Search

Purpose: To examine the relationship between remineralization of incipient root dentin lesions and the presence of polymicrobial biofilms, as well as examine changes in microbial composition., Methods: Bovine root dentin disks used as specimens for biofilm formation, were cultured using saliva from a single donor. Amsterdam Active Attachment biofilm model was used to grow biofilms. The culture medium was McBain 2005 with 0.2% sucrose and 0.4 ppm F as sodium fluoride. After cultivation for 48 hours to achieve demineralization, a control group (n=10) was obtained and the other specimens were further cultured for 336 hours in two types of remineralization culture medium, with sucrose (S+) and without sucrose (S-), through continuous anaerobic incubation (10% CO2,10% H2, 80% N2). Then half of the specimens cultured in the S- medium were transferred to the S+ medium for an additional 48 hours resulting in three experimental groups S(+) (n=10), S(-) (n=10), and S(-)de (n=10), respectively. Experiment 1: Transverse microradiography (TMR) analysis - Immediately after respective culture treatments, integrated mineral loss (IML) and lesion depth (LD) in the dentin specimens were analyzed by TMR. Experiment 2: Microbiome analysis - Sequence data of the 16S rRNA gene of each sample was obtained using MiSeq, and partial base sequences were determined. Next-generation sequencing was performed to determine the taxonomic groups of fungi present in the biofilm samples., Results: Experiment 1: In the control group, formation of dentin demineralization lesions by polymicrobial species biofilms was confirmed. The S(-) group showed significantly decreased IML and shallower LD compared to the control group. The S(-)de group showed a significant increase in IML and LD compared to the S(-) group. Experiment 2: There were statistically significant differences in microbiome between the control group and each of the three experimental groups, both at the genus and species levels. A significant difference in genus was observed between the S(-) group and the S(-)de group., Clinical Significance: The confirmation of the possibility of microbial shift occurring during the remineralization process of root caries will lead to the development of new remineralization therapies., Competing Interests: The authors declared no conflict of interest. This work was supported by MEXT/JSPS KAKENHI Grant Number 20K09965 Grant-in-Aid for Scientific Research (C), 2020-2023., (Copyright©American Journal of Dentistry.)

This review aims to present scientific knowledge regarding the demarcated opacities of molar incisor hypomineralisation (MIH) and factors that clarify the occurrence of posteruptive enamel breakdown. The demarcated opacities have distinct boundaries with the adjacent nonaffected enamel and may vary in color among white, creamy, yellow, and brownish. The hypomineralised enamel is more porous and less organized than the nonaffected enamel. As a result of the reduced mineral content and higher protein content, the hypomineralised enamel shows a progressive reduction in its mechanical properties according to the opacity feature. Chemically, the protein content of MIH opacities is abnormally high, mainly composed by albumin, which is a serum protein usually not found in mature enamel. The highest protein content is seen in brown opacities, followed by yellow and white opacities, both with higher protein content than nonaffected enamel. The fact that the hypomineralised enamel is more fragile than the nonaffected enamel is supported not only by laboratorial findings but also by clinical prospective studies that observed an aggravation of MIH over time, as well as the correlation between the color of the demarcated opacities and the risk of posteruptive enamel breakdown. A better understanding about the microstructure of the hypomineralised enamel has relevant implications for the clinical approach of the condition. In the clinic, besides a comprehensive assessment of anamnesis and clinical data, it is advisable to record the color and the location of the opacities by tooth surface in order to support the treatment decisions and estimate a prognosis for MIH patients., (© 2024 S. Karger AG, Basel.)

Molar incisor hypomineralisation (MIH) is a qualitative type of enamel defect, which occurs due to a failure in the biomineralisation process of the enamel organic matrix during amelogenesis. The tooth enamel affected by MIH shows changes in its chemical, structural, and mechanical properties, leading to different clinical repercussions. The color of MIH opacities varies from opaque white to yellow/brown, and elemental analyses of these lesions show a lower calcium and phosphate content, minerals that are more abundant in sound enamel. Furthermore, the incorporation of other molecules occurs, such as carbonate, a component that provides a greater degree of solubility, thus making hypomineralised enamel more susceptible to posteruptive fractures. At a structural level, the layer of hydroxyapatite crystals appears to be disorganized, with morphological changes, implying a greater degree of porosity in the structure. The increase in porosity of the structure may be associated with dental hypersensitivity, a common clinical repercussion among patients with MIH. Among the mechanical properties, a decrease in hardness and modulus of elasticity occurs, and this also makes the enamel more fragile. Deficiency in biomineralisation can be caused by changes in the function of ameloblasts or by failures at the intercellular junction that result in lower activity of proteases such as MMP-20 and KLK4. The increase in proteins in the organic matrix of enamel impairs the growth and incorporation of minerals into the hydroxyapatite crystals, so that the enamel becomes hypomineralised and has larger organic content, thus having an impact on its properties. These changes present in the enamel with MIH help to explain the clinical repercussions caused by this condition., (© 2024 S. Karger AG, Basel.)

Introduction: Visual imaging of subsurface caries lesions is of vital interest in dentistry, which can be obtained by invasive radiography technique as well as by available non-destructive imaging approaches. Thus, as a first step toward the development of a new innovative approach, Spectral-domain optical coherence tomography (SD-OCT) was applied to detect the lesion depth in comparison to the established reference technique (transverse microradiography [TMR])., Methods: Bovine enamel specimens were demineralized for 5 days, following previous studies. For OCT, the resulting artificial lesions were scanned three-dimensionally (SD-OCT) and semi-automated measured (CarLQuant). For TMR, specimens were sectioned and the lesion depth was manually determined (Inspektor Research System)., Results: The range of lesion depth detected with OCT was 24.0-174.0 μm (mouth rinse study), 18.0-178.0 μm (toothpastes study) and with TMR 59.2-198.0 μm (mouth rinse study), 33.2-133.4 μm (toothpastes study). We found a strong correlation between both methods in terms of lesion depth (Spearman rankwith outlierp < 0.001, Rho = 0.75, Spearman rankwithout outlierp = 0.001, Rho = 0.79). The two methods produce similar results (Passing-Bablok regression, 1.16). As deeper is the lesion, the smallest is the difference between both methods as indicated by Bland-Altman-plots., Conclusion: Especially in the case of deep lesions, the values obtained by both methods are in agreement, and OCT can potentially substitute TMR to detect and assess lesion depth with the benefit of being non-destructive., (© 2024 S. Karger AG, Basel.)

Objectives: This in vitro study investigated the ability of a blue protein-based hydroxyapatite porosity probe to selectively detect artificial enamel caries-like lesions of varying severities., Methods: Artificial caries-like lesions were formed in enamel specimens using a hydroxyethylcellulose-containing lactic acid gel for 4/12/24/72 or 168 h. One untreated group was used as a control. The probe was applied for 2 min and unbound probe rinsed off with deionized water. Surface color changes were determined spectrophotometrically (L*a*b* color space) and with digital photography. Lesions were characterized using quantitative light-induced fluorescence (QLF), Vickers surface microhardness, and transverse microradiography (TMR). Data were analyzed using one-way ANOVA., Results: Digital photography did not reveal any discoloration in unaffected enamel. However, all lesions stained blue with color intensity positively correlated with demineralization times. The color data reflected similar trends: lesions became significantly darker (L* decreased) and bluer (b* decreased), while overall color differences (ΔE) increased significantly after probe application (4-h lesion, mean±standard deviation: ΔL*=-2.6 ± 4.1/Δb*=0.1 ± 0.8/ΔE=5.5 ± 1.3 vs. 168-h lesion: ΔL*=-17.3 ± 1.1/Δb*=-6.0 ± 0.6/ΔE=18.7 ± 1.1). TMR analysis revealed distinct differences in integrated mineral loss (ΔZ) and lesion depth (L) between demineralization times (4-h lesion: ΔZ=391±190 vol%min × µm/L = 18.1 ± 10.9 µm vs. 168-h lesion: ΔZ=3606±499 vol%min × µm/L = 111.9 ± 13.9 µm). QLF and microhardness were also able to differentiate between demineralization times. L and ΔZ strongly correlated (Pearson correlation coefficient [r]) with Δb* (L vs. Δb*: r=-0.90/ΔZ vs. Δb*: r=-0.90), ΔE (r = 0.85/r = 0.81), and ΔL* (r=-0.79/r=-0.73)., Conclusion: Considering the limitations of this study, the blue protein-based hydroxyapatite-binding porosity probe appears to be sufficiently sensitive to distinguish between unaffected enamel and artificial caries-like lesions., Clinical Significance: Early detection of enamel caries lesions remains one of the most critical aspects in the diagnosis and management of dental caries. This study highlighted the potential of a novel porosity probe in detecting artificial caries-like demineralization by objective means., Competing Interests: Declaration of Competing Interest This work was support by Incisive Technologies Pty Ltd, Melbourne, Victoria, Australia. JEM and KAH are employees of and have equity in Incisive Technologies., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objective: To investigate the effect of fluoride-containing whitening products on sound enamel and on artificial caries lesions during a cariogenic challenge., Materials and Methods: Bovine enamel specimens (n = 120) with three areas [non-treated sound enamel (NSE), treated sound enamel (TSE), and treated artificial caries lesion (TACL)] were randomly assigned to the four groups: whitening mouthrinse (WM: 2.5% hydrogen peroxide-100 ppm F - ), placebo mouthrinse (PM: 0% hydrogen peroxide-100 ppm F - ), whitening gel (WG: 10% carbamide peroxide-1130 ppm F - ), and deionized water (negative control; NC). The treatments (2 min for WM, PM, and NC, and 2 h for WG) were carried out during a 28-day pH-cycling model (6 × 60 min demineralization/day). Relative surface reflection intensity (rSRI) and transversal microradiography (TMR) analyses were performed. Fluoride uptake (surface and subsurface) was measured in additional enamel specimens., Results: For TSE, a higher value of rSRI was observed in WM (89.99% ± 6.94), and a greater decrease in rSRI was observed for WG and NC, and no sign of mineral loss was verified for all groups (p > 0.05). For TACL, rSRI significantly decreased after pH-cycling for all experimental groups with no difference between them (p < 0.05). Higher amounts of fluoride were found in WG. WG and WM exhibited intermediate values of mineral loss, similar to PM., Conclusions: The whitening products did not potentialize the enamel demineralization under a severe cariogenic challenge, and they did not exacerbate mineral loss of the artificial caries lesions., Clinical Relevance: Low concentrated hydrogen peroxide whitening gel and mouthrinse containing fluoride do not intensify the progression of caries lesions., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

White spot lesions and enamel cracks are the 2 most prominent diseases that occur after orthodontic treatment and are caused by enamel demineralization from accumulated bacterial biofilms and/or enamel damage caused by the removal of residual adhesive after bracket debonding. Inspired by the self-assembled amelogenin nanoribbons in enamel, we developed an enamel coating with a self-assembling antimicrobial peptide, D-GL13K, to simultaneously reduce demineralization and residual adhesive. The self-assembled amphiphilic nanoribbons significantly increased the hydrophobicity of the etched enamel, which reduced the permeability of the coated enamel surfaces as desired. The antimicrobial activity of this coating was evaluated against Streptococcus mutans by colony-forming unit counting and live/dead assays. The anti-demineralization effect was demonstrated by the reduced demineralization depth analyzed by optical coherence tomography and the increased Vickers hardness. The coatings did not reduce the shear bond strength but significantly reduced the adhesive remnant index score. This bioinspired enamel coating may provide a new strategy for preventing white spot lesions and enamel cracks after orthodontic treatment.

This study compared the effect of topically applied fluoride products on dentine lesions in an in vitro experiment. Demineralized bovine dentine specimens were treated once with either SDF solution (35,400 ppm F), NaF varnish (22,600 ppm F), TiF4 solution (9,200 ppm F), SnF2 gel (1,000 ppm F), no treatment (control), or preserved as baseline lesions. After the application and subsequent removal of the fluoride products, the specimens were subjected to pH-cycling. Calcium loss and uptake in the de- and remineralization buffers were assessed daily. Fluoride release into the buffers was analyzed on days 1, 2, 3, 5, 8, and 13. After the pH-cycling period, mineral distribution throughout the lesion depth was analyzed using transversal microradiography (TMR). X-ray energy-dispersive spectroscopy (EDS) examined the deposition of silver, titanium, and tin after application of SDF, TiF4, and SnF2, respectively. Overall, calcium loss and uptake analysis in the de- and remineralization buffers revealed that the SDF product was the most effective in inhibiting lesion progression, followed by the TiF4, NaF, and SnF2 products. Fluoride analysis disclosed a steep reduction of the amount of fluoride released into de- and remineralization buffers with time. The fluoride effects on de- and remineralization continued beyond the days that fluoride was released into the buffers. TMR analysis showed significant remineralization in the outer zone of the dentine lesions for all fluoride products, with SDF giving hypermineralization in this zone. In the inner zone, lesions developed in all fluoride groups, with the smallest in the SDF group. EDS showed silver and titanium deposition in depth up to 85 μm and 8 μm, respectively, while no tin deposition was observed. The silver in the dentine lesions did not contribute significantly to the density of the TMR profiles in the SDF group. In conclusion, all topical fluoride products protected the dentine lesions against lesion progression, but at different degrees. SDF showed a superior effect in protection against further demineralization and enhancement of remineralization. This was probably attributed to its fluoride concentration that was the highest among the fluoride products., (© 2021 The Author(s)Published by S. Karger AG, Basel.)

This retrospective pilot study used a newly developed evaluation tool to assess the prevalence and incidence of White Spot Lesions (WSL) before and after multibracket appliance (MB) therapy. Digital photographs of 121 adolescent patients (63 ♂, 58 ♀) with metal brackets were analyzed retrospectively before and after MB therapy. The labial surfaces of anterior teeth, canine teeth, and premolars in the upper (UJ) and lower jaws (LJ) were evaluated using the Enamel Decalcification Index (EDI) by Banks and Richmond (Eur J Orthod, 16(1):19-25, 1994, levels 0-3) and a specially developed digitally scaled graticule with concentric circles to quantify the extent of WSL (in %). The statistical data analysis was based on crosstabulations and logistic regression. Before MB, 69.4% of the patients presented at least one WSL and 97.5% after, an increase of 28.1%. Before MB, 18.4% of the tooth surfaces (TS) showed an EDI level of 1-3. After MB, 51.8% of the TS featured WSL. 18.2% of the TS showed a WSL to the extent of ≥ 20-100% before and 52.3% after MB. The incidence in the UJ (71-79%) as well as the LJ (64-76%) was highest for the first and second premolars and lowest for LJ incisors (22-35%). The probability for developing a new distal WSL is higher than developing gingival, mesial or occlusal WSL. Labial MB therapy drastically increases the risk of developing WSL. We verified a concise quantification of the extent of labial WSL with the evaluation index., (© 2021. The Author(s).)

Objective: To compare the ability of two calcium-releasing restorative materials to inhibit root dentin demineralization in an artificial caries model., Methods and Materials: Preparations were made at the cementum-enamel junction of extracted human molars (40, n=10/material) and restored with two calcium-releasing materials (Experimental composite, Pulpdent Corporation and Cention N, Ivoclar Vivadent), a resin composite (Filtek Supreme Ultra, 3M Oral Care), and a resin-modified glass ionomer (RMGI) (Fuji II LC, GC). All materials (other than the RMGI) were used with an adhesive (Scotchbond Universal Adhesive, 3M Oral Care) in the self-etch mode, which was light cured for 10 seconds. All restorative materials were light cured in 2-mm increments for 20 seconds and then finished with a polishing disc. Teeth were incubated (37°C) for 24 hours in water. An acid-resistant varnish was painted onto the teeth around the restoration, leaving a 2-mm border of uncovered tooth. A demineralization solution composed of 0.1 M lactic acid, 3 mM Ca3(PO4)2, 0.1% thymol, and NaOH (to adjust pH=4.5), and a remineralization solution composed of 1.5 mM Ca, 0.9 mM P, and 20 mM Tris(hydroxymethyl)-aminomethane (pH=7.0) were prepared. Specimens were placed in the demineralization solution for 4 hours, followed by the remineralization solution for 20 hours and cycled daily for 30 days. The specimens were embedded, sectioned into 100-μm sections, and the interface between the restorative material and root dentin was viewed with polarized light microscopy. A line was drawn parallel with the zone of demineralization for each tooth. The area of "inhibition" (defined as the area external to the line) or "wall lesion" (defined as the area internal to the line) was measured with image evaluation software. Areas of inhibition were measured as positive values, and areas of wall lesions were measured as negative areas., Results: A one-way analysis of variance (ANOVA) found significant differences between materials for "inhibition/wall lesion" areas in root dentin (p<0.001). Tukey post hoc analysis ranked materials (μm2, mean ±SD): Fuji II LC (5412±2754) > Cention N (2768±1576) and experimental composite (1484±1585) > Filtek Supreme Ultra (-1119±1029)., Conclusion: The experimental composite and Cention N materials (used with an adhesive) showed net areas of inhibition greater than a reference resin composite, albeit at a lower level than a reference RMGI material (used with no adhesive)., (©Operative Dentistry, 2021.)

To evaluate the effectiveness of a calcium silicate/phosphate fluoridated tooth paste and a serum compared with a toothpaste containing hydroxyapatite on protecting the enamel after interproximal reduction against demineralization. 3 sets of eleven incisors were created. The teeth underwent interproximal enamel reduction (IER) of 0.5 mm. Each set was allocated to one of three groups: (1) Brushing without toothpaste (control group); (2) Vitis toothpaste + Remin Pro; (3) Regenerate toothpaste + Regenerate Serum. The agents were applied three times a day and specimens subjected to demineralization cycles for 30 days. The weight percentages of calcium (Ca) and phosphorous (P) were quantified by X-ray microfluorescence spectroscopy. Surface microhardness measurements and electron scanning microscopy (SEM) observations were made. Ca data and the Ca/P ratio were significantly higher in Group 3 than the other groups (p < 0.017), while P was significantly lower in Group 3 (p < 0.017). No significant differences were found between Groups 1 and 2 (p > 0.017). Group 3 showed significantly higher microhardness values (p < 0.05) than Group 1. No significant differences were found for other comparisons between groups (p < 0.05). SEM images showed less demineralization in Group 3. The application of a calcium silicate/phosphate fluoridated tooth paste (Regenerate advance) and a dual serum (Regenerate advance enamel serum) protect the enamel with interproximal reduction against demineralization. Therefore, this treatment could be used to prevent the dissolution of hydroxyapatite after IER.

Developing artificial caries lesions with varying characteristics is needed to adequately study caries process in vitro. The objective of this study was to investigate artificial caries lesion characteristics after secondary demineralization protocol containing theobromine and fluoride. Sixty bovine enamel slabs (4 × 3 mm) were demineralized using a Carbopol-containing protocol for 6 days. A baseline area (2 × 3 mm) was protected with acid-resistant nail varnish, after which specimens were exposed for 24 h to a secondary demineralization protocol containing acetic acid plus one of four fluoride/theobromine combinations ( n = 15): theobromine (50 or 200 ppm) and fluoride (0 or 1 ppm). Specimens were sectioned and analyzed using transverse microradiography for changes in mineral content, lesion depth, and surface layer mineralization. Data was analyzed using paired t-test and analysis of variance followed by Bonferroni test at 0.05 significance level. After secondary demineralization, fluoride-containing groups had significantly deeper lesions ( p = 0.002 and 0.014) compared to the group with 0 ppm fluoride and 50 ppm theobromine. Mineral content and lesion depth were significantly different compared to baseline for all groups. Theobromine did not show an added effect on mineral uptake. Theobromine-containing groups exhibited particularly deep lesions with a more uniform mineral profile in the presence of fluoride.

Objective: evaluation of oral hygiene to enhance the prophylaxis of complications specifically of dental caries inchildren with disturbances in tooth formation (DTF) who live on radiologically contaminated territories after theChornobyl NPP (ChNPP) accident., Materials and Methods: Children aged 6-14 years (n = 1470) with DTF were the study subjects examined in 2012-2016. Among them there were (n = 528) residents of the zones III and IV of radiological contamination after theChNPP accident with 137Cs soil contamination density 1-15 Ci/km2. The effective radiation dose in them was notexceeding 1 mSv/year. The Green-Vermillion and Silness-Loe oral hygiene indices were assesses within clinicalexamination methods., Results: The worth oral hygiene was revealed in children having got the DTF, compensated chronic diseases oforgans and systems, and in those with burdened radiation history. The Green-Vermilion and Silness-Loe indices were1.7 ± 0.51 and 1.65 ± 0.46 respectively., Conclusions: A significant deterioration in oral hygiene confirmed by the highest values of the Green-Vermillion index (1.7 ± 0.51; p < 0.001 - «unsatisfactory oral hygiene» criterion) and Silness-Loe index (1.65 ± 0.46; p < 0.001 -«poor oral hygiene» criterion) was found in children with DTF aged 6-14 years being residents of contaminated areasas a result of the Chernobyl accident. The revealed deterioration may be due to a set of negative factors, includingthe impact of ionizing radiation in low doses and peculiarities of social status. Results of the questioning of surveyed pediatric contingents both having got an DTF and with no defects of the hard tissues of teeth indicate aninsufficient level of knowledge and skills in hygienic care of oral cavity regardless of the area of residence.Development of a set of measures to prevent the DTF complications in children should be carried out taking intoaccount the state of oral hygiene, level of knowledge and skills in oral care, and include the use of hygiene products, namely toothpastes and anti-caries mouthwashes., (S. F. Liubarets, V. V. Kaminskyi, T. F. Liubarets, D. E. Afanasyev, G. I. Shapovalova, O. V. Savychuk.)

This study was undertaken to assess in vivo the corrosion in two commercial nickel-titanium (NiTi) orthodontic archwires removed from the oral cavity of patients using fluoride mouthwashes. Five volunteers took part in this study on the corrosion behavior of two brands of NiTi archwires (3M and AO (brand of archwire)) during use of two mouthwashes with neutral sodium fluoride 1.1%, one with acidulated fluoride 1.1%, and one with placebo and a control group. Each patient used one mouthwash in three different periods of time for 1 min a day for 30 days. The archwires were assessed with scanning electron microscopy and atomic force microscopy for qualitative and quantitative analysis. The values obtained with atomic force microscopy (AFM) were submitted to normality test, two-way analysis of variance, and Tukey's test at a significance level of 5%. The AFM images showed a gradual qualitative increase in the roughness of both types of wire between the treatments: control < placebo < neutral fluoride < acidulated fluoride. The arithmetic average of the roughness and root mean square of the roughness were similar. As for 3M archwires, only the acidulated fluoride group differed statistically from the others. As for AO archwires, the control and placebo groups did not differ from each other, but differed from the other fluoride treatments. The group using neutral fluoride also differed significantly from the acidulated fluoride group. 3M archwires were not affected by daily oral challenges. AO archwires were not affected by daily oral challenges either; their association with fluoride, either neutral or acidulated, increased their roughness., (© 2020 Wiley Periodicals, Inc.)

Amelogenesis imperfecta (AI) is a heterogeneous group of genetic diseases characterised by dental enamel malformation. Pathogenic variants in at least 33 genes cause syndromic or non-syndromic AI. Recently variants in RELT, encoding an orphan receptor in the tumour necrosis factor (TNF) superfamily, were found to cause recessive AI, as part of a syndrome encompassing small stature and severe childhood infections. Here we describe four additional families with autosomal recessive hypomineralised AI due to previously unreported homozygous mutations in RELT. Three families carried a homozygous missense variant in the fourth exon (c.164C>T, p.(T55I)) and a fourth family carried a homozygous missense variant in the 11th exon (c.1264C>T, p.(R422W)). We found no evidence of additional syndromic symptoms in affected individuals. Analyses of tooth microstructure with computerised tomography and scanning electron microscopy suggest a role for RELT in ameloblasts' coordination and interaction with the enamel matrix. Microsatellite genotyping in families segregating the T55I variant reveals a shared founder haplotype. These findings extend the RELT pathogenic variant spectrum, reveal a founder mutation in the UK Pakistani population and provide detailed analysis of human teeth affected by this hypomineralised phenotype, but do not support a possible syndromic presentation in all those with RELT-variant associated AI., (© 2020 The Authors. Clinical Genetics published by John Wiley & Sons Ltd.)

Objective: The objective of this study is to evaluate the effects of radiotherapy doses on mineral density and percentage mineral volume of human permanent tooth enamel., Materials and Methods: Synchrotron radiation Xray microcomputed tomography (SRμCT) and microhardness testing were carried out on 8 and 20 tooth samples, respectively. Enamel mineral density was derived from SRμCT technique using ImageJ software. Microhardness samples were subjected to Vickers indentations followed by calculation of microhardness and percentage mineral volume values using respective mathematical measures. Data were analyzed using paired t-test at a significance level of 5%. Qualitative analysis of the enamel microstructure was done with two-dimensional projection images and scanned electron micrographs using μCT and field emission scanning electron microscopy, respectively., Results: Vickers microhardness and SRμCT techniques showed a decrease in microhardness and an increase in mineral density, respectively, in postirradiated samples. These changes were related to mineral density variation and alteration of hydroxyapatite crystal lattice in enamel surface. Enamel microstructure showed key features such as microporosities and loss of smooth homogeneous surface. These indicate tribological loss and delamination of enamel which might lead to radiation caries., Conclusions: Tooth surface loss might be a major contributing factor for radiation caries in head-and-neck cancer patients prescribed to radiotherapy. Such direct effects of radiotherapy cause enamel abrasion, delamination, and damage to the dentinoenamel junction. Suitable measures should, therefore, be worked out to protect nontarget oral tissues such as teeth while delivering effective dosages to target regions., Competing Interests: None

Hypophosphatasia (HPP) is a rare and intractable metabolic bone disease caused by mutations in the ALPL gene. Here, we undertook a nationwide survey of HPP in Japan, specifically regarding the prominent genetic and dental manifestations of odonto (n = 16 cases) and other (termed "non-odonto") (n = 36 cases) types. Mean serum alkaline phosphatase (ALP) values in odonto-type patients were significantly greater than those of non-odonto-type patients (P<0.05). Autosomal dominant and autosomal recessive inheritance patterns were detected, respectively, in 89% of odonto-type and 96% of non-odonto-type patients. The ALPL "c.1559delT" mutation, associated with extremely low ALP activity, was found in approximately 70% of cases. Regarding dental manifestations, all patients classified as odonto-type showed early exfoliation of the primary teeth significantly more frequently than patients classified as non-odonto-type (100% vs. 56%; P<0.05). Tooth hypomineralisation was detected in 42% of non-odonto-type patients, but not in any odonto-type patients (0%; P<0.05). Collectively, these results suggest that genetic and dental manifestations of patients with odonto-type and non-odonto-type HPP are significantly different, and these differences should be considered during clinical treatment of patients with HPP., Competing Interests: The authors have declared that no competing interests exist.

Objectives: This study investigates vitamin D deficiency patterns in individuals from birth to the beginning of adolescence. Microscopic computed tomography (micro-CT) evaluation of interglobular dentine (IGD) in teeth provides information on the age of disease onset and the number of deficient periods per individual, which will increase our understanding of factors influencing vitamin D deficiency prevalence, including sociocultural practices and latitude., Materials and Methods: Beemster and Hattem, two Dutch 17th-19th century communities, yielded relatively high prevalences of rickets (15-24%) and residual rickets (15-24%). From the affected individuals, a subsample of 20 teeth were selected for micro-CT scanning. Thin sections were made of 17 teeth, consisting of 6 teeth with and 11 teeth without observable IGD on micro-CT that were included for method comparison., Results: About 19 out of 29 (65.5%) individuals (one tooth was deemed unobservable) presented with IGD on micro-CT. Eight of the 11 (72.7%) individuals without IGD on micro-CT demonstrated histologically visible IGD. In 40.7% (11/27) of the affected individuals (combined micro-CT and histology results), vitamin D deficiency was recurrent, and in four individuals, some episodes occurred at approximately annual intervals suggesting vitamin D deficiency was seasonal. In three individuals, IGD occurred in the dentine formed around birth, suggesting maternal vitamin D deficiency., Discussion: Micro-CT analysis of IGD is found to be a valuable non-destructive method that can improve our understanding of the influence of sociocultural practices and latitude on disease development within age and sex groups in past communities., (© 2019 The Authors. American Journal of Physical Anthropology published by Wiley Periodicals, Inc.)

Purpose: Early diagnosis of tooth enamel demineralization, and dentin caries lesions, present a valuable solution to avoid or decrease their deleterious effect. The aim of this study was to design a simple, effective, and non-invasive technique, employing a novel laser wavelength to classify and differentiate between various tooth abnormalities in-vitro, by estimating wavelengths, showing distinctive appearance for each tooth class., Methods: This study implies a fluorescence hyperspectral imaging system employing a 395-nm laser diode source, irradiating a pre-diagnosed 12 molars and premolars teeth. The obtained reconstructed images were displayed and processed by HSAnalysis2XL, accompanied by a custom made digital, and image signal processing algorithms, revealing the exact wavelengths, characterizing the fluorescence of each tooth pre-diagnosed class., Results: The proposed hyperspectral imaging system was able to discriminate between normal, and abnormal dental classes for the entire specimens. Furthermore, a series of wavelengths, noting each lesion individually were obtained from the spectroscopic hyperspectral output. The root calculus, white spot, dentin caries, and enamel caries have a bright visual appearance at λ 3 = 702 nm, λ 5 = 771 nm, and λ 6 = 798 nm respectively. Consequently, these abnormalities exhibit a dark appearance at λ 1 = 421 nm, λ 2 = 462 nm, and λ 4 = 734 nm. The wavelength selections were confirmed by the grayscale image outcomes., Conclusions: This study provides a set of wavelengths that can be employed by dentists to diagnose white spot, root calculus, and enamel dentin caries lesions under the irradiation of a new UV-vis laser illumination source without, any hazardous thermal or mechanical effects., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Dental caries is associated with plaque dysbiosis, leading to an increase in the proportions of acidogenic and aciduric bacteria at the expense of alkali-generating commensal species. Stannous fluoride (SnF 2 ) slows the progression of caries by remineralization of early lesions but has also been suggested to inhibit glycolysis of aciduric bacteria. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) promotes fluoride remineralization by acting as a salivary biomimetic that releases bioavailable calcium and phosphate ions, and the peptide complex has also been suggested to modify plaque composition. We developed a polymicrobial biofilm model of caries using 6 bacterial species representative of supragingival plaque that were cultured on sound human enamel and pulsed with sucrose 4 times a day to produce a high cariogenic challenge. We used this model to explore the mechanisms of action of SnF 2 and CPP-ACP. Bacterial species in the biofilms were enumerated with 16S rRNA gene sequence analyses, and mineral loss and lesion formation were determined in the enamel directly under the polymicrobial biofilms via transverse microradiography. The model tested the twice-daily addition of SnF 2 , CPP-ACP, or both. SnF 2 treatment reduced demineralization by 50% and had a slight effect on the composition of the polymicrobial biofilm. CPP-ACP treatment caused a similar inhibition of enamel demineralization (50%), a decrease in Actinomyces naeslundii and Lactobacillus casei abundance, and an increase in Streptococcus sanguinis and Fusobacterium nucleatum abundance in the polymicrobial biofilm. A combination of SnF 2 and CPP-ACP resulted in a greater suppression of the acidogenic and aciduric bacteria and a significant 72% inhibition of enamel demineralization.

Aim: Silver diammine fluoride (SDF) is an anticaries agent that binds to tooth tissue. The aim of the present study was investigate the dose-response effect of SDF on demineralized dentine in basal medium mucin (BMM) saliva substitute and human saliva., Methods: Dentine discs stored in saliva substitute, BMM, and human unstimulated whole saliva (UWS) were chemically demineralized, and 10 μL of 10 wt%, 24 wt%, or 38 wt% SDF was applied and then stored in its respective saliva (BMM or UWS) for 5 days. Dentine disc samples were digested in 70% HNO 3 , and silver was quantitatively detected by inductively coupled plasma mass spectroscopy., Results: For both the BMM and UWS groups, the presence of silver increased proportionally with increasing concentrations of SDF (P < 0.05). For the 38 wt% SDF application, the mean absorption percentage of silver in dentine was 3.90% for the UWS group and 6.61% for the BMM group. The BMM group was found to yield a significantly higher amount of silver compared to UWS at 10 wt% and 38 wt% SDF application (α = 0.05); 38 wt% SDF interacts more with BMM with higher silver deposition than UWS., Conclusion: The amount of silver found in dentine was in proportion to the concentration of SDF. Extrapolation of in vitro investigations using artificial saliva for SDF tooth interactions should be interpreted with caution., (© 2018 John Wiley & Sons Australia, Ltd.)

Objectives: This prospective longitudinal study aimed to evaluate if the occurrence of post-eruptive breakdown of demarcated opacities in hypomineralized teeth is influenced by the color or location of the opacity., Materials and Methods: Patients diagnosed with molar-incisor hypomineralization (MIH) between 2012 and 2014 were eligible. Two calibrated examiners performed the initial and follow-up evaluations according to European Academy of Paediatric Dentistry (EAPD) criteria. Sixty-five patients were included. Fifty-eight (89.2%), with a mean age of 8.8 years (SD: 1.4), were reassessed after 1 year. Two hundred and nine of 1155 tooth surfaces were considered for the study: 86 with white opacity (OP-W), 91 with yellow opacity (OP-Y), and 32 with enamel breakdown (EB)., Results: From the OP-W, OP-Y, and EB, 14, 27.5, and 46.9% worsened to breakdown exposing dentin, atypical restoration, or extraction (DB + RA or EXT), respectively. Yellow opacities tended to be more prone to breakdown than white opacities. The occurrence of EB, DB + AR, or EXT was not influenced by the location (p = 0.25)., Conclusions: The color of the opacity seems to play an important role on the occurrence of fracture and should be considered as a potential predictor., Clinical Relevance: Dentists should be aware that demarcated opacities related to MIH tend to fracture over time. Moreover, children with MIH should be seen at shorter intervals.

As laboratory models are bridges to in vivo caries studies, they must mirror clinical conditions, where demineralization and remineralization alternate constantly (i.e., pH cycling) and are only interrupted during the very short period of application of investigational products, such as toothpaste or mouth rinse. In view of this, models have been developed, based on pH cycling, to study the anticaries or caries remineralizing effects of substances. The pH cycling models have long been accepted and utilized by the scientific community and the toothpaste industry as an appropriate alternative to animal caries testing, particularly for ionic fluoride-based dentifrices. Several pH cycling models have been developed and described in the literature over the years. However, in this chapter, we crudely categorize them into two types: according to what the investigational product is tailored to achieve, i.e., prevention of caries development (net demineralization) or remineralization of early caries (net remineralization). Thus the models are termed "demineralization" or "remineralization" models and are described in details here together with their disadvantages and applications.

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The purpose of this study was to evaluate the bonding interface between a self-adhesive resin cement to in vitro eroded dentin. Seventy-two third molars were used and divided into two groups: sound dentin and in vitro eroded dentin. The in vitro erosion was performed following a demineralization protocol, in which the specimens were immersed in a demineralizing solution for 2 minutes per cycle and remineralizing solution for 10 minutes per cycle for 9 days. Both groups were submitted to four dentin surface treatments: control group (without any treatment), 2% chlorhexidine, 20% polyacrylic acid, and 0.1 M EDTA (n = 9). Blocks of resin-based composite were bonded with RelyX U200 self-adhesive resin cement applied on the pretreated dentin surfaces. The teeth were sectioned into beams (1mm2) and submitted to microtensile bond strength testing to evaluate the bond strength of self-adhesive resin cement to dentin after 24 hours and 8 months of immersion in artificial saliva. Three specimens of each group were longitudinally cut and evaluated using confocal laser scanning microscopy to analyze the dentin/cement interface. Eroded dentin showed higher bond strength values when compared to sound dentin for the 2% chlorhexidine group (p = 0.03), 24 hours after adhesion. When considering eroded dentin, the 0.1M EDTA group showed higher bond strength values with a statistically significant difference only for the control group (p = 0.002). After 8 months of storage, the present results showed that there was no statistically significant difference between the two substrates for all experimental groups (p>0.05). Analysis of the microscopy confocal showed different types of treatments performed on dentin generally increased tags formation when compared to the control group. The eroded dentin showed a significant increase in density and depth of resinous tags when compared to sound dentin. The storage of samples for 8 months seems to have not caused significant degradation of the adhesive interface., Competing Interests: The authors have declared that no competing interests exist.

Objective: To assess Scanning Electron Microscopy in Back-Scattered Emission mode (BSE-SEM) for measurement of lesion mineral content as a function of depth. Direct comparison is made with Transverse Micro-Radiography (TMR) and Surface Micro-Hardness (SMH) on carious and erosive lesions., Design: Caries lesions prepared from sound bovine enamel at 37 °C and pH 4.6 in unsaturated (7d) or part-saturated (8d, 4.1 mM Ca 2+ , 8 mM Pi) lactic acid /methyl cellulose gel system, followed by TMR analysis. Erosive lesions prepared from sound bovine enamel (1% citric acid, pH3.8, room temperature) for 5, 10, 15 or 20 min at n = 10 per treatment group. SMH readings (Vickers diamond, 1.9 N, 20 s) were taken from acid-treated and reference areas of each sample. BSE-SEM performed on polished cross-sections of lesioned samples (Jeol JSM6490LV SEM; high vacuum, 10 keV beam voltage, magnification x500 with constant working distance of 10 mm). Under identical SEM conditions, polished standards i.e. MgF 2 , alumina, Mg, Al and Si provided a calibration plot of BSE-SEM signal vs. atomic number (z¯). Mineral content vs. depth plots were derived from the cross-sectional BSE-SEM data., Results: Cross-sectional BSE-SEM images clearly differentiate between caries and erosive lesions. Comparison of caries lesion mineral loss from BSE-SEM with TMR data showed good correlation (R 2  = 0.98). Similarly, comparison of BSE-SEM data from erosive lesions showed good correlation (R 2  = 0.99) with hardness loss data from SMH., Conclusion: BSE-SEM provides a relatively rapid and cost-effective method for the assessment of mineral content in demineralised tooth enamel and is applicable to both caries and erosive lesions., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Using micro-computed tomography (micro-CT), this study assessed the inhibitory effect of three different types of restorations on root-caries formation under a cariogenic challenge. Bovine-root dentin blocks with a cylindrical cavity were divided into three restoration groups: a fluoride-free self-etch adhesive and a resin composite (SE-ES); a self-etch adhesive and a resin composite with multi-ion release (FL-BF); and a glass ionomer cement (Fuji-VII). After the restorative procedures, the specimens were stored in artificial saliva for 1 d, then subjected to a demineralization solution for 4 d and a remineralization solution for 28 d. Mineral density and mean mineral loss of dentin around the restorations were measured using micro-CT. The mean mineral loss values of dentin around the restorations were highest among SE-ES restorations and lowest among Fuji-VII restorations. Fuji-VII showed the highest mineral density and the lowest mineral loss after 28 d of remineralization. The observations made by scanning electron microscopy demonstrated that Fuji-VII created the smallest outer lesion followed by FL-BF and SE-ES. Fuji-VII has a larger inhibitory effect on root caries around the restorations and enhances remineralization more effectively than either FL-BF or SE-ES. Multi-ion and fluoride release from the restorative materials may be beneficial for inhibition of root-dentin caries around the restorations., (© 2018 Eur J Oral Sci.)

This study investigated the effect of SnCl 2 /AmF pretreatment on short- and long-term bond strength of resin composite to eroded dentin mediated by two self-etch, MDP-containing adhesive systems. 184 dentin specimens were produced from extracted human molars. Half the specimens ( n = 92) were artificially eroded, and half were left untreated. For both substrates, half the specimens were pretreated with SnCl 2 /AmF, and half were left untreated. The specimens were treated with Clearfil SE Bond or Scotchbond Universal prior to application of resin composite. Microtensile bond strength ( μ TBS) was measured after 24 h or 1 year. Failure mode was detected and EDX was performed. μ TBS results were statistically analyzed ( α = 0.05). μ TBS was significantly influenced by the dentin substrate (eroded < noneroded dentin) and storage time (24 h > 1 year; p < 0.0001) but not by pretreatment with SnCl 2 /AmF or adhesive system. The predominant failure mode was adhesive failure at the dentin-adhesive interface. The content of Sn was generally below detection limit. Pretreatment with SnCl 2 /AmF did not influence short- and long-term bond strength to eroded dentin. Bond strength was reduced after storage for one year, was lower to eroded dentin than to noneroded dentin, and was similar for the two adhesive systems.

Aim: The aim of the present study was to compare the effectiveness and practicality of two commonly-used protocols for white spot lesion creation., Methods: Sound posterior human teeth were sectioned into halves and randomly allocated into two groups (n=20). Using the FluoreCam system, the enamel surfaces were imaged under standardized conditions, and baseline data (area, intensity, and impact) were recorded. Specimens were covered with an acid-resistant polish, leaving a 2×6-mm enamel window. Group 1 was immersed in Queiroz solution (64 hours/37°C/pH5 with agitation); Group 2 was immersed in Buskes solution (14 days/37°C/pH5 with agitation). The specimens then were imaged again. Within-group changes that occurred over time were evaluated using the Wilcoxon signed-rank test. Between-group differences were analyzed using the Mann-Whitney U-test. Polarized light microscopy (PLM) was used to calculate the mean lesion depth of representative specimens., Results: Each protocol showed significant changes in the surface area, intensity and impact of demineralization over time (P<.05). There were no statistically-significant differences (P=.53, P=.2, P=.74) between the two groups. PLM showed that the mean lesion depths of representative specimens were 51.4 and 73.0 μm for groups 1 and 2, respectively., Conclusions: While both protocols produce similar amounts of demineralization, the Queiroz solution is more practical, as it requires less time., (© 2017 John Wiley & Sons Australia, Ltd.)

Objectives: The immediate aim of this study was to develop and test a method to record, visualize and quantify real-time demineralization (or remineralization) of teeth in vitro using X-ray microtomography (XMT or micro-CT). The longer term objective is to improve understanding of demineralization processes, allowing the creation of better artificial models of dental caries and better quantification of the efficacy of caries treatment and prevention regimes., Methods: For demineralization studies, the tooth is mounted in a plastic container through which the demineralizing solution is circulated during simultaneous repeated scanning. Key features of the method are safe circulation of the demineralizing solution, periodic interruption to adjust X-ray filament current and re-focus, accurate beam-hardening correction and calibration, alignment of reconstructed scans, and normalization of grey-levels to compensate for changes in the X-ray spectrum. The method was tested by scanning an extracted third molar during 8days of demineralization., Results: From the reconstructed 3D images, the progression of an artificial carious lesion could be visualized and quantified. The lesion progressed at approximately 0.1mm per day and appeared to be more erosive in nature., Conclusions: A successful method has been developed to monitor real-time demineralization in 3 dimensions. Further work is now needed to create better models of true carious lesions., Clinical Significance: Observation of the dynamics of demineralization and remineralization will aid in the development of therapies to treat and prevent dental caries., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objective: Ultraviolet (UV) and infrared (IR) lasers can be used to specifically target protein, water, and mineral, respectively, in dental hard tissues to produce varying changes in surface morphology, permeability, reflectivity, and acid resistance. The purpose of this study was to explore the influence of laser irradiation and topical fluoride application on the surface morphology, permeability, reflectivity, and acid resistance of enamel and dentin to shed light on the mechanism of interaction and develop more effective treatments., Methods: Twelve bovine enamel surfaces and twelve bovine dentin surfaces were irradiated with various combinations of lasers operating at 0.355 (Freq.-tripled Nd:YAG (UV) laser), 2.94 (Er:YAG laser), and 9.4 μm (CO 2 laser), and surfaces were exposed to an acidulated phosphate fluoride gel and an acid challenge. Changes in the surface morphology, acid resistance, and permeability were measured using digital microscopy, polarized light microscopy, near-IR reflectance, fluorescence, polarization sensitive-optical coherence tomography (PS-OCT), and surface dehydration rate measurements., Results: Different laser treatments dramatically influenced the surface morphology and permeability of both enamel and dentin. CO 2 laser irradiation melted tooth surfaces. Er:YAG and UV lasers, while not melting tooth surfaces, showed markedly different surface roughness. Er:YAG irradiation led to significantly rougher enamel and dentin surfaces and led to higher permeability. There were significant differences in acid resistance among the various treatment groups., Conclusion: Surface dehydration measurements showed significant changes in permeability after laser treatments, application of fluoride and after exposure to demineralization. CO 2 laser irradiation was most effective in inhibiting demineralization on enamel while topical fluoride was most effective for dentin surfaces. Lasers Surg. Med. 49:913-927, 2017. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

Background: The decalcification of enamel leads to an increase in tissue roughness. The development of carious lesions may be halted or these lesions may heal, which results in a smoother surface., Objectives: The objective of the study was to assess the impact of selected agents on the roughness of demineralized enamel on the smooth surfaces of teeth., Material and Methods: A total of 25 samples with exposed enamel surfaces were examined in the study. The samples were randomized into groups and they were exposed to a demineralizing solution, after which the following agents were applied: Icon (Ic), Elmex gel (El), GC Tooth Mousse (TM), Colgate Total (K+) - a positive control group, and distilled water as a negative control group (K-). The impact of selected agents on the roughness of demineralized enamel was assessed with a profilometer. The parameters Ra, Rz and Rq were analyzed., Results: Following infiltration and demineralization, the Ra coefficient declined in groups Ic, TM, K+ and K-, Rz decreased in groups Ic, TM and Rq was lower in groups Ic, TM and Kafter using agents when compared with demineralization. The lowest Ra, Rz and Rq values, i.e. the highest degree of surface smoothness, were observed in group Ic. An increase in all coefficients (Ra, Rz and Rq) was noted in group El., Conclusions: Demineralized enamel can be smoothed out through the one-time application of Icon resin or the long-term application of GC Tooth Mousse. The least effective agents turned out to be those with a high fluoride content.

Objective: Fluoride excess of 0.05-0.07mgF/kgbw/day in water or food additives like salt is the principal cause of endemic dental fluorosis. How fluoride causes these defects is not clear yet. Recent studies in rodents suggest that development of enamel fluorosis is associated with insufficient neutralization of protons released during the formation of hypermineralized lines., Design: Here we examined whether hypermineralization could also be assessed by MicroCT in developing molar enamel of humans exposed to fluoride., Result: Micro-CT analysis of hypomineralized enamel from human fluorotic molars graded by the Thylstrup-Fejerskov (TF) Index as III-IV showed weak hypermineralized lines and hypermineralized patches not seen in TF-I/II grade enamel. The mesio-distal sides of these molar teeth were significantly smaller (∼18%, p=0.02) than in TF-I/II teeth., Conclusion: The patterns of changes observed in human fluorotic teeth were similar to those in fluorotic rodent incisors. The data are consistent with the hypothesis that also in developing human teeth fluoride-stimulated local acidification of enamel could be a mechanism for developing fluorotic enamel., (Copyright © 2017. Published by Elsevier Ltd.)

The progress of caries has conventionally been evaluated by checking changes in mineral density using transverse microradiography (TMR). Recent advances have seen development of a new measurement system, using in-air micro proton induced X-ray/gamma-ray emission (PIXE/PIGE). PIXE/PIGE enables analysis of distributions and concentrations of multiple mineral elements in a carious lesion. The aim of this study was to evaluate the effectiveness of PIXE/PIGE for investigating the development of root caries. In summary, we successfully established a multi-elemental sequential measuring method using in-air micro-PIXE/PIGE to identify the dynamic distributions and concentrations of Ca and F in human root dentin. The PIXE/PIGE potentially offers a useful advantageous technique for studying carious development by using as a combination with conventional techniques such as TMR and Micro-computed tomography (µCT).

Background: Resin infiltration may alter the mechanical properties of enamel hypomineralised lesions (HL); however, variable surface layer (SL) thickness may affect resin penetration., Aims: To determine the thickness of the SL of HL and to investigate the effect of resin infiltration on the mechanical properties of HL., Design: The thickness of the SL over HL was determined using polarised light microscopy. Etching time using 15% HCl gel to remove the SL of 52 samples was determined. Selected HL and control areas of 21 teeth were infiltrated with Icon ® resin infiltrant, and cross-sectional Knoop microhardness (KHN) measurements were recorded before and after resin infiltration., Results: Ninety-six point five per cent of HL had a detectable SL with mean thickness of 58 ± 29 μm. Application of HCl for 120 s produced a mean MIH erosion depth of 58 ± 12 μm. Eleven of 21 samples had evidence of infiltration using visual examination. The infiltrant penetrated some of the HL leading to an increase in KHN (111 ± 75 KHN) which, when compared to adjacent non-infiltrated HL (96 ± 52 KHN), was not statistically significantly different (P = 0.56)., Conclusions: There was marked variation in the SL thickness of HL. Resin infiltration of HL did not increase microhardness significantly., (© 2016 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

A white spot lesion is the first clinical sign of a caries lesion and represents mineral loss from the enamel subsurface. The purpose of this study was to evaluate the microhardness and surface roughness of white spot lesions after application of a resin infiltrant and subjection to different challenges. Caries-like lesions were induced in bovine enamel discs (n=50), and the specimens were randomly divided into five study groups (n=10): demineralized enamel (negative control, G1), infiltrated enamel (G2), infiltrated enamel submitted to brushing (G3), infiltrated enamel submitted to pH cycling (G4), and infiltrated enamel submitted to artificial aging (G5). Half of each enamel surface was used as its own positive control. Roughness data were analyzed using the Kruskal-Wallis test followed by the Dunn test. Results from microhardness were analyzed by two-way analysis of variance, followed by the Tukey test for multiple comparisons. The level of significance was set at 5%. Microhardness and roughness values obtained from the test side of the specimens were significantly lower compared with the sound enamel for all groups. Microhardness values obtained for G2, G3, and G5 were not significantly different. Values found for G1 were significantly lower compared with those for G2, G3, and G5. The lowest microhardness values were observed for G4, which was significantly different from the other groups. Surface roughness was not significantly different between G2 and G3. The resin infiltrant presented superiority over the unprotected white spot lesions, as they were more resistant to mechanical and aging challenges. However, resin infiltration was not able to reestablish the properties of sound enamel and was not resistant to a new cariogenic challenge.

Molar Incisor Hypomineralisation (MIH) and Hypomineralised Second Primary Molars (HSPM) involve prevalent qualitative structural developmental anomalies of tooth enamel affecting the first permanent molars (and often incisors) and the second primary molars, respectively. These demarcated hypomineralised lesions of enamel manifest as white-cream or yellow-brown opacities, with possible post-eruptive localised loss of enamel. Aetiological hypotheses have involved contemporary life factors (i.e. environmental pollutant exposure or early childhood medications) in contrast to factors not limited to a specific time period (i.e. hypoxia at birth or genetic predisposition). Evidence of MIH in ancient populations would reinforce aetiological factors present for many centuries. By means of microtomographic and X-ray fluorescence analyses the present study provides evidence that (i) two archaeological specimens: "S407" (Sains-en-Gohelle, France, 12 th -16 th centuries) and "B335" (Beauvais, France, 15 th -18 th centuries) were MIH-affected, and (ii) one individual "S323" was affected by HSPM and MIH (Sains-en-Gohelle, France, 7 th -11 th centuries).

Introduction: Radiographic methods have poor sensitivity for occlusal lesions and by the time the lesions are radiolucent they have typically progressed deep into the dentin. New more sensitive imaging methods are needed to detect occlusal lesions. In this study, cross-polarization optical coherence tomography (CP-OCT) and near-IR imaging were used to image questionable occlusal lesions (QOC's) that were not visible on radiographs but had been scheduled for restoration on 30 test subjects., Methods: Near-IR reflectance and transillumination probes incorporating a high definition InGaAs camera and near-IR broadband light sources were used to acquire images of the lesions before restoration. The reflectance probe utilized cross-polarization and operated at wavelengths from 1,500 to 1,700 nm where there is an increase in water absorption for higher contrast. The transillumination probe was operated at 1,300 nm where the transparency of enamel is highest. Tomographic images (6 × 6 × 7 mm 3 ) of the lesions were acquired using a high-speed swept-source CP-OCT system operating at 1,300 nm before and after removal of the suspected lesion., Results: Near-IR reflectance imaging at 1,500-1,700 nm yielded significantly higher contrast (P < 0.05) of the demineralization in the occlusal grooves compared with visible reflectance imaging. Stains in the occlusal grooves greatly reduced the lesion contrast in the visible range yielding negative values. Only half of the 26 lesions analyzed showed the characteristic surface demineralization and increased reflectivity below the dentinal-enamel junction (DEJ) in 3D OCT images indicative of penetration of the lesion into the dentin., Conclusion: This study demonstrates that near-IR imaging methods have great potential for improving the early diagnosis of occlusal lesions. Lasers Surg. Med. 49:215-224, 2017. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

Objective: Molar Incisor Hypomineralisation (MIH) is a structural anomaly that affects the quality of tooth enamel and has important consequences for oral health. The developmentally hypomineralised enamel has normal thickness and can range in colour from white to yellow or brown with or without surface breakdown. The possibility of finding MIH in 'ancient populations' could downplay several current aetiological hypotheses (e.g., dioxin derivatives, bisphenols, antibiotics) without excluding the possible multifactorial aspect of the anomaly. In an archaeological context, chemical elements contained in the burial ground can stain teeth yellow or brown and therefore might create a taphonomic bias. The purpose of the present study is to test a proposed diagnostic guide enabling determination of the pathological or taphonomic cause of enamel discolouration and defects that resemble MIH present on 'ancient teeth'., Design: Two sample groups including MIH discoloration (n=12 teeth) from living patients, taphonomic discoloration (n=9 teeth) and unknown discoloration (n=2 teeth) from medieval specimens were tested. Three non-destructive methods-Raman spectroscopy, X-ray micro-computed tomography and X-ray fluorescence were utilised., Results: Hypomineralised enamel has decreased mineral density (p<0.0001) and increased phosphate/β-carbonate ratio (p<0.01) compared to normal enamel whereas relative concentrations of manganese, copper, iron and lead are similar. In taphonomic discoloration, relative concentrations of these elements are significantly different (p<0.05) to normal enamel whereas mineral density and Raman spectra profile are comparable., Conclusions: Enamel hypomineralisation can be distinguished from taphonomic staining in archaeological teeth., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Objective: To report (1) the caries experience prevalence and mean, and the caries severity and distribution patterns, expressed clinically and combined with radiographs with the conventional and ICCMS™ systems in young children from Bogotá, Colombia; (2) the contribution of including radiographs to the clinical caries scoring and (3) in which surfaces the radiograph adds to the clinical caries registration., Material and Methods: Six hundred children from kindergartens/schools were enrolled: Cohort A: 2-year (n = 200), Cohort B: 4-year (n = 200) and Cohort C: 6-year (n = 200) olds. Radiographs were taken of the 4- and 6- year olds. Children were examined clinically using the Clinical (C) and Radiographic (R) ICCMS™-epi Caries Scoring Systems, staging caries lesions (d) as: Initial (C epi /R A ), Moderate (C M /R B ) or Extensive (C E /R C ). Caries experience including missing (m) and filled (f) surfaces was expressed as follows: clinical conventional (Cd ME mfs); clinical ICCMS™ (Cd epiME mfs); combined conventional (C + Rd ME mfs) and combined ICCMS™ (C + Rd epiME mfs)., Results: The prevalence of Cd ME mfs was: Cohort A: 32%; Cohort B: 59%; Cohort C: 67.5%, increasing to 73.5%, 99.8% and 100%, respectively, with the C + R d epiME mfs. The Cd ME mfs means doubled when initial caries lesions (Cd epi ) and radiographs (R) were included. The d component corresponded to over two-thirds of the caries experience. Findings on the radiographs significantly raised caries experience prevalence and means (p < .02), detecting primarily approximal lesions. Surfaces with highest caries frequency were occlusal/approximal of molar teeth and buccal of upper incisor teeth., Conclusion: Participants' caries experience was high. The radiographic assessment significantly contributed to caries experience. Molar and upper incisor teeth were most prone to caries.

The article presents the results of spectrophotometric tooth enamel scanning. Research was conducted in vitro on extracted teeth. Numeric color parameters measurements were made by VITA EasyShade spectrophotometer after the 37% phosphoric acid enamel conditioning, that was made for an artificial tooth enamel demineralization reconstruction. Then transverse section of this enamel part was evaluated by scanning electronic microscopy. The scale of enamel demineralization depth in mm/microns and its' lightness parameter compliance was created.

Objective: To investigate the relationship between degree of dentin demineralization with both lesion activity and morphology of the occlusal carious cavity., Design: Occlusal sites (n=138) were identified by visual examination (Nyvad's scores 0-6) in 67 extracted teeth which were scanned in a high energy micro-CT. After 3D reconstruction, each stack was resliced in the mesio-distal direction and tooth mineral density (MD) was measured along a path from enamel to the deepest part of dentin in the slice showing the most severe carious involvement. Each site was classified in "open" or "closed" (if cavitated) depending on the morphology of the surrounding enamel walls as measured using micro-CT and as active or inactive in enamel or dentin by a clinical scoring system., Results: Lesions showing dentin cavitation presented higher demineralization degree compared to non-cavitated, or enamel cavitated lesions. Inactive lesions presented lower demineralization degree compared to active lesions, although with a low effect size. According to the morphological aspect of the carious cavity, open enamel lesions showed lower dentin demineralization degree than closed lesion environments., Conclusion: Active lesions showed higher dentin demineralization degree than inactive ones, while lesions showing closed cavitation resulted in higher dentin demineralization degree only for enamel lesions. Including those parameters in treatment decisions may help to improve prognosis and increase effectiveness of the caries diagnostic systems in the clinical setting., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Objective: With respect to the unknown aetiology of molar incisor hypomineralisation (MIH), it is unclear whether this phenomenon was overlooked in the last century as a result of a high number of caries in children or if this developmental disorder was not present until then. Therefore, this study determined the presence of MIH in historical dentitions and teeth., Materials and Methods: Dental remains from late medieval (n = 191, twelfth-sixteenth century, Regensburg, Germany), post-medieval (n = 33, sixteenth-eighteenth century, Passau, Germany) and modern age archaeological skeletal series (n = 99, nineteenth-twentieth century, Altdorf, Germany) were examined for MIH. In addition, linear enamel hypoplasia (LEH), diffuse opacities, hypoplasia and Turner's teeth were documented., Results: MIH-related demarcated opacities or enamel breakdowns were found in only 15 (0.4 %) of the 3891 examined permanent teeth. Ten cases (3.1 %) from a total of 323 dentitions were classified as having MIH. In contrast, 98 individuals (30.3 %) showed LEH. Other enamel disorders were recorded in 64 individuals (19.8 %)., Conclusion: With respect to the low number of affected dentitions and teeth, MIH most likely did not exist or was at least rarely present in the investigated archaeological case series., Clinical Relevance: This study supports the hypothesis that MIH may be linked to contemporary living conditions or other health-related factors.

Purpose: To evaluate the bonding behavior of resin composite and different adhesives applied to demineralized or remineralized enamel., Methods: Bovine tooth crowns were polished to prepare a 5 mm2 enamel bonding area, and divided into five groups (n= 48) according to the surface treatment: CONT (sound enamel control), DEM (demineralized with acid to create white spot lesions), REMS (DEM remineralized with artificial saliva), REMF (DEM remineralized with sodium fluoride) and INF (DEM infiltrated with Icon resin infiltrant). The surface-treated teeth were divided into two subgroups (n= 24) according to adhesive type: ER (etch-and-rinse; Single Bond Universal) and SE (self-etching; Clearfill S3 Bond), and further subdivided into two categories (n= 12) according to aging process: Thermo (thermocycling) and NA (no aging). Composite blocks were made over bonded enamel and sectioned for microtensile bond strength (MTBS) testing. Data were analyzed with three-way ANOVA and post-hoc Tukey's test (α= 0.05)., Results: Significant differences were observed for enamel surface treatment (P< 0.0001), adhesive type (P< 0.0001) and aging (P< 0.0001). CONT and INF groups had higher MTBS than the other groups; Single Bond Universal had higher MTBS than Clearfil S3 Bond; thermo-aging resulted in lower MTBS irrespective of adhesive type and surface treatment condition. The predominant failure mode was mixed for all groups. Enamel surface infiltrated with Icon does not interfere with adhesive resin bonding procedures., Clinical Significance: Treatment of enamel surface containing white spot lesions or cavities with cavosurface margins in partially-demineralized enamel can benefit from infiltration with a low viscosity resin infiltrant prior to adhesive bonding of resin composites., Competing Interests: The authors declared no conflict of interest. Research supported by Fundação de Amparo a Pesquisa do Estado de São Paulo - FAPESP - Proc 2011/21917-4 - 2013/11745-7.

Background: The aim of the infiltration technique is to penetrate demineralized enamel with a low viscosity resin. Icon® (DMG) is the first ever and so far the only dental infiltrant. Bacteriostaticity is one of the properties that should be inherent in dental infiltrants, but Icon lacks this feature., Objectives: The aim of the preliminary study was to properly choose a dye which would allow us to assess the penetrating abilities of our own, experimental preparation with features of a dental infiltrant with bacteriostatic properties and to compare using an optical microscope the depth of infiltration of the designed experimental preparation with the infiltrant available on the market., Material and Methods: The preparation is supposed to infiltrate decalcified human enamel and be assessed with an optical microscope. Eosin, neutral fuchsine and methylene blue were added to experimental preparation with dental infiltrant features and to Icon® (DMG) in order to assess the depth of penetration of the experimental solution into the decalcified layers of enamel., Results: The experimental solution mixes well with eosin, neutral fuchsine, and methylene blue., Conclusions: During the preliminary study, the authors concluded that the experimental solution mixes well with methylene blue, neutral fuchsine, and eosin. An addition of eosin to a preparation which infiltrates inner, demineralized enamel layers, facilitates the assessment of such a preparation with an optical microscope. A designed experimental solution with the main ingredients, i.e., 2-hydroxyethyl methacrylate (HEMA) and tetraethylene glycol dimethacrylate (TEGDMA) with a ratio of 75% to 25% penetrates the demineralized (decalcified) inner parts of the enamel and polymerizes when exposed to light. In order to assess the infiltration of the experimental solution into the demineralized enamel layers, it is required to improve the measurement techniques that utilize optical microscopy.

Early detection of the most prevalent oral disease worldwide, i.e., dental caries, still remains as one of the major challenges in dentistry. The current dental standard of care relies on caries detection methods, such as visual inspection and x-ray radiography, which lack the sufficient specificity and sensitivity to detect caries at early stages of formation when they can be healed. We report on the feasibility of early caries detection in a clinically and commercially viable thermophotonic imaging system. The system incorporates intensity-modulated laser light along with a low-cost long-wavelength infrared (LWIR; 8 to 14???m) camera, providing diagnostic contrast based on the enhanced light absorption of early caries. The LWIR camera is highly suitable for integration into clinical platforms because of its low weight and cost. In addition, through theoretical modeling, we show that LWIR detection enhances the diagnostic contrast due to the minimal LWIR transmittance of enamel and suppression of the masking effect of the direct thermal Planck emission. Diagnostic performance of the system and its detection threshold are experimentally evaluated by monitoring the inception and progression of artificially induced occlusal and smooth surface caries. The results are suggestive of the suitability of the developed LWIR system for detecting early dental caries.

The purpose of this study is to show that optical clearing agents can be used to increase the visibility of deeply penetrating occlusal lesions that have reached the underlying dentin and spread laterally under the enamel. Previous studies have shown that high refractive index fluids can increase the contrast of caries lesions. Extracted teeth with natural occlusal lesions were imaged with optical coherence tomography (OCT) with and without the addition of a transparent vinyl polysiloxane impression material (VPS) currently used in vivo. The relative intensity of the reflectivity from the underlying lesion area for each sample was measured before and after application of the VPS. Lesion presence was confirmed with polarized light microscopy and microradiography. Application of VPS significantly increased (P < 0.0001) the integrated reflectivity of subsurface dentinal lesions. This study shows that optical clearing agents can be used to increase the optical penetration and the visibility of subsurface lesions and the dentinal–enamel junction under sound and demineralized enamel in OCT images.

Enamel demineralization adjacent to pit and fissure sealants leads to the formation of marginal caries, which can necessitate the replacement of existing sealants. Dental materials with bioactive glass, which releases ions that inhibit dental caries, have been studied. The purpose of this study was to evaluate the enamel surface adjacent to sealants containing 45S5 bioactive glass (BAG) under simulated microleakage between the material and the tooth in a cariogenic environment. Sealants containing 45S5BAG filler were prepared as follows: 0% 45S5BAG + 50.0% glass (BAG0 group), 12.5% 45S5BAG + 37.5% glass (BAG12.5 group), 25.0% 45S5BAG + 25.0% glass (BAG25.0 group), 37.5% 45S5BAG + 12.5% glass (BAG37.5 group), and 50.0% 45S5BAG + 0% glass (BAG50.0 group). A cured sealant disk was placed over a flat bovine enamel disk, separated by a 60-µm gap, and immersed in lactic acid solution (pH 4.0) at 37 °C for 15, 30, and 45 d. After the storage period, each enamel disk was separated from the cured sealant disk, and the enamel surface was examined with optical 3-dimensional surface profilometer, microhardness tester, and scanning electron microscopy. The results showed a significant increase in roughness and a decrease in microhardness of the enamel surface as the proportion of 45S5BAG decreased (P< 0.05). In the scanning electron microscopy images, enamel surfaces with BAG50.0 showed a smooth surface, similar to those in the control group with distilled water, even after prolonged acid storage. Additionally, an etched pattern was observed on the surface of the demineralized enamel with a decreasing proportion of 45S5BAG. Increasing the 45S5BAG filler contents of the sealants had a significant impact in preventing the demineralization of the enamel surface within microgaps between the material and the tooth when exposed to a cariogenic environment. Therefore, despite some marginal leakage, these novel sealants may be effective preventive dental materials for inhibiting secondary caries at the margins., (© International & American Associations for Dental Research 2016.)

Purpose: To compare two different enamel pretreatments and their effect on the efficiency of penetration of a one-component adhesive into natural carious lesions., Methods: Eight extracted human molars and premolars with non-cavitated interproximal lesions were selected. ICDAS code 1-2 was assessed by visual, microscopic, X-ray and Diagnocam record analysis. Samples were cut vertically across the demineralization to obtain two symmetrical lesions, (n=16). After isolating the cut surfaces with nail varnish, paired lesion halves' surfaces were pretreated with two different techniques: Group 1: surfaces were firstly abraded with fine diamond-coated metallic strips (Steelcarbo) and then etched with 37% H₃PO₄ acid (Omni-etch, 120 seconds); Group 2: lesion surfaces were etched with 15% HCl acid (Icon-etch, 120 seconds). All teeth were stained with rhodamine isothiocyanate (RITC) solution (12 hours) and subsequently stored in dry chamber (3 hours). All samples were penetrated with a one-component adhesive (Scotchbond Universal) for 180 seconds and coated with a thin layer of flowable composite (Tetric Flow). After light curing, un- encapsulated dye was bleached by immersion in 30% hydogen peroxide for 12 hours at 37°C. Remaining lesion pores were stained with sodium fluorescein solution. Thin cuts of the teeth were observed with confocal microscopy and computer image analysis was performed (ImageJ). The percentage of penetration (area of resin penetration/area of total demineralization x100) was calculated., Results: Pretreatment with fine aluminum oxide-coated metallic strip followed by 37% H₃PO₄ acid showed a larger infiltration area (51.7% ± 12.2) in almost all samples compared to pretreatment with 15% HCl acid alone (22.1% ± 13.2). Statistical analysis using t-test showed a significant difference between the two groups (P = 0.011).

Objectives: This study evaluates the effect of self-assembling peptide P11-4 (Curodont Repair, CDR) on bovine enamel remineralization by measuring changes in ultrasonic propagation velocity., Methods: Six specimens per group were prepared by sectioning bovine teeth into enamel blocks. These blocks were then immersed in lactic acid buffer solution (pH = 4.75) for 10 min twice a day and stored in artificial saliva. Other specimens were first treated with CDR, followed by a 10-min immersion in the lactic acid buffer solution twice a day, before storage in artificial saliva. The propagation time of longitudinal ultrasonic waves was measured using a pulser/receiver. Six specimens were used for each treatment protocol. The obtained data were statistically analyzed using ANOVA followed by Tukey's honestly significant difference tests (α = 0.05). Specimens were observed using laser scanning microscopy and scanning electron microscopy., Results: Sonic velocity was found to decrease with time for specimens stored in the demineralizing solution. On the other hand, increases in sonic velocity were found for specimens treated with CDR. These specimens also exhibited signs of mineral deposition., Conclusions: By measuring the ultrasonic propagation velocity, it can be concluded that CDR application has an ability to promote bovine enamel remineralization.