Harnessing antimicrobial peptides in endodontics.

Endodontic therapy includes various procedures such as vital pulp therapy, root canal treatment and retreatment, surgical endodontic treatment and regenerative endodontic procedures. Disinfection and tissue repair are crucial for the success of these therapies, necessitating the development of therapeutics that can effectively target microbiota, eliminate biofilms, modulate inflammation and promote tissue repair. However, no current endodontic agents can achieve these goals. Antimicrobial peptides (AMPs), which are sequences of amino acids, have gained attention due to their unique advantages, including reduced susceptibility to drug resistance, broad-spectrum antibacterial properties and the ability to modulate the immune response of the organism effectively. This review systematically discusses the structure, mechanisms of action, novel designs and limitations of AMPs. Additionally, it highlights the efforts made by researchers to overcome peptide shortcomings and emphasizes the potential applications of AMPs in endodontic treatments.

Penetration of Calcium Silicate and Epoxy Resin Sealers Into the Lateral Canals.

OBJECTIVES: The aim of this research was to compare the penetration ability of calcium silicate-based sealers (iRoot SP and TotalFill BC HiFlow) and an epoxy resin-based sealer (AH Plus) into the lateral canals using the single-cone (SC) or continuous wave compaction (CW) obturation techniques. METHODS: Ninety single-rooted human teeth received canal instrumentation and diaphanisation before artificial lateral canals were created at 3 different root levels. The specimens were randomly allocated into 6 groups based on the combination of obturation technique and sealer used. Radiographic and photographic measurements evaluated the percentage of sealer penetration. Statistical analysis was performed to compare the penetration ability amongst different types of sealers, obturation methods, and root levels. RESULTS: Although TotalFill BC HiFlow sealer combined with the CW technique demonstrated greater sealer penetration at the coronal level (P < .05), the overall penetration ability of iRoot SP, TotalFill BC HiFlow, and AH Plus was not significantly different. The deepest sealer penetration was observed at the apical root level. CONCLUSIONS: All sealers showed similar penetration abilities into the lateral canals using the SC or CW techniques in vitro. Calcium silicate-based sealers have comparable penetration ability into lateral canals compared to epoxy resin-based sealers using SC or CW obturation techniques.

Mouthwashes: Alternatives and Future Directions.

This narrative review summarises "alternative" or "natural" over-the-counter (OTC) mouthwashes not covered elsewhere in this supplement and newly emerging products, as potential mouthwashes of the future. The "natural" mouthwashes reviewed include saltwater, baking soda, coconut oil, charcoal, propolis, seaweeds, and probiotics. Other than essential oils, it is apparent that their clinical effectiveness is still under debate, but there is some evidence to suggest that propolis reduces plaque and gingivitis. This review also covers the host immune response, via novel anti-inmmunomodulant mouthwashes, such as erythropoietin to reduce inflammation with oral mucositis (OM) after radiotherapy. The emerging concept of nanoparticle-containing mouthwashes, such as iron oxide, is further discussed for OM, this agent having the potential for more targeted delivery of chemical antimicrobials. Unfortunately, there are impacts on the environment of widening mouthwash use with more new products, including increased use of packaging, antimicrobial resistance, and possible detrimental effects on marine life. Further, there are roadblocks, relating to regularly approvals and side effects, that still need to be overcome for any OTC deivered immunomodulant or nanoformulation mouthwashes. Despite these caveats, there are many new mouthwashes under development, which could help manage major oral diseases such as caries, gingivitis, and periodontal disease.

Tissue Stabilization, Bacterial Adhesion, and Stem Cell Viability in Trans-cinnamaldehyde-conditioned Dentin.

INTRODUCTION: This laboratory study aimed to evaluate the effect of trans-cinnamaldehyde (TC) conditioning on dentin tissue stabilization, bacterial adhesion, and stem cell toxicity. METHODS: Dentin beams (n = 204) from extracted human molars were demineralized in phosphoric acid and treated with TC (2.5, 5, and 7.5%), 50% ethanol-water mixture (vehicle control) or 2.5% glutaraldehyde (GA) (positive control) for 30 minutes. Demineralized but untreated specimens served as the negative control. After treatment, collagen crosslinking was characterized by measuring the elastic modulus (Er) and hardness (n = 5). Biodegradation resistance was examined by determining the loss of dry mass (n = 8), hydroxyproline release (n = 4) and scanning electron microscopy (n = 2), after exposure to bacterial collagenase. Inhibition of bacterial adhesion was investigated by colony counting assay (n = 12) and scanning electron microscopy (n = 2). Viability of stem cells of the apical papilla on TC-conditioned dentin was determined using the Cell Counting Kit-8 assay (n = 8). Data were statistically analyzed using one-way analysis of variance (ANOVA) test followed by Dunnett's multiple comparisons at a significance level of 5%. RESULTS: TC-conditioned dentin showed a concentration-dependent increase in Er and hardness. The Er and hardness of 5% and 7.5% TC-conditioned dentin were significantly greater than that of the negative control and vehicle control groups (P < .05). There was no significant difference in the biodegradation resistance between GA and 5% TC-conditioned dentin (P > .05). TC-conditioned dentin showed a well-preserved collagen fibril network with clear cross-banding, comparable to GA-conditioned dentin. All concentrations of TC inhibited bacterial adhesion on dentin, significantly greater than the negative control (P < .05). There was no reduction in viability of stem cells of the apical papilla viability on TC-conditioned dentin compared to the negative control (P > .05). CONCLUSIONS: TC conditioning stabilized the dentin and protected it from enzymatic degradation. TC prevented bacterial adhesion on the dentin but maintained stem cell viability.

Effect of Dentine Conditioning with Silver Diamine Fluoride on Wettability of Root Canal Sealers.

OBJECTIVE: To test the null hypothesis that dentine treatment with silver diamine fluoride (SDF), a potent antimicrobial agent, following use of proteolytic and chelating agents does not influence the wettability of an epoxy resin (AH Plus, Dentsply Sirona, Charlotte, NC, USA) and a tricalcium silicate sealer (BioRoot RCS, Septodont, Saint-Maur-des-Fossés, France). METHODS: Seventy-two intraradicular dentine specimens were divided into six groups based on the final irrigation solutions used: 2.5% sodium hypochlorite (NaOCl) and 17% ethylenediaminetetraacetic acid (EDTA) (NaOCl-EDTA) (group 1); NaOCl-EDTA-NaOCl (group 2); NaOCl-EDTA followed by 3.8% SDF, NaOCl-EDTA-SDF (group 3); NaOCl-EDTA-NaOCl-SDF (group 4); SDF (group 5) and saline (group 6). After irrigation, the specimens were divided into subgroups according to the sealer used, AH Plus or BioRoot RCS. Contact angles were measured using a contact angle analyser. The data were analysed using an independent t test, one-way analysis of variance (ANOVA) and Tamhane T2 post hoc test, with the level of significance set at P < 0.05. RESULTS: In the epoxy resin sealer group, dentine surfaces treated with only SDF showed the lowest contact angle. This was significantly less than the groups in which NaOCl was used as the final irrigant (P < 0.05). In the tricalcium silicate-based sealer group, the groups treated with SDF showed significantly greater contact angles when compared to the control group (P < 0.05). CONCLUSION: It was concluded that SDF conditioning of dentine favours the wettability of epoxy resin sealer but is detrimental to the wettability of tricalcium silicate sealer.

Effect of cryotherapy duration on experimentally induced connective tissue inflammation in vivo.

Objectives: This study tested the hypothesis that cryotherapy duration influences lipopolysaccharide (LPS)-induced inflammation in a rat model. Materials and Methods: Six Wistar rats (Rattus norvegicus albinus) were used. Five sites were selected per animal and divided into 5 groups: a negative control group (NC), 2 positive control groups (PC1 and PC2), and 2 experimental groups (E1 and E2). Cryotherapy was applied for 1 minute (E1) or 5 minutes (E2). An acute inflammatory response was induced in the PC and E groups via subcutaneous administration of 0.5 mL/kg. In the PC2 group, a catheter was inserted without additional treatment. For the E1 and E2 groups, 2.5°C saline solution was administered through the implanted catheters for 1 and 5 minutes, respectively. The rats were sacrificed, and samples were obtained and processed for histological analysis, specifically examining the presence of polymorphonuclear neutrophils and hemorrhage. The χ2 test was used to compare the presence of acute inflammation across groups. Dependent variables were compared using the linear-by-linear association test. Results: Inflammation and hemorrhage varied significantly among the groups (p = 0.001). A significantly higher degree of acute inflammation was detected (p = 0.0002) in the PC and E1 samples than in the E2 group, in which cryotherapy was administered for 5 minutes. The PC and E1 groups also exhibited significantly greater numbers of neutrophils (p = 0.007), which were essentially absent in both the NC and E2 groups. Conclusions: Cryotherapy administration for 5 minutes reduced the acute inflammation associated with LPS and catheter implantation.

A laboratory investigation on the effect of biguanide- and pyridine-derived antiseptics on the adhesion of resin composites to dentin.

Optimal bonding of adhesive restorations to dentin is crucial to prevent microleakage and enhance the survival of root-filled teeth. The aim of this study was to investigate the effect of chlorhexidine (CHX), alexidine (ALX) and octenidine dihydrochloride (OCT) on the bond strength of resin composites to coronal dentin. Human coronal dentin specimens were treated with 2% CHX, 0.1% ALX, 0.1% OCT or saline then restored with traditional or bulk-fill resin composites. The adhesion strength between the resin and dentin was measured using the microtensile bond strength and failure mode was determined using a stereomicroscope. Treatment with ALX and OCT resulted in significantly greater µTBS compared with CHX and saline, irrespective of the resin composite used. Alexidine treatment predominantly resulted in mixed failure, while adhesive failures were frequently observed in CHX and saline-treated dentin. In conclusion, final irrigation with ALX or OCT improved the bonding of resin composites to dentin.

Intracellular bacterial eradication using a novel peptide in vitro.

AIM: To determine the effect of a novel antimicrobial peptide (AMP; OP145) and cell-penetrating peptide (Octa-arginine/R8) conjugate on the killing of intracellular Enterococcus faecalis, compared to OP145 and an antibiotic combination recommended for regenerative endodontic procedures. METHODOLOGY: The biocompatible concentrations of OP145 and OP145-R8 were determined by assessing their cytotoxicity against human macrophages and red blood cells. Spatiotemporal internalization of the peptides into macrophages was investigated qualitatively and quantitatively by confocal laser scanning microscopy and flow cytometry respectively. Killing of extracellular and intracellular E. faecalis OG1RF by the peptides was determined by counting the colony-forming units (CFU). Intracellular antibacterial activity of the peptides was compared to a double antibiotic combination. Confocal microscopy was used to confirm the intracellular bacterial eradication. Significant differences between the different test groups were analysed using one-way analysis of variance. p < .05 was considered to be statistically significant. RESULTS: Peptides at a concentration of 7.5 µmol/L were chosen for subsequent experiments based on the results of the alamarBlue™ cell viability assay and haemolytic assay. OP145-R8 selectively internalized into lysosomal compartments and the cytosol of macrophages. Conjugation with R8 improved the internalization of OP145 into macrophages in a temporal manner (70.53% at 1 h to 77.13% at 2 h), while no temporal increase was observed for OP145 alone (60.53% at 1 h with no increase at 2 h). OP145-R8 demonstrated significantly greater extracellular and intracellular antibacterial activity compared to OP145 at all investigated time-points and concentrations (p < .05). OP145-R8 at 7.5 µmol/L eradicated intracellular E. faecalis after 2 h (3.5 log reduction compared to the control; p < .05), while the antibiotics could not reduce more than 0.5 log CFU compared to the control (p > .05). Confocal microscopy showed complete absence of E. faecalis within the OP145-R8 treated macrophages. CONCLUSIONS: The results of this study demonstrated that the conjugation of an AMP OP145 to a cell-penetrating peptide R8 eradicated extracellular and intracellular E. faecalis OG1RF without toxic effects on the host cells.

Accumulated Hard Tissue Debris and Root Canal Shaping Profiles Following Instrumentation with Gentlefile, One Curve, and Reciproc Blue.

INTRODUCTION: This laboratory study aimed to evaluate the accumulated hard tissue debris (AHTD) and shaping performance following root canal instrumentation with a novel stainless steel rotary system (Gentlefile; MedicNRG, Kibbutz Afikim, Israel) compared with two single-file nickel-titanium instruments of different kinematics through micro-computed tomography (micro-CT) analysis. METHODS: Thirty isthmus-containing mesial roots of human mandibular molars with a curvature of 15°-35° were scanned with micro-CT at an isotropic resolution of 12 µm and randomly assigned to three groups (n = 10) according to the instruments used for canal preparation: Gentlefile (GF; #23/.04), One Curve (OC; #25/.06; Micro-Mega, Besancon, France), and Reciproc Blue R25 (RB; #25/.08; VDW, Munich, Germany). The AHTD, unprepared canal surface area, volume changes, surface area changes, and transportation were evaluated by comparing preoperative and postoperative micro-CT images. In addition, the time required for canal preparation was recorded. Data were statistically analyzed using the one-way analysis of variance (ANOVA) test or the Kruskal-Wallis test at a significance level of 5%. RESULTS: No significant differences were observed in the percentage of AHTD or in the unprepared canal surface area between the three groups (P > .05). The RB group demonstrated a significantly greater percentage increase in volume and surface area than the GF group (P < .05). The GF and OC groups showed significantly less transportation at 3 mm from the apex than the RB group (P < .001). The OC group required significantly less time for instrumentation than the RB and GF groups (P < .05). CONCLUSIONS: Instrumentation with GF (#23/.04), OC, and RB yielded similar levels of AHTD and unprepared surface area in isthmus-containing curved canals. GF achieved this with less dentin removal and apical transportation, at the expense of a longer preparation time.

Revisiting ESKAPE Pathogens: virulence, resistance, and combating strategies focusing on quorum sensing.

The human-bacterial association is long-known and well-established in terms of both augmentations of human health and attenuation. However, the growing incidents of nosocomial infections caused by the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter sp.) call for a much deeper understanding of these organisms. Adopting a holistic approach that includes the science of infection and the recent advancements in preventing and treating infections is imperative in designing novel intervention strategies against ESKAPE pathogens. In this regard, this review captures the ingenious strategies commissioned by these master players, which are teamed up against the defenses of the human team, that are equally, if not more, versatile and potent through an analogy. We have taken a basketball match as our analogy, dividing the human and bacterial species into two teams playing with the ball of health. Through this analogy, we make the concept of infectious biology more accessible.

Evaluation of machine-assisted irrigation on removal of intracanal biofilm and extrusion of sodium hypochlorite using a three-dimensionally printed root canal model.

PURPOSE: This study aimed to compare the biofilm removal and apical extrusion of sodium hypochlorite (NaOCl) following machine-assisted irrigation using a three-dimensionally (3D) printed dentin-insert model. METHODS: Multispecies biofilms were formed in a 3D-printed curved root canal model with dentin insert. The model was then placed in a container that was filled with 0.2% agarose gel containing 0.1% m-Cresol purple. Root canals were irrigated with 1% NaOCl using syringe irrigation, sonically agitated (EndoActivator or EDDY) or ultrasonically activated (Endosonic Blue) irrigation. Samples were photographed and the color-changed area was measured. Biofilm removal was assessed using colony-forming unit counting, confocal laser scanning microscopic analysis and scanning electron microscopic observations. The data were analyzed using one-way ANOVA, followed by Tukey test (P < 0.05). RESULTS: EDDY and Endosonic Blue demonstrated significantly greater reduction of biofilms compared to other groups. No significant differences were observed in the remaining biofilm volume in syringe irrigation and EndoActivator groups. Furthermore, EDDY and Endosonic Blue presented with numerous exposed dentinal tubules. EDDY showed significantly greater NaOCl extrusion compared to other groups. CONCLUSION: Ultrasonic activation with a small-sized nickel-titanium file irrigation system may be beneficial in intracanal biofilm removal avoiding extrusion of NaOCl beyond the root apex.

A heptadeca amino acid peptide subunit of cathelicidin LL-37 has previously unreported antifungal activity.

Yeasts such as Candida albicans, albeit being ubiquitous members of the skin, oral and vaginal microbiome, can cause superficial to life-threatening infections. Human cathelicidin LL-37-based peptides have antibacterial activity and yet, their antifungal activity remains to be thoroughly characterized. The aim of this study was to comprehensively investigate the activity of LL-37-based peptides against C. albicans. LL-37 and its derivatives were tested for their ability to kill C. albicans planktonic cells in the presence of various biological matrices (serum, plasma, saliva and urine), that have been reported to inactivate peptides. The antibiofilm activity, resistance development and biocompatibility were investigated for the lead peptide. GK-17, a 17 amino acid peptide, showed remarkable stability to fungal aspartyl proteases and rapidly killed planktonic C. albicans despite the presence of biological matrices. GK-17 also inhibited adhesion to biotic and abiotic substrates, inhibited biofilm formation and eradicated preformed biofilms in the presence of biological matrices. Compared to nystatin, GK-17 had a lower propensity to allow for resistance development by C. albicans. The peptide showed concentration-dependent biocompatibility to red blood cells, with only 30% hemolysis even at 4× the fungicidal concentration. Taken together, GK-17 is a novel antifungal peptide with promising effects against C. albicans.

The presence of Enterococcus faecalis in saliva as a risk factor for endodontic infection.

Aim: The aim of the present study was to investigate and correlate the prevalence of Enterococcus faecalis in saliva and in root canals with different pulpal and periapical conditions. Methodology: Sixty-seven patients were divided into five groups based on pulpal and periapical tissue status: healthy vital teeth (HVT, n=7), healthy treated teeth without lesion (HTT, n=9), irreversible pulpitis (IP, n=13), necrosis (N, n=18), and post-treatment apical periodontitis (PTAP, n=20). Saliva, rubber dam, sterility control and pre-treatment root canal samples were collected and microbiologically processed by culture method. The phylogenetic relationship of E. faecalis isolates collected from root canals and saliva were investigated by whole genome sequencing. Fisher's exact test was used to correlate the presence of E. faecalis in root canals or saliva with clinical and/or radiographic findings. Linear/logistic regression analyses were performed to establish the relationship between the presence of E. faecalis in root canals, saliva, and the status of periapical tissues. Results: E. faecalis was found in 18 root canal and saliva samples. E. faecalis root canal isolates were recovered with the highest frequency from post-treatment apical periodontitis. The occurrence of E. faecalis in saliva was strongly associated with its detection in the root canals (P < 0.001). The pretreatment presence of E. faecalis in root canals was associated with significantly higher odds of having periapical lesions (OR=11.03; 95% CI, 1.27-95.70; p < 0.05). Saliva and root canal isolates from the same patient were highly correlated at the phylogenetic level (Jaccard index >0.95). Conclusion: This pilot study confirms the role of E. faecalis in developing peri-radicular lesions in secondary endodontic infections and suggests that saliva could be the main source of infection. Further studies are needed to investigate the exact origin of this bacteria and its true role in the pathogenesis of secondary/persistent endodontic infections.

A systematically biosynthetic investigation of lactic acid bacteria reveals diverse antagonistic bacteriocins that potentially shape the human microbiome.

BACKGROUND: Lactic acid bacteria (LAB) produce various bioactive secondary metabolites (SMs), which endow LAB with a protective role for the host. However, the biosynthetic potentials of LAB-derived SMs remain elusive, particularly in their diversity, abundance, and distribution in the human microbiome. Thus, it is still unknown to what extent LAB-derived SMs are involved in microbiome homeostasis. RESULTS: Here, we systematically investigate the biosynthetic potential of LAB from 31,977 LAB genomes, identifying 130,051 secondary metabolite biosynthetic gene clusters (BGCs) of 2,849 gene cluster families (GCFs). Most of these GCFs are species-specific or even strain-specific and uncharacterized yet. Analyzing 748 human-associated metagenomes, we gain an insight into the profile of LAB BGCs, which are highly diverse and niche-specific in the human microbiome. We discover that most LAB BGCs may encode bacteriocins with pervasive antagonistic activities predicted by machine learning models, potentially playing protective roles in the human microbiome. Class II bacteriocins, one of the most abundant and diverse LAB SMs, are particularly enriched and predominant in the vaginal microbiome. We utilized metagenomic and metatranscriptomic analyses to guide our discovery of functional class II bacteriocins. Our findings suggest that these antibacterial bacteriocins have the potential to regulate microbial communities in the vagina, thereby contributing to the maintenance of microbiome homeostasis. CONCLUSIONS: Our study systematically investigates LAB biosynthetic potential and their profiles in the human microbiome, linking them to the antagonistic contributions to microbiome homeostasis via omics analysis. These discoveries of the diverse and prevalent antagonistic SMs are expected to stimulate the mechanism study of LAB's protective roles for the microbiome and host, highlighting the potential of LAB and their bacteriocins as therapeutic alternatives. Video Abstract.

One-Pot Preparation of Cetylpyridinium Chloride-Containing Nanoparticles for Biofilm Eradication.

Quaternary ammonium compounds (QACs) have been widely used due to their excellent antimicrobial activity. However, using the technology where nanomaterials are employed as drug carriers to deliver QAC drugs has not been fully explored. In this study, mesoporous silica nanoparticles (MSNs) with short rod morphology were synthesized in a one-pot reaction using an antiseptic drug cetylpyridinium chloride (CPC). CPC-MSN were characterized via various methods and tested against three bacterial species (Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis), which are associated with oral infections, caries, and endodontic pathology. The nanoparticle delivery system used in this study prolonged the release of CPC. The manufactured CPC-MSN effectively killed the tested bacteria within the biofilm, and their size allowed them to penetrate into dentinal tubules. This CPC-MSN nanoparticle delivery system demonstrates potential for applications in dental materials.

Fracture Resistance of Endodontically Treated Maxillary Premolars with Non-carious Cervical Lesions Restored with Different Post Systems.

OBJECTIVE: To test the hypothesis that the (i) presence of non-carious cervical lesions (NCCLs) and (ii) type of post system have no effect on the fracture resistance and pattern in endodontically treated maxillary premolars. METHODS: Human maxillary first premolars (n=60) with two root canals were randomly allocated into four groups (n=15). Buccal wedge-shaped NCCLs were prepared in 45 teeth specimens. Following root canal treatment, the specimens were randomly divided into (i) composite resin core (CRC); (ii) NCCLs + composite resin core (NCCL+CRC); (iii) NCCLs+prefabricated fibre-reinforced composite post + composite resin core (NCCL+PFRC+CRC); (iv) NCCLs+custom fibre posts + composite resin core (NCCL+CFP+CRC). All specimens were subjected to thermocycling (5°C to 55°C/5000 cycles). The compressive load was applied non-axially to the palatal cusp with a universal testing machine at a crosshead speed of 0.5 mm/min at a 30° angle until fracture. Fracture patterns were examined using a loupe magnification (2.5×) under transillumination. Statistical analyses were performed using non-parametric tests and pairwise comparisons of the load-to-fracture among the groups. Chi-square test was used to analyse the fracture patterns (P=0.05). RESULTS: Fracture resistance of NCCL+PFRC+CRC was significantly higher than NCCL+CRC (P=0.011), while NCCL+CFP+CRC did not show any significant difference when compared to NCCL+CRC (P=0.089). No statistical difference was found between CRC, NCCL+PFRC+CRC and NCCL+CFP+CRC (P=1.000). The frequencies of favourable fracture patterns in descending orders were as follows: CRC (80%), NCCL+CFP+CRC (73%), NCCL+PFRC+CRC (60%), and NCCL+CRC (40%). Chi-square test did not show significant differences in fracture patterns among all groups (P=0.110). CONCLUSION: Restoration of the endodontically treated maxillary premolars with NCCLs, with or without post, resulted in similar fracture resistance as their counterparts without NCCLs. Placement of a prefabricated fibre-reinforced composite post exhibited greater fracture resistance to the maxillary premolars with restored NCCLs than those without a post. (EEJ-2022-06-077).

Influence of Coronal Flaring on the Shaping Ability of Two Heat-Treated Nickel-Titanium Endodontic Files: A Micro-Computed Tomographic Study.

Nickel-titanium (NiTi) usage is associated in endodontics with some complications including canal transportation. Centering ability of a NiTi file is the ability to stay centered in the root canal system during instrumentation. Any undesirable deviation from the natural canal path is indicated as canal transportation. A possible strategy to improve the centering ability of NiTi instruments is the pre-enlargement of the coronal third of the root canal to minimize coronal interferences. This procedure is known as coronal flaring. The aim of this study was to perform a micro-computed tomographic (micro-CT) evaluation of the effect of coronal flaring on canal transportation and centering ability of two heat treated nickel-titanium rotary instruments, 2Shape (Micro Mega, Besançon, France) and HyFlex CM (Coltène Whaledent, Altstätten, Switzerland). Thirty extracted mandibular molars with two independent mesial canals were selected and randomly instrumented (n = 15 canals) with One Flare (Micro Mega, Besançon, France) before HyFlex CM, HyFlex CM (without coronal flaring), One Flare before 2Shape and 2Shape (without coronal flaring). One Flare (Micro Mega, Besançon, France) was introduced 4 mm below the canal entrance for canals prepared with coronal flaring. HyFlex CM and 2Shape were used accordingly to manufacturers' instructions. New files were used for each canal. During and after instrumentation, irrigation procedures were performed. Micro-CT images were obtained pre- and post-preparation to measure and record root canal transportation and centralization. They were reconstructed from root apex to canal orifices, generating approximately 1000 sections per specimen. The anatomical thirds were determined by dividing the number of cross-sectional slices by three. Root canal transportation and centralization were determined by Gambil method, and the mean values were analyzed by repeated measures analysis of variance followed by multiple comparisons of Bonferroni to compare the different instrumentations procedures and the root thirds (p < 0.05). As for root canal transportation, 2Shape reported significantly higher values compared to HyFlex CM in the cervical region independently from the coronal flaring. In the apical region, 2Shape caused significantly minor canal transportation when used with coronal flaring with compared with the absence of coronal flaring. Regarding the centralization, HyFlex CM showed higher values than 2Shape in the cervical, independently from coronal flaring. In the apical region, 2Shape with coronal flaring exhibited significant major centering ratio, compared with not. Within the limitations of this study, coronal flaring reduced canal transportation and improved centralization of the 2Shape files in the apical section while it had no significant influence on shaping ability of the HyFlex CM instruments. Coronal flaring could represent a valid strategy to improve the shaping ability of NiTi files knowing that its benefit could be influenced by the shaping file used.

Effects of L-Chg10-Teixobactin on Viability, Proliferation, and Osteo/Odontogenic Differentiation of Stem Cells from Apical Papilla.

INTRODUCTION: Intracanal medicament is one of the essential steps for ensuring success in regenerative endodontic procedures. L-Chg10-teixobactin is a novel antimicrobial agent that exhibited potent antibacterial and antibiofilm effects against Enterococcusfaecalis at low concentrations compared with ampicillin. At the same time, its cytotoxicity on dental stem cells has not been studied. This study aimed to investigate the effects of L-Chg10-teixobactin on the viability, proliferation, migration, and osteo/odontogenic differentiation of stem cells from apical papilla (SCAPs). MATERIALS AND METHODS: SCAPs isolated from immature human third molars were treated with various concentrations of L-Chg10-teixobactin, calcium hydroxide, and dimethyl sulfoxide. The viability and proliferation of SCAPs were assessed using the LIVE/DEAD Viability/Cytotoxicity Kit and Cell Counting Kit-8. A scratch wound healing test was used to evaluate the lateral migration capacity of SCAPs. Alkaline phosphatase (ALP) activity, calcium mineralization ability tests -ie, ALP staining and alizarin red S staining, and quantitative real-time polymerase chain reaction were performed to assess the osteo /odontogenic differentiation of SCAPs. RESULTS: The tested concentrations of L-Chg10-teixobactin (0.01, 0.02, and 0.03 mg/mL), 1 mg/mL calcium hydroxide, and 0.03% dimethyl sulfoxide had no significant cytotoxic effect on SCAPs at any time point (P > .05). Besides, there were no significant differences between the control and experimental groups in SCAPs' viability, proliferation, and migration. L-Chg10-teixobactin upregulated the gene expression of osteo/odontogenic markers in SCAPs, while no significant difference was found in the ALP activity and alizarin red S staining. CONCLUSIONS: L-Chg10-teixobactin demonstrated excellent biocompatibility on SCAPs at concentrations from 0.01 to 0.03 mg/mL and potentially enhance the osteo/odontogenic differentiation of SCAPs; suggesting its promising role as root canal medicament for regenerative endodontic procedures.

Effect of magnifying loupes on tooth preparation of complete coverage crown: A quantitative assessment using a digital approach.

OBJECTIVE: The purpose of this study was to evaluate the use of magnifying loupes (×2.5) on the quality of tooth preparation for complete coverage crowns; performed by predoctoral students using an objective and quantitative digital method. MATERIALS AND METHODS: Forty-two predoctoral students were randomly assigned into 2 groups to perform tooth preparation for a complete coverage crown on a mandibular first molar in a manikin, with and without the use of magnifying loupes. All preparations were digitally evaluated by PrepCheck 3.0 (Dentsply Sirona). Parameters including tooth reduction, total occlusal convergence (TOC), undercut, margin quality and surface quality were assessed. Continuous data were analysed using Paired t-tests or Wilcoxon Signed Ranks tests. Ordinal data were analysed by McNemer's tests. To further detect the majority pattern ( > 50%) in each group (with and without loupes), one-sample t-test or one-sample Wilcoxon Signed Rank test was performed. The level of significance was set at p = .001 after Bonferroni adjustments for multiple testing. RESULTS: No significant differences in the measured outcomes were found between the groups with or without the use of magnifying loupes (p > .002). The majority (>50%) of both groups had no undercuts (99.3% and 99.4% both p < .001) and the preparation type was within tolerance (81.6% and 85.3%, both p < .001) with acceptable margin (86.4% and 86.3%, both p < .001) and acceptable surface quality (99.0% and 99.1%, both p < .001). However, the majority of both groups underprepared occlusally (96.0% and 95.4%, both p < .001) and axially (65.3% and 67%, both p < .001). Only 30.0%-42.1% of the participants achieved the TOC within 0-20°. CONCLUSIONS: Within the limitations of this study, the use of magnification loupes does not appear to significantly improve the quality of tooth preparation for complete coverage crown. The TOC was also found to be overprepared and occlusally underprepared.

PROMs Following Root Canal Treatment and Surgical Endodontic Treatment.

The FDI is currently working on developing a tool to encompass patient-reported outcome measures (PROMs) within the overall assessment of outcomes of endodontic treatment. The outcome of endodontic treatment has traditionally been determined by various clinical and radiographic criteria. However, these parameters do not address the impact of treatment on a patient's oral health-related quality of life (OHRQoL). OHRQoL, a crucial PROM, can be used to understand treatment outcome from a patient-centred perspective, thus improving clinician-patient communication whilst guiding decision-making. This focussed review aims to recount the OHRQoL of patients following nonsurgical root canal treatment and surgical endodontic treatment, with a specific focus on the minimal important difference (MID; the minimum score changes of an outcome instrument for a patient to register a clinically significant change in their OHRQoL and/or oral condition) and the methods used to determine it. The current evidence indicates that the OHRQoL of patients requiring root canal treatment is poorer than those without such need. Accordingly, the literature suggests that OHRQoL improves following nonsurgical or surgical endodontic treatment. However, study methodologies vary widely, and conclusions cannot be drawn with high confidence, nor can MID recommendations be provided. Well-designed clinical studies with baseline measurements and appropriate follow-up time frames are therefore needed. Despite that the literature is rife with outcome studies, research on PROMs is an area that deserves greater attention, particularly in relation to the MID. Determining the MID will facilitate the understanding of changes in outcome scores from the patients' perspective, thus allowing for more informed decision-making in clinical practice.