\u0026ndash;33.42\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.651\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.529 NS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Demineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e18.95\u0026ndash;32.08\u003c/p\u003e \u003cp\u003e22.64\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;4.02\u003c/p\u003e \u003cp\u003e1.27\u003c/p\u003e \u003cp\u003e19.76\u0026ndash;25.51\u003c/p\u003e \u003cp\u003e19.52\u0026ndash;23.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e16.38\u0026ndash;24.42\u003c/p\u003e \u003cp\u003e20.86\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.69\u003c/p\u003e \u003cp\u003e0.85\u003c/p\u003e \u003cp\u003e18.93\u0026ndash;22.78\u003c/p\u003e \u003cp\u003e20.27\u0026ndash;22.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e16.07\u0026ndash;23.84\u003c/p\u003e \u003cp\u003e20.16\u003csup\u003e\u003cb\u003eY,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;2.59\u003c/p\u003e \u003cp\u003e0.82\u003c/p\u003e \u003cp\u003e18.31\u0026ndash;22.01\u003c/p\u003e \u003cp\u003e18.70-22.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.625\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.216 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Remineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e21.68\u0026ndash;36.41\u003c/p\u003e \u003cp\u003e29.49\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;3.99\u003c/p\u003e \u003cp\u003e1.26\u003c/p\u003e \u003cp\u003e26.64\u0026ndash;32.35\u003c/p\u003e \u003cp\u003e28.18\u0026ndash;31.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e25.45\u0026ndash;31.77\u003c/p\u003e \u003cp\u003e29.14\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;2.39\u003c/p\u003e \u003cp\u003e0.76\u003c/p\u003e \u003cp\u003e27.43\u0026ndash;30.85\u003c/p\u003e \u003cp\u003e26.51\u0026ndash;30.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.58\u0026ndash;25.85\u003c/p\u003e \u003cp\u003e22.66\u003csup\u003e\u003cb\u003ec,Y,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;2.14\u003c/p\u003e \u003cp\u003e0.68\u003c/p\u003e \u003cp\u003e21.13\u0026ndash;24.19\u003c/p\u003e \u003cp\u003e21.18\u0026ndash;25.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;16.936\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOne-way repeated measures analysis\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eF\u003csub\u003e(GG)(df=1.260)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;16.179\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;71.922\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF\u003csub\u003e(GG)(df=1.186)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;44.448\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Demineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e-41.15\u0026ndash;3.45\u003c/p\u003e \u003cp\u003e-27.56\u0026thinsp;\u0026plusmn;\u0026thinsp;12.49\u003c/p\u003e \u003cp\u003e3.95\u003c/p\u003e \u003cp\u003e-36.50 - -18.63\u003c/p\u003e \u003cp\u003e-34.88 - -23.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e-44.43 - -13.81\u003c/p\u003e \u003cp\u003e-30.26\u0026thinsp;\u0026plusmn;\u0026thinsp;10.14\u003c/p\u003e \u003cp\u003e3.21\u003c/p\u003e \u003cp\u003e-37.51 - -23.01\u003c/p\u003e \u003cp\u003e-36.79 - -21.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e-57.69 - -19.24\u003c/p\u003e \u003cp\u003e-33.77\u0026thinsp;\u0026plusmn;\u0026thinsp;12.26\u003c/p\u003e \u003cp\u003e3.88\u003c/p\u003e \u003cp\u003e-42.54 - -25.00\u003c/p\u003e \u003cp\u003e-44.48 - -23.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.710\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.501 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e-30.09\u0026ndash;19.97\u003c/p\u003e \u003cp\u003e-5.61\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e \u0026plusmn;12.43\u003c/p\u003e \u003cp\u003e3.93\u003c/p\u003e \u003cp\u003e-14.50\u0026ndash;3.29\u003c/p\u003e \u003cp\u003e-10.20 - -1.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e-17.20\u0026ndash;14.24\u003c/p\u003e \u003cp\u003e-2.56\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;9.83\u003c/p\u003e \u003cp\u003e3.11\u003c/p\u003e \u003cp\u003e-9.59\u0026ndash;4.47\u003c/p\u003e \u003cp\u003e-8.99\u0026ndash;3.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e-37.01 - -16.49\u003c/p\u003e \u003cp\u003e-26.24\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;7.21\u003c/p\u003e \u003cp\u003e2.28\u003c/p\u003e \u003cp\u003e-31.39 - -21.08\u003c/p\u003e \u003cp\u003e-31.94 - -17.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;16.412\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (After Demineralization vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e-32.42\u0026ndash;66.41\u003c/p\u003e \u003cp\u003e34.20\u003csup\u003e\u003cb\u003ea,b,c\u003c/b\u003e\u003c/sup\u003e \u0026plusmn;27.93\u003c/p\u003e \u003cp\u003e8.83\u003c/p\u003e \u003cp\u003e14.21\u0026ndash;54.18\u003c/p\u003e \u003cp\u003e27.05\u0026ndash;49.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e23.36\u0026ndash;61.84\u003c/p\u003e \u003cp\u003e40.88\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;12.23\u003c/p\u003e \u003cp\u003e3.87\u003c/p\u003e \u003cp\u003e32.13\u0026ndash;49.63\u003c/p\u003e \u003cp\u003e32.54\u0026ndash;51.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e1.73\u0026ndash;60.86\u003c/p\u003e \u003cp\u003e13.94\u003csup\u003e\u003cb\u003ea,c\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;18.18\u003c/p\u003e \u003cp\u003e5.75\u003c/p\u003e \u003cp\u003e0.93\u0026ndash;26.94\u003c/p\u003e \u003cp\u003e4.71\u0026ndash;14.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;4.686\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.018*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMin. \u0026ndash; Max. : Minimum \u0026ndash; Maximum S.D.: Standard Deviation\u003c/p\u003e \u003cp\u003eSE: Standard error CI: Confidence interval\u003c/p\u003e \u003cp\u003edf: degree of freedom\u003c/p\u003e \u003cp\u003e*: Statistically significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.05) NS: Statistically not significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;.05)\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters for intergroup comparison\u003c/strong\u003e \u003cp\u003eBiomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eBonferroni Pairwise multiple comparison\u003c/strong\u003e \u003cp\u003emean of groups that is labeled with similar superscript letter are statistically not significantly different\u003c/p\u003e \u003c/p\u003e \u003cp\u003eGG: Greenhouse-Geisser correction\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters for intragroup comparison\u003c/strong\u003e \u003cp\u003eBaseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(4): Comparison of Phosphorus level (% mass) at different repeated points of the measurement interval in the three studied groups\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabd\" border=\"1\"\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eP (% mass)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003eProgression\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003cp\u003eof significance\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBiomin\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBiomin and Laser\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBaseline\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.93\u0026ndash;17.32\u003c/p\u003e \u003cp\u003e16.38\u003csup\u003eX,Z\u003c/sup\u003e\u0026plusmn;0.84\u003c/p\u003e \u003cp\u003e0.27\u003c/p\u003e \u003cp\u003e15.78\u0026ndash;16.98\u003c/p\u003e \u003cp\u003e16.04\u0026ndash;17.01\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.29\u0026ndash;17.23\u003c/p\u003e \u003cp\u003e15.90\u003csup\u003eX,Z\u003c/sup\u003e\u0026plusmn;0.88\u003c/p\u003e \u003cp\u003e0.28\u003c/p\u003e \u003cp\u003e15.27\u0026ndash;16.52\u003c/p\u003e \u003cp\u003e15.75\u0026ndash;16.39\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.00-17.62\u003c/p\u003e \u003cp\u003e16.21\u003csup\u003eX\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96\u003c/p\u003e \u003cp\u003e0.30\u003c/p\u003e \u003cp\u003e15.52\u0026ndash;16.90\u003c/p\u003e \u003cp\u003e16.10-16.59\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.752\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.529 NS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Demineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.40-12.78\u003c/p\u003e \u003cp\u003e10.77\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96\u003c/p\u003e \u003cp\u003e0.30\u003c/p\u003e \u003cp\u003e10.09\u0026ndash;11.46\u003c/p\u003e \u003cp\u003e10.35\u0026ndash;10.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.14\u0026ndash;13.48\u003c/p\u003e \u003cp\u003e11.34\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003c/p\u003e \u003cp\u003e0.59\u003c/p\u003e \u003cp\u003e10.02\u0026ndash;12.67\u003c/p\u003e \u003cp\u003e9.23\u0026ndash;12.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.28\u0026ndash;12.56\u003c/p\u003e \u003cp\u003e10.76\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27\u003c/p\u003e \u003cp\u003e0.40\u003c/p\u003e \u003cp\u003e9.85\u0026ndash;11.67\u003c/p\u003e \u003cp\u003e10.29\u0026ndash;11.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.555\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.581 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Remineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.47\u0026ndash;18.38\u003c/p\u003e \u003cp\u003e16.72\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;0.82\u003c/p\u003e \u003cp\u003e0.26\u003c/p\u003e \u003cp\u003e16.13\u0026ndash;17.31\u003c/p\u003e \u003cp\u003e16.29\u0026ndash;16.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.06\u0026ndash;17.11\u003c/p\u003e \u003cp\u003e16.01\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;1.23\u003c/p\u003e \u003cp\u003e0.39\u003c/p\u003e \u003cp\u003e15.13\u0026ndash;16.90\u003c/p\u003e \u003cp\u003e15.44\u0026ndash;16.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.30-15.68\u003c/p\u003e \u003cp\u003e14.04\u003csup\u003e\u003cb\u003ec,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;1.18\u003c/p\u003e \u003cp\u003e0.37\u003c/p\u003e \u003cp\u003e13.20-14.89\u003c/p\u003e \u003cp\u003e13.44\u0026ndash;15.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;16.056\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOne-way repeated measures analysis\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;140.679\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;39.902\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;50.919\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Demineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-45.73 - -23.06\u003c/p\u003e \u003cp\u003e-34.07\u0026thinsp;\u0026plusmn;\u0026thinsp;6.69\u003c/p\u003e \u003cp\u003e2.12\u003c/p\u003e \u003cp\u003e-38.86 - -29.28\u003c/p\u003e \u003cp\u003e-38.86 - -28.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-49.19 - -15.06\u003c/p\u003e \u003cp\u003e-28.27\u0026thinsp;\u0026plusmn;\u0026thinsp;13.24\u003c/p\u003e \u003cp\u003e4.19\u003c/p\u003e \u003cp\u003e-37.74 - -18.80\u003c/p\u003e \u003cp\u003e-43.29 - -17.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-49.05 - -18.79\u003c/p\u003e \u003cp\u003e-33.23\u0026thinsp;\u0026plusmn;\u0026thinsp;10.02\u003c/p\u003e \u003cp\u003e3.17\u003c/p\u003e \u003cp\u003e-40.39 - -26.07\u003c/p\u003e \u003cp\u003e-37.67 - -24.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.919\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.411 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-4.25\u0026ndash;14.59\u003c/p\u003e \u003cp\u003e2.24\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;5.81\u003c/p\u003e \u003cp\u003e1.84\u003c/p\u003e \u003cp\u003e-1.91-6.39\u003c/p\u003e \u003cp\u003e-1.99-3.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-24.20\u0026ndash;18.33\u003c/p\u003e \u003cp\u003e1.26\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e\u0026plusmn;11.96\u003c/p\u003e \u003cp\u003e3.78\u003c/p\u003e \u003cp\u003e-7.30\u0026ndash;9.82\u003c/p\u003e \u003cp\u003e-3.62\u0026ndash;7.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-24.47 - -2.93\u003c/p\u003e \u003cp\u003e-13.15\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;8.21\u003c/p\u003e \u003cp\u003e2.60\u003c/p\u003e \u003cp\u003e-19.02 - -7.27\u003c/p\u003e \u003cp\u003e-20.89 - -4.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;9.111\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (After Demineralization vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28.72\u0026ndash;88.72\u003c/p\u003e \u003cp\u003e56.52\u0026thinsp;\u0026plusmn;\u0026thinsp;18.02\u003c/p\u003e \u003cp\u003e5.70\u003c/p\u003e \u003cp\u003e43.63\u0026ndash;69.41\u003c/p\u003e \u003cp\u003e45.26\u0026ndash;62.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.18\u0026ndash;89.68\u003c/p\u003e \u003cp\u003e43.88\u0026thinsp;\u0026plusmn;\u0026thinsp;21.26\u003c/p\u003e \u003cp\u003e6.72\u003c/p\u003e \u003cp\u003e28.67\u0026ndash;59.10\u003c/p\u003e \u003cp\u003e31.41\u0026ndash;44.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.62\u0026ndash;81.28\u003c/p\u003e \u003cp\u003e32.60\u0026thinsp;\u0026plusmn;\u0026thinsp;23.26\u003c/p\u003e \u003cp\u003e7.35\u003c/p\u003e \u003cp\u003e15.96\u0026ndash;49.24\u003c/p\u003e \u003cp\u003e14.29\u0026ndash;47.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;3.259\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.054 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMin. \u0026ndash; Max. : Minimum \u0026ndash; Maximum S.D.: Standard Deviation\u003c/p\u003e \u003cp\u003eSE: Standard error CI: Confidence interval\u003c/p\u003e \u003cp\u003edf: degree of freedom\u003c/p\u003e \u003cp\u003e*: Statistically significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.05) NS: Statistically not significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;.05)\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters\u003c/strong\u003e \u003cp\u003eBiomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eBonferroni Pairwise multiple comparison\u003c/strong\u003e \u003cp\u003emean of groups that is labeled with similar superscript letter are statistically not significantly different\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters for intragroup comparison\u003c/strong\u003e \u003cp\u003eBaseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eTable\u0026nbsp;(5): Comparison of Fluoride level (% mass) at different repeated points of the measurement interval in the three studied groups\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabe\" border=\"1\"\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eF (% mass)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003eProgression\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003cp\u003eof significance\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBiomin\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBiomin and Laser\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;10)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBaseline\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.63\u0026ndash;1.44\u003c/p\u003e \u003cp\u003e0.94\u003csup\u003eX,Z\u003c/sup\u003e \u0026plusmn; 0.27\u003c/p\u003e \u003cp\u003e0.08\u003c/p\u003e \u003cp\u003e0.75\u0026ndash;1.13\u003c/p\u003e \u003cp\u003e0.76\u0026ndash;1.18\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.52\u0026ndash;1.38\u003c/p\u003e \u003cp\u003e0.87\u003csup\u003eX,Z\u003c/sup\u003e\u0026plusmn; 0.28\u003c/p\u003e \u003cp\u003e0.09\u003c/p\u003e \u003cp\u003e0.67\u0026ndash;1.07\u003c/p\u003e \u003cp\u003e0.61\u0026ndash;1.04\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.53\u0026ndash;1.17\u003c/p\u003e \u003cp\u003e0.78\u003csup\u003eX\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;.20\u003c/p\u003e \u003cp\u003e0.06\u003c/p\u003e \u003cp\u003e0.63\u0026ndash;0.92\u003c/p\u003e \u003cp\u003e0.61\u0026ndash;0.90\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.098\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.348 NS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Demineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.29\u0026ndash;1.17\u003c/p\u003e \u003cp\u003e0.62\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;.29\u003c/p\u003e \u003cp\u003e0.09\u003c/p\u003e \u003cp\u003e0.42\u0026ndash;0.83\u003c/p\u003e \u003cp\u003e0.37\u0026ndash;0.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.20\u0026ndash;0.98\u003c/p\u003e \u003cp\u003e0.61\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e \u003cp\u003e0.09\u003c/p\u003e \u003cp\u003e0.41\u0026ndash;0.82\u003c/p\u003e \u003cp\u003e0.35\u0026ndash;0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.50\u0026ndash;0.80\u003c/p\u003e \u003cp\u003e0.64\u003csup\u003e\u003cb\u003eY\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003cp\u003e0.03\u003c/p\u003e \u003cp\u003e0.58\u0026ndash;0.70\u003c/p\u003e \u003cp\u003e0.61\u0026ndash;0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.037\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.967 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAfter Remineralization\u003c/b\u003e\u003c/p\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.39\u0026ndash;1.40\u003c/p\u003e \u003cp\u003e0.97\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e\u0026plusmn; 0.29\u003c/p\u003e \u003cp\u003e0.09\u003c/p\u003e \u003cp\u003e0.76\u0026ndash;1.17\u003c/p\u003e \u003cp\u003e0.77\u0026ndash;1.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.45\u0026ndash;1.50\u003c/p\u003e \u003cp\u003e0.98\u003csup\u003e\u003cb\u003ea,b,X,Z\u003c/b\u003e\u003c/sup\u003e \u0026plusmn; 0.30\u003c/p\u003e \u003cp\u003e0.09\u003c/p\u003e \u003cp\u003e0.77\u0026ndash;1.19\u003c/p\u003e \u003cp\u003e0.72\u0026ndash;1.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u0026ndash;0.84\u003c/p\u003e \u003cp\u003e0.63\u003csup\u003e\u003cb\u003ec,Z\u003c/b\u003e\u003c/sup\u003e \u0026plusmn; 0.14\u003c/p\u003e \u003cp\u003e0.04\u003c/p\u003e \u003cp\u003e0.53\u0026ndash;0.73\u003c/p\u003e \u003cp\u003e0.56\u0026ndash;0.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;6.241\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.006*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOne-way repeated measures analysis\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;9.465\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u0026thinsp;=\u003c/em\u003e\u0026thinsp;.002*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;17.450\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;4.719\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.023*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Demineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-61.84\u0026ndash;31.75\u003c/p\u003e \u003cp\u003e-33.32\u0026thinsp;\u0026plusmn;\u0026thinsp;27.54\u003c/p\u003e \u003cp\u003e8.71\u003c/p\u003e \u003cp\u003e-53.02 \u0026ndash; -13.62\u003c/p\u003e \u003cp\u003e-54.32 \u0026ndash; -19.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-65.57\u0026ndash;27.63\u003c/p\u003e \u003cp\u003e-31.71\u0026thinsp;\u0026plusmn;\u0026thinsp;25.42\u003c/p\u003e \u003cp\u003e8.04\u003c/p\u003e \u003cp\u003e-49.89 \u0026ndash; -13.53\u003c/p\u003e \u003cp\u003e-41.57 \u0026ndash; -28.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-32.43\u0026ndash;28.30\u003c/p\u003e \u003cp\u003e-13.59\u0026thinsp;\u0026plusmn;\u0026thinsp;20.26\u003c/p\u003e \u003cp\u003e6.41\u003c/p\u003e \u003cp\u003e-28.08\u0026ndash;0.90\u003c/p\u003e \u003cp\u003e-30.00\u0026ndash;1.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.985\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.157 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (Baseline vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-58.51\u0026ndash;43.21\u003c/p\u003e \u003cp\u003e5.53\u0026thinsp;\u0026plusmn;\u0026thinsp;28.57\u003c/p\u003e \u003cp\u003e9.04\u003c/p\u003e \u003cp\u003e-14.91\u0026ndash;25.97\u003c/p\u003e \u003cp\u003e-7.32\u0026ndash;22.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-26.23\u0026ndash;97.37\u003c/p\u003e \u003cp\u003e16.59\u0026thinsp;\u0026plusmn;\u0026thinsp;35.21\u003c/p\u003e \u003cp\u003e11.13\u003c/p\u003e \u003cp\u003e-8.60\u0026ndash;41.77\u003c/p\u003e \u003cp\u003e-6.96\u0026ndash;32.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-61.11\u0026ndash;21.31\u003c/p\u003e \u003cp\u003e-14.99\u0026thinsp;\u0026plusmn;\u0026thinsp;24.63\u003c/p\u003e \u003cp\u003e7.79\u003c/p\u003e \u003cp\u003e-32.61\u0026ndash;2.63\u003c/p\u003e \u003cp\u003e-29.67\u0026ndash;5.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;2.892\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.073 NS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePercentage Change (%) (After Demineralization vs After Remineralization)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e- Min-Max\u003c/p\u003e \u003cp\u003e- Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003cp\u003e- SE of Mean\u003c/p\u003e \u003cp\u003e- 95.0% CI for Mean\u003c/p\u003e \u003cp\u003e- 25th Percentile \u0026ndash; 75th Percentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-39.06\u0026ndash;213.51\u003c/p\u003e \u003cp\u003e83.08\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e \u0026plusmn; 90.41\u003c/p\u003e \u003cp\u003e28.59\u003c/p\u003e \u003cp\u003e18.41\u0026ndash;147.75\u003c/p\u003e \u003cp\u003e9.47\u0026ndash;185.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.39\u0026ndash;260.00\u003c/p\u003e \u003cp\u003e83.91\u003csup\u003e\u003cb\u003ea,b\u003c/b\u003e\u003c/sup\u003e \u0026plusmn; 69.61\u003c/p\u003e \u003cp\u003e22.01\u003c/p\u003e \u003cp\u003e34.11\u0026ndash;133.71\u003c/p\u003e \u003cp\u003e35.80\u0026ndash;97.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-44.44\u0026ndash;19.35\u003c/p\u003e \u003cp\u003e-1.79\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;18.65\u003c/p\u003e \u003cp\u003e5.90\u003c/p\u003e \u003cp\u003e-15.13\u0026ndash;11.54\u003c/p\u003e \u003cp\u003e-12.33\u0026ndash;5.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF\u003csub\u003e(df=2)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;5.442\u003c/p\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;.010*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMin. \u0026ndash; Max. : Minimum \u0026ndash; Maximum S.D.: Standard Deviation\u003c/p\u003e \u003cp\u003eSE: Standard error CI: Confidence interval\u003c/p\u003e \u003cp\u003edf: degree of freedom\u003c/p\u003e \u003cp\u003e*: Statistically significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;.05) NS: Statistically not significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;.05)\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters for intergroup comparison\u003c/strong\u003e \u003cp\u003eBiomin group assigned letter (a), Biomin and laser group assigned letter (b), and control group assigned letter (c)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eBonferroni Pairwise multiple comparison\u003c/strong\u003e \u003cp\u003emean of groups that is labeled with similar superscript letter are statistically not significantly different\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSuperscript letters for intragroup comparison\u003c/strong\u003e \u003cp\u003eBaseline assigned letter (x), After Demineralization assigned letter (y), and after Remineralization assigned letter (z)\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eMineral content (Ca,P,F) using EDX\u003c/b\u003e:\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ea. Comparison between baseline and (After demineralization\u003c/b\u003e \u003cem\u003e)\u003c/em\u003e:\u003c/p\u003e \u003cp\u003eAt baseline, there was no statistical significant difference between groups A,B and C regarding to mass% value of Ca, P and F.\u003c/p\u003e \u003cp\u003eAfter demineralization, it was shown that calcium level (% mass) significantly decreased in the three studied groups .Calcium level decreased from a mean of 31.34 ,29.98 and 30.91 (%mass) in groups A,B and C respectively at baseline to a mean of 22.64 ,20.86 and 20.16% mass after demineralization (Fig.\u0026nbsp;7) (table 3).\u003c/p\u003e \u003cp\u003ePhosphorus level % mass also significantly decreased from a mean of 16.38,15.90 and 16.21 to 10.77,11.34 and 10.76%mass in groups A,B and C respectively (Fig.\u0026nbsp;8) (table4).\u003c/p\u003e \u003cp\u003eRegarding to fluoride level (%mass), there was significant decrease from 0.94,0.87 and 0.78 to 0.62,0.61and 0.64 in groups A,B and C respectively (p\u0026lt;0.05) (Fig.\u0026nbsp;9) (table 5).\u003c/p\u003e \u003cp\u003eThere was no statistical significant difference among the three studied groups in Ca,P,F level after demineralization (p\u0026thinsp;\u0026ge;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cb\u003eb. Comparison between (after demineralization) and (after remineralization)\u003c/b\u003e:\u003c/p\u003e \u003cp\u003eAfter the application of the remineralizing agents, calcium ions gained a significant increase in groups A and B. Group B showed the highest gain of ions as percentage change (%) was 40.88\u0026thinsp;\u0026plusmn;\u0026thinsp;12.23. There was no statistical difference between group A and B after remineralization (p\u0026thinsp;\u0026ge;\u0026thinsp;0.05) and both were significantly higher than the control group (p˂0.05) (Table\u0026nbsp;3) (Fig.\u0026nbsp;7).\u003c/p\u003e \u003cp\u003eAs regards the phosphorus ions, a significant increase was observed in all groups. Group A showed the highest gain as P level percentage change(%mass) was 56.52\u0026thinsp;\u0026plusmn;\u0026thinsp;18.02 and group C showed the least gain as P level percentage change was 32.60\u0026thinsp;\u0026plusmn;\u0026thinsp;23.26 .There was no significant difference between group A and group B but both were significantly higher than group C after remineralization (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;8) (Table\u0026nbsp;4).\u003c/p\u003e \u003cp\u003eFluoride ions increased significantly in groups A and B (p˂0.05) but decreased significantly in group C as shown in percentage change formula (-1.78\u0026thinsp;\u0026plusmn;\u0026thinsp;18.65). There was no statistical difference between group A and B (p\u0026thinsp;\u0026ge;\u0026thinsp;.05) in fluoride level after remineralization (Fig.\u0026nbsp;9) (Table\u0026nbsp;5). Both groups A and B were significantly higher than group C (p\u0026thinsp;=\u0026thinsp;.006).\u003c/p\u003e \u003cp\u003e \u003cb\u003eC. Comparison between baseline and after remineralization\u003c/b\u003e \u003c/p\u003e \u003cp\u003eWhen comparing the mean values of minerals % mass between the baseline and the last stage after application of the remineralizing agents, it was obvious that there was no significant difference in minerals percentage (Ca,P,F) between baseline and after remineralization in groups A and B (p\u0026thinsp;\u0026ge;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eWhile in the control group, there was significant decrease in the Ca,P and F %mass between baseline and after remineralization process.\u003c/p\u003e \u003cp\u003e \u003cb\u003e2. Environmental scanning electron microscope (ESEM) analysis\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe sample surface characteristics at each stage were described using ESEM at a magnification of x2000. At base line, micrographs showed smooth enamel surface. After demineralization, samples showed honey comb appearance which represent areas of minerals dissolution. After application of the remineralizing agent, samples showed partial restoration of enamel surface structure. (Figs.\u0026nbsp;10\u0026ndash;14)\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThis study was conducted to evaluate the impact of diode laser and Biomin F toothpaste on the remineralization of WSLs.\u003c/p\u003e \u003cp\u003eThe null hypothesis was accepted as there was no significant difference between the effect of Biomin F toothpaste coupled with diode laser and Biomin F alone on the remineralization of white spot lesions.\u003c/p\u003e \u003cp\u003eThe natural physiological processes of demineralization and remineralization in tooth structure may be disrupted when there is an imbalance between pathogenic and protective factors [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. From a scientific standpoint, it has been recognized that salivary dysfunction, fermentable carbohydrates, and cariogenic bacteria play significant roles as pathogenic factors [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Calcium and phosphate ions presence in a supersaturated state within human saliva enables it to potentially facilitate the remineralization of enamel [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. However, if acid challenges exceed this physiological remineralization process, alternative therapeutic interventions are needed to strengthen remineralization [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThroughout orthodontic treatment, tooth enamel faces an elevated risk of developing white spot lesions due to food accumulation triggered by appliances and insufficient oral hygiene practices. When treating visible white spot lesions using concentrated fluoride agents (hypermineralization), the lesion is arrested at the surface instead of allowing saliva to promote remineralization in the deeper areas [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Presently, there are numerous methods available to halt or reverse the advancement of WSLs utilizing low levels of fluoride, such as casein phosphopeptides\u0026ndash;amorphous calcium phosphate (CPP-ACP) [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], Nano-hydroxyapatite, Trimetaphosphate ion and Bioactive glasses [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTherefore, Biomin F was utilized in this study as it is a bioactive glass material containing a low level of fluoride. Ali et al [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] noted in their study that Biomin F toothpastes exhibited lower total fluoride content compared to the values asserted by their respective companies (approximately 400 ppm). Biomin F possesses the characteristic of prolonged fluoride delivery over a 12-hour period through gradual dissolution of the glass [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. This characteristic is due to the polymer that enhances the bond between the calcium in the bioglass material and the calcium on the enamel. This bonding reduces the leaching of bioactive glass material [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Biomin F contains small bioglass particles that aid in the infiltration of remineralizing agents into subsurface lesions [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. With its high phosphate content, Biomin F facilitates rapid apatite formation (within 6 hours) and contains fewer carbon impurities, thereby rendering enamel less soluble in acid [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe majority of studies conducted on Biomin F have explored its efficacy in occluding dentinal tubules and addressing hypersensitivity [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. A systematic review focused on investigating the role of bioactive glass in enamel remineralization highlighted the significant contribution of fluoride-containing bioactive glass dentifrice, specifically Biomin F, in enamel regeneration [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAidaros et al [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] carried out an in-vitro investigation utilizing SEM and elemental analysis to assess the mineral composition of extracted permanent third molars prior to and following the application of remineralizing agents, which included Biomin F. Their study involved comparing these agents, employing a similar application regimen to that of the current study (two minutes, twice daily for two weeks). They utilized the materials in the form of toothpaste and concluded that the combination of fluoride with bioactive glass technology, as seen in Biomin F toothpaste, had the most significant impact on the demineralized enamel surface. This finding aligned with our study, which demonstrated that Biomin F possessed the ability to remineralize white spot lesions (WSLs) to restore them to their baseline mineral content.\u003c/p\u003e \u003cp\u003eIn this current study, laser was examined in conjunction with Biomin F, given that numerous studies have evaluated various types of lasers, including CO2, Nd:YAG, Er:YAG, and diode lasers, utilizing different parameters for caries prevention and enamel remineralization, either with or without fluoride-containing agents [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe present study revealed that the mineral content following remineralization reached the baseline mineral level in both test groups. This finding contradicted the results reported by Omran et al. [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] as the mean calcium mass percentage after remineralization was significantly lower compared to the baseline calcium mass percentage in the Biomin group. Notably, both studies employed the same Biomin F application protocol (two minutes twice daily for two weeks). This variation might be explained by the fact that Omran-T applied Biomin F as toothpaste slurries and subjected the samples to a shorter demineralization period (72 hours), while the present study utilized it in the form of toothpaste, as commonly used by orthodontic patients. Nonetheless, both studies concurred on the high phosphate content of Biomin F.\u003c/p\u003e \u003cp\u003eAn intriguing observation emerged from the study comparing laser treatment with a bioactive glass material (Novamin) using SEM: laser therapy did not provide additional benefits to Novamin in the process of remineralizing the enamel surface [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. In the current study, where Biomin F demonstrated the capability to remineralize the enamel surface, the diode laser did not exhibit a synergistic effect in enamel remineralization, as indicated by the insignificant difference between group A and B. Similarly, comparable outcomes were observed when Novamin was utilized alongside laser therapy.\u003c/p\u003e \u003cp\u003eThe potential rationale for the findings of this study could be that the efficacy of the bioactive glass material (Biomin F) relies on its interaction with physiological aqueous solutions, leading to the release of calcium, phosphorus, and fluoride. However, when a diode laser is employed, a certain degree of heat is generated within the treated surface, typically ranging from 1 to 6 degrees Celsius [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. As a consequence, this leads to some dryness and removal of moisture from the paste, which is essential for mineral release.\u003c/p\u003e \u003cp\u003eThe analysis of SEM micrographs enabled us to observe the notable regeneration of the enamel structure and the deposition of mineral crystals, a result that aligns with findings reported by Bakrey et al., who observed the deposition of mineral crystals blocking the dentinal tubules [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e] after using Biomin F.\u003c/p\u003e \u003cp\u003eThis study has a limitation that we must consider the dynamic complex system in oral environment which may differ from the in-vitro study employed in the present work.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eWithin the limitation of the present study, we concluded that Biomin F toothpaste is promising in repairing the white spot lesions on the surface of the demineralized enamel. Diode laser didn\u0026rsquo;t affect the remineralizing ability of Biomin F toothpaste which add hazard without benefit.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRecommendation\u003c/b\u003e \u003c/p\u003e \u003cp\u003eClinical studies are needed for more evaluation of the benefits of these approaches. It is recommended to evaluate the synergistic effect between laser and biomin F in decreasing the white spot lesions depth with a variation of laser protocols and exposure time.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe present study was approved by the Research Ethics Committee of the Faculty of Dentistry, Alexandria University (IRB:00010556– IORG:0008839).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data and materials used to support the findings of this study are available from the corresponding authors upon request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis clinical research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNT, AM Guide the correct methodology for the trial and supervise the work\u0026nbsp;. Help the student in interpreting results and revising the manuscript. AIE contributed to the concept, methodology and the design of the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eOgaard B. Prevalence of white spot lesions in 19-year-olds: a study on untreated and orthodontically treated persons 5 years after treatment. Am J Orthod Dentofacial Orthop. 1989;96:423-7.\u003c/li\u003e\n\u003cli\u003ePalamara J, Phakey PP, Rachinger WA, Orams HJ. Ultrastructure of the intact surface zone of white spot and brown spot carious lesions in human enamel. J Oral Pathol. 1986;15:28-35.\u003c/li\u003e\n\u003cli\u003eFeatherstone JD. The science and practice of caries prevention. J Am Dent Assoc. 2000;131:887-99.\u003c/li\u003e\n\u003cli\u003eOgaard B, R\u0026oslash;lla G, Arends J, ten Cate JM. 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Some evidence that fluoride during orthodontic treatment reduces occurrence and severity of white spot lesions. Evid Based Dent. 2004;5:98-9.\u003c/li\u003e\n\u003cli\u003eLinton JL. Quantitative measurements of remineralization of incipient caries. Am J Orthod Dentofacial Orthop. 1996;110:590-7.\u003c/li\u003e\n\u003cli\u003eAidaros N, Eliwa ME, Kamh R. Remineralization Efficiency of Different Toothpastes on Human Enamel Subjected to Acid Challenge: in Vitro Study. Al-Azhar DJ Girls. 2022;9:61-72.\u003c/li\u003e\n\u003cli\u003eGoswami M, Saha S, Chaitra TR. Latest developments in non-fluoridated remineralizing technologies. J Indian Soc Pedod Prev Dent. 2012;30:2-6.\u003c/li\u003e\n\u003cli\u003eElkabbany SMH, Mosleh AA, Metwally NI. Remineralization effect of diode laser, Nanoseal\u0026reg;, and Zamzam water on initial enamel carious lesions induced around orthodontic brackets. J Nat Sci Med. 2021;4:50-7.\u003c/li\u003e\n\u003cli\u003eFarhadian N, Rezaei-Soufi L, Jamalian SF, Farhadian M, Tamasoki S, Malekshoar M, et al. Effect of CPP-ACP paste with and without CO2 laser irradiation on demineralized enamel microhardness and bracket shear bond strength. Dental Press J Orthod. 2017;22:53-60.\u003c/li\u003e\n\u003cli\u003eTaha AA, Patel MP, Hill RG, Fleming PS. The effect of bioactive glasses on enamel remineralization: A systematic review. J Dent. 2017;67:9-17.\u003c/li\u003e\n\u003cli\u003eOmran TM, Mostafa MH, El-Raouf A, Eman M. Assessment of Remineralizing Effect of Bioactive Glass Based Toothpastes: an In-Vitro Comparative Study. Al-Azhar DJ Girls. 2021;8:491-9.\u003c/li\u003e\n\u003cli\u003eCurran-Everett D. Evolution in statistics: P values, statistical significance, kayaks, and walking trees. Adv Physiol Educ. 2020;44:221-4.\u003c/li\u003e\n\u003cli\u003eAdel SM, Marzouk ES, El-Harouni N. 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London, California, New Delhi: SAGE Publications Ltd; 2013.\u003c/li\u003e\n\u003cli\u003eGreenland S, Senn SJ, Rothman KJ, Carlin JB, Poole C, Goodman SN, et al. Statistical tests, P values, confidence intervals, and power: a guide to misinterpretations. Eur J Epidemiol. 2016;31:337-50.\u003c/li\u003e\n\u003cli\u003eMontgomery D. Experiments with a Single Factor. In: The Analysis of Variance. Design and Analysis of Experiments. I. Ch 3: John Wiley \u0026amp; Sons; 2001.\u003c/li\u003e\n\u003cli\u003eLowry R. One Way ANOVA\u0026ndash;Independent Samples. Vassar. edu. Retrieved on December 4th. 2008 Retrieved on December 4th. 2008.\u003c/li\u003e\n\u003cli\u003eRuxton GD, Beauchamp G. Time for some a priori thinking about post hoc testing. Behav Ecol. 2008;19:690-3.\u003c/li\u003e\n\u003cli\u003eGueorguieva R, Krystal JH. Move over ANOVA: progress in analyzing repeated-measures data and its reflection in papers published in the Archives of General Psychiatry. 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An in-vitro evaluation of fluoride content and enamel remineralization potential of two toothpastes containing different bioactive glasses. Biomed Mater Eng. 2020;30:487-96.\u003c/li\u003e\n\u003cli\u003eRamadoss R, Padmanaban R, Subramanian B. Role of bioglass in enamel remineralization: Existing strategies and future prospects-A narrative review. J Biomed Mater Res B Appl Biomater. 2022;110:45-66.\u003c/li\u003e\n\u003cli\u003eBakry AS, Abbassy MA, Alharkan HF, Basuhail S, Al-Ghamdi K, Hill R. A Novel Fluoride Containing Bioactive Glass Paste is Capable of Re-Mineralizing Early Caries Lesions. Materials (Basel). 2018;11.\u003c/li\u003e\n\u003cli\u003eBakry AS, Al-Harbi N, Al-Hadeethi Y, Abbassy MA, Katturi N, Xin B, et al. Invitro evaluation of new treatment for dentin hypersensitivity using BioMin F and BioMin C. J Non Cryst Solids. 2023;602:122072.\u003c/li\u003e\n\u003cli\u003eArshad S, Zaidi SJA, Farooqui WA. Comparative efficacy of BioMin-F, Colgate Sensitive Pro-relief and Sensodyne Rapid Action in relieving dentin hypersensitivity: a randomized controlled trial. BMC Oral Health. 2021;21:498.\u003c/li\u003e\n\u003cli\u003eYavagal CM, Chavan VV, Yavagal PC. Laser induced enamel remineralization: A systematic. Int J Appl Dent Sci. 2020;6:168-73.\u003c/li\u003e\n\u003cli\u003eAlqahtani MA, Almosa NA, Alsaif KA, Alsaif NM, Aljaser YJ. Effect of topical fluoride application and diode laser-irradiation on white spot lesions of human enamel. Saudi Dent J. 2021;33:937-43.\u003c/li\u003e\n\u003cli\u003eAoki A, Sasaki KM, Watanabe H, Ishikawa I. Lasers in nonsurgical periodontal therapy. Periodontol 2000. 2004;36:59-97.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Biomin, laser therapy, WSL, tooth remineralization","lastPublishedDoi":"10.21203/rs.3.rs-3990752/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3990752/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: White-spot lesions are considered an initial carious stage characterized by an outer enamel layer with significantly reduced mineralization. This study was conducted to assess the combined effect of Biomin F toothpaste and Diode laser on remineralization of white spot lesions\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials and methods\u003c/strong\u003e:\u003cstrong\u003e \u003c/strong\u003eAn invitro study conducted on a total of 30 premolars divided into three groups; Group A (Biomin F Tooth paste), Group B (Biomin F with laser application for 30 sec), Group C (Negative control).The three groups were submitted to three stages; stage 1:Baseline,stage 2:After demineralization ,and stage 3:After remineralization .In each stage, elemental analysis(calcium, phosphorus, and fluoride)was measured quantitatively using Energy Dispersive X-ray (EDX) analysis and qualitatively by micrographs using scanning electron microscope. The data were tested to find significant difference between mineral changes during stages by using (ANOVA) test and Bonferroni test.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e:\u003cstrong\u003e \u003c/strong\u003eCalcium, phosphorus and fluoride ions decreased in all groups after demineralization. In stage 3, after application of remineralizing agents, Calcium ions increased significantly in groups A and B where P˂.05. As regards to the phosphorus ions, a significant increase was observed in all groups with group A showed the highest gain as phosphorus level percentage change (%mass) was 56.52±18.02 . Fluoride ions increased significantly in groups A and B (p˂0.05) but decreased significantly in group C. There was no statistical significant difference between group A and B (p≥.05) in calcium, phosphorus, and fluoride level after remineralization\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: Within the limitation of the present study, we concluded that Biomin F toothpaste is promising in the repairing of white spot lesions on the surface of the demineralized enamel. Diode laser didn’t affect the remineralizing ability of Biomin F toothpaste which add hazard without benefit\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","manuscriptTitle":"Effect of Biomin F Toothpaste and Diode Laser on Remineralization of White Spot Lesions (In Vitro Study)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-07 18:17:59","doi":"10.21203/rs.3.rs-3990752/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-04-11T11:39:59+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-04-09T13:02:48+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-04-07T15:31:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"b2ec2c4d-2853-4755-b7bd-deb93ab40a5f","date":"2024-03-30T13:16:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"2bb45aae-c415-4149-83ad-56c6e3e84b06","date":"2024-03-28T08:20:27+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-03-10T08:33:01+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-10T06:12:10+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-03-04T20:12:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-03-04T20:10:12+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Oral Health","date":"2024-02-26T11:07:33+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e1e632f6-8d08-4daa-b1ef-065fe1cf9e04","owner":[],"postedDate":"March 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-05T16:02:47+00:00","versionOfRecord":{"articleIdentity":"rs-3990752","link":"https://doi.org/10.1186/s12903-024-04589-9","journal":{"identity":"bmc-oral-health","isVorOnly":false,"title":"BMC Oral Health"},"publishedOn":"2024-07-30 15:57:33","publishedOnDateReadable":"July 30th, 2024"},"versionCreatedAt":"2024-03-07 18:17:59","video":"","vorDoi":"10.1186/s12903-024-04589-9","vorDoiUrl":"https://doi.org/10.1186/s12903-024-04589-9","workflowStages":[]},"version":"v1","identity":"rs-3990752","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3990752","identity":"rs-3990752","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}