Randomized controlled clinical trial on the efficacy of a novel antimicrobial chewing gum in reducing plaque and gingivitis in adolescent orthodontic patients.

OBJECTIVES: Chewing gums containing antiseptics or other antimicrobial substances may be effective in reducing plaque and gingivitis. Therefore, the aim of this randomized placebo-controlled clinical trial was to investigate the efficacy of a novel antimicrobial chewing gum containing essential oils (cinnamon, lemon, peppermint) and extracts on reduction of dental plaque and gingivitis as well as on oral health-related quality of life (OHRQoL) in adolescent orthodontic patients. MATERIALS: 52 patients (11-22 years of age) were randomly assigned to use a test chewing gum (COVIDGUM, Clevergum) or a commercially available control chewing gum over a period of 10 days. Approximal plaque index (API), papillary bleeding index (PBI) and an OHRQoL questionnaire for children (COHIP-G19) were assessed at baseline (BL), after 10 days (10d) and 30 days (30d). In addition, oral health and oral hygiene related questions of the COHIP-G19 questionnaire were evaluated separately in subscales at each timepoint. Data were analyzed using non-parametrical statistical procedures (α = 0.05). RESULTS: API and PBI decreased significantly over time from BL to 10d and from BL to 30d in both groups, without significant differences between the groups. In both groups, the COHIP-G19 score, oral health subscale and oral hygiene subscale decreased significantly over time. Regarding the oral hygiene subscale, the test group showed significantly better scores at 30d (p = 0.011). CONCLUSION: Both chewing gums performed similarly effective in terms of reducing plaque accumulation and gingival inflammation and improving OHRQoL. CLINICAL RELEVANCE: Chewing gums without antimicrobial ingredients may be sufficient to decrease plaque accumulation and gingival inflammation.

In-vitro-cytotoxicity of self-adhesive dental restorative materials.

OBJECTIVES: Although the introduction of self-adhesive composites in restorative dentistry is very promising, the innovation of new materials also presents challenges and unknowns. Therefore, the aim of this study was to investigate the cytotoxicity of four different self-adhesive composites (SAC) in vitro and to compare them with resin-modified glass ionomer cements (RM-GIC), a more established group of materials. METHODS: Samples of the following materials were prepared according to ISO 7405/10993-12 and eluted in cell culture medium for 24 h at 37 °C: Vertise Flow, Fusio Liquid Dentin, Constic, Surefil One, Photac Fil and Fuji II LC. Primary human pulp cells were obtained from extracted wisdom teeth and cultured for 24 h with the extracts in serial dilutions. Cell viability was evaluated by MTT assay, membrane disruption was quantified by LDH assay and apoptosis was assessed by flow cytometry after annexin/PI staining. RESULTS: Two SAC (Constic and Vertise Flow) and one RM-GIC (Photac Fil) significantly reduced cell viability by more than 30% compared to the untreated control (p < 0.001). Disruptive cell morphological changes were observed and the cells showed signs of late apoptosis and necrosis in flow cytometry. Membrane disruption was not observed with any of the investigated materials. CONCLUSION: Toxic effects occurred independently of the substance group and need to be considered in the development of materials with regard to clinical implications. CLINICAL SIGNIFICANCE: SAC have many beneficial qualities, however, the cytotoxic effects of certain products should be considered when applied in close proximity to the dental pulp, as is often required.

An in vitro coculture approach to study the interplay between dental pulp cells and Streptococcus mutans.

AIM: To develop a new coculture system that allows exposure of dental pulp cells (DPCs) to Streptococcus mutans and dentine matrix proteins (eDMP) to study cellular interactions in dentine caries. METHODOLOGY: Dental pulp cells and S. mutans were cocultured with or without eDMP for 72 h. Cell proliferation and viability were assessed by cell counting and MTT assays, while bacterial growth and viability were determined by CFU and LIVE/DEAD staining. Glucose catabolism and lactate excretion were measured photometrically as metabolic indicators. To evaluate the inflammatory response, the release of cytokines and growth factors (IL-6, IL-8, TGF-ß1, VEGF) was determined by ELISA. Non-parametric statistical analyses were performed to compare all groups and time points (Mann-Whitney U test or Kruskal-Wallis test; α = .05). RESULTS: While eDMP and especially S. mutans reduced the number and viability of DPCs (p ≤ .0462), neither DPCs nor eDMP affected the growth and viability of S. mutans during coculture (p > .0546). The growth of S. mutans followed a common curve, but the death phase was not reached within 72 h. S. mutans consumed medium glucose in only 30 h, whereas in the absence of S. mutans, cells were able to catabolize glucose throughout 72 h, resulting in the corresponding amount of l-lactate. No change in medium pH was observed. S. mutans induced IL-6 production in DPCs (p ≤ .0011), whereas eDMP had no discernible effect (p > .7509). No significant changes in IL-8 were observed (p > .198). TGF-ß1, available from eDMP supplementation, was reduced by DPCs over time. VEGF, on the other hand, was increased in all groups during coculture. CONCLUSIONS: The results show that the coculture of DPCs and S. mutans is possible without functional impairment. The bacterially induced stimulation of proinflammatory and regenerative cytokines provides a basis for future investigations and the elucidation of molecular biological relationships in pulp defence against caries.

Shear Bond Strength of Adhesives Placed following Selective Removal of Red-Fluorescing Carious Dentine in vitro.

Red-fluorescing dentine indicates bacterial contamination [Caries Res 2002; 36: 315-319]. We investigated effect of removal of red fluorescent dentine caries on shear bond strength and fracture mode of 4 adhesive approaches. Sixty-five carious teeth and 50 noncarious controls were distributed into 4 groups: Clearfil™ self-etch (CSE), OptiBond™ FL total etch (OTE), Scotchbond™ Universal total etch (STE) and self-etch (SSE). Samples were excited at 405 nm and viewed through 530 nm filter. Carious samples were ground flat exposing strongly red-fluorescing (StrongRF) dentine, on which a composite cylinder was placed, using one of 4 adhesives. After 22 h in water, shear bond strength and fracture mode were analysed. StrongRF was removed; composite cylinders were placed on weakly red-fluorescing (WeakRF) dentine and tested as described above. Finally, red-fluorescing dentine was removed, and composite cylinders were placed on non-fluorescing (NonRF) dentine and tested. Composites were placed at 3 corresponding heights in controls. After 22 h in water, shear bond strength testing and fracture mode analysis were performed. Differences were tested using Mann-Whitney or Wilcoxon tests (p ≤0.05). Median (Q1, Q3) shear bond strength on StrongRF was SSE 14.4 (9.2, 18.2) MPa >CSE 10.2 (6.4, 17.3) MPa >STE 9.1 (6.9, 11.2) MPa >OTE 6.8 (4.0, 10.8) MPa. Shear bond strength increased statistically significantly for all adhesives on WeakRF: SSE 19.8 (13.6, 24.3) MPa >STE 19.5 (12.7, 23.1) MPa >CSE 17.5 (12.0, 22.5) MPa >OTE 15.8 (11.9, 20.9) MPa. Only STE 25.6 (22.4, 29.1) MPa and CSE 22.1 (17.6, 24.6) MPa were significantly different on NonRF compared to WeakRF. For controls tested at corresponding depths, superficial shear bond strength was OTE 18.7 (16.0, 22.1) MPa >STE 18.4 (12.0, 25.9) MPa >CSE 18.1 (12.7, 20.7) MPa >SSE 13.0 (9.6, 17.8) MPa. This was significantly higher compared to StrongRF except for SSE. Central shear bond strength was not significantly different to WeakRF, deep shear bond strength was significantly lower for SSE and CSE but higher for OTE compared to carious. Conclusion: StrongRF dentine should be removed for higher shear bond strength, but WeakRF dentine can often be preserved without compromising adhesive bond strength.

Randomized Clinical Split-Mouth Study on Partial Ceramic Crowns Luted with a Self-adhesive Resin Cement with or without Selective Enamel Etching: Long-Term Results after 15 Years.

PURPOSE: This follow-up of a randomized clinical split-mouth study aimed to investigate the influence of selective enamel etching on the long-term clinical performance of partial ceramic crowns (PCCs) luted with a self-adhesive resin cement. MATERIALS AND METHODS: 43 patients received two PCCs (Vita Mark II; Cerec 3D) each for the restoration of extensive lesions with multiple cusp coverage, inserted with a self-adhesive resin cement (RelyX Unicem, RXU). Using a split-mouth design, one PCC received additional selective enamel etching (RXU+E) and one did not (RXU-E). Patients were clinically evaluated at baseline and after up to 15 years (median observation period 176 months) using modified USPHS and FDI criteria. The data were analyzed non-parametrically (chi-squared tests, α = 0.05). Clinical survival of all restorations after 15 years was evaluated using the Kaplan-Meier analysis. RESULTS: After 15 years, 19 patients were available for clinical assessment (recall rate: 56%). Kaplan-Meier analysis showed a cumulative survival of 78.1% for RXU+E and of 42.9% for RXU-E, indicating a significantly higher survival rate for RXU+E (p = 0.004). Regarding the clinical performance of PCCs available for the 15-year evaluation, no statistically significant differences were found between RXU+E and RXU-E using modified USPHS and FDI criteria. Both groups revealed significant deterioration over time regarding surface luster, marginal adaptation, and marginal discoloration. RXU+E resulted in significantly inferior anatomic form over time and a significant improvement in post-operative hypersensitivity compared to baseline. CONCLUSION: For posterior PCCs, selective enamel etching can be recommended based on higher survival rates after 15 years. Clinically, deterioration due to aging is similar in both groups.

General diseases and medications in 687 patients reporting on adverse effects from dental materials.

OBJECTIVES: Examination of patients claiming adverse effects from dental materials can be very challenging. Particularly, systemic aspects must be considered besides dental and orofacial diseases and allergies. Therefore, the aim of this study was to investigate a cohort of 687 patients reporting on adverse effects from dental materials focusing on findings related to known general diseases or conditions or medication-related findings with relevance to their subjective complaints. METHODS: Six hundred eighty-seven patients visiting a specialized consultation on claimed adverse effects from dental materials were retrospectively investigated for their subjective complaints, findings related to known general diseases or conditions, medication-related findings, dental and orofacial findings, or allergies with relevance to their subjective complaints. RESULTS: The most frequent subjective complaints were burning mouth (44.1%), taste disorders (28.5%), and dry mouth (23.7%). In 58.4% of the patients, dental and orofacial findings relevant to their complaints could be found. Findings related to known general diseases or conditions or medication-related findings were found in 28.7% or 21.0% of the patients, respectively. Regarding medications, findings related to antihypertensives (10.0%) and psychotropic drugs (5.7%) were found most frequently. Relevant diagnosed allergies toward dental materials were found in 11.9%, hyposalivation in 9.6% of the patients. In 15.1% of the patients, no objectifiable causes for the expressed complaints could be found. CONCLUSIONS: For patients complaining of adverse effects from dental materials, findings related to known general diseases or conditions and medications should be given particular consideration, while still in some patients, no objectifiable causes for their complaints can be found. CLINICAL RELEVANCE: For patients complaining about adverse effects from dental materials, specialized consultations and close collaboration with experts from other medical fields are eligible.

Preprocedural mouthwashes for infection control in dentistry-an update.

OBJECTIVES: Aerosols and splatter are routinely generated in dental practice and can be contaminated by potentially harmful bacteria or viruses such as SARS-CoV-2. Therefore, preprocedural mouthwashes containing antiseptic agents have been proposed as a potential measure for infection control in dental practice. This review article aims to summarize the clinical (and, if insufficient, preclinical) evidence on preprocedural mouthwashes containing antiseptic agents and to draw conclusions for dental practitioners. METHODS: Literature on preprocedural mouthwashes for reduction of bacterial or viral load in dental aerosols was searched and summarized. RESULTS: Preprocedural mouthwashes, particularly those containing chlorhexidine digluconate (CHX), cetylpyridinium chloride (CPC), or essential oils (EO), can significantly reduce the bacterial load in dental aerosols. With respect to viruses such as HSV-1, there are too little clinical data to draw any clear recommendations. On the other hand, clinical data is consolidating that CPC-containing mouthwashes can temporarily reduce the intraoral viral load and infectivity in SARS-CoV-2 positive individuals. Nevertheless, potential risks and side effects due to regular antiseptic use such as ecological effects or adaptation of bacteria need to be considered. CONCLUSIONS: The use of preprocedural mouthwashes containing antiseptics can be recommended according to currently available data, but further studies are needed, particularly on the effects on other viruses besides SARS-CoV-2. When selecting a specific antiseptic, the biggest data basis currently exists for CHX, CPC, EO, or combinations thereof. CLINICAL RELEVANCE: Preprocedural mouthwashes containing antiseptics can serve as part of a bundle of measures for protection of dental personnel despite some remaining ambiguities and in view of potential risks and side effects.

Elemental Compositions of Enamel or Dentin in Human and Bovine Teeth Differ from Murine Teeth.

Teeth with different chemical compositions can show vastly different physical properties, so knowledge of elemental composition is required to use animal teeth as substitutes for human teeth in research. In vitro, energy dispersive X-ray spectroscopy (EDX), improved by calibration standards and Si3N4-window material, enables determining local elemental compositions of inorganic and organic compounds without sample destruction. Six human molars, bovine incisors, murine incisors, and murine molars were analyzed. EDX-field scans and line scans were analyzed for elements Ca, P, O, C, N, F, Na, Mg, Fe, Cl, and S. Furthermore, Ca/P- and Ca/N-Ratios were calculated. The presence of iron in murine incisor enamel was investigated using additional wavelength dispersive X-ray spectroscopy measurements (WDX) near the enamel surface. Bovine and human enamel and dentin revealed close similarities regarding elemental composition. The median (25-75% percentiles) of At%Ca was 21.1 (20.8-21.3) in human enamel, 21.0 (20.7-21.2) in bovine enamel, and in murine enamel, 18.3 (17.85-18.88) for molars and 18.35 (18.00-18.75) for incisors. In dentin, murine teeth revealed a higher At%Ca compared to human and bovine teeth. Significant differences between human and bovine teeth were found for nitrogen in dentin, with a median of 4.5 (3.3-5) At%N for human dentin and 2.7 (2.3-3.2) At%N for bovine dentin. The Ca/P-Ratio was the highest in human and bovine enamel, which did not differ significantly. Enamel from murine molars had a higher Ca/P-Ratio than murine incisors and the highest Ca/P-Ratio in dentin was observed for human teeth and murine molars (1.49). WDX revealed iron in the outer 10 µm of pre- and post-eruptive enamel of murine incisors. Pre- and post-eruptive enamel on murine incisors only differed significantly in At%Ca (p = 0.041) and At%P (p = 0.026) with both At% higher in the pre-eruptive enamel. Murine teeth differ significantly from human and bovine teeth in terms of the elemental composition of enamel and dentin.

Impact of Endodontic Irrigant Activation on Smear Layer Removal and Surface Disintegration of Root Canal Dentine In Vitro.

The objective of this study was to compare the ability of different endodontic irrigation activation methods to enable irrigant penetration, remove the smear layer from root canal walls after preparation, and investigate surface effects on dentine. Root canals of 90 single-rooted teeth were prepared and irrigated with EDTA (17%) and sodium hypochlorite (5%), where both irrigants or sodium hypochlorite only were activated as follows: conventional needle irrigation, ultrasonic activation, sonic activation (EDDY), or laser-based activation (photon-induced photoacoustic streaming/PIPS). For the evaluation of irrigant penetration into dentinal tubules, methylene blue was injected and activated as well. Subsequently, teeth were sectioned horizontally, and dye penetration depths were measured. Alternating sections were split in halves and randomly selected for scanning electron microscopic analysis. Root canal dentine was assessed for smear layer removal and surface disintegration according to a defined scoring system. The data were analyzed statistically with nonparametric and chi-squared tests for whole teeth and separately for coronal, middle, and apical thirds. All the tested activation methods removed a thicker smear layer than needle irrigation only. Additional activation of EDTA improved penetration depths of the irrigants, but not the smear layer removal. Surface disintegration of root canal dentine was observed with the additional activation of EDTA and particularly after laser-based techniques. Additional activation of EDTA does not seem to offer any convincing advantages in terms of irrigant penetration or smear layer removal but disrupts the dentine surface. Especially laser-based activation resulted in undesirable destruction of root canal wall dentine.

Fluorescence Spectroscopy Shows Porphyrins Produced by Cultured Oral Bacteria Differ Depending on Composition of Growth Media.

Red fluorophores synthesized by oral bacteria are important for fluorescence-based diagnosis and treatment because they are used as markers for bacterially infected tissue, mature plaque, or calculus. A range of porphyrins have been identified as the source of this fluorescence in carious tissue. It is not clear which of these porphyrins are produced by individual oral bacteria or whether this ability depends on other factors. This study examined and compared the fluorescence spectra produced by selected cultured oral bacteria when grown on agars containing different nutrients with spectra for protoporphyrin IX, Zn-protoporphyrin IX, haematoporphyrin, and haematin. Actinomyces israelii (Deutsche Sammlung von Mikroorganismen [DSM], 43320), Actinomyces naeslundii (DSM 43013), Fusobacterium nucleatum (DSM, 20482), Lactobacillus casei (DSM, 20011), Prevotella intermedia (DSM, 20706), Streptococcus mutans (DSM, 20523), Streptococcus oralis (DSM, 20627), Streptococcus salivarius (DSM, 20560) and Streptococcus sobrinus (DSM, 20742) were rehydrated and grown anaerobically on caso, caso blood (containing 5% sheep blood), and caso chlorophyll (containing 5% spinach extract) agar for 3 days at 37°C in the dark. Colonies were harvested, transferred to ethanol, and centrifuged. Fluorescence emission spectra were recorded from the supernatant at 405 nm excitation (Fluorolog 3-22, Jobin Yvon-Spex ISA, Edison, NJ, USA). All Streptococci, L. casei, and F. nucleatum produced red fluorescence when grown on caso and caso chlorophyll agar but not on caso blood agar. A. naeslundii and P. intermedia emitted intense red fluorescence when grown on caso or caso blood agar but not on caso chlorophyll agar. Fluorescence emission spectra of A. naeslundii and P. intermedia grown on caso blood agar correlated exactly with both fluorescence peaks for protoporphyrin-IX at 632 and 701 nm. Most peaks observed could be correlated with at least one of the emission peaks of protoporphyrin IX, Zn-protoporphyrin IX, or haematoporphyrin. Oral bacteria emitted red fluorescence matching known porphyrins, but this depended on nutrients available in the agar.

Surface Accumulation of Cerium, Self-Assembling Peptide, and Fluoride on Sound Bovine Enamel.

The accumulation of caries-preventive compounds on sound enamel is crucial in order to improve the inhibition of carious lesion initiation. The aim of this research was to investigate the initial accumulation of cerium, oligopeptide p11-4, and fluoride from NaF or amine fluoride (AmF) on sound enamel in vitro by means of energy dispersive X-ray spectroscopy (EDX). Polished bovine enamel specimens (n = 120 from 60 teeth) were fabricated. Out of these, 12 specimens each were treated with CeCl3 (cerium(III) chloride heptahydrate 25%), oligopeptide p11-4 (Curodont Repair, Credentis), NaF (10,000 ppm F−), AmF (amine fluoride, Elmex Fluid, CP-GABA GmbH, 10,000 ppm F−), or Aqua demin (control). After rinsing with water, the surface elemental composition (Ce, N, F, Ca, P, O, Na, Mg) was measured (EDX; EDAX Octane Elect detector, APEX v2.0), expressed in atomic percent (At%) and analyzed (non-parametric statistics, α = 0.05, error rates method). Another 12 specimens per treatment group were fabricated and used for analyzing accumulation in cross-sections with EDX linescans and two-dimensional EDX-mappings. The surface median atomic percent of cerium (At%Ce) was 0.8 for CeCl3, but no Ce was found for any other group. N, specifically for oligopeptide p11-4, could not be detected. Fluorine could only be detected on fluoridated surfaces. The median atomic percent of fluorine (At%F) was 15.2 for NaF and 17.0 for AmF. The Ca/P ratio increased significantly compared to the control following the application of NaF and AmF (p < 0.001), but decreased significantly for CeCl3 (p < 0.001). In cross-sectioned specimens of the CeCl3-group, 12.5% of the linescans revealed cerium at the enamel surface, whereas 83.3% of the NaF linescans and 95.8% of the AmF linescans revealed fluorine at the enamel surface. Following the application of oligopeptide p11-4, no traces of N were detectable. In the depth of the samples, no signal was detected for any of the corresponding elements exceeding the background noise. Cerium and fluorine (from both NaF and AmF), but not the oligopeptide p11-4, precipitated on sound enamel.

Randomized clinical split-mouth study on a novel self-adhesive bulk-fill restorative vs. a conventional bulk-fill composite for restoration of class II cavities - results after three years.

OBJECTIVES: This randomized prospective split-mouth study evaluated the clinical performance of a novel, tooth-colored, self-adhesive bulk-fill restorative (SABF, 3M) for restoration of class II cavities as compared to a conventional bulk-fill composite (Filtek One, 3M; FOBF) over 36 months. The null-hypothesis was that both materials perform equally regarding clinical success and performance according to the FDI clinical criteria and scoring system. METHODS: 30 patients received one SABF and one FOBF restoration each. For FOBF, Scotchbond Universal (3M) was used as adhesive (self-etch mode), whereas SABF was applied without adhesive. Two blinded examiners evaluated the restorations at baseline, 24 and 36 months using FDI criteria. Data were analyzed non-parametrically (χ2-tests; α=0.05). RESULTS: 29 patients were available for the 24- and 36-month examinations. Clinical success rate was 96.6% for both materials at 36-mo (one restoration failure due to secondary caries each). All other restorations revealed clinically acceptable FDI scores at all recalls. FOBF performed significantly better than SABF at all time points regarding surface lustre (p<0.001) and color match and translucency (p<0.001) and regarding marginal staining at 36-months (p=0.008). Marginal staining and marginal adaptation deteriorated significantly over time for both materials (both p<0.001). CONCLUSIONS: The null-hypothesis could only partially be rejected. Both materials performed similarly regarding clinical success and performance within 36 months of clinical service, but SABF exhibited significantly inferior, but clinically fully acceptable esthetic properties as compared to FOBF. Both restorative materials showed clinically fully acceptable results over 36 months of clinical service and thus may be recommended for clinical use. CLINICAL SIGNIFICANCE: The novel tooth-colored self-adhesive bulk-fill restorative exhibited clinically fully acceptable results over 36 months of clinical service, similarly to a conventional bulk-fill restorative used with a universal adhesive, but with slight shortcomings in esthetic properties. Therefore, both restorative materials may be recommended for clinical use.

Ecological Effects of Daily Antiseptic Treatment on Microbial Composition of Saliva-Grown Microcosm Biofilms and Selection of Resistant Phenotypes.

Antiseptics are widely used in dental practice and included in numerous over-the-counter oral care products. However, the effects of routine antiseptic use on microbial composition of oral biofilms and on the emergence of resistant phenotypes remain unclear. Microcosm biofilms were inoculated from saliva samples of four donors and cultured in the Amsterdam Active Attachment biofilm model for 3 days. Then, they were treated two times daily with chlorhexidine digluconate (CHX) or cetylpyridinium chloride (CPC) for a period of 7 days. Ecological changes upon these multiple antiseptic treatments were evaluated by semiconductor-based sequencing of bacterial 16S rRNA genes and identification of amplicon sequence variants (ASVs). Furthermore, culture-based approaches were used for colony-forming units (CFU) assay, identification of antiseptic-resistant phenotypes using an agar dilution method, and evaluation of their antibiotic susceptibilities. Both CHX and CPC showed only slight effects on CFU and could not inhibit biofilm growth despite the two times daily treatment for 7 days. Both antiseptics showed significant ecological effects on the microbial compositions of the surviving microbiota, whereby CHX led to enrichment of rather caries-associated saccharolytic taxa and CPC led to enrichment of rather gingivitis-associated proteolytic taxa. Antiseptic-resistant phenotypes were isolated on antiseptic-containing agar plates, which also exhibited phenotypic resistance to various antibiotics. Our results highlight the need for further research into potential detrimental effects of antiseptics on the microbial composition of oral biofilms and on the spread of antimicrobial resistance in the context of their frequent use in oral healthcare.

Retrospective Cohort Study on Potential Risk Factors for Repeated Need of Dental Rehabilitation under General Anesthesia in a Private Pediatric Dental Practice.

The need for dental rehabilitations under general anesthesia (DRGAs) is continuously increasing, particularly for dental treatment of children. The present retrospective cohort study aimed to investigate potential risk factors for repeated need of DRGA in a cohort of patients from a private pediatric dental practice. Demographic and anamnestic data, dental status, and treatments performed during DRGA were retrospectively analyzed from the electronic dental charts of 1155 children that received at least one DRGA between October 2016 and December 2021. The median age of all children was 5 years at time of their first DRGA. The rate of repeated DRGAs was 9%. Patients with repeated need of DRGA were significantly younger at time of their first DRGA and revealed significantly more often a history of preterm birth and current use of a baby bottle as compared to patients with only one DRGA. There were significantly fewer treatments (regardless of type) in the second DRGA than at the first. Within the limitations of this study, young age at first DRGA, a history of preterm birth, and current use of a baby bottle may be risk factors for repeated need of DRGA. The search for effective strategies to minimize the repeated need for DRGA in children remains critical.

Impact of access cavity cleaning on the seal of postendodontic composite restorations in vitro.

AIM: The aim of the study was to investigate the influence of cavity cleaning and conditioning on marginal integrity of directly placed post-endodontic composite class-I-restorations in vitro. METHODOLOGY: A total of 168 fully intact teeth without caries or fillings received pre-endodontic composite restorations (class-II) after their extraction. Occlusal endodontic access-cavities were prepared, and root canals were instrumented and filled with gutta-percha and an epoxy resin-based sealer. Prior to post-endodontic class-I-restoration, access cavities were completely contaminated with sealer, cleaned with alcohol and pre-treated as follows: cleaner only (alcohol), glycine-polishing, Al2 O3 sandblasting, carbide bur (immediate as well as delayed restoration). A positive control (not contaminated with sealer and adhesive used) and negative control (cleaner used but no adhesive) were established. Half of the teeth from each group were subjected to thermocycling and mechanical loading (TCML). Marginal integrity of post-endodontic restoration was evaluated in oro-vestibular or mesio-distal sections after AgNO3 dye penetration (DP) by standardized photomacroscopic imaging and expressed in per cent of margin length along all segments and separately for enamel, dentine and composite, respectively. Results were analysed non-parametrically (α = .05). RESULTS: No restorations or teeth fractured or debonded completely. Without TCML, the median DP of all segments was significantly higher for the negative control compared with all other groups in oro-vestibular cutting direction (53%; p = .002) and in mesio-distal cutting direction (51%; p ≤ .041). The other groups without TCML revealed 16%-24% DP (oro-vestibular) and 12%-24% DP (mesio-distal). With TCML, the median DP in oro-vestibular cutting direction for all segments ranged between 48% and 62% for all groups, a significant difference was only observed between glycine-polishing and carbide bur (p = .041). In mesio-distal cutting direction, the median DP in negative control was 69% with TCML and significantly higher compared with all other groups (p = .002). For all other groups, the median DP of all segments ranged between 28% and 40% with TCML without significant differences. Error rates method (k = 7) revealed a significant influence of TCML in general on penetration of all segments in both oro-vestibular and mesio-distal cutting directions. CONCLUSION: Additional access cavity pre-treatment after alcohol cleaning did not improve the marginal integrity of post-endodontic composite restorations. Thorough cleaning of the access cavity with alcohol seems to assure an acceptable marginal integrity to the tooth and restorative composite.

Phenotypic Adaptation to Antiseptics and Effects on Biofilm Formation Capacity and Antibiotic Resistance in Clinical Isolates of Early Colonizers in Dental Plaque.

Despite the wide-spread use of antiseptics in dental practice and oral care products, there is little public awareness of potential risks associated with antiseptic resistance and potentially concomitant cross-resistance. Therefore, the aim of this study was to investigate potential phenotypic adaptation in 177 clinical isolates of early colonizers of dental plaque (Streptococcus, Actinomyces, Rothia and Veillonella spp.) upon repeated exposure to subinhibitory concentrations of chlorhexidine digluconate (CHX) or cetylpyridinium chloride (CPC) over 10 passages using a modified microdilution method. Stability of phenotypic adaptation was re-evaluated after culture in antiseptic-free nutrient broth for 24 or 72 h. Strains showing 8-fold minimal inhibitory concentration (MIC)-increase were further examined regarding their biofilm formation capacity, phenotypic antibiotic resistance and presence of antibiotic resistance genes (ARGs). Eight-fold MIC-increases to CHX were detected in four Streptococcus isolates. These strains mostly exhibited significantly increased biofilm formation capacity compared to their respective wild-type strains. Phenotypic antibiotic resistance was detected to tetracycline and erythromycin, consistent with the detected ARGs. In conclusion, this study shows that clinical isolates of early colonizers of dental plaque can phenotypically adapt toward antiseptics such as CHX upon repeated exposure. The underlying mechanisms at genomic and transcriptomic levels need to be investigated in future studies.

Transcriptomic Stress Response in Streptococcus mutans following Treatment with a Sublethal Concentration of Chlorhexidine Digluconate.

Despite the widespread use of antiseptics such as chlorhexidine digluconate (CHX) in dental practice and oral care, the risks of potential resistance toward these antimicrobial compounds in oral bacteria have only been highlighted very recently. Since the molecular mechanisms behind antiseptic resistance or adaptation are not entirely clear and the bacterial stress response has not been investigated systematically so far, the aim of the present study was to investigate the transcriptomic stress response in Streptococcus mutans after treatment with CHX using RNA sequencing (RNA-seq). Planktonic cultures of stationary-phase S. mutans were treated with a sublethal dose of CHX (125 µg/mL) for 5 min. After treatment, RNA was extracted, and RNA-seq was performed on an Illumina NextSeq 500. Differentially expressed genes were analyzed and validated by qRT-PCR. Analysis of differential gene expression following pathway analysis revealed a considerable number of genes and pathways significantly up- or downregulated in S. mutans after sublethal treatment with CHX. In summary, the expression of 404 genes was upregulated, and that of 271 genes was downregulated after sublethal CHX treatment. Analysis of differentially expressed genes and significantly regulated pathways showed regulation of genes involved in purine nucleotide synthesis, biofilm formation, transport systems and stress responses. In conclusion, the results show a transcriptomic stress response in S. mutans upon exposure to CHX and offer insight into potential mechanisms that may result in development of resistances.

Influence of HEMA on LPS- and LTA-stimulated IL-6 release from human dental pulp cells.

OBJECTIVE: Dental pulp cells interact with immunogenic components such as LPS (lipopolysaccharide) or LTA (lipoteichoic acid) released from microorganisms in carious lesions. In the present investigation, the formation of the pro-inflammatory cytokines TNFα and IL-6 in LPS- or LTA-stimulated cells from the dental pulp interface and pulp fibroblasts was analyzed in the presence of the resin monomer 2-hydroxyethyl methacrylate (HEMA) under varying cellular redox conditions. METHOD: Human pulp fibroblasts (HPC) or cells from the dental pulp interface expressing an odontoblast phenotype (hOD-1) were exposed to LTA, LPS or HEMA for 1 h or 24 h. Redox homeostasis was modified by the prooxidant BSO (L-buthionine sulfoximine) or the antioxidant NAC (N-acetyl cysteine). Formation of TNFα or IL-6 was analyzed by ELISA, and cell survival was determined by a crystal violet assay. Statistical analyses were performed using the Mann-Whitney-U-test. RESULTS: Secretion of TNFα was not detected in LPS- or LTA-stimulated HPC or hOD-1, and IL-6 was not found after a short exposure (1 h). After a 24 h exposure, LPS induced a 3-fold increase in IL-6 formation in HPC, while LTA stimulated IL-6 release about 20-fold. Likewise, LTA was more effective than LPS in hOD-1 stimulating IL-6 levels about 50-fold. HEMA inhibited the LPS- and LTA-induced IL-6 release, and this effect was enhanced by BSO but counteracted by NAC in both cell types. IL-6 release was independent of cell survival rates. CONCLUSIONS: The protective immune response in odontoblasts and pulp fibroblasts is impaired by monomers such as HEMA through the disturbance of the redox homeostasis.

Molecular Biological Comparison of Dental Pulp- and Apical Papilla-Derived Stem Cells.

Both the dental pulp and the apical papilla represent a promising source of mesenchymal stem cells for regenerative endodontic protocols. The aim of this study was to outline molecular biological conformities and differences between dental pulp stem cells (DPSC) and stem cells from the apical papilla (SCAP). Thus, cells were isolated from the pulp and the apical papilla of an extracted molar and analyzed for mesenchymal stem cell markers as well as multi-lineage differentiation. During induced osteogenic differentiation, viability, proliferation, and wound healing assays were performed, and secreted signaling molecules were quantified by enzyme-linked immunosorbent assays (ELISA). Transcriptome-wide gene expression was profiled by microarrays and validated by quantitative reverse transcription PCR (qRT-PCR). Gene regulation was evaluated in the context of culture parameters and functionality. Both cell types expressed mesenchymal stem cell markers and were able to enter various lineages. DPSC and SCAP showed no significant differences in cell viability, proliferation, or migration; however, variations were observed in the profile of secreted molecules. Transcriptome analysis revealed the most significant gene regulation during the differentiation period, and 13 biomarkers were identified whose regulation was essential for both cell types. DPSC and SCAP share many features and their differentiation follows similar patterns. From a molecular biological perspective, both seem to be equally suitable for dental pulp tissue engineering.

Biology of selective caries removal: a systematic scoping review protocol.

INTRODUCTION: Primary goal of restorative caries therapy is to preserve pulp vitality and the dentition. Whereas the conventional approach of complete caries removal aims at the elimination of all affected substances without regard to losses of hard tissue or pulp vitality, the innovative concept of selective caries removal (SCR) is characterised by a targeted and non-invasive excavation. It presents a lower risk of accidental pulp exposure, which reportedly has a positive effect on tooth survival. Although clinical data show the benefits of SCR, knowledge about the biological processes during this procedure in the pulp-dentine complex of permanent teeth is scarce. Hence, the aim of this work is to systematically scope the existing literature and map the existing evidence according to the Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews guideline. This protocol details the scoping review's methodological and analytical approaches. METHODS AND ANALYSIS: First, a structure was established (phase I) as basis for a systematic scoping of literature (phase II). In the course of phase I, a total of 100 systematic reviews related to selective caries removal were searched in MEDLINE and information or theories on the biological processes were extracted. During the entire procedure, two reviewers independently screened the articles, and controversies were mediated by vote of a third reviewer. Eventually, it became apparent that different biological explanations can be organised into four categories: pulp response, cavity seal, remaining bacteria and cavity liner. Based on this structure, a search for original publications (phase II) will be performed and retrieved evidence will be assembled using a predefined conceptual framework. ETHICS AND DISSEMINATION: As primary data will not be included in this study, ethical approval is not required. Findings will be disseminated through peer-reviewed publications, conference presentations and summaries for key stakeholders.