A comparative evaluation of biomimetic remineralization potential of enamel white spot lesions: A Pilot in-vitro study

International Journal of Dental Science and Innovative Research (IJDSIR) IJDSIR : Dental Publication Service Available Online at:www.ijdsir.com Volume – 8, Issue – 4, July – 2025, Page No. : 01 - 07 Corresponding Author: Dr. Priyanka Mahajan, ijdsir, Volume – 8 Issue - 4, Page No. : 01 - 07 Page1 ISSN: 2581-5989 PubMed - National Library of Medicine - ID: 101738774 A comparative evaluation of biomimetic remineralization potential of enamel white spot lesions: A Pilot in-vitro study 1Dr. Priyanka Mahajan, Senior Lecturer, Department of Conservative Dentistry and Endodontics, Sudha Rustagi College of Dental Sciences & Research, Faridabad, Haryana. Corresponding Author: Dr. Priyanka Mahajan, Senior Lecturer, Department of Conservative Dentistry and Endodontics, Sudha Rustagi College of Dental Sciences & Research, Faridabad, Haryana. Citation of this Article: Dr. Priyanka Mahajan, “A comparative evaluation of biomimetic remineralization potential of enamel white spot lesions: A Pilot in-vitro study”, IJDSIRJuly – 2025, Volume – 8, Issue – 4, P. No. 01 – 07. Copyright: © 2025, Dr. Priyanka Mahajan, et al. This is an open access journal and article distributed under the terms of the creative common’s attribution non-commercial License. Which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given, and the new creations are licensed under the identical terms. Type of Publication: Original Research Article Conflicts of Interest: Nil Abstract Introduction: The remineralization potential of three recent remineralizing agents on white spot lesions were compared and evaluated using the Vicker’s microhardness testing machine. This short study aims to evaluate and compare the remineralization potential of recently introduced remineralizing agents like Curodont Protect Gel, GC Tooth Mousse Plus, Dente 91 on surface hardness of white spot lesions in human teeth1. Method: Thirty single rooted human premolars extracted due to periodontal reasons or orthodontic reasons were chosen for this study. The teeth were decoronated at Cemento-enamel junction level. Samples were then mounted in auto-polymerization acrylic resin. All the groups were subjected to demineralization process to induce white spot lesions. A post demineralization vicker hardness test was performed followed by random allocation to the three groups. The three groups were namely Self-assembling peptide (P114), Casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF) and Nanohydroxyapatite. A post remineralization Vickers hardness test was conducted to evaluate the remineralization potential of tested agents. Result: An analysis of variance (ANOVA) was performed to determine the significant difference of Vickers microhardness. Self-assembling peptide showed the highest microhardness value. Conclusion: Self-assembling peptide (Curodont Protect Gel) was more efficient than nanohydroxyapatite(Dente91) and CPP-ACP (GC Tooth Mousse Plus) for the prevention of white spot lesions(WSLs). Keywords: Tooth, Enamel, Health, Dental. Introduction Despite of implementing various oral and dental health promotion programs, dental caries still remains the most common and a unique infectious disease of dental hard tissues1.The most common initial clinical sign of the Dr. Priyanka Mahajan, et al. International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 Page2 dental caries is the appearance of ‘white spot lesions’ or areas of demineralization, which are reversible in nature. The bidirectional dynamic process of demineralization followed by remineralization affects the enamel surface during its life span2. At the stage of demineralization, dental caries progress can be prevented by shifting the reaction towards remineralization1,2. Today, in dentistry more emphasis is given on caries interception at an earliest by non-invasive remineralization of the lesion rather than opting for invasive preventive techniques1. Till date, many commercial remineralizing agents in market are being used for remineralization of white spot lesions such as fluoride compounds, tricalcium phosphate, casein phosphopeptide-amorphous calcium phosphate and a lot more. Recently, the use of biomimetic approach has been introduced as an innovative strategy for enamel remineralization4. This short study aims to evaluate and compare the remineralization potential of recently introduced remineralizing agents like Curodont Protect Gel, GC Tooth Mousse Plus, Dente 91 on surface hardness of white spot lesions in human teeth1. Materials & Methods Thirty sound single rooted premolars extracted were collected seeking exclusion and inclusion criteria. Non carious, sound and intact human single rooted premolar teeth with normal morphology were included. Teeth with any craze line or crack, caries as well as any mineralization defect were excluded5. The collected teeth were cleaned and polished with wet pumice slurry paste and were stored in 0.1 % thymol solution until further use6,7. The teeth were decoronated at Cemento-enamel junction level. The coronal half of teeth were then split vertically using a Isomet diamond saw(1-0.25micrometre), mounted on a contra-angle handpiece thereby, retaining the buccal portion of the coronal part of the teeth. The samples were then mounted in auto-polymerization acrylic resin with buccal surface facing the observer. These samples were stored in deionized water. All the samples were subjected to demineralization to induce white spot lesions with the help of demineralization medium. The demineralizing solution was prepared using composition as follows: 2.2 mM potassium dihydrogen phosphate, KH2PO4.7H2O; 2.2 mM calcium chloride, CaCl2..2H2O ; 0.05 mM lactic acid, C3H6O3. The pH of the demineralizing solution was checked during and after preparation of solution using a digital pH meter. The acetic acid buffer was added to adjust the ultimate pH at 4.5. All the samples were immersed into a glass container containing 50mL of the prepared demineralizing solution for a period of 96 hours at 37°C to produce an artificial subsurface enamel lesion in all samples. After demineralization, the teeth were washed with deionized water and dried5,6. Consequent to this process of demineralization, the 30 premolar tooth were allocated at random into one of the following three groups (10 in each group) depending on the remineralizing paste used for application: Group A: CURODONT PROTECT GEL (N=10) Group B: GC TOOTH MOUSSE PLUS (N=10) Group C: DENTE 91 (N=10) The samples from group A to group C were treated with respective remineralizing agent for 3minutes twice daily for 14 days. The samples were applied with respective remineralizing agent using a cotton applicator tip for 3 minutes twice daily, washed with deionized water and stored in artificial saliva. The artificial saliva was formulated with the subsequent substances in specific proportions—potassium dihydrogen phosphate Dr. Priyanka Mahajan, et al. International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 Page3 (KH2PO4) 0.9 mmol/L, potassium chloride (KCl) 50 mmol/L, calcium chloride (CaCl2) 1.5 mmol/L, and tris buffer 20 mmol/L6,9. After 14 days cycles of remineralization, the surface microhardness of the specimens was determined using the Vickers microhardness testing machine. A load of 100 g was applied to the surface of specimen for a period of 10seconds using Vickers elongated diamond pyramid indenter under a x40 objective lens. The accuracy of values of diagonal length of indentations was determined under high power magnification of x400. The depth of the indentation was measured, and the values were converted to Vickers microhardness values.Five indentations were placed on the surface and the mean value was considered for each specimen6-8. Statistical Analysis Descriptive and analytical statistics were done. The data were presented in mean and standard deviation. The paired sample t-test and one-way analysis of variance (ANOVA) test were analyzed with the help of SPSS (Statistical Package for Social Sciences) Version 24.0 (IBM Corporation, Chicago, USA). The level of significance was kept at p<0.05. Results Table 1 Comparison of mean Microhardness in HV at Demineralization and Remineralization stage for each of the three groups Groups Stages N Mean Std Deviation t Value pValue Group A self-assembling peptide-Curodont repair gel Demineralization 10 389.2 5.534 2.631 0.02 Remineralization 10 319.4 30.544 Group B CPP-ACP GC Tooth Mousse Plus Demineralization 10 368.8 8.039 1.76 0.01 Remineralization 10 271 26.34 Group C Nano-hydroxyapatite Dente-91 Demineralization 10 315.3 25.721 0.986 0.334 Remineralization 10 277.2 28.635 Significant at p< 0.05 Graph 1: Comparison of mean Microhardness in HV at Demineralization and Remineralization stages GROUP 1 GROUP2 GROUP 3 2.631 1.76 0.986 0 0.5 1 1.5 2 2.5 3 1 2 3 4 5 6 Groups t Value Dr. Priyanka Mahajan, et al. International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 Page4 The table 1 compares the mean microharness of each group at demineralization and remineralization stage (Xaxis Group A: self-assembling peptide-Curodont repair gel, Group B: CPP-ACP GC Tooth Mousse Plus, Group C: Nano-hydroxyapatite Dente-91 and on Yaxis t values) on demineralization and Remineralization. Group A shows the highest t-value (2.631) and a significant p-value (0.02) in demineralization, indicating its effectiveness compared to other groups. Notably, Group A also displays a higher mean in Remineralization (319.4) suggesting its potential in promoting tooth Remineralization. Conversely, Group B and Group C exhibit lower t-values and p-values. Overall, the self-assembling peptide-Curodont repair gel (Group A) stands out with promising Remineralization capabilities, emphasizing its potential dental health benefits. Table 2 Comparison of mean Microhardness in HV among the three groups at baselines, Demineralization and Remineralization stages Group N Mean Std Deviation F Value pValue Baseline self-assembling peptide-Curodont repair gel 10 350.7 23.348 1.046 0.342 CPP-ACP GC Tooth Mousse Plus 10 366 4.086 Nano-hydroxyapatite Dente-91 10 312 19.711 Demineralization self-assembling peptide-Curodont repair gel 10 389.2 5.534 0.465 0.928 CPP-ACP GC Tooth Mousse Plus 10 368.8 8.039 Nano-hydroxyapatite Dente-91 10 315.3 25.721 Remineralization self-assembling peptide-Curodont repair gel 10 319.4 30.544 1.216 0.507 CPP-ACP GC Tooth Mousse Plus 10 271 26.34 Nano-hydroxyapatite Dente-91 10 277.2 28.635 Dr. Priyanka Mahajan, et al. International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Page5 Graph 2: Comparison of mean Microhardness in HV among the three groups at baselines, Demineralization and Remineralization stages The Table 2 presents a comparison of stages of mean micro hardness (HV) among three dental treatment groups – self-assembling peptide-Curodont repair gel, CPP-ACP GC Tooth Mousse Plus, and Nanohydroxyapatite Dente-91 – on Xaxis Baseline, Demineralization, and Remineralization stages vs on Yaxis F value. Micro hardness reflects material strength. At Baseline, mean HV values were 350.7, 366, and 312 for the three groups, respectively. No significant differences emerged, as indicated by p-values above 0.05. Similarly, during Demineralization (mean HV: 389.2, 368.8, 315.3) and Remineralization (mean HV: 319.4, 271, 277.2) stages, p-values remained above 0.05, suggesting no substantial microhardness variation. The study implies that the examined materials exhibited consistent microhardness levels across these stages and did not exhibit statistically significant distinctions in their hardness properties. Discussion This study highlights the remineralization effects of different remineralizing agent on demineralized enamel surface of the extracted teeth. In this research, greatest mean remineralizing potential was noted in Curodont protect gel (GROUP A) for the lost enamel structure in induced white spot lesion. This result can be due to its hydrogel appearence formed by beta-sheet amyloids that creates a 3Dmatrix and promotes strength, toughness similar to muscle tissues. This biomimetic remineralization approach that spyron the biomineralization process is in gleam to replicate the natural mineralization process. The monomeric selfassembling peptide(P11-4) offered by curodont repair technology can generate new hydroxyapatite crystals as a foci and speed up the mineral crystal development in a cycle of biomimetic mineralization [10]. Dente-91 (GROUP C) has shown the second highest remineralization potential. The reason behind this can be attributed to the similar morphology, structure and crystallinity of this synthetic nano hydroxyapatite as that of a biological apatite. The nano-sized particles diffuse into the demineralized areas of enamel and form a new apatite layer. Being a potent biocompatible, bioactive substance with greater affinity for enamel it has its own bench mark in dentistry now days11. 1.046 0.465 1.216 0 0.2 0.4 0.6 0.8 1 1.2 1.4 123456789 Stages F Value Dr. Priyanka Mahajan, et al. International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 Page6 GC TOOTH MOUSSE PLUS (GROUP B) showed the third highest remineralization potential for white spot lesions in the present study. This result can be postulated due to its degradation of the phosphopeptides which in turn leads to dephosphorylation of Casein phosphopeptide-amorphous calcium phosphate by phosphatases. This results in substantially reduction of the peptide ability to bind calcium and phosphate ions thereby decreasing the remineralizing potential12. Conclusions The results of this in-vitro study suggest that among the three agents tested, self-assembling peptide (Curodont Protect Gel) is the most effective remineralizing agent followed by nanohydroxyapatite (Dente-91) and Casein phosphopeptide-amorphous calcium phosphate (GC Tooth Mousse) when tested under the conditions mentioned in this study. References 1. Behrouzi.P,Heshmat.H,Ganjkar.MH,Tabatabaei.SF, Kharazifard M: Effect of two methods of remineralization and resin infiltration on surface hardness of artificially induced enamel lesions. J Dent Shiraz Univ Med Sci.2020, 21(1): 12-17. 10.30476/DENTJODS.2019.77864 2. Dixit.A, Mampilly.M, Nallamothu.R, Gopakumar.R, Jayachandran.M, Terence.N: Analysis of Remineralization Potential of Three Different Remineralizing Pastes on Demineralized Enamel A Comparative Study. J Contemp Dent Pract.2021,Volume 22.10.5005/jp-journals-100243095. 3. 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International Journal of Dental Science and Innovative Research (IJDSIR) ©2025 IJDSIR, All Rights Reserved Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 Page7 peptide P11 -4 on enamel erosion AFM and SEM studies. Scanning 2016;38 (4): 344-51. doi: 10.1002/sca.21276. 11. Anil A, Ibraheem W.I, Meshni A.A, Preethanath R.S, Anil S: Nano-Hydroxyapatite (nHAp) in the Remineralization of Early Dental Caries: A Scoping Review. Int. J. Environ. Res. Public Health. 2022; 19, 5629. doi.org/10.3390/ijerph19095629. 12. Lata S, Varghese NO, Varughese JM: Remineralization potential of fluoride and amorphous calcium phosphate-casein phospho peptide on enamel lesions An in vitro comparative evaluation. J Conserv Dent. 2010;13(1):42-6. doi: 10.4103/0972-0707.62634.