Dentin Demineralization Research Articles

Inhibitory Effects of Toothpaste Containing Pyrrolidone Carboxylic Acid on Dentin Demineralization and Collagen Degradation

Inhibitory Effects of Toothpaste Containing Pyrrolidone Carboxylic Acid on Dentin Demineralization and Collagen Degradation

Testing the Clinical Applicability of Resin Infiltration of Developmental Enamel Hypomineralization Lesions Using an In Vitro Model.

ABSTRACTAimThe aim of this study is to investigate the penetration abilities of a commercially available low-viscosity resin infiltrant into developmentally hypomineralized teeth in vitro.Materials and methodsFour extracted third molars of a 17-year-old patient with signs of developmental enamel hypomineralization (discoloration, increased opacity, and surface roughness) were infiltrated with a low-viscosity resin mixed with a fluorescent dye, according to the manufacturer's standard protocol. Four extracted molars with sound enamel or showing only initial fissure caries were used as a control group. Specimens were embedded in polymethylmethacrylate, and grindings were prepared. High-resolution projectional radiography of the grindings was performed, and, for one specimen, quantitative micro-computed tomography was used to measure hydroxyapatite density in enamel and dentin lesions. After decalcification, the grindings were examined by reflected bright-field microscopy, wide-field fluorescence microscopy, and confocal laser scanning microscopy. Fluorescence micrographs were superimposed on the radiographs and analyzed correlatively.ResultsThe pattern of hypo-/demineralization in enamel and dentin in developmentally hypomineralized teeth showed a good congruence with the pattern of resin infiltration. Cavitations and dentin tubules up to a depth of 2 mm beyond cavitations were filled by the infiltrant. In control teeth, the penetration of the infiltrant was limited to decalcified enamel areas (initial fissure caries).ConclusionsIn vitro infiltration of developmentally hypomineralized enamel was successful.Clinical significanceResin infiltration might be considered as a routine procedure in the treatment of developmentally hypomineralized teeth. Further investigations with higher sample sizes, different degrees of severity, different stages of lesion extension, and modified treatment protocols are necessary.How to cite this articleSchnabl D, Dudasne-Orosz V, et al. Testing the Clinical Applicability of Resin Infiltration of Developmental Enamel Hypomineralization Lesions Using an In Vitro Model. Int J Clin Pediatr Dent 2019;12(2):126–132.

Caries protective effect of two Self-etch dentin bonding agents on denuded root dentin surface: In Vitro Study

Objective: To evaluate the caries protective effect of two self-etching dental adhesive agents on the denuded root surface. Materials and methods: The roots of 30 freshly extracted , non-carious human premolars were cleaned and denuded of cementum. The teeth surfaces were covered with acid-resistant nail varnish, exposing two rectangular windows (lingual and buccal surface) of 4 x 4 mm each. The window on the lingual furface served as the control , while the buccal window were treated with dentin bonding agent. All specimens were store in acidifed gel for 6 days for demineralization. The specimens were then divided among two experimental groups-Group A: Optibond all in one and Group B: Bond-1SF.In Group C (control group) bonding agent was not applied. The specimens were stored in 0.9% sodium chloride solution for 14 days. Plano parallel tooth sections (80 ± 20 μm) were obtained. Tooth sections were evaluated for caries‑like demineralized zone under a polarized microscope. The values obtained were analyzed using one‑way ANOVA and Tukey honesty tests. Results: In the Control group the mean depth of demineralization was 80.08µm .The depth of demineralization was significantly reduced in Group A and B. Conclusion: Self-etching dental adhesive agents with and without solvent is an effective method to prevent demineralization of root dentin.

Dentin-pulp complex reactions in conventional and radiation-related caries: A comparative study.

BackgroundRadiation-related caries (RRC) is one of the most significant oral toxicities of head and neck radiotherapy (HNRT); however, the potential of radiation to directly cause harmful dentin and pulpal effects and impair response to caries progression is controversial.Material and MethodsTherefore, the aim of this study was to characterize the reactions of the dentin-pulp complex in teeth affected by RRC. Patients and methods: Twenty-two carious teeth extracted from 22 head and neck cancer (HNC) patients were divided into control (conventional caries; n=11) and irradiated (RRC; n=11) groups and paired matched by dental homology, clinical patterns of caries progression following the Post-Radiation Dental Index (PRDI) and microscopic depth of carious invasion. Histopathological characteristics based on morphological hierarchy, cell populations of dental pulp, blood vessels, neural elements, extracellular matrix components, inflammation, patterns of carious invasion and reactionary dentin presence were evaluated by optical light microscopy and histomorphometry.ResultsMean PRDI scores were 3.2 for the control group and 3.8 for the irradiated group. Dentin demineralization patterns were also similar between the groups and the mean depths of demineralization were 1,158.58µm and 1,056.89µm for the control and irradiated groups, respectively.ConclusionsPulp histopathological changes and dentin reaction patterns were similar between groups and varied according to the PRDI scores and carious lesions depth. Dentin and pulp reactions are highly preserved in RRC teeth. Key words:Cancer, radiotherapy, radiation-related caries, teeth, pulp.

Root Dentin Demineralization under Varnish Containing Multi-ion Releasing Filler

Root Dentin Demineralization under Varnish Containing Multi-ion Releasing Filler

Demineralization, Collagen Modification and Remineralization Degree of Human Dentin after EDTA and Citric Acid Treatments.

The aim of the study was to investigate the effects of several decalcifying agents used as irrigant solutions in endodontic treatment on collagen and mineral components of dentin. Coronal dentin discs from five caries-free human third molars with a smear layer were treated for one minute with a chelating solution (1% Ethylenediaminetetraacetic acid (EDTA), 10% EDTA, 17% EDTA, 10% citric acid). Mineralization degree (Ca/N and P/N atomic ratios, IR Iapatite/Iamide II and I1410(carbonate)/I554(phosphate) spectroscopic ratios) and possible collagen rearrangements (collagen infrared (IR) amide II e III shifts) were evaluated by environmental scanning electron microscopy (ESEM)/energy dispersive X-ray spectroscopy (EDX) and IR spectroscopy before and after treatment (T0) and after ageing (T24h and T2m) in simulated body fluid (SBF). At T0, analysis showed that the highest demineralizing effect was achieved using a 10% citric acid solution and 10% EDTA, while the smallest effect was observed when using 17% EDTA. No significant collagen modifications were detected upon treatment with 1% EDTA, while subtle changes were observed after the other treatments. At T24h or T2m, analyses showed the highest remineralization values for 1% EDTA and the lowest for 10% citric acid, mainly at T2m. The samples treated with 17% EDTA showed slight collagen rearrangements upon remineralization. In conclusion, the highest demineralizing effect was observed for 10% EDTA and 10% citric acid. Collagen rearrangement was found for all the treatments except for 1% EDTA. The highest remineralization capability in SBF values was recorded for 1% EDTA and the lowest for 10% citric acid. A slight collagen rearrangement upon remineralization was still present in 17% EDTA-treated samples. Clinical use as a chelating agent in the endodontic therapy of citric acid and concentrated EDTA solutions should be reconsidered.

Candida albicans Increases Dentine Demineralization Provoked by Streptococcus mutans Biofilm

Streptococcus mutans are considered the most cariogenic bacteria, but it has been suggested that Candida albicans could increase their cariogenicity. However, the effect of this dual-species microorganisms’ combination on dentine caries has not been experimentally evaluated. Biofilms of C. albicans, S. mutans and C. albicans + S. mutans (n = 12/biofilm) were grown in ultra-filtered tryptone yeast extract broth culture medium for 96 h on root dentine slabs of known surface hardness and exposed 8 times per day for 3 min to 10% sucrose. The medium was changed 2 times per day (after the 8 cariogenic challenges and after the overnight period of famine), and aliquots were analyzed to determinate the pH (indicator of biofilm acidogenicity). After 96 h, the biofilms were collected to determine the wet weight, colony-forming units, and the amounts of extracellular polysaccharides (soluble and insoluble). Dentine demineralization was assessed by surface hardness loss (% SHL). The architecture of the biofilms was analyzed by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Data were analyzed by ANOVA followed by Tukey’s test (α = 0.05). The dual-species C. albicans + S. mutans biofilm provoked higher % SHL on dentine (p < 0.05) than the S. mutans and C. albicans biofilm. This was supported by the results of biofilm acidogenicity and the amounts of soluble (6.4 ± 2.14 vs. 4.0 ± 0.94 and 1.9 ± 0.97, respectively) and insoluble extracellular polysaccharides (24.9 ± 9.22 vs. 18.9 ± 5.92 and 0.7 ± 0.48, respectively) (p < 0.05). The C. albicans biofilm alone presented low cariogenicity. The images by CLSM and TEM, respectively, suggest that the C. albicans + S. mutans biofilm is more voluminous than the S. mutans biofilm, and S. mutans cells interact with C. albicans throughout polysaccharides from the biofilm matrix. These findings show that C. albicans enhances the cariogenic potential of the S. mutans biofilm, increasing dentine demineralization.

Evaluation of discoloration of sound/demineralized root dentin with silver diamine fluoride: In-vitro study.

The objective was to evaluate the effect of application of silver diamine fluoride (SDF) on discoloration of demineralized dentin over time. Dentin specimens were divided into four groups according to time of dentin demineralization. A 38% SDF solution was then applied to the dentin surfaces. Half of the specimens were placed in light-proof boxes while the remainder were exposed to light. Both groups were maintained at 37˚C. Color change was determined using a spectrophotometer at different time intervals. SEM/EDS analysis were also undertaken. The 13 h EDTA demineralized group showed the highest values for color change among different time intervals, with the control being the lowest. The light exposed groups showed more color change compared to the unexposed groups. We concluded that the degree of dentin demineralization leads to a significant increase of the rate of dentin color change after application of SDF.

Chitosan-Based Extrafibrillar Demineralization for Dentin Bonding

Instability of resin-dentin bonds is the Achilles’ heel of adhesive dentistry. To address this problem, a chelate-and-rinse extrafibrillar dentin demineralization strategy has been developed that keeps intrafibrillar minerals within collagen fibrils intact to prevent activation of endogenous proteases that are responsible for collagen degradation within hybrid layers. The objective of the present study was to evaluate the potential of using chitosan >40 kDa as an antimicrobial extrafibrillar dentin-chelating agent to enhance bond durability. Transmission electron microscopy provided evidence for retention of intrafibrillar minerals and smear plugs in dentin conditioned with 1 wt% chitosan. Analyzed by Kruskal-Wallis analysis of variance, Dunn’s statistic, and separate Mann-Whitney tests, tensile bond strengths to wet- and dry-bonded dentin indicated that chelating dentin with chitosan for 60 s prior to bonding did not result in a significant decline in resin-dentin bond strength when compared with that of phosphoric acid etching (P > 0.05). Gelatinolytic activity within the hybrid layers was examined via in situ zymography after 24-h storage or after thermomechanical cycling and analyzed with 3-factor analysis of variance. After 24 h, enzymatic activity was detected only within completely demineralized phosphoric acid–etched dentin, with values derived from dry bonding significantly higher than those derived from wet bonding (P < 0.05). Negligible fluorescence was detected within hybrid layers when dentin was conditioned with chitosan, even after thermomechanical cycling, as compared with the controls. Reduction in water permeability in chitosan-conditioned dentin, attributed to smear plug retention, also fostered long-term bond stability. Antibacterial testing performed with live/dead staining indicated that the acetic acid–solubilized chitosan possessed antibacterial activities against 3 single-species biofilms: Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis. Taken together, the new chitosan-based extrafibrillar demineralization strategy retains intrafibrillar minerals, reduces endogenous protease-initiated collagen degradation, prevents water permeation within hybrid layers, and kills bacteria on dentin surfaces, which are crucial factors for enhancing resin-dentin bond durability.

One-step phosphorylated poly(amide-amine) dendrimer loaded with apigenin for simultaneous remineralization and antibacterial of dentine.

Dental caries, starting from demineralization of enamel and dentine, is closely related with acid-producing bacteria in oral cavity, for example, Streptococcus mutans. Remineralization is an efficient way to prevent the disease progression and facilitate the therapy of incipient caries. Therefore, for the purpose of effective dentine repair, remineralization and antibacterial should be combined simultaneously. However, most of the literatures only focus on one single aspect, while combing remineralization and antibacteria for dentine repair in one system is rarely reported. Here in this work, phosphoryl-terminated poly(amide-amine) dendrimers were loaded with apigenin, a water-nonsoluble drug antibacterial agains Streptococcus mutans. The apigenin-loaded dendrimers bind strongly with dentine, which further induce dentine tubules occlusion through mineralization in artificial saliva, and the release of apigenin can prevent further erosion of dentine by bacteria. Meanwhile, the phosphorylated dendrimers are easily prepared by one-step modification of poly(amide-amine) and exhibit good cytocompatibility. This strategy developed here can provide reference for the design of effective anti-caries materials.

Mineral density in carious dentine after treatment with calcium silicates and polyacrylic acid-based cements.

To test the hypothesis that polyacrylic acid-based and calcium silicate materials are able to increase mineral density of artificially demineralized dentine produced by a microcosm model for caries formation. Forty-five dentine cavities were prepared in nine sound human third molars (five in each tooth). Specimens received a bacterial inoculum and were incubated in BHI plus 5% of sucrose for 96h to allow biofilm formation. After that, the specimens were scanned in a high energy micro-CT (baseline stack) and then, each cavity was filled with different cements: zinc polycarboxylate (Poly Zinc™, Prevest DenPro, Jammu, India), conventional glass-ionomer (Ketac Molar, 3M ESPE, Seefeld, Germany) and two calcium silicate-based materials (MTA, Angelus, Londrina, Brazil and Biodentine™, Septodont, Saint Maur des Fosses, France) whilst the central cavity was filled with utility wax (negative control). Specimens were kept for 45days under simulated intrapulpal pressure using simulated body fluid and after, they were again scanned (treated stacks). Reconstruction of the acquired projections was accomplished using standardized parameters, and the percentage mineral density change was calculated around each cavity. Data were tested for normality using the Shapiro-Wilk test, followed by nonparametric methods. A microcosm artificial caries model was able to cause dentine demineralization (±90% related to sound dentine). Significant increases in dentine mineral density were achieved using MTA or Poly Zinc™ (43.56% and 41.64% remineralization), followed by Biodentine™ (36.40%) and Ketac Molar (32.54%), P<0.05. All cements increased mineral density in demineralized dentine, but zinc polycarboxylate cement and MTA had greater bioactive potential. Micro-CT proved to be an effective method to evaluate changes in mineral density within dentine in contact with bioactive materials.

Demineralization capacity of commercial 10-methacryloyloxydecyl dihydrogen phosphate-based all-in-one adhesive

ObjectiveWe determined the amounts of calcium salt of 10-methacryloyloxydecyl dihydrogen phosphate (MDP-Ca salt) and dicalcium phosphate dihydride (DCPD) with an amorphous phase developed during the application of commercial MDP-based all-in-one adhesives to enamel and dentin. This is because the demineralization by MDP and following calcium salt formation of MDP may be limited by an ionic bond formation of MDP to hydroxyapaptite in the enamel and dentin and following intermediary layer formation of MDP, since MDP forms a chemically-stable adsorption layer. MethodsScotchbond Universal Adhesive, Clearfil Tri–S Bond ND, Clearfil Tri–S Bond ND Quick, G–Bond Plus and our designed MDP-based all-in-one adhesive were used. Enamel and dentin reactant residues of each adhesive were prepared by varying the adhesive application periods: 1, 30 and 60min, and were analyzed using phosphorous-31 nuclear magnetic resonance and X-ray diffraction. ResultsIncreasing the adhesive application period to enamel and dentin led to the increased amount of MDP-Ca salt in contrast to amorphous DCPD. In the dentin, each adhesive showed a saturated value on the production amount of MDP-Ca salt when the adhesive was applied more than 30min. In contrast, in the enamel, each adhesive showed an intermediate value on the saturated production amount of MDP-Ca salt that the respective adhesive exhibited. This is due to MDP employed demineralizes the enamel and dentin until MDP was completely consumed yielding MDP-Ca salt. ConclusionCommercial MDP-based all-in-one adhesives would not form an intermediary layer of MDP on hydroxyapatite throughout their application period to enamel and dentin. Clinical relevanceThe rate of MDP-Ca salt produced by the demineralization of enamel and dentin depends on the components that constitute commercial adhesive more strongly than on the concentrations of MDP and water in the respective adhesive. This is because HEMA-containing adhesive shows a slower production rate of MDP-Ca salt than HEMA-free adhesive in the enamel and dentin samples.

Cariogenicity of a Milk-Based Drink Used as a Dietary Supplement for Older Adults Using a Root Caries Experimental Model

The aim of this study was to evaluate the cariogenicity of a milk-based drink intended for older adults that was used as part of a governmental initiative in Chile to improve their nutritional conditions. This drink contains a high concentration of sugars, which can contribute to root caries development. To test this hypothesis, an experimental biofilm/caries model was used. Dentin slabs were used to grow biofilms of Streptococcus mutans UA159. Slabs/biofilms were exposed 3× per day to bovine milk with different fat content, the milk-based drink, and the milk-based drink supplemented with 10 g of sucrose added per serving. Slabs exposed to 10% sucrose or 0.9% NaCl were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial counts and acidogenicity. Dentin demineralization was estimated by the loss of surface microhardness and integrated mineral loss. Results were compared by analysis of variance and Tukey’s test. The milk-based drink showed higher acidogenicity than milk with its entire (whole) or reduced total fat content (skim). The milk-based drink supplemented with ­sucrose had similar acidogenicity as the 10% sucrose positive control (p = 0.506). Whole milk exposure elicited lower bacterial counts than the positive control, the milk-based drink, and the milk-based drink supplemented with sucrose (p = 0.002; 0.006 and 0.014 respectively). Although skim milk induced higher demineralization than whole milk, both milk types produced lower demineralization than the milk-based drink. Regarding integrated mineral loss, demineralization induced by the milk-based drink and the milk-based drink supplemented with sucrose was similar to that induced by the positive control and skim milk (p > 0.05). Sugar-containing milk-based drinks used as dietary supplements for older adults may be highly cariogenic and could represent a potential risk for root caries.

Potensi Ekstrak Buah Delima Merah (Punica granatum Linn) terhadap Penurunan Jumlah Koloni Streptococcus mutans

Streptococcus mutans is a bacteria that initiates the formation of plaque on the tooth surface. S. mutans works by fermenting carbohydrates to produce an acidic atmosphere, so the plaque pH becomes low, this condition can lead to demineralization of enamel and dentin commonly called caries. One of alternative treatment to reduce the population of S. mutans by using herbal plants, namely red pomegranate. Red pomegranate contains flavonoids, tannins, and alkaloids as antibacterial. This study aimed to determine the inhibition of red pomegranate extract on growth of S. mutans. The method used is by counting the number of S. mutans using a colony counter. This method with 4 samples in each study group. The study group consists of 4 treatment groups (25%, 50%, 75% and 100% red pomegranate extract), positive control group (chlorhexidine), and negative control group (sterile aquades). Data were analyzed using Kruskal-Wallis Test and Mann-Whitney test. Red pomegranate extract has the ability to inhibit the growth of S. mutans. The concentration of red pomegranate extract that has the greatest inhibitory effect on S. mutans growth is 100%.&#x0D; Keyword: Antibacterial, Streptococcus mutans, caries, red pomegranate fruit extract

In-office tooth whitening

Peroxides have been used to whiten teeth for over 100 years and among all whitening therapies the oldest is in-office bleaching. In this article, we will present the summary of the step-by-step procedure and side effects of this bleaching. Some comments regarding the number of clinical appointments for teeth whitening, concentration of the bleaching products, the effects of dentin dehydration and demineralization on the final outcome as well as bleaching-induced tooth sensitivity are given. At the end, some references for further reading can be found.

Starch Combined with Sucrose Provokes Greater Root Dentine Demineralization than Sucrose Alone

Since there is no consensus about whether starch increases the cariogenic potential of sucrose, we used a validated 3-species biofilm model to evaluate if starch combined with sucrose provokes higher root dentine demineralization than sucrose alone. Biofilms (n = 18) composed by Streptococcus mutans (the most cariogenic bacteria), Actinomces naeslundii (which has amylolytic activity), and Streptococcus gordonii (which binds salivary amylase) were formed on root dentine slabs under exposure 8 ×/day to one of the following treatments: 0.9% NaCl, 1% starch, 10% sucrose, or a combination of 1% starch and 10% sucrose. Before each treatment, biofilms were pretreated with human whole saliva for 1 min. The pH of the culture medium was measured daily as an indicator of biofilm acidogenicity. After 96 h of growth, the biofilms were collected, and the biomass, bacteria viability, and polysaccharides were analyzed. Dentine demineralization was assessed by surface hardness loss (% SHL). Biofilm bioarchitecture was analyzed using confocal laser scanning microscopy. Treatment with a starch and sucrose combination provoked higher (p = 0.01) dentine demineralization than sucrose alone (% SHL = 53.2 ± 7.0 vs. 43.2 ± 8.7). This was supported by lower pH values (p = 0.007) of the culture medium after daily exposure to the starch and sucrose combination compared with sucrose (4.89 ± 0.29 vs. 5.19 ± 0.32). Microbiological and biochemical findings did not differ between biofilms treated with the combination of starch and sucrose and sucrose alone (p > 0.05). Our findings give support to the hypothesis that a starch and sucrose combination is more cariogenic for root dentine than sucrose alone.

Correction: Correlation between ICDAS and histology: Differences between stereomicroscopy and microradiography with contrast solution as histological techniques.

[This corrects the article DOI: 10.1371/journal.pone.0183432.].

Cariogenicity induced by commercial carbonated beverages in an experimental biofilm-caries model.

Objectives:Frequent consumption of sugars-containing carbonated beverages has been associated with caries, but the consequences on the dental biofilm remain unclear. The aim was to evaluate the effect of commercial carbonated beverages and their sugar-free version on enamel and dentine demineralization and on the cariogenic properties of Streptococcus mutans biofilms.Materials and Methods:Biofilms of S. mutans UA159 were grown on enamel and dentin slabs and exposed 3 times/day for 5 min, to a commercial cola or orange-flavored carbonated beverage or to their sugar-free version. Biofilms/slabs were recovered to assess biomass, viable microorganisms, protein content and polysaccharides. Demineralization was estimated by the variation of Knoop surface microhardness.Results:Exposures to the biofilm with sugars-containing carbonated beverages resulted in similar biomass, viable microorganisms, proteins, and polysaccharides than sucrose (P < 0.05). The sugar-free cola and orange-flavored drink showed lower effect on the biofilm, as compared with sucrose or their sugared version (P < 0.05). All of the products tested, included the sugar-free, showed higher demineralization than the negative control (P < 0.05).Conclusions:Sugars-containing carbonated beverages enhance cariogenic activity of S. mutans biofilms, comparable with sucrose. Sugar-free carbonated beverages also have a high demineralizing potential, without affecting biofilm properties.

Pathophysiology of Dental Caries.

Carious lesion dynamics are dependent predominantly on the availability of fermentable sugars, other environmental conditions, bacteria, and host factors. Our current understanding of the microorganisms involved in the initiation and progression of caries is still rather incomplete. The most relevant acidogenic-aciduric bacterial species known to date are Streptococcus mutans, bifidobacteria, and lactobacilli. Whereas mutans streptococci are initiators, bifidobacteria and lactobacilli are more enhancers for progression. Boosters for microbial activity are specific environmental conditions, such as the presence of fermentable dietary sugars and the absence of oxygen. Based on these conditions, the necrotic and/or contaminated zone fulfils all criteria for disease progression and has to be removed. For those deep lesions where the pulp vitality is not affected, a selective removal of the contaminated leathery dentine should take place as this approach lowers the risk of regrowth of the few embedded microbial cells here. In repelling the microbial attack and repairing damage, the host has developed several ingenious strategies. A major resistance to carious lesion progression is mounted by the dentine-pulp tissues. The signalling molecules and growth factors released upon dentine demineralisation upregulate the odontoblast activity and act as sensor cells. After carious stimulation, odontoblasts initiate an inflammatory reaction by producing chemokines and synthesise a protective tertiary dentine. After the destruction of these cells, the pulp still has a high capacity to synthesise this tertiary dentine thanks to the presence of adult stem cells within the pulp. Also, in addition to the systemic regulation, the pulp which is located within inextensible the confines of the dentine walls has a well-developed local regulation of its inflammation, regeneration, and vascularisation. This local regulation is due to the activity of different pulp cell types, mainly the fibroblasts, which secrete soluble molecules that regulate all these processes.

Etching Treatment Effect on Surface Morphology of Dental Structures

This study examined and compared surface of human dentine after acidic etching with hydrogen peroxide, phosphoric acid liquid and gel. Surface demineralization of dentin is necessary for a strong bond of adhesive at dental surface. Split human teeth were used. After application of mentioned substances at dentin level measures of the contact angle and surface morphology were employed. Surface morphology was analyzed with the help of scanning electron microscopy and atomic force microscopy. Liquid phosphoric acid yielded highest demineralization showing better hydrophobicity than the rest, thus having more contact surface. Surface roughness are less evident and formed surface micropores of 4 �m remained open after wash and air dry providing better adhesive canalicular penetration and subsequent bond.