Search

Background: The main cause of pulpal and periapical diseases is bacterial infection, but mechanical and chemical preparation in root canal therapy is difficult to completely remove the bacterial microorganism. Antibacterial photodynamic therapy (aPDT) is a medical method that kills microorganisms by activating a photoactive agent or photosensitizer by exposure to visible light of a specific wave-length in the presence of oxygen. The present study aimed to evaluate the killing in vitro effect of aPDT with 0.01% methylene blue (MB) against young and mature Enterococcus faecalis (E. faecalis) biofilms in bovine and human dentin with the long-term exposure using confocal laser scanning microscopy (CLSM)., Methods: Prepared bovine and human dentin blocks and their structure were observed by scanning electron microscopy (SEM). Semicylindrical bovine dentin blocks and human root canal dentin blocks were inoculated with E. faecalis and incubated in air to form 1- and 3-week-old biofilms. The biofilms in dentin were subjected to aPDT with 0.01% MB, 5% NaOCl and saline with the exposure of 3, 12 and 30 min. The dead portions of bacterial cells in E. faecalis biofilms were analyzed with using LIVE/DEAD bacteria viability staining and CLSM., Results: A clean dentin surface in bovine dentin blocks were verified with SEM. In bovine and human dentin blocks, significantly more bacteria were dead when aPDT with MB and 5% NaOCl were used with the long exposure time (12 and 30 min) than with 3 min (P < 0.05). The speed of killing was fastest during the first 3 min, and few more bacterial cells were killed after 12 min in the disinfection groups. 5% NaOCl exhibited the highest effectiveness of bacterial killing in dentin at each time point than aPDT with MB groups (P < 0.05). The proportion of killed bacteria was higher in young biofilms than in mature biofilms in aPDT with MB and NaOCl groups (P < 0.05). Moreover, there were no clearly visible changes in structure of bovine dentin surfaces subjected to aPDT with MB for 30 min., Conclusion: aPDT with 0.01% MB has the capability to kill bacterial cells in E. faecalis biofilms on bovine and human dentin blocks. Young E. faecalis biofilms in dentin canals were more susceptible to disinfection approaches than mature biofilms., Competing Interests: Declarations. Ethics approval and consent to participate: Ethical approval The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Science and Ethics Commission of the First Affiliated Hospital of Zhengzhou University. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. Informed consent: Not applicable., (© 2025. The Author(s).)

Researching disinfection strategies is pivotal because effectively eliminating bacteria and their byproducts during root canal treatment (RCT) remains a challenge. This study investigated the antimicrobial efficacy of natural antimicrobial compounds, propolis (PRO) and copaiba oil-resin (COR), compared to conventional agents in Endodontics. Antimicrobials were tested against endodontic pathogens via macrodilution with standardized inoculums to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). Biofilm killing efficacy were performed using two dual-species biofilms: Enterococcus faecalis (ATCC 29212) and Streptococcus mutans (ATCC 20523) and Streptococcus oralis (J22) and Actinomyces naeslundii (T14V-J1) grown on dentine discs. At the intratubular level (dentine cylinders), dentine tubule contamination was performed with E. faecalis and S. mutans. The specimens were exposed to antimicrobials to simulate their use at different sets of RCT and bacterial viability was quantified using Live/Dead staining via confocal laser scanning microscopy (CLSM). Biofilm characteristics and immediate removal of S. oralis and A. naeslundii biofilm model were evaluated employing optical coherence tomography (OCT) and CFU/mL counting. Statistical tests were applied according to data distribution for each analysis (α=0.05). Macrodilutions showed different effects against endodontic pathogens. Direct contact and intratubular analysis showed that PRO and COR promoted disinfection like conventional agents (p > 0.05). According to OCT analysis, PRO and COR showed similar biofilm reduction after immediate contact (p < 0.05). CFU/mL counting showed decontamination (p < 0.05) after using natural and conventional agents. PRO and COR showed antimicrobial effects, indicating their suitability as complementary approaches in RCT to eliminate as much microbial load as possible., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Post-space disinfectants methylene blue loaded with silver (Ag) and Quartz nanoparticles (NPs) and MB alone activated by photodynamic therapy (PDT) on the survival of Enterococcus faecalis, smear layer (SL) removal efficacy, and extrusion bond strength (EBS) of fiber posts to canal dentin. Hundred mandibular premolars underwent root canal treatment using the rotary ProTaper system. The canals were obturated and post-space was prepared up to a length of 8 mm, maintaining a 5 mm seal. To assess antibacterial efficacy E. faecalis were inoculated in the canal (n = 20). The samples were then randomly allocated into four groups according to the canal disinfectant used to sterilize the canals. Group 1: 2.5% NaOCl+17% EDTA, Group 2: MB-PDT, Group 3: MB@QP-PDT, and Group 4: MB@AgNP-PDT (n = 25). SEM analysis was conducted on five samples from each disinfectant group to assess the removal of the SL. Survival rates were calculated (n = 5 from each group). Glass fiber post (GFP) was cemented to the root dentin of the remaining samples followed by artificial aging. Sectioning of the specimens was performed in all three-thirds of the canals. PBS was assessed followed by failure evaluation. ANOVA and Tukey post hoc test were used to compare the E. faecalis survival rate and PBS of fiber post among different investigated groups. Group 4 (MB@AgNP-PDT) treated canals exhibited the minimum survival rate (0.30 ± 0.04 CFU/mL) of E. faecalis and maximum PBS. However, the highest survival rate and minimum bond strength of GFP were observed in Group 1 (NaOCl+17% EDTA) and Group 2 (MB-PDT) treated teeth respectively. The highest SL removal was recorded in the coronal section of the samples of Group-4 disinfected with MB@AgNP-PDT. The lowest removal of SL was recorded in Group 2 samples sterilized with MB-PDT at apical one-third. Silver nanoparticles when utilized as nano-carriers to enhance the efficiency of MB activated by PDT, have been shown to exhibit the highest antimicrobial potency, improved capacity for SL removal and improved PBS., (© 2024 Wiley Periodicals LLC.)

Background: Disinfection of the root canal system is a challenge to all clinicians, calcium hydroxide Ca(OH) 2, one of the most popular intracanal medications used for this purpose, has some unwanted effects on dentine. This study aimed to investigate the antibiofilm effect of Nanochitosan (CSNPs) and Calcium hydroxide Ca(OH) 2 intra canal medications and their effect on the microhardness and chemical structure of radicular dentine., Methodology: A total of 52 extracted human mandibular premolars were used. Eighteen premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis (E. faecalis) then divided randomly into 2 groups based on the intracanal medication used: group (A) 2% CSNPs gel and group (B) Ca(OH) 2 paste. Antibiofilm effect was evaluated using bacterial counts. For the effect on microhardness, 56 specimens were subjected to Vicker's microhardness test before and after 1 week, and 4 weeks of medication application. Twelve cervical radicular dentine specimens (6 samples per group) were examined using Fourier transform infrared spectroscopy and scanning electron microscope., Results: Both groups showed a significant reduction in the bacterial count at (p = 0.008) with no significant difference between them (P = 0.605). Ca(OH) 2 showed the highest reduction in the microhardness compared to CSNPs groups after 1 week (P = 0.0495) and after 4 weeks (P = 0.0495). FTIR spectrum results revealed that the control group (no treatment) showed the highest significant Phosphate: amide ratio compared to both CSNPs and Ca(OH) 2 at p = 0.006. SEM images revealed absence of discernible smear layer in CSNPs treated samples after 4 weeks and all the dentinal tubules were open., Conclusions: Nanochitosan gel could be considered as a viable option as an intra canal medication., Competing Interests: Declarations. Ethics approval and consent to participate: This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of the Egyptian Russian University no FD-ERU-REC 6). The need for consent was waived by the Ethics Committee of the Egyptian Russian University. Consent for publication: N/A. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Introduction: To evaluate the bacterial biofilm, smear layer and debris removal efficacy of a hydro-dynamic cavitation system with physiological saline using a new ex vivo model., Methods: Seventy-five dentin discs were prepared from fifty-four extracted teeth. Seventy-five artificial root sockets were prepared. Sixty dentin discs were used to grow 3-week-old bacterial biofilms, while smear layer and debris were produced on fifteen dentin discs. These dentin discs were adhered to the middle third of the artificial root canals. The sixty ex vivo models with biofilm-covered dentin discs were divided into six groups: control, needle with physiological saline, ultrasonic with physiological saline, Odne™ Clean with physiological saline, needle with 3% NaOCl, and ultrasonic with 3% NaOCl. Biofilm removal efficacy was evaluated using confocal laser scanning microscopy. The fifteen ex vivo models with smear layer and debris-covered dentin discs were divided into three groups: control, Odne™ Clean with physiological saline, and 5% NaOCl followed by 17% EDTA. Smear layer and debris removal efficacy was evaluated using scanning electron microscopy. Statistical analysis was performed using one-way analysis of variance for comparisons involving more than two groups. Post-hoc pairwise comparisons were conducted using the Tukey test., Results: Odne TM Clean with physiological saline (98%) performed significantly better than needle irrigation (47%) or ultrasonic activation (54%) with physiological saline (P < 0.05). Odne TM Clean with physiological saline removed biofilms as effectively as needle irrigation (97%) or ultrasonic activation (98%) with 3% NaOCl (P > 0.05). Additionally, 5% NaOCl followed by 17% EDTA (score: 1.33) removed the smear layer significantly better than Odne TM Clean with physiological saline (score: 4.47) (P < 0.05). However, Odne TM Clean with physiological saline (score: 1.27) removed debris as effectively as 5% NaOCl followed by 17% EDTA (score: 1.13) (P > 0.05)., Conclusions: Odne TM Clean with physiological saline can effectively remove bacterial biofilm and debris from the dentin surface but cannot effectively remove the smear layer. Utilizing Odne TM Clean during the final irrigation may enhance root canal cleaning efficacy., Competing Interests: Declarations. Ethics approval and consent to participate: The clinical Research Ethics committee of the University of British Columbia (certificate H1202430) approved the protocol of this study. All participants provided signed informed consent, authorizing the use of their extracted premolars for research purposes. The authors ensure that all procedures were conducted in line with relevant guidelines and regulations. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

This study assessed the effect of composite resins, aggregated or not with S-PRG particles, and the use of toothpaste in controlling demineralization and bacterial growth. Human molars were distributed into 3 groups: control (CT) - sound teeth, Beautifil Bulk Restorative System (aggregated with S-PRG) (BB), Filtek One Bulk Fill (without S-PRG) (FB). Teeth destined for groups BB and FB previously received Class I preparations (4 × 4 × 4 mm), followed by single-increment restorations. All teeth were sectioned mesiodistally, with all specimens subjected to cariogenic challenge for 5 days, including microcosm biofilm formation. Half of each tooth was exposed to toothpaste (CTF, BBF, FBF). The loss of microhardness was assessed considering the initial microhardness as 100% on enamel, dentin, and composite resin substrates. Colony Forming Units (CFU/mL) were counted in 3 media. Data analysis used one-way ANOVA, Tukey HSD test, and paired t-test (α = 0.05). Toothpaste significantly reduced CFU/mL for total bacteria and genus Streptococcus (p < 0.05), with no significant difference for Streptococcus mutans. Enamel microhardness was positively affected by toothpaste. Both restorative systems controlled enamel demineralization, with FB and FBF outperforming BB and BBF. There was minor degradation of both composite resins, between 10% and 22%. Toothpaste effectively reduced microorganisms, irrespective of the composite resin. Regarding demineralization control, both restorative systems, with and without S-PRG particles, were effective on enamel., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval: All procedures performed in this study involving human teeth were in accordance with the ethical standards of the institutional and national human research committee (CEP/UFMS - Federal University of Mato Grosso do Sul (MS/Brazil); protocol: 3.678.506, CAAE: 21527119.6.0000.0021) and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. Informed consent: Informed consent was obtained from all individual participants included in the study., (© 2024. The Author(s).)

Objectives: This cross-sectional study aimed to evaluate the occurrence of Streptococcus spp., Streptococcus mutans, its serotypes (c, e, f, and k), collagen-binding genes (cnm/cbm), and Candida albicans in medium deep (D2) and deep (D3) dentin carious lesions of permanent teeth., Materials and Methods: Carious dentin was collected from D2 (n = 23) and D3 (n = 24) lesions in posterior teeth from 31 individuals. DNA was extracted and analyzed using polymerase chain reaction (PCR)., Results: Streptococcus spp. exhibited a high prevalence in both D2 and D3 lesions, with a 100% occurrence rate in D3. Although S. mutans was prevalent in both lesion types, serotype distribution highlighted differences in complexity. Serotype e was the most frequently detected in D2 samples. Serotype c was absent in D2 but present in 19.0% of D3; serotype k was exclusively found in multi-serotype D3 alongside f, c, and e. The cbm gene and undetermined serotypes were detected only in D3. Candida albicans was not found in any samples, regardless of lesion depth., Conclusion: Dentinal lesions exhibited a complex array of S. mutans serotypes, with a notable prevalence of the genus Streptococcus and an overall S. mutans prevalence of approximately 35%. Candida albicans was absent. In addition, a high diversity of serotypes was observed, including multiple and indeterminate serotypes, along with the presence of the collagen-binding cbm gene., Clinical Relevance: The identification of diverse Streptococcus mutans serotypes and the cbm gene in carious dentin from permanent teeth offers insights into microbial factors potentially linked to dentinal caries lesions. Additionally, the absence of Candida albicans suggests a limited role in this context., Competing Interests: Declarations. Ethical approval: The study was approved by the Research Ethics Committee of the Federal University of Ceará - CEP/UFC/PROPESQ (CAAE 54451316.7.0000.5054) and (CAAE 46268821.6.0000.5054). Informed consent: Written informed consent was obtained from all participants included in the study. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Enterococcus faecalis (E. faecalis) is the most common microorganism responsible for post-endodontic infections, which can penetrate deep into dentinal tubules. This study aimed to compare the survival of E. faecalis in dentinal tubules following root canal therapy with AH Plus (AHP), Endoseal MTA (ESM), and cold ceramic (CC) sealers., Methods: In this in vitro experimental study, 80 single-canal human teeth were decoronated at the cementoenamel junction. The root canals were cleaned and shaped, and auto-clave sterilized after smear layer removal. The roots were then inoculated with E. faecalis, and incubated for 3 weeks. The teeth were randomly assigned to four groups (n = 20) for root canal obturation with AHP, ESM, and CC sealers and 2% gutta-percha with the cold lateral compaction technique. The fourth group served as the control group. After 30 days, dentin chips were collected from the canals, and cultured on blood agar. The colony count was reported and analyzed by the Kruskal-Wallis test (alpha = 0.05)., Results: The mean colony count was 23.73 ± 13.84 in the AHP, 34.78 ± 18.75 in the ESM, and 28.47 ± 13.73 in the CC group after 30 days. The difference in this regard was not significant among the three experimental groups (P = 0.102)., Conclusions: Within the limitations of this in vitro study, the results indicated comparable antibacterial activity of AHP, ESM, and CC sealers against E. faecalis in infected dentinal tubules after 30 days., Competing Interests: Declarations. Ethics approval and consent to participate: The need for informed consent was waived by the ethics committee of Yazd Shahid Sadoughi University of Medical Sciences due to the use of anonymized extracted teeth. This study did not involve any human subjects directly and no identifiable patient information was used.”. The study protocol was approved by the ethics committee of Yazd Shahid Sadoughi University of Medical Sciences (IR.SSU.DENTISTRY.REC.1401.073). All methods were carried out in accordance with relevant guidelines and regulations. Consent for publication: Identified images or other personal or clinical details of participants are not presented in this manuscript. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

This study aimed to evaluate the antibiofilm activity of different concentrations of silver nanoparticles (AgNPs) in combination with calcium hydroxide [Ca(OH) 2 ] against Enterococcus faecalis biofilm. On an E. faecalis biofilm on dentin discs, the following medicaments were applied for 7 days (n = 13/group): 0.005% AgNPs+Ca(OH) 2 , 0.01% AgNPs+Ca(OH) 2 , 0.02% AgNPs + Ca(OH) 2 , Ca(OH) 2 and saline/control. Specimens were stained with LIVE/DEAD® BacLight™ dye and analysed with confocal laser scanning microscopy. Proportion of dead bacteria was calculated and analysed. There was a significant reduction in E. faecalis biofilm in all medicament groups (43.5%, 49.1%, 69.1%, 48.7%) respectively, compared with control group (2.54%) (p < 0.001). The 0.02% AgNPs + Ca(OH) 2 group demonstrated the most significantly superior antibiofilm effect, with no significant difference between remaining groups. In conclusion, combining 0.02% AgNPs enhanced the antibiofilm effect of Ca(OH) 2 on E. faecalis biofilm compared with lower AgNPs concentrations., (© 2024 Australian Society of Endodontology Inc.)

As one of the global concerns, antimicrobial resistance crisis increases the clinical importance of Enterococcus species. Enterococcus faecalis (E. faecalis) specifically penetrates the dentinal tubules and remains prevalent even after endodontic treatment. It has also biofilm forming character as well as the development of resistance to antibiotics. Sodium hypochlorite (NaOCl) is considered the gold standard among antibacterial washing solutions. However, due to its toxic effects, its usage limitations have led to the search for natural, non-toxic alternatives. Phages can be considered an important alternative because of their effects on specific bacteria. The objective of this study was to compare the effect of isolated active vB&#95;Ef1 phage on the removal of E. faecalis biofilm in dentin, together and separately with the chemical irrigation solution NaOCl. As a result of study, the optimal NaOCl solution concentration to be applied with vB&#95;Ef1 phage is 0.5 %, and the use of solution at this value reduces the biofilm mass by 84 %, reaching the highest biofilm mass reduction value. It was found that the combination of phage and NaOCl at appropriate concentrations had the strongest biofilm disruption effect., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Purpose: To evaluate and compare: (1) the effect of the bacterial biofilm on the dentin mineral density at the restoration-tooth interface and (2) the mineralization potential of three resin-based restorative materials (RBRM)., Methods: 16 extracted human molars free of caries and cracks were collected and stored for disinfection. Each tooth received two standardized Class II preparations with the cervical margin placed in dentin. Teeth were secured into a dentiform with adjacent natural teeth to ensure interproximal contact. All tooth preparations were hybridized using a three-step etch-and-rinse adhesive system (OptiBond FL) and assigned randomly to three experimental groups according to the RBRM (n= 8): Group A - a nanofill resin composite (Filtek Supreme Ultra); Group B - a high-viscosity bulk-fill resin composite (Tetric Powerfill); Group C - a low-viscosity bulk-fill resin composite (SureFil SDR flow+ bulk-fill); and a positive control: Group D - bioactive resin composite (Activa Bioactive-Restorative). All materials were used according to manufacturers' instructions. All specimens were subjected to two distinct challenges: first, thermomechanical cycling was performed within 24 hours of restoring the specimens to simulate 1 year of masticatory function. Subsequently, the specimens were stored for 18 days in a laboratory biofilm model to promote biofilm formation and to mimic the effects of tooth demineralization. Two sessions of micro-CT imaging were conducted: the first immediately after the thermomechanical cycling and the second post-exposure to the biofilm model. All data on mineral profile measurements reconstructed in the Perkin-Elmer Quantum GX-II CT were transferred to Image J software for analysis and interpretation. The ANOVA test (P< 0.05) was used to analyze the mineral density values and mean mineral loss values for each group., Results: No statistically significant difference in mean mineral loss value (mean ΔZ) was found between the groups (P= 0.209). Regardless, increased mean ΔZ variation was found between SureFil SDR flow+ (-56.95) and the remaining groups, Filtek Supreme Ultra, Tetric Powerfill and Activa Bioactive (-1.17, -1.41, and -7.97, respectively), showing, within the limits of the present laboratory study, the remineralization potential of SureFil SDR flow+. All tested RBRM demonstrated some remineralization capacity under caries risk conditions., Clinical Significance: The mineralization potential of some resin-based composites under caries-risk conditions can represent a paradigm shift in restorative material selection for moderate-to-high-risk patients., Competing Interests: The authors of this article certify that they have no proprietary, financial, or other personal interest of any nature or kind in any product, service, and/or company presented in this article. Dr. Shamieh was supported by a graduate student scholarship award sponsored by the Sturdevant Research Fund, managed by the Dental Foundation of North Carolina., (Copyright©American Journal of Dentistry.)

This in vitro study aimed to verify the influence of chlorhexidine (CHX) (gel and solution) in association with different activation protocols on disinfection of root canals contaminated with Enterococcus faecalis. In total, 120 lower premolar roots were selected, contaminated and divided into 12 groups according to irrigation technique and substance. Samples were collected before and after each irrigation technique and analysed for colony-forming units (CFU). Three dentin discs were obtained for quantification of viable intratubular cells by using confocal laser scanning microscopy (CLSM). CFU results showed that sonic and ultrasonic were more effective than conventional irrigation. Ultrasonic activation was more effective than sonic for CHX gel (p < 0.05). CLSM showed that either activation was better than conventional in all groups and root canal thirds (p < 0.05). It was concluded that ultrasonic/sonic activation was more effective than conventional techniques and CHX enhanced intratubular dentin decontamination across all irrigation methods., (© 2024 Australian Society of Endodontology Inc.)

Assessment of the antimicrobial, micro tensile bond strength (μTBS), and degree of conversion (DC) of fifth-generation adhesive modified using photoactivated 0.5% rose bengal (RB) and photoactivated RB-doped titanium dioxide nanoparticles (TiO 2 NPs) in different concentrations (2% and 5%) as compared with the unmodified adhesive bonded to the carious affected dentin (CAD). Forty mandibular molars with caries progression up to the middle third of the dentin, as per the International Caries Detection and Assessment System (ICDAS) score of 4 and 5 were included. Specimens were divided into four groups based on etch and rinse adhesive (ERA) modification group 1: unmodified ERA, group 2: photoactivated 0.5% RB photosensitizer (PS) modified ERA, group 3: photoactivated RB-doped 2 wt% TiO 2 NPs adhesive, group 4: photoactivated RB-doped 5 wt% TiO 2 NPs adhesive. Followed by adhesive and composite restoration on the CAD surface. All the specimens were thermocycled and an assessment of μTBS and failure pattern analysis was performed. The antibacterial potency of RB and RB-doped TiO 2 NPs (2% and 5%) followed by their activation using visible light against Streptococcus mutans (S.mutans) were tested. The survival rate of S.mutans was assessed using the Kruskal-Wallis test. The analysis of μTBS involved the use of ANOVA, followed by a post-hoc Tukey honestly significant difference (HSD) multiple comparisons test. Group 1 (Unmodified ERA) (0.52 ± 0.31 CFU/mL) treated samples unveiled the highest means of bacterial survival and lowest μTBS (11.32 ± 0.63 MPa). Nevertheless, group 4: photoactivated RB-doped 5 wt% TiO2NPs adhesive displayed the lowest outcomes of S.mutans survival (0.11 ± 0.02 CFU/mL) and highest bond strength (18.76 ± 1.45 MPa). The photoactivated RB-doped 2 wt% TiO 2 NPs in adhesive demonstrated promising enhancements in both μTBS and antibacterial efficacy against S.mutans. However, it is noteworthy that this modification led to a decrease in the DC of the adhesive. RESEARCH HIGHLIGHTS: Unmodified ERA-treated samples unveiled the highest bacterial survival and the lowest μTBS. Photoactivated RB-doped 5 wt% TiO 2 NPs adhesive displayed the lowest S.mutans survival rate and highest bond strength. DC decreased with an increase in concentration of TiO 2 ., (© 2024 Wiley Periodicals LLC.)

Objective: This study aimed to evaluate the effectiveness of two ion-releasing, resin-based composites (RBCs), ACTIVA BioACTIVE-Restorative (BIO1) and ACTIVA Presto (BIO2), in preventing demineralization around restorations., Methods: Class I and II cavities were prepared on bovine (n = 4) and human teeth (n = 8) and restored with BIO1, BIO2, a conventional composite (RBC, Filtek Supreme XTE), and a resin-modified glass ionomer cement (RMGIC, Ionolux). Following restorations (class I, n= 16/material; class II, n= 8/material), the specimens were exposed to Streptococcus mutans biofilm in a continuous-flow bioreactor over two weeks. Micro-computed tomography (micro-CT) assessed demineralization depths at restoration margins, and supernatant pH changes were measured after a 24-h acidic challenge. Statistical analyses included one and two-way ANOVA and Tukey's test (p < 0.05)., Results: On enamel surfaces, RMGIC showed no demineralization, followed by BIO1 (≈50 μm), BIO2 (≈125 μm), and RBC (≈150 μm). No difference between human or bovine enamel was observed. In dentin, RMGIC showed the least demineralization (≈190 μm), followed by BIO1 (≈230 μm), BIO2 (≈280 μm), and RBC (≈400 μm). pH buffering was highest in RMGIC (+ 0.24 pH), while BIO1, BIO2, and RBC showed similar buffering capacities (∼ +0.1 pH). Gaps were found at several interfaces for BIO1 and RMGIC., Conclusions: Ion-releasing RBCs varied in effectiveness for reducing demineralization of surrounding tissues, with limited pH buffering capacity. RMGIC exhibited better performance. Gaps between cavity walls and BIO1/RMGIC raised concerns about long-term adhesion., Clinical Significance: ACTIVA BioACTIVE Restorative (BIO1) and ACTIVA Presto (BIO2) resin-based composites promoted slight inhibition of demineralization in an in-vitro biofilm model, suggesting that further modifications in their chemical composition are necessary., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Ltd.)

This study evaluated the antibiofilm efficacy of a calcium silicate-based intracanal medicament (Bio-C Temp) against Fusobacterium nucleatum (F. nucleatum). Dentin slices (n = 88) were inoculated with F. nucleatum biofilms and divided into experimental groups (n = 24 each) treated with Bio-C Temp, triple antibiotic paste (TAP), or Ca(OH) 2 for 1 or 2 weeks. Untreated dentin slices exposed to saline served as a positive control, while sterile dentin slices incubated anaerobically served as a negative control. Bacterial viability was assessed using confocal laser scanning microscopy after staining with LIVE/DEAD ® BacLight ™ dye. Samples treated with TAP demonstrated the highest percentage of dead bacteria (95%). The antibiofilm effect of TAP and Bio-C Temp significantly increased from Week 1 to Week 2. At Week 2, samples treated with Bio-C Temp showed a significantly higher percentage of dead bacteria (75.25%) than those treated with Ca(OH) 2 (58.65%). TAP showed superior efficacy against F. nucleatum biofilms. After an application time of 2 weeks, the antibiofilm efficacy of Bio-C Temp was superior to that of Ca(OH) 2 ., (© 2024. The Author(s).)

Objective: This work aimed to evaluate the in vitro effect of zinc oxide-eugenol paste (ZOE) on planktonic aggregates (EfPA) and biofilm (EfBio) of Enterococcus faecalis, focusing on their morphological aspects observed and analyzed using atomic force microscopy (AFM)., Design: The eugenol and paste were characterized by Gas Chromatography coupled with Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FTIR), respectively. The effect of ZOE on EfPA and EfBio was evaluated by a direct-contact test through colony counting and crystal violet staining protocol. AFM images of untreated and treated EfPA and EfBio growth on bovine dentin were obtained to analyze the morphological damage caused by the treatments., Results: The characterization showed high purity in the eugenol composition and chemical interaction between the components of the paste. A bactericidal effect on aggregates was observed after 6 h of exposure, and on biofilm after 24 h of treatment (p < 0.001). A disruptive effect on the biofilm was also evident. AFM images revealed the formation of EfPA, with a notable presence of an exopolysaccharide matrix. After 6 h of ZOE treatment, there was a significant increase in the size and surface roughness profile of treated cells (p < 0.05). Loss of typical cell morphology was observed after 24 h. The effect on the biofilm showed a tendency towards a less condensed biofilm pattern in the treated group, with no differences in surface roughness., Conclusion: ZOE presents bactericidal action on EfPA and EfBio, promoting significant morphological changes after treatment, especially in the aggregates., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study evaluated conventional and alternative irrigation solutions combined with ultrasonic irrigant activation (UIA) against Fusobacterium nucleatum ( F. nucleatum ) and its endotoxin (LPS) in main root canal and dentinal tubules, using a new intratubular infection model. Seventy dentin cylinders were infected with F. nucleatum for seven days under anaerobic conditions and treated with 2.5% sodium hypochlorite (NaOCl), limewater + 2.5% NaOCl, and 10% ethanolic propolis extract (EEP), administered by syringe irrigation (SI) or UIA. Microbiological samples were collected before and after irrigation to determine CFU ml -1 and LPS levels. Confocal microscopy assessed bacterial membrane damage with Live/Dead staining. Irrigation solutions effectively reduced CFU ml -1 . UIA caused greater damage to the bacterial membranes and reduced LPS levels. The ultrasonic activation of 10% EEP and limewater + 2.5% NaOCl were comparable to 2.5% NaOCl ( p  > 0.05). UIA improved the effectiveness of solutions, suggesting potential for alternative substances. Randomized clinical trials using these protocols are recommended.

Phosphate-based glasses are known for their excellent biocompatibility and adjustable degradation rates. In this study, we fabricated a rapidly soluble zinc-ion-releasing phosphate-based glass (RG) specifically designed for use in dental cavity liners. The aim of this study was to evaluate the ion-releasing properties and antibacterial effects of RG. Additionally, a dental resin incorporating RG was fabricated to serve as a cavity liner, and its effectiveness was investigated in vitro. The RG formulation exhibited high solubility and released high concentrations of Zn 2+ at various pH values. To assess the antibacterial properties of RG, six bacterial species detected in deep carious regions were incubated in the presence of RG. In vitro antibacterial testing against six bacterial species revealed that RG exhibited strong bactericidal effects against these prevalent bacteria. Furthermore, using a dentin model infected with Lactobacillus casei or Streptococcus mutans , the experimental resin containing RG demonstrated an effective bactericidal effect in the dentinal tubules, highlighting its potential as a promising material for cavity liners or pulp-capping applications.

Introduction: The co-existence of S. mutans and C. albicans is frequently detected in root caries and early child caries and is reported to be associated with recurrent caries. The aim of this study was to investigate the effects of potassium iodide (KI) in combination with toluidine blue O-mediated antimicrobial photodynamic therapy (aPDT) on S. mutans and C. albicans mixed-species biofilm, as well as the antibiofilm mechanisms involved., Methods: Mixed-species biofilm was constructed of S. mutans and C. albicans on dentin blocks. The antibiofilm efficacy, cytotoxicity and antibiofilm mechanism of KI in combination with aPDT were determined and evaluated., Results: KI+TBO-aPDT treatment caused reduction in microorganism counts, metabolic activity, and biofilm biomass of mixed-species biofilm without inducing cytotoxicity to hDPCs (human dental pulp cells). Observations such increased ROS (reactive oxygen species) levels, impaired cell membrane function, cell apoptosis and reduced expression in several genes seem to be artifacts of reduced growth and general killing by KI+TBO-aPDT treatment., Discussion: These data suggested that KI in combination with aPDT as an innovative approach to combat S. mutans and C. albicans biofilm, and thus as an optional treatment for caries., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Li, Huang, Du, Wang, Cai and Huang.)

Introduction: The objective of this study is to investigate the shear bonding strength of a glycoalkaloid, also a novel matrix metalloproteinase enzyme known as α-tomatine, on two different surfaces of dentin (sound & caries-affected) and its efficacy against cariogenic microorganisms using in vitro and in silico methods., Methods: The effect of a-tomatine at different concentrations (0.75 / 1 / 1.5 µM) on shear bonding strength in caries-affected and sound dentin was also investigated (n = 10; each per subgroup). The analysis of shear bonding and failure tests was conducted after a 24-hour storage period. Fracture surfaces were examined under a scanning electron microscope. A stock solution 3 mM of a-tomatine was prepared for antimicrobial evaluation. Antimicrobial activities of the agents against Streptococcus mutans ATCC 25175, Lactobacillus casei ATCC 4646, and Candida albicans ATCC 10231 standard strains were investigated by microdilution method. In addition, through the method of molecular docking and dynamic analysis, the affinity of a-tomatine for certain enzymes of these microorganisms was examined., Results: The pretreatment agent and dentin type significantly influenced shear bonding strength values (p < 0.05). As the molarity of a-tomatine increased, the bonding value decreased in sound dentin, while the opposite was true in caries-affected dentin. According to molecular docking and dynamic analysis, the highest affinity was observed in L. casei's signaling protein. Microdilution assays revealed a-tomatine to exhibit fungicidal activity against C. albicans and bacteriostatic effects against S. mutans. No antimicrobial effect was observed on L. casei., Conclusion: a-tomatine demonstrates a positive impact by serving as both a pretreatment agent for bonding strength and an inhibitor against certain cariogenic microorganisms., (© 2024. The Author(s).)

This study aimed to assess the antibiofilm effects of dentin desensitizers using a modified Robbins device flow cell system. The test desensitizers were Saforide, Caredyne Shield, and Clinpro White Varnish. Standardized dentin specimens were prepared from human single-rooted premolars, treated with one of the materials, and mounted on the modified Robbins device flow cell system. Streptococcus mutans biofilms were developed for 24 h at 37 °C under anaerobic conditions. Scanning electron microscopy, fluorescence confocal laser scanning microscopy, viable and total cell counts, acid production, and gene expression analyses were performed. A wavelength-dispersive X-ray spectroscopy electron probe microanalyzer was used to analyze the ion incorporations. Clinpro White Varnish showed the greatest inhibition, suggesting its suppression of bacterial adherence and transcription of genes related to biofilm formation. Saforide reduced only the number of viable bacteria, but other results showed no significant difference. The antibiofilm effects of Caredyne Shield were limited. The uptake of ions released from a material into dentin varies depending on the element. Clinpro White Varnish is effective for the short-term treatment of tooth sensitivity due to dentin demineralization. It prioritizes remineralization by supplying calcium and fluoride ions while resisting biofilm formation.

Background and Objectives: The purpose of this study was to evaluate the antimicrobial activity of methylene blue dye with 660-nm diode laser in cavity disinfection and to compare the total bacterial count in dentinal samples preexcavation, postexcavation, and postdisinfection. The study design was experimental in vivo., Materials and Methods: Fifteen children aged 5-12 years with 15 primary molars requiring atraumatic restorative treatment (ART) were selected. Dentinal samples were collected from the carious tooth with a sterile spoon excavator three times, i.e., before caries excavation (preexcavation), after hand excavation of caries (postexcavation), and after doing laser photodynamic therapy with methylene blue dye and 660-nm diode laser (postdisinfection) and sent for microbiological analysis. The samples were transported in reduced transport fluid. The total bacterial count was carried out by an experienced microbiologist immediately after reporting., Results: A significant number of bacteria was present after hand excavation of caries using ART. The bacterial count significantly reduced postlaser disinfection. The total viable bacterial count was reduced to 87.17% with hand excavation of caries, whereas it was reduced to 98.59% postlaser disinfection using methylene blue dye with 660-nm diode laser., Conclusion: It was seen that ART cannot eliminate all the microorganisms. Methylene blue dye with a 660-nm diode laser was able to reduce the total bacterial count and can be effectively used as an antimicrobial cavity disinfectant., (Copyright © 2025 Journal of Indian Society of Pedodontics and Preventive Dentistry.)

Objectives: To compare, in vitro, the efficacy of three proprietary silver diamine fluoride (SDF) products in mitigating progression of dentinal caries induced by an inter-kingdom, dual-species, bacterial-yeast biofilm., Methods: Human dentin blocks were demineralized to create artificial caries lesions and randomized into three SDF test groups: Saforide, Topamine, T-SDF, and an aqueous control (n = 26 per group). After application of foregoing SDF variants, the blocks were incubated with Streptococcus mutans and Candida albicans for 24 h for biofilm development, and subsequently subjected to a microbe-induced, pH-cycling process for 7 days, to mimic the oral eco-system. The biofilm cell viability and surface topography were assessed by colony-forming units (CFUs) and scanning electron microscopy respectively. The lesion depth and mineral density were evaluated by micro-computer tomography. SDF precipitate and matrix-to-mineral ratio were evaluated by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. Standard, accepted methodology was used for all these evaluations and procedures., Results: After pH cycling, the SDF groups demonstrated comparable inhibition of the biofilm relative to the control. the log CFU of S. mutans for Saforide, Topamine, T-SDF, and control were 6.69±0.73, 6.48±0.56, 6.63±0.66, and 8.01±0.45, respectively. For C. albicans, the log CFU were 4.86±0.44, 4.72±0.53, 4.92±0.29, and 5.60±0.27, respectively. The log CFU of S. mutans and C. albicans in the SDF groups were significantly lower than the control group (p<0.001). Further, the lesion depth decreased by approximately 14.79±7.00% in the SDF groups, while it increased by 11.07±8.61% in the control (p<0.001), and the mineral density increased by 16.36±4.58% in the SDF group, as opposed to a 5.59±2.64% reduction in the control (p<0.001) implying their caries mitigating effect. These findings were corroborated by SEM images of the lesions., Conclusion: SDF significantly mitigated dentin caries due to an assault by a polymicrobial plaque biofilm whilst arresting mineral loss and lesion growth. There was no difference in the caries-arresting efficacy of the compared SDF variants., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kaewkamchai et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. To measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study., Methodology: adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity., Results: although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains' site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated., Conclusion: although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

Purpose/Aim: Acrylamides are hydrolytically stable at pH lower than 2, and were shown to preserve bonded interface integrity with two-step, total etch adhesives. The objective of this study was to leverage those two characteristics in self-etching primers containing the acidic monomer 10-MDP and test the microtensile bond strength before and after incubation with S. mutans incubation. Materials and Methods: Acidic primers (10 wt % 10-methacryloyloxydecyl dihydrogen phosphate─10-MDP; 45 wt % N , N -diethyl-1,3-bis(acrylamido)propane─DEBAAP, or 2-hydroxyethyl methacrylate─HEMA; 45 wt %, glycerol-dimethacrylate─GDMA) and adhesives (DEBAAP or HEMA/10-MDP/UDMA 45/10/45 wt %) were made polymerizable by the addition of 0.2 wt % camphorquinone, 0.8 wt % ethyl-4-dimethylaminobenzoate, 0.4 wt % diphenyliodonium hexafluorophosphate, and 0.1 wt % butylhydroxytoluene. Nonsolvated materials were characterized for flexural strength (FS), modulus (E), toughness, water sorption/solubility (WS/SL), contact angle, and vinyl conversion (DC). Viscosity was evaluated after adding 20 and 40 vol % ethanol to the primer and adhesive, respectively. The experimental materials or Clearfil SE Bond (CC─commercial control) were used to bond a commercial composite (Filtek Supreme) to the flat surface of human dentin. Microtensile bond strength (MTBS) was tested in 1 mm 2 sticks for the 5 primer/bond combinations: CC (Clearfil Bond Primer and Bond), HH (HEMA/HEMA), DD (DEBAAP/DEBAAP), HD (HEMA/DEBAAP), and DH (DEBAAP/HEMA). Prior to testing, sticks were stored in water or biofilm-inducing culture medium with S. mutans for 1 week. Confocal images and FTIR-ATR evaluation evaluated the hybrid layer of the adhesives. Results were analyzed using Student's t -test (WS, SL, DC, contact angle, FS, E, toughness), one-way ANOVA/Tukey's test for viscosity, and two-way ANOVA/Tukey's test for MTBS (95%). Results: HEMA-based materials had lower contact angle ( p = 0.004), higher WS ( p < 0.001), and similar SL values compared to DEBAAP ( p = 0.126). FS ( p = 0.171) and E ( p = 0.065) dry values were similar, but after one week of water storage, FS/E dropped more significantly for HEMA materials. Dry and wet toughness was greater for DEBAAP ( p < 0.001), but it also had the greatest drop (46%). Clearfil bonds had the highest viscosity, followed by DEBAAP and HEMA, respectively ( p = 0.002). For the primers, HEMA had the lowest viscosity ( p = 0.003). As far as MTBS, all groups tested in water were statistically different when compared with HH ( p < 0.001). After storage in biofilm, DH had the highest MTBS value, being statistically different from HH ( p = 0.002), CC ( p = 0.015), and DD ( p = 0.027). Conclusions: The addition of a diacrylamide and its association with HEMA in self-etching adhesive systems provided greater bonding stability after bacterial challenge.

Aim: To investigate the dynamic recovery of biofilms within dentinal tubules after primary irrigation with different protocols, and to evaluate the efficacy of various re-irrigation protocols on recovered biofilm, considering factors such as smear layer, nutrient conditions, and primary irrigants., Methodology: A total of 416 mono or multi-species biofilms samples were prepared from human teeth and incubated for 3 weeks. After inducing a smear layer on half of the samples, all specimens were irrigated with one of the following irrigant sequences: (1) 6% NaOCl +17% EDTA, (2) 6% NaOCl +8.5% EDTA, (3) 6% NaOCl and (8.5% EDTA +10 μg/mL DJK-5 antimicrobial peptide), or (4) sterile water. Thirty-two samples were used to assess immediate effect, whilst the rest were re-incubated to assess biofilms recovery. Nutrient conditions were defined based on whether culture media were changed (nutrient-rich) or not (nutrient-poor) during re-incubation. After 16 weeks, recovered biofilms underwent re-irrigation using four additional protocols, with or without DJK-5 peptide, based on primary irrigants. Confocal laser scanning microscopy was employed to evaluate immediate irrigant effects, biofilms recovery intervals (1, 3, 5, 8, 12, and 16 weeks after primary irrigation), and re-irrigation effects at the 16-week. Statistical analysis included one-way anova and two-way mixed anova tests., Results: The DJK-5 peptide irrigation protocols demonstrated the highest killing rates during primary irrigation and resulted in a longer biofilms recovery time of 16 weeks compared to non-peptide protocols (p < .001). Both primary irrigation type and smear layer presence significantly influenced biofilms recovery (p < .001). In the absence of smear layer, re-irrigation efficacy didn't significantly differ from primary irrigation, regardless of primary irrigation type or nutrient conditions. However, with a smear layer present, re-irrigation led to significantly higher proportion of dead bacteria compared to primary irrigation (p < .05). Inclusion of the DJK-5 peptide into the re-irrigation protocol displayed superior killing rate compared to other protocols (p < .001)., Conclusions: Biofilms exhibited susceptibility to both peptide and non-peptide protocols during re-irrigation, irrespective of nutrient conditions or primary irrigation protocols. The DJK-5 peptide irrigation protocols consistently displayed superior effectiveness compared to non-peptide protocols., (© 2024 The Author(s). International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society.)

The purpose of this study was to evaluate the antibacterial effect of silver nanoparticles (AgNPs) against Enterococcus faecalis and compare it with different irrigation solutions. This study was performed using 64 dentin blocks. E. faecalis suspension was dispensed to each sample and incubated under anaerobic conditions at 37°C throughout 21 days. After the inoculation period, the following solutions were added to each group and kept for 5 min: Group 1, 5.25% sodium hypochlorite (NaOCl); Group 2, 2.5% NaOCl; Group 3, 1% NaOCl; Group 4, 2% chlorhexidine (CHX); Group 5, 200 ppm hypochlorous acid (HOCl); and Group 6, AgNPs. The samples of positive control were treated with sterile saline. Biofilm viability assay was performed using the LIVE/DEAD BacLight Bacterial Viability Kit. Samples were examined using confocal laser scanning microscopy, respectively. There was no significant difference between the 5.25% NaOCl, 2.5% NaOCl, and 1%NaOCl groups (p > .05). However, these groups showed statistically higher antibacterial activity than the 2% CHX, 200 ppm HOCl, and AgNP groups. Also, 2% CHX showed greater percentage of dead cells compared with the AgNP and HOCl groups. While AgNPs group showed lower dead cell rate than all NaOCl groups and 2% CHX, it caused higher dead cells than 200 ppm HOCl group. The 200 ppm HOCl group showed the lowest percentage of dead cells (p < .05) Although the antibacterial effect of AgNPs is not as high as NaOCl and CHX, it has considerable bactericidal activity against E. faecalis and can be improved by further studies. RESEARCH HIGHLIGHTS: New antimicrobial approaches for root canal irrigation. Antimicrobial effect of silver nanoparticles against E. faecalis. Elimination of the biofilm layer for the success of endodontic treatment., (© 2024 The Authors. Microscopy Research and Technique published by Wiley Periodicals LLC.)

The aim of the study was to measure the degree of dentine surface roughness caused by five distinct lasers used to treat dentine hypersensitivity, as well as to evaluate the subsequent bacterial colonization on these irradiated surfaces. Sixty human maxillary premolar teeth without caries or restoration which were extracted for periodontal reasons were used in this study. Five different types of lasers were applied to the root dentin surface. Tested samples were divided into six groups of 10 samples each; control, diode (810 nm), diode (980 nm), Nd: YAG, Er: YAG, and Er, Cr: YSGG laser groups. The arithmetic mean of the surface roughness values (Ra) and the average roughness over a measurement area (Sa) were measured pre- and post-application using any of the laser types. Swab samples were then collected from the dentin surface. Following a 24-hour incubation period at 37 °C, the colony forming units were counted using a stereoscope. The results demonstrated a statistically significant difference in the surface roughness values pre- and post-application (Ra and Sa, respectively) in the Er, Cr: YSGG laser group (p = 0.037,p = 0.007). No significant difference was observed in the other groups (p > 0.05). There was no statistically significant difference in the number of bacterial colonies observed between the test and control groups. Diode and Nd: YAG lasers showed either a decrease or no change in surface roughness; however, the hard tissue lasers (Er: YAG, Er, Cr: YSGG) showed an increase. The Er: YAG and Nd: YAG laser groups exhibited decreased bacterial adhesion compared to the other groups., (© 2024. The Author(s).)

Background: This study aimed to engineer and optimise a dysbiotic biofilm model to develop in vitro root caries for investigating microbial modulation strategies. The model involved growing complex biofilms from a saliva inoculum collected from four volunteers using two strategies. In the first strategy ("pre-treatment strategy"), bovine root slabs were used, and two natural compounds were incorporated at time 0 of the 10-day biofilm experiment, which included sucrose cycles mimicking the cariogenic environment. In the second strategy ("post-treatment strategy"), mature biofilms were grown in a modified Calgary biofilm device coated with collagen and hydroxyapatite for 7 days and then were exposed to the same natural compounds. The metatranscriptome of each biofilm was then determined and analysed. Collagenase activity was examined, and the biofilms and dentine were imaged using confocal and scanning electron microscopy (SEM). Mineral loss and lesion formation were confirmed through micro-computed tomography (μ-CT)., Results: The pH confirmed the cariogenic condition. In the metatranscriptome, we achieved a biofilm compositional complexity, showing a great diversity of the metabolically active microbiome in both pre- and post-treatment strategies, including reads mapped to microorganisms other than bacteria, such as archaea and viruses. Carbohydrate esterases had increased expression in the post-treated biofilms and in samples without sugar cycles, while glucosyltransferases were highly expressed in the presence of sucrose cycles. Enrichment for functions related to nitrogen compound metabolism and organic cyclic component metabolism in groups without sucrose compared to the sucrose-treated group. Pre-treatment of the roots with cranberry reduced microbial viability and gelatinase (but not collagenase) activity (p < 0.05). SEM images showed the complexity of biofilms was maintained, with a thick extracellular polysaccharides layer., Conclusions: This root caries model was optimized to produce complex cariogenic biofilms and root caries-like lesions, and could be used to test microbial modulation in vitro. Pre-treatments before biofilm development and cariogenic challenges were more effective than post-treatments. The clinical significance lies in the potential to apply the findings to develop varnish products for post-professional tooth prophylaxis, aiming at implementing a strategy for dysbiosis reversal in translational research. Video Abstract., (© 2024. The Author(s).)

Objective: To evaluate the effect of the association of potassium iodide to antimicrobial photodynamic therapy on human carious dentin produced with a microcosm biofilm model., Methods: A microcosm biofilm model was used to generate a caries lesion on human dentin. Pooled human saliva diluted with glycerol was used as an inoculum on specimens immersed on McBain artificial saliva enriched with 1 % sucrose (24 h at 37 °C in 5 % CO 2 ). After refreshing culture media for 7 days, the dentin specimens were divided in 5 groups (3 specimens per group, in triplicate; n = 9): C (NaCl 0.9 %), CX (2 % chlorhexidine), PKI (0.01 % methylene blue photosensitizer+50 mM KI), L (laser at 15 J, 180 s, 22.7 J/cm 2 ), and PKIL (methylene blue + KI + Laser). After the treatments, dentin was collected, and a 10-fold serial dilution was performed. The number of total microorganisms, total lactobacilli, total streptococci, and Streptococcus mutans was analyzed by microbial counts (CFU/mL). After normality and homoscedasticity analysis, the Welch's ANOVA and Dunnett's tests were used for CFU. All tests used a 5 % significance level., Results: CX and PKIL groups showed significant bacterial decontamination of dentin, compared to group C (p < 0.05) reaching reductions up to 3.8 log 10 for CX for all microorganisms' groups and PKIL showed 0.93, 1.30, 1.45, and 1.22 log 10 for total microorganisms, total lactobacilli, total streptococci, and S. mutans, respectively., Conclusion: aPDT mediated by the association of KI and methylene blue with red laser reduced the viability of microorganisms from carious dentin and could be a promising option for cavity decontamination., Competing Interests: Declaration of Competing Interest The authors have no potential conflict of interest to declare., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study assessed canal preparation effects on disinfection and dentin preservation. Thirty mandibular incisors were paired into two experimental groups (n = 10). Following contamination, the initial microbial sample was collected. Instruments 30/0.03 (Group 1) and 30/0.05 (Group 2) were employed and a second sample was obtained. Canals were enlarged using instruments 40/0.03 and 40/0.05, respectively, and a third sample was collected. Final irrigation was performed, and sample S4 obtained. A final scan evaluated volume, surface area, unprepared areas, removed dentin and dentin thickness. Data were analysed using Student t-test, Mann-Whitney, Kruskal-Wallis and Dunn tests. A significant difference was observed between S1 and other time points (p < 0.05). Comparison between groups showed no differences in bacterial loads and in the percentage of microbial reduction (p > 0.05). Group 2 exhibited greater reduction in dentin thickness than group 1 in the mesial aspect of the root (p < 0.05). Instrument 30/0.03 might provide effective disinfection and safety during mandibular incisors canal preparation., (© 2024 Australian Society of Endodontology Inc.)

The aim of this study was to assess whether calcium silicate root fillings prevent bacterial penetration and to determine how bacteria penetrate roots. Extracted single-rooted, single-canal human teeth were decoronated, prepared and filled with ProRootMTA or Biodentine (n = 12 each). Positive and negative (n = 2 each) controls were not filled. A two-chamber model was used with Streptococcus gordonii. The lower compartment was evaluated for turbidity over 150 days. Roots were split and examined for bacteria via SEM. The chi-squared test was used for comparisons (α = 0.05). Experimental groups had bacteria in their coronal thirds. Tubules contained bacteria in 90.9% and 91.7% of areas examined in the Biodentine and ProRootMTA groups, respectively, with no significant difference (p = 0.914). Experimental and negative roots had no turbidity with no significant difference between Biodentine and ProRootMTA (p = 1.000). Positive controls had turbidity. Bacteria penetrate roots via dentine tubules of root-filled teeth. Biodentine was comparable to ProRoot MTA., (© 2024 The Authors. Australian Endodontic Journal published by John Wiley & Sons Australia, Ltd on behalf of Australian Society of Endodontology Inc.)

To incorporate different concentrations of Al 2 O 9 Zr 3 (1%, 5%, and 10%) nanoparticles (NP) into the ER adhesive and subsequently assess the impact of this addition on the degree of conversion, μTBS, and antimicrobial efficacy. The current research involved a wide-ranging examination that merged various investigative techniques, including the application of scanning electron microscopy (SEM) for surface characterization of NP coupled with energy-dispersive x-ray spectroscopy (EDX), Fourier-transform infrared (FTIR) spectroscopy, μTBS testing, and microbial analysis. Teeth were divided into four groups based on the application of modified and unmodified three-step ER adhesive primer. Group 1 (0% Al 2 O 9 Zr 3 NPs) Control, Group 2 (1% Al 2 O 9 Zr 3 NPs), Group 3 (5% Al 2 O 9 Zr 3 NPs), and Group 4 (10% Al 2 O9Zr3 NPs). EDX analysis of Al 2 O 9 Zr 3 NPs was performed showing elemental distribution in synthesized NPs. Zirconium (Zr), Aluminum (Al), and Oxides (O 2 ). After primer application, an assessment of the survival rate of Streptococcus mutans was completed. The FTIR spectra were analyzed to observe the characteristic peaks indicating the conversion of double bonds, both before and after the curing process, for the adhesive Etch and rinse containing 1,5,10 wt% Al 2 O 9 Zr 3 NPs. μTBS and failure mode assessment were performed using a Universal Testing Machine (UTM) and stereomicroscope respectively. The μTBS and S.mutans survival rates comparison among different groups was performed using one-way ANOVA and Tukey post hoc (p = .05). Group 4 (10 wt% Al 2 O 9 Zr 3 NPs + ER adhesive) specimens exhibited the minimum survival of S.mutans (0.11 ± 0.02 CFU/mL). Nonetheless, Group 1 (0 wt% Al 2 O 9 Zr 3 NPs + ER adhesive) displayed the maximum surviving S.mutans (0.52 ± 0.08 CFU/mL). Moreover, Group 2 (1 wt% Al 2 O 9 Zr 3 NPs + ER adhesive) (21.22 ± 0.73 MPa) samples displayed highest μTBS. However, the bond strength was weakest in Group 1 (0 wt% Al 2 O 9 Zr 3 NPs + ER adhesive) (14.13 ± 0.32 MPa) study samples. The etch-and-rinse adhesive exhibited enhanced antibacterial activity and micro-tensile bond strength (μTBS) when 1% Al 2 O 9 Zr 3 NPs was incorporated, as opposed to the control group. Nevertheless, the incorporation of Al 2 O 9 Zr 3 NPs led to a decrease in DC. RESEARCH HIGHLIGHTS: 10 wt% Al 2 O 9 Zr 3 NPs + ER adhesive specimens exhibited the minimum survival of S.mutans. 1 wt% Al 2 O 9 Zr 3 NPs + ER adhesive samples displayed the most strong composite/CAD bond. The highest DC was observed in Group 1: 0 wt% Al 2 O 9 Zr 3 NPs + ER adhesive., (© 2024 Wiley Periodicals LLC.)

Background: The global rise in the elderly population has increased the prevalence of root caries. Streptococcus mutans, Lactobacilli and Actinomyces are considered the primary pathogens of dental caries in culture-based studies. This study aimed to investigate bacterial profiles in coronal and root caries lesions and determine the association of specific bacterial genera at each site., Methods: Dentine samples from carious lesions were collected from 22 extracted teeth using an excavator. Microbial DNA was extracted from the samples using a protocol developed for this study. 16S rRNA gene amplicon sequencing was employed for microbial analysis. PCR amplification targeted the V3-V4 region of the bacterial 16S rRNA, and the amplicon sequencing used an Illumina MiSeq system (2 × 300 bp paired-end reads). Statistical analysis was performed by the Phyloseq and DESeq2 packages in R., Results: In coronal caries, Olsenella, Lactobacillus and Prevotella were the most prevalent genera, comprising approximately 70% of the microbiome community. In the root caries, however, although Olsenella, Prevotella and Lactobacillus remained the dominant genera, they accounted for only half of the microbiome community. This study identified significant differences in alpha diversity indices between the coronal and root caries. LEfSE analysis revealed several unique genera in each caries lesion., Conclusion: The microbiome of root caries lesions was richer and more complex than the coronal caries microbiota. The results suggest that lesion-related variations in the oral microflora may be detected in carious dentine., (© 2024. The Author(s).)

The purpose of this study was to evaluate the antibacterial efficacy of using 2.5% NaOCl, 2% chlorhexidine (CHX), Irritrol, and chitosan-coated silver nanoparticles (AgCNPs) alone or in combination with deoxyribonuclease I (DNase I) and trypsin pre-enzyme applications in dentin samples contaminated with Enterococcus faecalis (E. faecalis) by CLSM. 144 dentin blocks with confirmed E. faecalis biofilm formation were divided randomly according to the irrigation protocol (n = 12): NaOCl, CHX, Irritrol, AgCNPs, trypsin before NaOCl, CHX, Irritrol, AgCNPs, and DNase I before NaOCl, CHX, Irritrol, AgCNPs. Dentin blocks were stained with the Live/Dead BacLight Bacterial Viability Kit and viewed with CLSM after irrigation applications. The percentage of dead and viable bacteria was calculated using ImageJ software on CLSM images. At a significance level of p < 0.05, the obtained data were analyzed using one-way Anova and post-hoc Tukey tests. In comparison with NaOCl, CHX had a higher percentage of dead bacteria, both when no pre-enzyme was applied and when DNase I was applied as a pre-enzyme (p < 0.05). There was no difference in the percentage of dead bacteria between the irrigation solutions when trypsin was applied as a pre-enzyme (p > 0.05). AgCNPs showed a higher percentage of dead bacteria when trypsin was applied as a pre-enzyme compared to other irrigation solutions (p < 0.05), while the pre-enzyme application did not affect the percentage of dead bacteria in NaOCl, CHX, and Irritrol (p > 0.05). No irrigation protocol tested was able to eliminate the E. faecalis biofilm. While the application of trypsin as a pre-enzyme improved the antimicrobial effect of AgCNPs, it did not make any difference over other irrigation solutions., (© 2024. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

The aim of this systematic review and meta-analysis (SRM) was to evaluate whether bioceramic sealers have better penetration capacity in dentinal tubules and antimicrobial activity when compared to AH Plus® sealer. This SRM was recorded in the Open Science Framework database and followed the guidelines of the PRISMA 2020. Five databases were searched by two independent reviewers. Only in vitro studies that evaluated the effects of bioceramic sealers on dentinal tubule penetration and antimicrobial activity outcomes compared to AH Plus® sealer were included. Meta-analysis was conducted using R software, using the effect measure of the standardized mean difference (SMD) and inverse variance method. A modified Joanna Briggs Institute's Checklist was used for the risk of bias assessment. A total of 1486 studies were identified, and only 54 studies that fulfilled our eligibility criteria were included in this review. There was no statistical difference between the sealers evaluated for dentinal tubule penetration, in the thirds evaluated: coronal SMD 0.58 [0.14; 1.31], p = 0.12; middle SMD 0.07 [0.54; 0.39], p = 0.75; and apical SMD 0.08 [0.73; 0.56], p = 0.80. Both sealers demonstrated similar antimicrobial action (SMD [3.42; 5.32], p = 0.67 and SMD 0.67 [1.89; 0.55], p = 0.28). The studies presented a low risk of bias. Based on the in vitro studies included and according to the limitations of the present review, the data suggest that bioceramic and AH Plus® sealers present similar penetration capacity in dentinal tubules and antimicrobial effect, making them suitable materials to be considered in clinical practice., (© 2024. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

This study aimed to evaluate the antimicrobial activity of calcium hypochlorite (Ca (OCl)2) and sodium hypochlorite (NaOCl) using confocal laser scanning microscopy (CLSM) and dentin organic matrix alteration by picrosirius staining and light microscopy (LM). Samples of human extracted teeth were infected with Enterococcus faecalis by centrifugation of the bacterial suspension and were treated with Ca(OCl)2 or NaOCl at 0.5%, 2.5%, and 6% for 15, 30, and 60 seconds. CLSM and viability staining were used to quantitatively analyze the proportions of dead/live bacteria in the canal lumen and border of the root canal. The data were analyzed by ANOVA and Fisher test. For LM analysis, one hundred bovine teeth were randomly divided into 10 test groups (n=10): G1- Without treatment; G2- 17% EDTA; G3- 6% NaOCl; G4- 6% NaOCl + EDTA; G5- 0.5% Ca(OCl)2; G6- 0.5% Ca(OCl)2 + EDTA; G7- 2.5% Ca(OCl)2; G8- 2.5% Ca(OCl)2 + EDTA; G9- 6% Ca(OCl)2; G10- 6% Ca(OCl)2 + EDTA. The samples were fragmented and stained with Picrosirius. Data were analyzed by Kruskal-Wallis and Dunn (P<0.05). There was a strong correlation between the results of the canal lumen and the border of the root canal (r=0.962). Both hypochlorites at a concentration of 0.5% showed less microbial reduction compared to 2.5% and 6% (P<0.05). There was less antimicrobial activity at 15 seconds compared to 30 and 60 seconds (P<0.05). Ca(OCl)2 and NaOCl showed similar results at the same concentrations (P>0.05). In conclusion, Ca(OCl)2 caused fewer alterations to the dentin organic matrix at concentrations of 0.5% and 2.5%. Ca(OCl)2 presents antimicrobial activity similar to NaOCl, and collagen damage is concentration-dependent.

Objective: To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions., Methodology: bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters., Results: All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Aims: To assess the impact of various cavity disinfectants PC-PDT (Phycocyanin activated by Photodynamic therapy), PC@AgNPs-PDT (Phycocyanin and silver nanoparticles activated by PDT), and Nd: YAG laser on the survival rate of S.mutans and the bond integrity of composite restoration METHODS: Sixty human mandibular molars that scored 4 and 5 based on ICDAS criteria were included. The infected dentin was removed while the CAD was preserved based on visual, tactile, and staining assessment. S.mutans were cultured on the CAD of twenty samples. All the specimens were indiscriminately distributed into four groups based on cavity disinfection (n=20 each includes n = 5 each group incubated with S.mutans) Group 1: CHX, Group 2: Nd:YAG laser, Group 3: PC-PDT and Group 4: PC@AgNPs-PDT. S.mutans survival rate was assessed for each group(n = 5). Forty samples underwent composite bonding for SBS and failure mode assessment using universal testing machine (UTM) and stereomicroscope. The calculations for the mean and standard deviation (SD) and their comparison among different groups were performed using a one-way analysis of variance (ANOVA) and the Tukey post hoc test (p ≤ 0.05) RESULTS: CAD surface treated disinfected with PC@AgNPs-PDT yielded the lowest survival rates (0.13 ± 0.05 CFU/ml) and highest SBS (17.23 ± 1.45 MPa). Group 1 (CHX) unveiled the highest survival rate of S.mutans (0.33 ± 0.12 CFU/ml). However, Group 2 (Nd:YAG Laser) (11.87 ± 0.67 MPa) presented the lowest SBS CONCLUSION: The combination of Phycocyanin loaded with silver nanoparticles and activated with Photodynamic therapy demonstrates the highest antimicrobial potential and bond strength of composite restorations., (Copyright © 2024. Published by Elsevier B.V.)

Introduction: Bacterial fluorescence methods are of interest in endodontics for informing endpoints for debridement. This study explored potential fluorescence quenching reversal effects of a water-soluble vitamin E conjugate (d-α-Tocopherol polyethylene glycol 1000 succinate, TPGS) when applied to polymicrobial biofilms grown on dentine that had been exposed to sodium hypochlorite (NaOCl) to cause quenching., Method: Extracted human teeth were debrided, embedded in transparent acrylic resin and sectioned. After smear layer removal, tooth dentine sections were inoculated with a polymicrobial inoculum, and cultured for 7 days to create biofilms. Samples (n = 8 per group) were exposed to 1 % or 4 % NaOCl for 2 or 4 min, and then treated with TPGS. Bacterial fluorescence readings under laser excitation at 655 nm were assessed over 10 min using a calibrated DIAGNOdent device. All data were assessed for normality (Kolmogorov-Smirnov test) and analysed with ANOVA followed by Bonferroni post-hoc tests., Results: NaOCl at both concentrations quenched fluorescence readings of biofilms grown on dentine samples, with a maximal reduction of 40.4 % at 5 min after 4 % NaOCl. Treatment with TPGS gave faster recovery of fluorescence readings compared to the control at 5 and 10 min., Conclusion: The water-soluble antioxidant TPGS partially reversed fluorescence quenching caused by NaOCl. This agent may have value clinically for reducing the time needed for fluorescence readings to recover when NaOCl is used as an irrigant. This will facilitate more accurate assessment of endpoints for canal debridement., Competing Interests: Declaration of competing interest The authors report no conflicts of interest., (Copyright © 2024. Published by Elsevier B.V.)

The role of bacteria in the etiology of dental caries is long established, while the role of fungi has only recently gained more attention. The microbial invasion of dentin in advanced caries especially merits additional research. We evaluated the fungal and bacterial community composition and spatial distribution within carious dentin. Amplicon 16S rRNA gene sequencing together with quantitative PCR was used to profile bacterial and fungal species in caries-free children (n = 43) and 4 stages of caries progression from children with severe early childhood caries (n = 32). Additionally, healthy (n = 10) and carious (n = 10) primary teeth were decalcified, sectioned, and stained with Grocott's methenamine silver, periodic acid Schiff (PAS) and calcofluor white (CW) for fungi. Immunolocalization was also performed using antibodies against fungal β-D-glucan, gram-positive bacterial lipoteichoic acid, gram-negative endotoxin, Streptococcus mutans, and Candida albicans. We also performed field emission scanning electron microscopy (FESEM) to visualize fungi and bacteria within carious dentinal tubules. Bacterial communities observed included a high abundance of S. mutans and the Veillonella parvula group, as expected. There was a higher ratio of fungi to bacteria in dentin-involved lesions compared to less severe lesions with frequent preponderance of C. albicans, C. dubliniensis, and in one case C. tropicalis. Grocott's silver, PAS, CW and immunohistochemistry (IHC) demonstrated the presence of fungi within carious dentinal tubules. Multiplex IHC revealed that fungi, gram-negative, and gram-positive bacteria primarily occupied separate dentinal tubules, with rare instances of colocalization. Similar findings were observed with multiplex immunofluorescence using anti-S. mutans and anti-C. albicans antibodies. Electron microscopy showed monomorphic bacterial and fungal biofilms within distinct dentin tubules. We demonstrate a previously unrecognized phenomenon in which fungi and bacteria occupy distinct spatial niches within carious dentin and seldom co-colonize. The potential significance of this phenomenon in caries progression warrants further exploration., Competing Interests: The authors have no competing interests relating to this work., (Copyright: © 2024 Sulyanto et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objectives: To assess the growth of a multispecies biofilm on root canal dentin under different radiotherapy regimens., Materials and Methods: Sixty-three human root dentin cylinders were distributed into six groups. In three groups, no biofilm was formed (n = 3): NoRT) non-irradiated dentin; RT55) 55 Gy; and RT70) 70 Gy. In the other three groups (n = 18), a 21-day multispecies biofilm (Enterococcus faecalis, Streptococcus mutans, and Candida albicans) was formed in the canal: NoRT + Bio) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. The biofilm was quantified (CFUs/mL). Biofilm microstructure was assessed under SEM. Microbial penetration into dentinal tubules was assessed under CLSM. For the biofilm biomass and dentin microhardness pre- and after biofilm growth assessments, 45 bovine dentin specimens were distributed into three groups (n = 15): NoRT) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm., Results: Irradiated specimens (70 Gy) had higher quantity of microorganisms than non-irradiated (p = .010). There was gradual increase in biofilm biomass from non-irradiated to 55 Gy and 70 Gy (p < .001). Irradiated specimens had greater reduction in microhardness after biofilm growth. Irradiated dentin led to the growth of a more complex and irregular biofilm. There was microbial penetration into the dentinal tubules, regardless of the radiation regimen., Conclusion: Radiotherapy increased the number of microorganisms and biofilm biomass and reduced dentin microhardness. Microbial penetration into dentinal tubules was noticeable., Clinical Relevance: Cumulative and potentially irreversible side effects of radiotherapy affect biofilm growth on root dentin. These changes could compromise the success of endodontic treatment in oncological patients undergoing head and neck radiotherapy., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: To compare ultrasonic scaler prototypes based on a planar piezoelectric transducer with different working frequencies featuring a titanium (Ti-20, Ti-28, and Ti-40) or stainless steel (SS-28) instrument, with a commercially available scaler (com-29) in terms of biofilm removal and reformation, dentine surface roughness and adhesion of periodontal fibroblasts., Materials and Methods: A periodontal multi-species biofilm was formed on specimens with dentine slices. Thereafter specimens were instrumented with scalers in a periodontal pocket model or left untreated (control). The remaining biofilms were quantified and allowed to reform on instrumented dentine slices. In addition, fibroblasts were seeded for attachment evaluation after 72 h of incubation. Dentine surface roughness was analyzed before and after instrumentation., Results: All tested instruments reduced the colony-forming unit (cfu) counts by about 3 to 4 log 10 and the biofilm quantity (each p < 0.01 vs. control), but with no statistically significant difference between the instrumented groups. After 24-hour biofilm reformation, no differences in cfu counts were observed between any groups, but the biofilm quantity was about 50% in all instrumented groups compared to the control. The attachment of fibroblasts on instrumented dentine was significantly higher than on untreated dentine (p < 0.05), with the exception of Ti-20. The dentine surface roughness was not affected by any instrumentation., Conclusions: The planar piezoelectric scaler prototypes are able to efficiently remove biofilm without dentine surface alterations, regardless of the operating frequency or instrument material., Clinical Relevance: Ultrasonic scalers based on a planar piezoelectric transducer might be an alternative to currently available ultrasonic scalers., (© 2024. The Author(s).)

Objectives: This study aimed to compare the antimicrobial action, cytotoxicity, cleaning ability, and erosion of dentine of hypochlorous acid (HClO) obtained from an electrolytic device at two different concentrations (Dentaqua) and three concentrations of sodium hypochlorite (NaOCl)., Methods: Microbiological test-The root canals of sixty single-rooted extracted human teeth were inoculated with Enterococcus faecalis and divided into 6 groups (n = 10), according to decontamination protocol: DW (control); 1% NaOCl; 2.5% NaOCl; 5.25% NaOCl; 250 ppm HClO and 500 ppm HClO. The colony-forming units were counted to evaluate the decontamination potential of each group, calculating the reduction in bacterial percentage. Cytotoxicity test-Cytotoxicity was evaluated after inoculation of the same tested protocols in fibroblastic cells for 3 min, calculating the cell viability percentages. Specifical statistical analysis was performed (α = 5%). Cleaning ability and erosion-Fifty-six single-rooted bovine lower incisors were divided into seven groups of 8 roots each, being the test groups 1% NaOCl; 2.5% NaOCl; 5,25% NaOCl; 250 ppm HClO and 500 ppm HClO, and a negative and positive control. Negative control was not contaminated, and the other groups were inoculated with Enterococcus faecalis. SEM images were ranked as from the cleanest to the least clean. Erosion was also assessed, being ranked from the least to the most eroded dentine., Results: The highest bacterial reduction was observed in experimental groups, with no statistical differences between them (p > 0.05). The highest number of viable cells was observed in control group, followed by 250 ppm HClO and 500 ppm HClO groups, with statistical differences between them (p < 0.05). 1% NaOCl; 2.5% NaOCl; 5.25% NaOCl and 500 ppm HClO displayed the cleanest areas. All sodium hypochlorite groups displayed erosion with higher ranks with greater concentration, while hypochlorous acid did not display any erosion regardless the concentration., Conclusions: It is possible to conclude that HClO obtained from an electrolytic device presented high antimicrobial activity and low cytotoxicity in both tested concentrations. 500 ppm HClO did not display erosion and showed great cleaning ability., Clinical Relevance: The use of 500 ppm hypochlorous acid may reduce unfavorable behavior of sodium hypochlorite whilst maintaining its antimicrobial action., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: Dental caries results from an ecologic shift within the dental biofilm from a balanced population of microorganisms to an acidogenic, aciduric, and cariogenic microbiological population developed and maintained by frequent consumption of fermentable dietary carbohydrates. Total caries removal (TCR) of deep lesion may result in pulpal exposure requiring more invasive treatment. Hence, current pediatric dentistry has shifted to minimally invasive treatment that avoids more complex, time-consuming procedure, and the child's discomfort., Aim: The aim of this study is to evaluate and compare clinical performance and radiographic changes after complete and incomplete caries removal procedures., Materials and Methods: The study was conducted on 60 primary molars in children aged 6-9 years. Selected 60 primary molars were randomly divided into two groups. Group 1 (PCR): infected dentin was removed, while the affected dentin was maintained on the pulpal wall. Group 2 (TCR): both infected and affected dentin were removed through low-speed carbide bur and hand excavator. Teeth were evaluated at 4 and 6 months clinically and radiographically., Results: The proportion was compared using Fisher's exact test. The Statistical Package for the Social Sciences version 21 was used for analysis. The level of significance was kept at 5%., Conclusion: The clinical and radiographic success rates of ICR and CCR in primary teeth with deep carious lesions were high and did not differ significantly, indicating that the retention of carious dentin does not interfere with pulp vitality. Thus, ICR is a reliable minimally invasive approach that might replace the CCR in primary teeth when correctly indicated., (Copyright © 2024 Copyright: © 2024 Journal of Indian Society of Pedodontics and Preventive Dentistry.)

This study aimed to evaluate the effectiveness of two sodium hypochlorite concentrations at different exposure times and temperatures against Enterococcus faecalis biofilms of varying ages in human dentinal tubules. Dentin blocks were infected with E. faecalis for either 3 days or 3 weeks of incubation. Subsequently, the samples were exposed to sterile water, 2%, and 5.25% sodium hypochlorite for 3 and 10 min at 20 °C and 60 °C . Viability staining and confocal laser scanning microscopy were used to assess the proportion of killed bacteria in the dentinal tubules after exposure. There are no significant differences in the efficacy of E. faecalis killing between 2% sodium hypochlorite at 60 °C for various exposure times and 5.25% sodium hypochlorite at different temperatures or exposure times (P > 0.05). When both solutions were compared at the same temperatures with a 10-min exposure time, no significant differences in the effectiveness of E. faecalis killing between 2% and 5.25% sodium hypochlorite were observed (P > 0.05). To optimize the effectiveness of sodium hypochlorite in killing E. faecalis while minimizing potential damage to root dentin and soft tissue, clinicians should prioritize increasing the temperature or exposure time of sodium hypochlorite, rather than raising its concentration., (© 2023. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Introduction and Aim: The presence of host collagenases in the degradation of the protein matrix at later stages of carious dentin lesions development, as well as the potential involvement of bacterial collagenases, have been suggested but lack conclusive evidence. This study aims to conduct a systematic review to comprehensively assess the profile of host and bacterial-derived collagenolytic proteases in both root and coronal dentin carious lesions., Methods: The search was performed in eight databases and the grey literature. Studies evaluating ex vivo dentin, extracted teeth, or biofilms from natural caries lesions were included. The methodological quality of studies was assessed using the Joanna Briggs Institute tool. Synthesis of the results and the certainty of evidence were performed following the Synthesis without Meta-analysis (SWiM) checklist and GRADE approach for narrative synthesis, respectively., Results: From 935 recovered articles, 18 were included. Although the evidence was very uncertain, it was possible to suggest that 1) MMP-2, MMP-9, MMP-13, and CT-B may be increased in carious dentin when compared to sound dentin; 2) there is no difference in MMP-2 presence, while MMP-13 may be increased in root when compared to coronal carious dentin; 3) there is no difference of MMP-2 and MMP-9 expression/activity before and after cavity sealing; 4) MMP-8 may be increased in the dentin before cavity sealing compared to dentin after cavity sealing; 5) there is no difference of MMP-20 in irradiated vs. non-irradiated carious dentin. MMP-20 probably reduces in carious outer dentin when compared to carious inner dentin (moderate certainty). Genes encoding bacterial collagenolytic proteases and protein-degrading bacteria were detected in coronal and root carious lesions., Conclusion: Trends in the direction of the effect were observed for some collagenolytic proteases in carious dentin, which may represent a potential target for the development of new treatments. (Protocol register-PROSPERO: CRD42020213141)., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Barbosa, Monici Silva, Cena, Stefani and Dame-Teixeira.)

Aims: The conventional caries removal technique has been replaced with minimally invasive (MI) techniques to preserve healthy natural teeth and to provide durable dental restorations. Each of these MI caries removal protocols is reported to be favorable in dealing with different caries conditions. The current study aimed to trace the residual bacteria that may remain in a prepared cavity following a visual-tactile (VT), caries detection dye (CDD), and chemo-mechanical caries removal (CMCR) protocol., Materials and Methods: A total of 29 extracted human molar teeth with visible caries lesions were randomly divided into three groups. The cavity preparation and caries removal of each group was accomplished following one of the MI caries removal protocols. Swab samples (one from each specimen) were taken and inoculated onto a blood agar plate and incubated for 48 hours. The growth of the bacterial colony was observed under a microscope and the specific genome of the bacteria was identified by polymerase chain reaction (PCR) test., Results: The maximum number of traceable bacteria was observed following the chemo-mechanical caries removal group followed by the caries detection dye group and the least in the visual-tactile group. The PCR test revealed the presence of Streptococcus mutans in all the observed colonies; however, Streptococcus sobrinus was absent completely. The Chi-square test reveals a statistically insignificant ( p = 0.646) difference among the tested groups., Conclusion: All of the MI caries removal protocols used in this study showed a trace of microbes in certain teeth. The cavity prepared following a visual tactile protocol showed the least amount of traceable bacteria in the prepared cavity., Clinical Significance: Cavity that is prepared following individual MI protocol has a risk of leaving microbes in it.

Aims: The present study aimed to investigate the bond integrity and disinfection efficacy of Methylene blue(MB) alone, MB-PDT (Photodynamic therapy), MB@ carbon nanoparticles (CP)-PDT, and Cr, Cr: YSGG (ECL) against lactobacilli in Caries-affected dentin (CAD) MATERIAL AND METHODS: Methods consisted of Shear bond strength (SBS), Scanning electron microscopy (SEM), energy dispersive X-ray (EDX), methods of disinfection, and failure analysis. CAD samples were prepared and biofilm formed on the specimens randomly allocated into five groups based on disinfection. Group 1: CHX; Group 2: MB; Group 3: MB-PDT: group 4: MB@CP-PDT and group 5: ECL. After disinfection Colony forming units were measured and specimens were restored and positioned under a universal testing machine (UTM). Failure analysis was performed using a stereomicroscope. The difference in survival rate was assessed using the Kruskal-Walis test. Mean and standard deviation for bond values after different methods of disinfection was evaluated using analysis of variance (ANOVA) and Post Hoc Tukey. The significance level was p<0.05 RESULTS: Morphological analysis revealed that CPs under SEM are flat discs with edged irregular shapes. EDX analyses show a spike indicating carbon particles by more than 95%. MB@CP-PDT displayed the highest reduction in lactobacillus levels in comparison to the other disinfection methods. The highest SBS was exhibited by the CAD sample disinfected with ECL. The lowest SBS values in CAD specimens after cavity cleansing with MB alone. The predominant failure type in CAD disinfected with MB alone, ECL CHX, MB-PDT, and MB@CP-PDT was adhesive., Conclusion: The use of MB@CP-PDT showed high antibacterial potency against lactobacillus but demonstrated bond values similar to CHX. Use of Er, Cr: YSGG showed considerable effectiveness against lactobacillus along with the highest bond values., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The prevalence of dental caries in the Mexican adult population aged 20 to 85 years is around 93.3%, and 50% in Mexican children and adolescents. Worldwide, it is the most common non-communicable disease. One of the main etiological factors for dental caries is the oral microbiome and changes in its structure and function, with an expansion of pathogenic bacteria like Streptococcus mutans . The exposed dental pulp tissue triggers an innate immune response to counteract this bacterial invasion. The relation between oral dysbiosis and innate immune responses remains unclear. We aimed to understand the relationship between innate immune response and the oral microbiota by quantifying the expression of Toll-like receptors (TLRs) and proinflammatory markers (cytokines and a chemokine) in dental pulp tissue, either exposed or not to carious dentin, and to correlate this information with the oral microbiome found in healthy teeth and those with moderate caries. RNA was purified from pulp tissue, subjected to RT-qPCR and analysed with the ΔΔCt method. Supragingival dental plaque of non-carious teeth and dentin of carious teeth were subjected to 16S targeted sequencing. Principal coordinate analysis, permutational multivariate ANOVA, and linear discriminant analysis were used to assess differences between non-carious and carious teeth. Correlations were assessed with Spearman´s test and corrected for multiple comparisons using the FDR method. The relative abundance (RA) of Lactobacillus, Actinomyces, Prevotella , and Mitsuokella was increased in carious teeth; while the RA of Haemophilus and Porphyromonas decreased. Olsenella and Parascardovia were only detected in carious teeth. Significant overexpression of interleukin 1 beta (IL1 β), IL6, and CXCL8 was detected in pulp tissue exposed to carious dentin. IL1β correlated positively with TLR2 and Actinomyces ; yet negatively with Porphyromonas. These findings suggest that immune response of pulp tissue chronically exposed to cariogenic microbiome is triggered by proinflammatory cytokines IL1β and IL6 and the chemokine CXCL8., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gómez-García, López-Vidal, Pinto-Cardoso and Aguirre-García.)