Microbial shift of oral biofilm associated with remineralization of root dentin lesions.

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.

Detection of artificial enamel caries-like lesions with a blue hydroxyapatite-binding porosity probe.

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.

Effect of whitening products on sound enamel and on artificial caries lesions during a cariogenic challenge.

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.

Bioinspired Anti-demineralization Enamel Coating for Orthodontics.

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.

The Effect of Various Fluoride Products on Dentine Lesions during pH-Cycling.

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.

A novel method quantifying caries following orthodontic treatment.

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.

Artificial Caries Lesion Characteristics after Secondary Demineralization with Theobromine-Containing Protocol.

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.

Effectiveness of a toothpaste and a serum containing calcium silicate on protecting the enamel after interproximal reduction against demineralization.

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.

Demineralization Inhibition by Two Calcium-releasing Restorative Materials.

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).

ORAL HYGIENE IN CHILDREN WITH DISTURBANCES IN TOOTH FORMATION WHO WERE AFFECTED AS A CONSEQUENCE OF THE ChNPP ACCIDENT. / STAN GIGIIeNY POROZhNYNY ROTA U DITEI Z PORUShENNIaMY FORMUVANNIa ZUBIV, POSTRAZhDALYKh VNASLIDOK AVARIÏ NA ChAES.

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 hygiene¼ criterion) and Silness-Loe index (1.65 ± 0.46; p < 0.001 -«poor oral hygiene¼ 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.

Demineralization of tooth enamel following radiation therapy; An in vitro microstructure and microhardness analysis.

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.

In vivo assessment of the corrosion of nickel-titanium orthodontic archwires by using scanning electron microscopy and atomic force microscopy.

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.

New missense variants in RELT causing hypomineralised amelogenesis imperfecta.

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.

Effect of fermented milk with kefir grains on the in vitro demineralization of bovine tooth enamel / Efecto de la leche fermentada con granos de kéfir sobre la desmineralización in vitro del esmalte dental bovino

The dental caries is a progressive destruction of the teeth tissue due to the disbalance in the normal molecule interactions between the enamel and the bio!lm, which alters the demineralization-remineralization process. Milk fermentation produces caseinphosphopeptides with proved remineralizing capacity of the enamel. The presence of these peptides in fermented milk with ke!r grains has been described. The purpose of this work was to evaluate in vitro the capacity of milk ke!r to prevent the demineralization of dental enamel. Bovine incisors (n=68, 17 per group) were treated for 72 h with different solutions: I: artificial saliva at pH 7.2 , II: demineralizing solution at pH 4.5, III: supernatant of kefir fermented milk at pH 4.5, IV: milk supernatant at pH 4.5. The effects of treatments were evaluated by the change in the weight of the specimens, calcium concentration in the solution and by scanning electron microscopy (SEM) of the enamel. Kefir milk supernatant prevented the demineralization process, that was evidenced by a change in weight and calcium concentration that were not different from group I, although the pH was 4.5. In contrast, group IV showed a decrease in weight and an increase in calcium concentration, compared with group I (one way ANOVA, p<0.05). Images of SEM agree with the values of weight and calcium concentration. These results indicate that kefir milk supernatant has a protective effect on enamel demineralization in vitro. (AU)

La caries dental es una patología debido a un desequilibrio en las interacciones moleculares normales entre el esmalte y la biopelícula, que altera el proceso de desmineralización remineralización. La fermentación de la leche produce fosfopéptidos de caseína con probada capacidad remineralizante del esmalte, y se ha descripto la presencia de estos péptidos en la leche fermentada con granos de kéfir. El propósito de este trabajo fue evaluar in vitro la capacidad del kéfir de leche para prevenir la desmineralización del esmalte dental. Sesenta y ocho incisivos bovinos (17 por grupo) fueron tratados durante 72 h con diferentes soluciones: I: saliva artificial, pH 7.2, II: solución desmineralizante, pH 4.5, III: sobrenadante de leche fermentada con kefir, pH 4.5, IV: sobrenadante de leche, pH 4.5. El proceso de desmineralización se evaluó mediante el cambio en el peso de las muestras, la concentración de calcio en la solución y microscopía electrónica de barrido (SEM) del esmalte. El sobrenadante de leche fermentada con kéfir impidió el proceso de desmineralización, que se evidenció por un cambio en el peso y la concentración de calcio que no discreparon del grupo I, a pesar de haber tenido un pH de 4.5. En contraste, el grupo IV mostró una disminución en el peso y un aumento en la concentración de calcio, en comparación con el grupo I (ANOVA a un criterio, p<0.05). Las imágenes SEM concuerdan con los cambios en el peso y la concentración de calcio en los grupos estudiados. Los datos obtenidos demuestran que el sobrenadante de la leche tratada con kéfir tiene un efecto protector sobre la desmineralización del esmalte in vitro, inducida por el pH ácido. (AU)

Japanese nationwide survey of hypophosphatasia reveals prominent differences in genetic and dental findings between odonto and non-odonto types.

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.

Micro-CT assessment of dental mineralization defects indicative of vitamin D deficiency in two 17th-19th century Dutch communities.

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.

Protocols to Study Dental Caries In Vitro: pH Cycling Models.

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.

Breakdown of demarcated opacities related to molar-incisor hypomineralization: a longitudinal study.

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.

Classification of dental diseases using hyperspectral imaging and laser induced fluorescence.

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 λ3 = 702 nm, λ5 = 771 nm, and λ6 = 798 nm respectively. Consequently, these abnormalities exhibit a dark appearance at λ1 = 421 nm, λ2 = 462 nm, and λ4 = 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.

CPP-ACP Promotes SnF2 Efficacy in a Polymicrobial Caries Model.

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 (SnF2) 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 SnF2 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 SnF2, CPP-ACP, or both. SnF2 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 SnF2 and CPP-ACP resulted in a greater suppression of the acidogenic and aciduric bacteria and a significant 72% inhibition of enamel demineralization.