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.

The damage and remineralization strategies of dental hard tissues following radiotherapy.

OBJECTIVES: This study pursued two main purposes. The first aim was to expound on the microscopic factors of radiation-related caries (RRC). Further, it aimed to compare the remineralization effect of different remineralizing agents on demineralized teeth after radiotherapy. METHODS: The enamel and dentin samples of bovine teeth were irradiated with different doses of radiation. After analysis of scanning electron microscope (SEM), X-Ray diffraction (XRD), and energy dispersive spectrometer (EDS), the samples irradiated with 50 Gy radiation were selected and divided into the demineralization group, the double distilled water (DDW) group, the Sodium fluoride (NaF) group, the Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) group, the NaF + CPP-ACP group, and the Titanium tetrafluoride (TiF4) group. After demineralization, remineralizing agents treatment, and remineralization, the samples were evaluated using SEM, atomic force microscope (AFM), EDS, and transverse microradiography (TMR). RESULTS: A radiation dose of 30 Gy was sufficient to cause damage to the dentinal tubules, but 70 Gy radiation had little effect on the microstructure of enamel. Additionally, the NaF + CPP-ACP group and the TiF4 group significantly promoted deposit formation, decreased surface roughness, and reduced mineral loss and lesion depth of demineralized enamel and dentin samples after radiation. CONCLUSIONS: Radiation causes more significant damage to dentin compared to enamel. NaF + CPP-ACP and TiF4 had a promising ability to promote remineralization of irradiated dental hard tissues. ADVANCES IN KNOWLEDGE: This in vitro study contributes to determining a safer radiation dose range for teeth and identifying the most effective remineralization approach for RRC.

In vitro Effect of Occlusal Loading on Cervical Wall Lesion Development in a Class II Composite Restoration.

The aim of this study was to determine the effect of simulated occlusal loading on wall lesion development in cervical gaps of class II composite restorations in vitro. Sixty-four extracted human molars received standardized (4.0 × 4.2 × 3.0 mm) box preparations. The teeth were randomly assigned to one of two restoration groups: restoration with a normal or a low E-modulus composite material (CLEARFIL AP-X: E-modulus 16.8 GPa or CLEARFIL MAJESTY ES Flow: E-modulus 6.6 GPa). A metal matrix was placed at the bottom of the box for each restoration, creating a cervical gap of about 100 µm wide. Samples were exposed to simulated caries lesion development in a lactic acid solution (pH 4.8) for 8 weeks in a Rub&Roll device. Half of the samples were subjected to 90 N cyclic loading. After demineralization, the teeth were sectioned. Wall lesion development was measured using microradiography (transversal wavelength-independent microradiography) in two different locations (location 1: 1,000 µm and location 2: 1,600 µm from the gap entrance) and recorded in lesion depth (LD) (µm) and mineral loss (µm × vol%). Linear regression modeling was used to estimate the effect of loading and material on wall lesion development. Mean wall LD in location 1 across all groups was 150.83 µm with a standard deviation (SD) of 61.83 µm. In location 2, mean overall wall LD was 102.98 µm with an SD of 64.92 µm. Linear regression showed no significant effect of either loading or material on wall lesion development. Occlusal loading had no significant effect on secondary caries lesion development in composite class II restoration in this in vitro study.

Dentin anti-demineralization potential of surface reaction-type pre-reacted glass-ionomer filler containing self-adhesive resin cement.

OBJECTIVE: To investigate the anti-demineralization potential of a newly developed surface reaction-type pre-reacted glass-ionomer (S-PRG) filler containing self-adhesive resin cement against acidic attacks on the dentin surface. MATERIALS AND METHODS: A total of 32 bovine teeth were used. Cavities were prepared on crown dentin slaps and filled with three self-adhesive resin cement: (1) S-PRG-based cement, (2) Si-based cement, and (3) RelyX cement. Specimens were then subjected to pH cycling for 28 days, and the depth of demineralization was assessed using swept-source optical coherence tomography (SS-OCT) after 7, 14, 21, and 28 days. Sixty-four root dentin blocks were divided into four groups and then subjected to a pH cycling procedure with the aforementioned three material blocks and one negative control. The mineral loss was observed using transverse microradiography (TMR), and the surface microhardness (SMH) test was conducted to investigate the mechanical properties of treated dentin surfaces. RESULTS: The depth of demineralization for the S-PRG-based cement was significantly lower than that of the Si-based cement after 7, 21, and 28 days. Conversely, the RelyX cement was not significantly different from the Si-based cement after 7, 14, and 21 days (p < 0.05). Regarding the TMR and SMH test, the S-PRG-based cement showed the least mineral loss with the highest resistance to acidic challenge. CONCLUSION: The S-PRG filler containing resin cement can reduce mineral loss and promote remineralization of dentin substrate and has the potential to preserve dentin integrity and resist acidic attack. Clinical significance Self-adhesive resin cement containing S-PRG fillers maintained the surface integrity of dentin after exposure to 28 days of acidic challenge with a significant anti-demineralization effect.

Do bleaching gels affect the stability of the masking and caries-arresting effects of caries infiltration-in vitro.

OBJECTIVES: The aim of this study was to evaluate the influence of different bleaching gels on the masking and caries-arresting effects of infiltrated and non-infiltrated stained artificial enamel caries lesions. MATERIALS AND METHODS: Bovine enamel specimens (n = 240) with each two sound areas (SI and SC) and each two lesions (DI and DC) were infiltrated (DI and SI), stained (1:1 red wine-coffee mixture,70 days), and randomly distributed in six groups to be bleached with the following materials: 6%HP (HP-6), 16%CP (CP-16), 35%HP (HP-35), 40%HP (HP-40), and no bleaching (NBl,NBl-NBr). Subsequently, specimens were pH-cycled (28 days, 6 × 60 min demineralization/day) and all groups except NBl-NBr were brushed with toothpaste slurry (1.100 ppm, 2×/day, 10 s). Differences in colorimetric values (ΔL, ΔE) and integrated mineral loss (ΔΔZ) between baseline, infiltration, staining, bleaching, and pH cycling were calculated using photographic and transversal microradiographic images. RESULTS: At baseline, significant visible color differences between DI and SC were observed (ΔEbaseline = 12.2; p < 0.001; ANCOVA). After infiltration, these differences decreased significantly (ΔEinfiltration = 3.8; p < 0.001). Staining decreased and bleaching increased ΔL values significantly (p ≤ 0.001). No significant difference in ΔΔE was observed between before staining and after bleaching (ΔEbleaching = 4.3; p = 0.308) and between the bleaching agents (p = 1.000; ANCOVA). pH-cycling did not affect colorimetric values (ΔEpH-cycling = 4.0; p = 1.000). For DI, no significant change in ΔZ during in vitro period was observed (p ≥ 0.063; paired t test). CONCLUSIONS: Under the conditions chosen, the tested materials could satisfactorily bleach infiltrated and non-infiltrated stained enamel. Furthermore, bleaching did not affect the caries-arresting effect of the infiltration. CLINICAL RELEVANCE: The present study indicates that bleaching is a viable way to satisfactorily recover the appearance of discolored sound enamel and infiltrated lesions.

Effects of Additional Use of Bioactive Glasses or a Hydroxyapatite Toothpaste on Remineralization of Artificial Lesions in vitro.

OBJECTIVES: This in vitro study aimed to evaluate and compare the effect of two different bioactive glasses, a hydroxyapatite-containing, fluoride-free toothpaste (HTP) and a fluoride toothpaste (FTP) on the remineralization behavior of initial caries lesions. MATERIALS AND METHODS: A total of 100 bovine enamel samples were randomly allocated to five groups of 20 samples each: NC = negative control group (artificial saliva); HTP = HTP group (Karex); FTP = FTP group (Elmex caries protection, 1,400 ppm); FTP + BGnano = FTP followed by Actimins bioactive glass; FTP + BGamorph = FTP followed by Schott bioactive glass. Radiographic documentation (advanced transversal microradiography; aTMR) was applied before and after all samples were exposed to a demineralizing gel for 10 days. Over a period of 28 days, samples were covered twice a day (every 12 h) with a toothpaste slurry of the respective test group or with artificial saliva in NC for 60 s and brushed with 15 brushing strokes. Samples in FTP + BGnano and FTP + BGamorph were additionally treated with the respective bioactive glass slurry for 30 s after brushing with the FTP. In the meantime, all samples were stored in artificial saliva. After 28 days, the structure of all samples was assessed again using aTMR and compared to the values measured after demineralization. The statistical evaluation of the integrated mineral loss was performed using Kruskal-Wallis test followed by a post hoc Conover test. RESULTS: The FTP revealed the significantly highest increase of mineral content while the HTP showed the significantly lowest remineralization. Compared to artificial saliva, the use of the HTP or the combined application of FTP followed by bioactive glasses (FTP + BGnano and FTP + BGamorph) showed no significant remineralization. CONCLUSION: Under remineralizing in vitro conditions, brushing with 1,400 ppm FTP induced significantly more remineralization compared to storage in artificial saliva. The additional administration of both bioactive glasses as well as the substitutional brushing with an HTP resulted in significantly less remineralization compared to brushing with 1,400 ppm FTP.

Effect of different acid etchants on the remineralization process of white-spot lesions: An in vitro study.

PURPOSE: To investigate the effect of acid etchants with different low concentrations on remineralization of white spot lesion (WSL). METHODS: WSL were prepared on buccal surfaces of 100 intact premolars using the methyl cellulose gel/lactic acid method. The samples were then placed in a remineralizing solution in addition to fluoride application twice daily for 5 minutes. The changes were quantified weekly using the Quantitative Light-induced Fluorescence (QLF) system. When changes in fluorescence radiance approached zero, each sample was etched with one of the following acids; 5% phosphoric acid, 10% phosphoric acid, 5% polyacrylic acid or 10% polyacrylic acid for 15 seconds, washed, dried, and placed again in the remineralizing solution. Two samples were randomly selected from each group for transverse microradiography (TMR) and scanning electron microscopy (SEM) analysis. RESULTS: The 10% polyacrylic acid group showed the most significant improvement in fluorescence gain over the second phase of remineralization. It also showed partial loss of surface minerals without affecting enamel thickness as the phosphoric acid did. Additionally, 10% polyacrylic acid created the largest number of pores and smallest in size when compared to phosphoric acid, thus enhancing remineralization more efficiently than phosphoric acid without compromising the enamel outermost layer. CLINICAL SIGNIFICANCE: The findings of this study may improve the remineralization of WSL from the bottom of the lesion instead of precipitation on the outermost layer of the lesion leaving a better quality of enamel. 10% polyacrylic acid enhanced remineralization more efficiently than phosphoric acid without compromising the enamel outermost layer.

Microvascularization of the human central and peripheral nervous system: A new microcomputed tomography method.

Microcomputed X-ray tomography (microCT), developed since the late 1990s, is a miniaturized version of the tomographs used daily in medical imaging. It produces vascular images that are different from those obtained by microradiography, in particular by facilitating the vision in space, thus understanding microvascularisation. The anatomical specimens, once treated with formalin, are injected with a mixture made of gelatin containing a contrast product (barium) and then analyzed by microCT. The acquisition times that can exceed 24hours and metal sheets used for X-ray filtering vary according to the sample. The projection images are reconstructed to produce 2D sections. These are combined for the reconstruction of 3D models using a volume rendering software. Four examples will allow the imaging of microvascularization: the inferior alveolar nerve, the cerebral cortex and pia-mother, brain stem, central gray nuclei (ganglia at the base of the brain). Small capillaries are highlighted using high-end software for reconstruction. Conventional software or freeware cause a considerable loss of information on small vessels that are not visualized. The VGStudio max high-end software allows the production of videos that are particularly useful for 3D exploration and teaching (four videos are provided with this article).

Comparison between micro-computed tomography and transverse microradiography of sound dentine treated with fluorides and demineralized by microcosm biofilm.

The study aimed to apply micro-computed tomography (micro-CT) and transverse microradiography (TMR) to measure dentine demineralization and to test the preventive effect of titanium tetrafluoride (TiF4 ) under microcosm biofilm. Sound dentine specimens from bovine root were treated for 6 h with: (i) 4.0% titanium tetrafluoride (TiF4 ) varnish [pH 1.0, 2.45% fluoride (F-); (ii) 5.42% sodium fluoride (NaF) varnish (pH 5.0, 2.45% F); (iii) 2% chlorhexidine (CHX) gel (pH 7.0); (iv) placebo varnish (pH 5.0); or (v) no agent (untreated). Dentine specimens were then exposed to human saliva mixed with McBain saliva for 8 h. Thereafter, McBain saliva containing 0.2% sucrose was applied daily, for 5 d, onto dentine specimens to stimulate formation of microcosm biofilm. Although a high correlation was found between the results of both methods regarding integrated mineral loss, the results of the methods did not show good agreement in Bland-Altman plots, with significant biases in calculations of lesion depth. Fluoride varnishes were able to reduce dentine demineralization (P < 0.05), while CHX failed to do so. Fluorides are still the best option to reduce dentine demineralization. Micro-CT may be used to measure dentine mineral loss, but not the lesion depth, for which TMR is superior.

Chlorhexidine, a Matrix Metalloproteinase Inhibitor and the Development of Secondary Caries Wall Lesions in a Microcosm Biofilm Model.

This study investigated the role of a matrix metalloproteinase (MMP) inhibitor (CHX 2%) in the development of secondary caries wall lesions in different interface conditions with small (run 1) and wider gaps (run 2). Dentin discs were restored and pretreated with or without CHX 2%. In run 1, interfaces were made with gaps of 30, 60, or 90 µm. Interfaces with composite placed directly onto the dentin were either bonded (Adper Single Bond 2) or not bonded. In run 2, interfaces were made with gaps of 100 µm, with or without adhesive on the composite side (CLEARFIL SE Bond). Interfaces were either bonded or not bonded, as in run 1. Microcosm biofilms were grown on dentin-composite samples for 14 days. Caries lesion outcomes were analyzed by transversal wavelength-independent microradiography at 3 locations: the outer surface, and the interface wall at a distance of 200 and 500 µm from the gap entrance. Linear regression analyses showed that pretreatment with MMP inhibitor did not influence progression of the wall lesion at any location (p ≥ 0.218). Interfaces with intentional gaps showed positive and significant effect on the wall lesion progression at 200 µm from the gap entrance (p ≤ 0.005). A small trend of increase in wall lesion development was observed at the 200-µm location when bonding was present on the composite side. In conclusion, the dentin pretreatment with CHX 2% was not able to slow down the development of secondary caries wall lesions in small and wide gaps in this biofilm model.

Bleaching of simulated stained-remineralized caries lesions in vitro.

OBJECTIVE: Non-invasive esthetic treatment options for stained arrested caries lesions have not been explored. This study aimed to develop laboratory models to create stained-remineralized caries-like lesions (s-RCLs) and to test the efficacy of bleaching on their esthetic treatment. MATERIALS AND METHODS: One hundred twelve enamel/dentin specimens were prepared from human molars, embedded, and had their color measured spectrophotometrically at baseline and after demineralization. They were randomly divided into four groups (n = 14) based on the staining/remineralization protocols for a total of 5 days: G1, no staining/no remineralization; G2, no staining/remineralization in artificial saliva (AS); G3, non-metallic staining/remineralization with sodium fluoride/AS; and G4, metallic staining/remineralization with silver diamine fluoride/AS. The lesion mineral loss (ΔZ) and depth (L) were measured using transverse microradiography along with color change (ΔE). Specimens were bleached and color was re-evaluated. Data were analyzed using ANOVA models followed by Fisher's PLSD tests (α = 0.05). RESULTS: s-RCLs in G4 were significantly (p < 0.001) darker than G3, G2, and G1 regardless of substrate type and condition. s-RCLs in G2, G3, and G4 showed significantly lower ΔZ and L than G1 (all p < 0.001), confirming occurrence of remineralization. G4 exhibited significantly lower ΔZ and L compared to G2 (p < 0.001). Bleaching was more effective in non-metallic than in metallic stained lesions regardless of substrate type (p < 0.001). CONCLUSION: The proposed models created distinct s-RCLs. Non-metallic s-RCLs were lighter and more responsive to bleaching compared to metallic s-RCLs. CLINICAL RELEVANCE: The developed experimental models allow the further investigation of the efficacy and safety of different clinical strategies for the esthetic management of s-RCLs.

Methods of assessment of early dentine erosion: a review.

Dentine erosion is an increasingly recognised problem, especially in aging population, and various methods have been utilised for its assessment. This narrative review was planned to summarise the methods for the assessment of the early stages of dentine erosion. Relevant original articles published in the English language from 2013 to 2017 were reviewed. Laboratory techniques and methods with in vivo potential were separately studied. It is evident that the assessment of early dentine erosion is complex and requires a combination of methods. For clinical evaluation, chemical analysis and optical methods show great potential but are in need of more validation.

Accuracy of the DIAGNOcam and bitewing radiographs in the diagnosis of cavitated proximal carious lesions in primary molars.

AIMS: To evaluate the diagnostic abilities of near-infrared light transillumination (using the DIAGNOcam) and bitewing radiographs in detecting cavitated proximal carious lesions in primary molars. SUBJECTS AND METHODS: The study was a cross-sectional analytical, clinical study. The proximal surfaces of primary molars of healthy 5- to 8-year-old children were radiographically screened for the presence of carious lesions in the enamel or outer third of dentin (D1). Two trained and calibrated examiners evaluated the depth of caries in bitewing radiographs and DIAGNOcam images and then verified the presence of cavitation by direct visual examination using the "International Caries Detection and Assessment System" after temporary tooth separation. RESULTS: A total of 236 proximal lesions were included in the study. Most of the clinically cavitated lesions (51.9%) were D1 radiographically and in outer dentin lesions (scores 3 and 4) by the DIAGNOcam (37% and 48.1%, respectively). Although DIAGNOcam showed higher sensitivity (0.852) compared to the radiographs (0.519), it showed slightly less specificity (0.569) compared to the radiographs (0.579). However, DIAGNOcam showed higher value of the area under the curve (AUC = 0.722; P < 0.001) compared to the radiographic method (AUC = 0.561; P = 0.308). CONCLUSIONS: The DIAGNOcam showed higher sensitivity and better accuracy than bitewing radiographs in diagnosing cavitated proximal lesions in primary molars and can be generally considered as an alternative to radiographs to detect cavitation without the hazards of ionizing radiation in children.

Mineral features of connective dental hard tissues in hypoplastic amelogenesis imperfecta.

OBJECTIVE: To explore the mineral features of dentin and cementum in hypoplastic Amelogenesis imperfecta AI teeth. MATERIALS AND METHODS: Forty-four (44) teeth cleaned and free of caries were used: 20 control and 24 affected by hypoplastic amelogenesis imperfecta. Thirty-two teeth were studied by pQCT, cut in sections, and analyzed under microradiography, polarized light microscopy, and confocal Raman spectroscopy. Eight teeth were observed under scanning electron microscope. Four teeth were used for an X-ray diffraction. The mineral density data were analyzed statistically with the Mann-Whitney U test, using GraphPad InStat software. RESULTS: Both coronal dentin and radicular dentin were less mineralized in AI teeth when compared to control (respectively 6.2% and 6.8%; p < .001). Root dentinal walls were thin and irregular, while the cellular cementum layers were thick, reaching sometimes the cervical region of the tooth. Regular dentinal tubules and sclerotic dentin areas were noticed. Partially tubular or cellular dysplastic dentin and hyper-, normo-, or hypomineralized areas were noticed in the inter-radicular areas of hypoplastic AI teeth. The main mineral component was carbonate hydroxyapatite as explored by Raman spectroscopy and X-ray diffraction. CONCLUSIONS: Dentin and cementum in hypoplastic AI teeth are (i) hypomineralized, (ii) constituted of carbonate hydroxyapatite, and (iii) of non-homogenous structure.

In-office bleaching for the remineralization of enamel lesions filled with organic components of red wine.

PURPOSE: To investigate the effects of in-office bleaching on the remineralization of enamel lesions filled with organic components of red wine. METHODS: Enamel specimens were exposed to 0.1% NaF solution for 1 minute immersed in red wine for 5 days at 37°C, and subjected to in-office bleaching followed by remineralization in 1.5 mM CaCl2, 0.9 mM KH2PO4, 130 mM KCl, 20 mM HEPES, pH 7.0, at 37°C for 28 days. The presence of organic substances on the enamel surface was detected by Raman spectroscopy. The specimens were also subjected to transverse microradiography (TMR). RESULTS: Raman spectroscopy of baseline lesions showed characteristic peaks at 1,300-1,600 cm-1 which disappeared in bleached specimens. TMR showed that red wine formed subsurface lesions with surface content at approximately 22 mineral volume %. The integrated mineral loss (IML) was significantly lower in unbleached remineralized specimens than at baseline (P< 0.05). The IML of bleached remineralized specimens was lower than that of unbleached specimens, although not significantly (P> 0.05). Lesion depth was significantly lower in the bleached than in the unbleached group (P< 0.05). CLINICAL SIGNIFICANCE: In-office bleaching can enhance the remineralization of enamel lesions filled with organic components of red wine.

Type 1 primary hyperoxaluria: A case report and focus on bone impairment of systemic oxalosis.

Primary hyperoxaluria is a rare genetic disorder characterized by oxalate overproduction, leading to kidney failure due to nephrocalcinosis, and is eventually responsible for systemic oxalosis. Bone impairment, secondary to oxalate deposits, is one of the many complications that may occur. Skeletal involvement can be difficult to diagnose because of lack of clinical symptoms and therefore needs to be confirmed by invasive testing, such as transiliac bone biopsy. If confirmed, bone oxalosis is the proof of disease severity and that combined liver-kidney transplantation should be performed.

Effect of topical fluoride application on enamel after in-office bleaching, as evaluated using a novel hardness tester and a transverse microradiography method.

This study evaluated the effect of topical fluoride application on enamel hardness after in-office bleaching. Twelve human incisors were cut along the long axis, resulting in 24 halves used in four treatment groups (n = 6 in each group): (i) untreated group (C); (ii) in-office bleaching material (B); (iii) treatment with surface reaction-type prereacted glass-ionomer varnish after in-office bleaching (B+PRG); and (iv) treatment with acidulated phosphate fluoride solution after bleaching (B+F). All specimens were subjected to pH-cycling for 4 wk. Knoop hardness was measured using a Cariotester. The decalcification of enamel was assessed quantitatively by measuring the integrated mineral loss (ΔIML). Games-Howell analysis was used to assess statistical significance of between-group differences. The Knoop hardness decreased significantly after bleaching for all groups. In treatment groups B+PRG and B+F, the Knoop hardness returned to the original unbleached values after the first pH cycle and did not change afterwards. In treatment groups C and B there was a gradual decrease in the Knoop hardness until the fourth pH cycle. The integrated mineral loss, ΔIML, was significantly higher in treatment group B+F after 2 wk than in the other treatment groups. After 4 wk, the ΔIML in treatment group B was significantly higher than in treatment group B+PRG. The application of fluoride-containing materials after bleaching results in recuperation of hardness to levels similar to those of unbleached enamel.

Minimal Gap Size and Dentin Wall Lesion Development Next to Resin Composite in a Microcosm Biofilm Model.

This in vitro study investigated the development of dentin wall lesions next to resin composite containing very small gap sizes using an in vitro biofilm model, and evaluated whether a relevant threshold for the gap size could be established. Microcosm biofilms were grown for 14 days within small interfacial gaps between dentin-resin composite discs under intermittent cariogenic challenge. The factor under study was gap size: samples were either restored with composite resin without adhesive procedure (no intentional gap; no bonding [NB] group) or with intentional gaps of 30, 60, or 90 µm, or with complete adhesive procedure (no gap; bonding [B] group). Secondary caries wall lesion progression was measured in lesion depth (LD) and mineral loss (ML) using transversal wavelength independent microradiography at 3 locations: outer surface lesion and wall lesions at 200 and 500 µm distance from gap entrance. Results from linear regression analysis showed that the presence of an intentional gap (30, 60, and 90 µm) affected the secondary caries progression at 200 µm from the gap entrance (p ≤ 0.013). The NB group did not show significant wall lesion development (ML and LD, p ≥ 0.529). At 500 µm distance almost no wall caries development was observed. In conclusion, dentin wall lesions developed in minimal gap sizes, and the threshold for secondary wall lesion development was a gap of around 30 µm in this microcosm biofilm model.

Evidence of an in vitro Coupled Diffusion Mechanism of Lesion Formation within Microcosm Dental Plaque.

The purpose of this study was to determine whether or not the dual constant-depth film fermenter (dCDFF) is able to produce caries-like enamel lesions and to ascertain further information regarding the performance of this fully functional biological caries model. Conditions were defined by the continuation (CF) or cessation (FF) of a saliva-type growth medium supply during 50-mM sucrose exposures (8 times daily). Hydroxyapatite (n = 3) and bovine enamel (n = 3) substrata were included within each condition and samples extracted after 2, 4, 8, and 16 days. Community profiles were generated for fastidious anaerobes, Lactobacillus spp., Streptococcus spp., mutans streptococci (MS), and Veillonella spp. using selective culture techniques and enamel demineralisation assessed by transverse microradiography. Results demonstrated that the dCDFF model is able to produce caries-like enamel lesions with a high degree of sensitivity where reduced ionic strength within the FF condition increased surface layer mineral deposition. Between conditions, biofilm communities did not differ significantly, although MS in the biofilms extracted from the FF condition rose to a higher proportion (by 1.5 log10 units), and Veillonella spp. were initially greater within the CF condition (by 2.5 log10 units), indicating an enhanced ability for the clearance of low-pKa acids following exposures to sucrose. However, both conditions retained the ability for caries-like lesion formation.

PARP1-dependent recruitment of KDM4D histone demethylase to DNA damage sites promotes double-strand break repair.

Members of the lysine (K)-specific demethylase 4 (KDM4) A-D family of histone demethylases are dysregulated in several types of cancer. Here, we reveal a previously unrecognized role of KDM4D in the DNA damage response (DDR). We show that the C-terminal region of KDM4D mediates its rapid recruitment to DNA damage sites. Interestingly, this recruitment is independent of the DDR sensor ataxia telangiectasia mutated (ATM), but dependent on poly (ADP-ribose) polymerase 1 (PARP1), which ADP ribosylates KDM4D after damage. We demonstrate that KDM4D is required for efficient phosphorylation of a subset of ATM substrates. We note that KDM4D depletion impairs the DNA damage-induced association of ATM with chromatin, explaining its effect on ATM substrate phosphorylation. Consistent with an upstream role in DDR, KDM4D knockdown disrupts the damage-induced recombinase Rad51 and tumor protein P53 binding protein foci formation. Consequently, the integrity of homology-directed repair and nonhomologous end joining of DNA breaks is impaired in KDM4D-deficient cells. Altogether, our findings implicate KDM4D in DDR, furthering the links between the cancer-relevant networks of epigenetic regulation and genome stability.