Triple-blinded randomized clinical trial comparing efficacy and tooth sensitivity of in-office and at-home bleaching techniques.

OBJECTIVE: Our study aims to compare the efficacy and tooth sensitivity following in-office (35% hydrogen peroxide) or at-home (10% carbamide peroxide) bleaching treatments both preceded by 2% potassium nitrate (2%KF) desensitizing gel. METHODOLOGY: 130 volunteers were randomly allocated to a) in-office bleaching and a placebo at-home protocol; or b) in-office placebo and at-home bleaching treatment. 2% KF was applied for 10 min before both treatments. OBJECTIVE: color evaluation was performed (spectrophotometer CIEL*a*b* system and CIEDE2000) to calculate the color change (ΔE00). Subjective evaluation was performed using the VITA classical shade guide followed by shade variation (ΔSGU) at the beginning and end of bleaching treatment and 2 weeks post-bleaching. Tooth sensitivity was daily recorded using a Likert scale varying from 1 (no sensitivity) to 5 (severe sensitivity). Analysis was carried out using non-parametric tests. RESULTS: Regarding the color change, at-home bleaching resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.003) and Δa* (p=0.014). Two weeks post-bleaching, the at-home treatment resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.037) and ΔE00 (p=0.033). No differences were observed in either ΔSGU parameters. Concerning sensitivity, patients treated with in-office bleaching reported more tooth sensitivity than the at-home group only on the first day after bleaching started, without significant differences in the other periods evaluated (p>0.05). CONCLUSIONS: At-home and in-office bleaching, preceded by a desensitizing agent, were effective for vital teeth bleaching and 10% carbamide peroxide produced a higher whitening effect than 35% hydrogen peroxide in the short time evaluation. Tooth sensitivity rates were similar for the two techniques tested.

Influence of Isolation Technique on the Survival of Resin-Modified Glass-Ionomer Restorations in Primary Molars: A 9-Months Randomized Controlled Trial

ABSTRACT Objective: To compare the survival of occlusal and occlusal-proximal restorations performed with resin-modified glass-ionomer cement (RMGIC) in deciduous molars using rubber dam and cotton rolls isolation. Material and Methods: Ninety-two patients were included and 200 deciduous molars with cavitated occlusal or occlusoproximal dentin caries lesions were randomized into two groups: cotton rolls (n = 100) and rubber dam (n = 100) and RMGIC restorations were placed. At baseline and in the follow-up visit, presence, severity and activity of caries lesions were registered. Two independent, blinded examiners evaluated the treated teeth clinically using the USPHS criteria and radiographically after 9 months. Descriptive analysis, survival curve (log-rank test) and Cox regression were performed to assess risk factors related to failure. Results: Out of the 179 teeth (92 cotton rolls group and 87 rubber dam group) evaluated at 9-month follow-up period. No lesion progression was observed radiographically. The overall treatment success rate was 85.47% (83.47% for cotton rolls and 87.35 rubber dam group). No significant difference between isolation methods was observed in the log-rank test (p = 0.16). Cox regression showed no risk factors related to failure. Conclusion: No difference was found in the survival of occlusal and occlusal-proximal restorations performed with RMGIC in deciduous molars using a rubber dam and cotton rolls isolation after a 9-month follow-up period.

Triple-blinded randomized clinical trial comparing efficacy and tooth sensitivity of in-office and at-home bleaching techniques

Abstract Objective Our study aims to compare the efficacy and tooth sensitivity following in-office (35% hydrogen peroxide) or at-home (10% carbamide peroxide) bleaching treatments both preceded by 2% potassium nitrate (2%KF) desensitizing gel. Methodology 130 volunteers were randomly allocated to a) in-office bleaching and a placebo at-home protocol; or b) in-office placebo and at-home bleaching treatment. 2% KF was applied for 10 min before both treatments. Objective color evaluation was performed (spectrophotometer CIEL*a*b* system and CIEDE2000) to calculate the color change (ΔE00). Subjective evaluation was performed using the VITA classical shade guide followed by shade variation (ΔSGU) at the beginning and end of bleaching treatment and 2 weeks post-bleaching. Tooth sensitivity was daily recorded using a Likert scale varying from 1 (no sensitivity) to 5 (severe sensitivity). Analysis was carried out using non-parametric tests. Results Regarding the color change, at-home bleaching resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.003) and Δa* (p=0.014). Two weeks post-bleaching, the at-home treatment resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.037) and ΔE00 (p=0.033). No differences were observed in either ΔSGU parameters. Concerning sensitivity, patients treated with in-office bleaching reported more tooth sensitivity than the at-home group only on the first day after bleaching started, without significant differences in the other periods evaluated (p>0.05). Conclusions At-home and in-office bleaching, preceded by a desensitizing agent, were effective for vital teeth bleaching and 10% carbamide peroxide produced a higher whitening effect than 35% hydrogen peroxide in the short time evaluation. Tooth sensitivity rates were similar for the two techniques tested.

Hidden Microatelectases Increase Vulnerability to Ventilation-Induced Lung Injury.

Mechanical ventilation of lungs suffering from microatelectases may trigger the development of acute lung injury (ALI). Direct lung injury by bleomycin results in surfactant dysfunction and microatelectases at day 1 while tissue elastance and oxygenation remain normal. Computational simulations of alveolar micromechanics 1-day post-bleomycin predict persisting microatelectases throughout the respiratory cycle and increased alveolar strain during low positive end-expiratory pressure (PEEP) ventilation. As such, we hypothesize that mechanical ventilation in presence of microatelectases, which occur at low but not at higher PEEP, aggravates and unmasks ALI in the bleomycin injury model. Rats were randomized and challenged with bleomycin (B) or not (H = healthy). One day after bleomycin instillation the animals were ventilated for 3 h with PEEP 1 (PEEP1) or 5 cmH2O (PEEP5) and a tidal volume of 10 ml/kg bodyweight. Tissue elastance was repetitively measured after a recruitment maneuver to investigate the degree of distal airspace instability. The right lung was subjected to bronchoalveolar lavage (BAL), the left lung was fixed for design-based stereology at light- and electron microscopic level. Prior to mechanical ventilation, lung tissue elastance did not differ. During mechanical ventilation tissue elastance increased in bleomycin-injured lungs ventilated with PEEP = 1 cmH2O but remained stable in all other groups. Measurements at the conclusion of ventilation showed the largest time-dependent increase in tissue elastance after recruitment in B/PEEP1, indicating increased instability of distal airspaces. These lung mechanical findings correlated with BAL measurements including elevated BAL neutrophilic granulocytes as well as BAL protein and albumin in B/PEEP1. Moreover, the increased septal wall thickness and volume of peri-bronchiolar-vascular connective tissue in B/PEEP1 suggested aggravation of interstitial edema by ventilation in presence of microatelectases. At the electron microscopic level, the largest surface area of injured alveolar epithelial was observed in bleomycin-challenged lungs after PEEP = 1 cmH2O ventilation. After bleomycin treatment cellular markers of endoplasmic reticulum stress (p-Perk and p-EIF-2α) were positive within the septal wall and ventilation with PEEP = 1 cmH2O ventilation increased the surface area stained positively for p-EIF-2α. In conclusion, hidden microatelectases are linked with an increased pulmonary vulnerability for mechanical ventilation characterized by an aggravation of epithelial injury.

Optical magnification has no benefits on the detection of occlusal caries lesions in permanent molars using different visual scoring systems: An in vitro study.

BACKGROUND: Some studies have addressed the influence of optical magnification on the detection of caries lesions using a visual scoring system. However, there is a lack of research related to the use of the CAST and ADA-CCS visual scoring systems. In addition, the reliability and accuracy of ADA-CCS index in permanent teeth were not studied yet. So, the aim of this study was to evaluate, in vitro, the influence of different levels of optical magnification on the detection of occlusal caries lesions in permanent molars using three visual scoring systems. MATERIAL AND METHODS: One occlusal site per tooth was analyzed in 120 extracted permanent molars. Two trained examiners inspected the teeth using ICDAS (International Caries Detection and Assessment System), CAST (Caries Assessment Spectrum and Treatment), and ADA-CCS (American Dental Association-Caries Classification System) visual criteria, twice with each scoring system, with a one-week interval between examinations. The study was conducted in three phases: (A) without optical magnification, (B) using a binocular lens (3.5× magnification), and (C) using an operating microscope (16× magnification). Then, the teeth were sectioned longitudinally through the center of the selected site and the section with the more severe lesion was histological evaluated considering the D1 (lesions in enamel and dentin) and D3 (dentin lesions) thresholds. RESULTS: Kappa values for intra- and inter-examiner reproducibility were good to excellent for all systems. At the D1 threshold, sensitivity, accuracy, and area under the ROC curve were high for ICDAS and CAST in all phases. However, this was not the case for the ADA-CCS in phase C (<0.05). At the D3 diagnostic threshold, there was no significant difference between the visual scoring systems during the study phases (>0.05). CONCLUSIONS: The magnification does not improve the accuracy of the visual scoring systems in the detection of occlusal caries lesions in permanent molars. Key words:Dental caries, caries detection, permanent teeth, visual examination, magnification.

Image analysis of mechanistic protein biomarkers for the characterization of genotoxicants: Aneugens, clastogens, and reactive oxygen species inducers.

The early detection of genotoxicity contributes to cutting-edge drug discovery and development, requiring effective identification of genotoxic hazards posed by drugs while providing mode of action (MoA) information in a high throughput manner. In other words, there is a need to complement standard genotoxicity testing according to the test battery given in ICH S2(R1) with new in vitro tools, thereby contributing to a more in-depth analysis of genotoxic effects. Here, we report on a proof-of-concept MoA approach based on post-translational modifications of proteins (PTMs) indicative of clastogenic and aneugenic effects in TK6 cells using imaging technology (with automated analysis). Cells were exposed in a 96-well plate format with a panel of reference (geno)toxic compounds and subsequently analyzed at 4 and 24 hr to detect dose-dependent changes in PTMs, relevant for mechanistic analysis. All tested compounds that interfere with the spindle apparatus yielded a BubR1 (S640) (3/3) and phospho-histone H3 (S28) (7/9) positive dose-response reflecting aneugenicity, whereas compounds inducing DNA double-strand-breaks were associated with positive FANCD2 (S1404) and 53BP1 (S1778) responses pointing to clastogenicity (2/3). The biomarker p53 (K373) was able to distinguish genotoxicants from non-genotoxicants (2/4), while the induction of reactive oxygen species (ROS), potentially causing DNA damage, was associated with a positive Nrf2 (S40) response (2/2). This work demonstrates that genotoxicants and non-genotoxicants induce different biomarker responses in TK6 cells which can be used for reliable classification into MoA groups (aneugens/clastogens/non-genotoxicants/ROS inducers), supporting a more in-depth safety assessment of drug candidates.

Performance of light-emitting diode device in detecting occlusal caries in the primary molars.

This in vitro study aimed to compare the performance of a light-emitting diode (LED) device (Midwest Caries I.D.: MID), International Caries Detection and Assessment System (ICDAS) visual criteria, and fluorescence-based devices (DIAGNOdent: LF; DIAGNOdent pen: LFpen; and Quantitative Light-induced Fluorescence: QLF) in detecting occlusal caries in the primary molars. Eighty-eight primary molars with sound occlusal surfaces or carious lesions at different stages were assessed twice, with a 1-week interval in between, by one examiner using all three methods. Subsequently, the teeth were sectioned and lesion depth was verified using stereomicroscopy as a gold standard. Sensitivity, specificity, and accuracy were calculated at D1 (all carious lesions-enamel and dentin) and D3 (dentin lesions) thresholds. Correlation with histological analysis was evaluated using Spearman's rank correlation coefficients (rho). Weighted Kappa and intraclass-correlation (ICC) coefficients were calculated to assess intra-examiner reproducibility. At D1 threshold, ICDAS and LFpen showed higher sensitivity than the other methods, whereas ICDAS, LF, and QLF showed higher specificity (p < 0.05), and MID showed lower accuracy. At D3 threshold, ICDAS, LFpen, and QLF showed higher sensitivity than MID, whereas ICDAS, LF, and MID showed higher specificity (p < 0.05). All methods, except MID, showed statistically similar accuracy values (p < 0.05). Correlations with histopathological analysis varied from 0.15 (MID) to 0.57 (ICDAS). Intra-examiner reproducibility varied from 0.30 (MID) to 0.92 (ICDAS, LF, and QLF). The MID device exhibited a poor performance in detecting occlusal carious lesions in the primary molars, and ICDAS visual criteria exhibited greater accuracy than LF, LFpen, and QLF devices.

Differentiation of Aneugens and Clastogens in the In Vitro Micronucleus Test by Kinetochore Scoring Using Automated Image Analysis.

The in vitro micronucleus test according to OECD Test Guideline 487 (TG 487) is widely used to investigate the genotoxic potential of drugs. Besides the identification of in vitro genotoxicants, the assay can be complemented with kinetochore staining for the differentiation between clastogens and aneugens. This differentiation constitutes a major contribution to risk assessment as especially aneugens show a threshold response. Thus, a novel method for automated MN plus kinetochore (k+) scoring by image analysis was developed based on the OECD TG 487. Compound-induced increases in MN frequency can be detected using the cytokinesis-block (cytochalasin B) method in V79 cells after 24 h in a 96-well format. Nuclei, MN, and kinetochores were labeled with nuclear counterstain and anti-kinetochore antibodies, respectively, to score MN in binuclear or multinuclear cells and to differentiate compound-induced MN by the presence of kinetochores. First, a reference data set was created by manual scoring using two clastogens and aneugens. After developing the automated scoring process, a set of 14 reference genotoxicants were studied. The automated image analysis yielded the expected results: 5/5 clastogens and 6/6 aneugens (sensitivity: 100%) as well as 3/3 non-genotoxicants (specificity: 100%) were correctly identified. Further, a threshold was determined for identifying aneugens. Based on the data for our internally characterized reference compounds, unknown compounds that induce ≥53.8% k+ MN are classified as aneugens. The current data demonstrate excellent specificity and sensitivity and the methodology is superior to manual microscopic analysis in terms of speed and throughput as well as the absence of human bias. Environ. Mol. Mutagen. 60:227-242, 2019. © 2018 Wiley Periodicals, Inc.

Classification of in vitro genotoxicants using a novel multiplexed biomarker assay compared to the flow cytometric micronucleus test.

Regulatory in vitro genotoxicity testing exhibits shortcomings in specificity and mode of action (MoA) information. Thus, the aim of this work was to evaluate the performance of the novel MultiFlow® assay composed of mechanistic biomarkers quantified in TK6 cells after treatment (4 and 24 hr): γH2AX (DNA double strand breaks), phosphorylated H3 (mitotic cells), translocated p53 (genotoxicity), and cleaved PARP1 (apoptosis). A reference dataset of 31 compounds with well-established MoA was studied using the MicroFlow® micronucleus assay. A positive call was raised following the earlier published criteria from Litron Laboratories. In the light of our data, these evaluation criteria should probably be adjusted since only 8/11 (73%) nongenotoxicants and 18/20 (90%) genotoxicants were correctly identified. Moreover, there is a need for new in vitro tools to delineate the predominant MoA as in the MicroFlow® assay only 5/9 (56%) aneugens and 4/11 (36%) clastogens were correctly classified. In contrast, the MultiFlow® assay provides more in-depth information about the MoA and therefore reliably discriminates clastogens, aneugens, and nongenotoxicants. By using a lab-specific, practical threshold for the aforementioned biomarkers, 10/11 (91%) nongenotoxicants and 19/20 genotoxicants (95%), 9/11 (82%) clastogens, and 8/9 (89%) aneugens were correctly categorized, suggesting a clear improvement over the MicroFlow® . Furthermore, the MultiFlow markers were benchmarked against established methods to assess the validity of the data. Altogether, these findings demonstrated good agreement between the MultiFlow® assay and the benchmarking methods. Finally, p21 may improve class discrimination given the correct identification of 4/4 (100%) aneugens and 2/5 (40%) clastogens. Environ. Mol. Mutagen. 58:662-677, 2017. © 2017 Wiley Periodicals, Inc.

Interlaboratory evaluation of a multiplexed high information content in vitro genotoxicity assay.

We previously described a multiplexed in vitro genotoxicity assay based on flow cytometric analysis of detergent-liberated nuclei that are simultaneously stained with propidium iodide and labeled with fluorescent antibodies against p53, γH2AX, and phospho-histone H3. Inclusion of a known number of microspheres provides absolute nuclei counts. The work described herein was undertaken to evaluate the interlaboratory transferability of this assay, commercially known as MultiFlow® DNA Damage Kit-p53, γH2AX, Phospho-Histone H3. For these experiments, seven laboratories studied reference chemicals from a group of 84 representing clastogens, aneugens, and nongenotoxicants. TK6 cells were exposed to chemicals in 96-well plates over a range of concentrations for 24 hr. At 4 and 24 hr, cell aliquots were added to the MultiFlow reagent mix and following a brief incubation period flow cytometric analysis occurred, in most cases directly from a 96-well plate via a robotic walk-away data acquisition system. Multiplexed response data were evaluated using two analysis approaches, one based on global evaluation factors (i.e., cutoff values derived from all interlaboratory data), and a second based on multinomial logistic regression that considers multiple biomarkers simultaneously. Both data analysis strategies were devised to categorize chemicals as predominately exhibiting a clastogenic, aneugenic, or nongenotoxic mode of action (MoA). Based on the aggregate 231 experiments that were performed, assay sensitivity, specificity, and concordance in relation to a priori MoA grouping were ≥ 92%. These results are encouraging as they suggest that two distinct data analysis strategies can rapidly and reliably predict new chemicals' predominant genotoxic MoA based on data from an efficient and transferable multiplexed in vitro assay. Environ. Mol. Mutagen. 58:146-161, 2017. © 2017 Wiley Periodicals, Inc.