International endodontic journal最新文献

Aim: Natural bioactive products have been tested as alternative antimicrobial agents. This study evaluated the effect of Punica granatum extract (PGE) on oral multispecies biofilms.

Methodology: Lyophilized extracts from pomegranate peel were prepared, and the punicalagin content was assessed by ultra-performance liquid chromatography (UPLC). Oral multispecies biofilms from 2 donors were grown on four collagen-coated hydroxyapatite discs. After incubation for 7 days or 3 weeks, the biofilms were exposed to water (control), 2% CHX, 10% PGE, 20% PGE or 30% PGE for 3 min. The proportions of dead bacteria were assessed by the live/dead staining and confocal microscopy. After the analysis, the best PGE concentration (30%) was combined with CHX. The experimental phases were repeated using water, 2% CHX, 30% PGE and 30% PGE + 2% CHX. Five random areas of the biofilm on each disc were scanned, resulting in 20 scanned areas for each group.

Results: Regarding the biofilm volume, no differences were found amongst solutions (p = .111). The PGE solution killed bacteria effectively in 1-week, 2-week and 3-week-old-plaque biofilms, ranging from 37 to 55.3%, depending on the PGE concentration. The 30% PGE (a) (p = .0009) had greater antibiofilm effectiveness than 2% CHX (b), which killed bacteria in the 25.2 to 48.7% range. The 10% and 20% PGE had intermediate values (ab), without significant differences from 30% PGE (p = 1.002). Water (c) had the lowest proportion of dead bacteria (p < .00001) in a range of 5 to 6.7% and lower effectiveness in killing bacteria (p < .05). The PGE alone or mixed with 2% CHX had greater anti-biofilm effectiveness than CHX (p < .05). The old plaque biofilms were more resistant than the 7-day-old plaque (p < .05).

Conclusions: The 30% PGE (alone or combined with CHX) exhibited a greater antibiofilm effect on oral multispecies biofilms grown on hydroxyapatite discs than 2% CHX.

Aim: The aim of this study is to investigate the expression of inflammatory biomarkers (TNF-α, IL-10, IL-1β) and the pulpitis-associated miRNA (miR-30a-5p and miR-128-3p) in pulp tissue samples from unrestored teeth with a vital normal pulp (NP), teeth with symptomatic irreversible pulpitis (IP) and in unrestored teeth with periodontal disease, unresponsive to periodontal therapy, and a vital pulp (EP).

Methodology: Thirty patients were included in this observational study (10 teeth with NP, 10 teeth with IP, 10 teeth with EP). Dental pulp tissues samples were collected from patients during root canal treatment (RCT). RNA was extracted and qRT-PCR of target genes (tumour necrosis factor [TNF]-α, interleukin [IL]-1β, IL-10) and miRNAs (has-miR-30a-5p, has-miR-128-3p) performed to assess the expression profile. Fold-change in expression was calculated using the formula 2^{-(ΔCt(Exp)-ΔCt(Ctrl))}. One-way anova with post-hoc Tukey's was used to determine significant differences between groups. The significance level was set at 5% (p < .05). All teeth were also followed up clinically for 1 year and evaluated for a range of endodontic and periodontal-related outcomes.

Results: All investigated genes significantly increased in expression and miRNAs significantly decreased in expression in the IP and EP groups compared with the NP group (p < .05). With regards to TNF-α and IL-1β there were no significant differences in expression between the IP and EP groups (p > .05), whereas IL-10 expression levels were significantly reduced in the EP compared with the IP group (p < .05). Both miR-30a-5p and miR-128-3p showed significantly reduced expression in both IP and EP lesions, compared with NP (p < .05); however, no significant differences in miRNA expression were observed between IP and EP groups (p > .05). One year after root canal treatment and periodontal maintenance, tooth mobility and probing depth were significantly reduced in the EP group (p < .05).

Conclusion: Pulp tissues from teeth with IP and EP presented similar levels of altered inflammatory markers compared with NP. TNF-α, IL-10, IL-1β cytokines and miRNAs (miR-30a-5p and miR-128-3p) are potential objective biomarkers to indicate pulpal inflammatory status, aiding diagnosis and directing clinical decision-making. RCT may be beneficial to improve stage III periodontitis unresponsive to non-surgical periodontal treatment, but further research is required.

Aim: To evaluate the 6-year outcome of root canal treatment irrigated with 0.5% or 3% sodium hypochlorite (NaOCl).

Methodology: The baseline trial was designed as a quasi-randomized clinical trial. Patients referred for root canal treatment to an endodontic specialist clinic were recruited to the study (n = 298). The concentration of NaOCl was allocated quasi-randomized to 271 subjects (0.5% [n = 139], 3% [n = 132]). Bacterial sampling was performed immediately before root canal filling. Samples were cultured and evaluated as growth or no growth. Patients were invited to a clinical and radiological follow-up >5 years postoperatively. The clinical outcome measurements were tooth survival, cumulative incidence of endodontic retreatments, patients' assessment of pain, clinical findings and radiological signs of apical periodontitis (AP).

Results: Tooth survival was 85.6% in the 0.5% NaOCl group and 81.1% in the 3% NaOCl group (p = .45). There was no record of retreatment in 94.4% in the 0.5% NaOCl group and in 92.2% in the 3% NaOCl group (p = .76). The percentage of asymptomatic cases were 87.8% in the 0.5% group and 85.3% in the 3% NaOCl group (p = .81). Absence of clinical signs of AP was seen in 86.6% in the 0.5% NaOCl group and in 83.6% in the 3% NaOCl group (p = .80). Absence of radiological signs of AP was seen in 74.0% in the 0.5% NaOCl group and 64.1% in the 3% NaOCl group (p = .20). Subjects with positive culture before root filling reported subjective pain with a significantly higher frequency as compared to negative-culture subjects (p = .014).

Conclusions: The use of 0.5% or 3% NaOCl for irrigation during root canal treatment resulted in similar clinical outcomes 5-7 years postoperatively. Persisting bacteria immediately before root filling may predict future episodes of subjective pain.

Aim: To evaluate M1 and M2 macrophage polarization in radicular cysts and periapical granulomas through an immunohistochemical analysis and the correlation between macrophage polarization and histopathological diagnosis, clinical characteristics and lesion volume using cone-beam computed tomography.

Methodology: Periapical biopsies diagnosed as radicular cysts (n = 52) and periapical granulomas (n = 51) were analysed by immunohistochemical method. Teeth with periapical lesion with no history of root canal treatment (primary lesion) and lesions persistent to root canal treatment (persistent lesions) were included. Pathological diagnosis, patients' age, gender and clinical characteristics were obtained from treatment records. A cone-beam computed tomographic periapical volume index (CBCTPAVI) score was assigned to each periapical lesion based on the volume of the lesion. Immuno-expressions of CD68 and CD163 were quantified. The CD68/CD163 ratio was adopted to represent M1 or M2 macrophage polarization. Mann-Whitney U test was used to determine the different CD68/CD163 ratio between groups of radicular cyst and periapical granuloma. Spearman's correlation test was performed to assess the correlation between the CD68/CD163 ratio and lesion volume and CBCTPAVI score.

Results: Radicular cysts and periapical granulomas had CD68/CD163 median of 2.05 (IQR = 1.33) and 1.26 (IQR = 0.81), respectively. A significantly higher CD68/CD163 ratio was observed in radicular cysts (p < .001). In contrast, periapical granulomas had significantly lower median of CD68/CD163 ratio. Larger lesions had a higher median of CD68/CD163 ratio, while smaller lesions had lower median of CD68/CD163 ratio (p = .007, r_s = .262). CD68/CD163 ratio was significantly correlated with the CBCTPAVI score in the overall periapical lesions (p = .002, r_s = .306). The higher CD68/CD163 ratio in larger lesions indicated a higher degree of M1 polarization compared to smaller lesions. Regarding the pathological diagnosis, there was a significant positive correlation between CBCTPAVI score and CD68/CD163 ratio in periapical granulomas (p < .001, r_s = .453), whereas the negative correlation was observed for radicular cysts (p < .001, r_s = -.471).

Conclusions: Periapical granulomas are characterized by a M2-dominant macrophage polarization, while radicular cysts have significantly higher M1 macrophages. The higher degree of M1 macrophage polarization was significantly correlated with larger volume and higher CBCTPAVI scores of overall periapical lesion and periapical granuloma.

Aim: The pathways to post-operative pain are complex and encompass factors that extend beyond the treatment protocol employed. This study aimed to identify patient-related predictors of post-operative pain following root canal treatment.

Methodology: A total of 154 patients received a single-visit root canal treatment for asymptomatic necrotic mandibular molars. Before treatment, dental anxiety, dental fear and sense of coherence (SOC) were measured as predictors for each patient using validated questionnaires. Other measured predictors included gender, age, previous negative experiences at the dental offices and prior root canal treatment. Post-operative pain was assessed using the Numeric Rating Scale at multiple time-points over 30 days. Structural equation analysis was employed to evaluate the direct and indirect effects of patient-related predictors on a theoretical model of post-operative pain. The irrigant solution was also included in the model, as it was the only aspect that varied in the treatment protocol (sodium hypochlorite 2.5% and 8.25%).

Results: Dental anxiety (coefficient 0.028; p < .01), dental fear (coefficient 0.007; p = .02) and irrigant solution (coefficient 0.004; p = .03) exerted a direct effect on post-operative pain. SOC exerted an indirect effect on post-operative (coefficient 0.006; p = .01) through dental anxiety and dental fear. Moreover, previous negative experiences (coefficient 0.048; p = .04) exerted an indirect effect on post-operative pain through dental anxiety.

Conclusions: Dental anxiety, dental fear, previous negative experiences and SOC are patient-related predictors of post-operative pain following root canal treatment. These factors should be taken into consideration in clinical practice, as patients with these characteristics may be at an increased risk of experiencing post-operative pain.

Objectives: The objective of this study is to analyse the gene expression profile of the dental pulp (DP) of human premolars subjected to 7 and 28 days of orthodontic force (OF) in vivo by using RNA sequencing. The maxillary and mandibular DP were additionally compared.

Methods: Healthy patients requiring orthodontic premolar extractions were randomly assigned to one of the three groups: control (CG) where no OF was applied, 7 and 28 days, where premolars were extracted either 7 or 28 days after the application of a 50-100 g OF. Total RNA was extracted from the DP and analysed via RNA-seq. Differentially expressed genes (DEGs) were identified using a false discovery rate and fold change threshold of <0.05 and ≥1.5, respectively. Functional analysis was performed.

Results: After 7 days of OF, pulp reaction indicates immune response, hypoxia, DNA damage and epigenetic regulation. After 28 days, cell adhesion, migration, organization and tissue repair are evident. The maxillary and mandibular pulp tissues react differently to OF. The maxilla exhibits minimal alterations, mostly related to immune response at 7 days and tissue repair at 28 days, whereas the mandible shows mostly DNA damage and epigenetic regulation at 7 days and return to the original state at 28 days.

Conclusions: This study demonstrates that the early reaction of the DP to OF is marked by immune response, hypoxia and DNA damage. In contrast, after 28 days, cell adhesion, migration, organization, tissue repair and dentine formation are observed. Maxillary and mandibular premolars react differently to OF: although the maxilla exhibits minimal alterations at both time points, the mandible mostly shows DNA damage, epigenetic regulation, and immune response at 7 days. These disparities could stem from different blood supplies or the lower maxillary bone density, potentially triggering faster biological changes. Our findings provide insights into the gene regulatory networks modulating DP response to OF.

Aim: Angiogenesis is a key event in the successful healing of pulp injuries, and hypoxia is the main stimulator of pulpal angiogenesis. In this study, we investigated the effect of hypoxia on the proangiogenic potential of human dental pulp stem cells (hDPSCs) and the role of miR-143-5p in the process.

Methodology: Human dental pulp stem cells were isolated, cultured and characterized in vitro. Cobalt chloride (CoCl₂) was used to induce hypoxia in hDPSCs. CCK-8 and Transwell assays were used to determine the effect of hypoxia on hDPSCs proliferation and migration. Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting (WB) and ELISA were performed to assess the mRNA and protein levels of HIF-1α and angiogenic cytokines in hDPSCs. The effect of hypoxia on hDPSCs proangiogenic potential was measured in vitro using Matrigel tube formation and chick chorioallantoic membrane (CAM) assays. Recombinant lentiviral vectors were constructed to stably overexpress or inhibit miR-143-5p in hDPSCs, and the proangiogenic effects were assessed using qRT-PCR, WB, and tube formation assays. miR-143-5p target genes were identified and verified using bioinformatics prediction tools, dual-luciferase reporter assays and RNA pull-down experiments. Finally, a subcutaneous transplantation model in nude mice was used to determine the effects of hypoxia treatment and miR-143-5p overexpression/inhibition in hDPSCs in dental pulp regeneration.

Results: Hypoxia promotes hDPSCs proliferation, migration and proangiogenic potential. The in vivo experiments showed that hypoxia treatment (50 and 100 μM CoCl₂) promoted pulp angiogenesis and dentine formation. In contrast to the levels of proangiogenic factors, miR-143-5p levels decreased with increasing CoCl₂ concentration. miR-143-5p inhibition significantly promoted proangiogenic potential of hDPSCs, whereas miR-143-5p overexpression inhibited angiogenesis in vitro. Dual-luciferase reporter assay identified retinoic acid receptor-related orphan receptor alpha (RORA) as an miR-143-5p target gene in hDPSCs. RNA pull-down experiments demonstrated that HIF-1α and RORA were pulled down by biotin-labelled miR-143-5p, and the levels of HIF-1α and RORA bound to miR-143-5p in the hypoxia group were lower than those in the normoxia group. Inhibition of miR-143-5p expression in hDPSCs promoted ectopic dental pulp tissue regeneration.

Conclusions: CoCl₂-induced hypoxia promotes hDPSCs-driven paracrine angiogenesis and pulp regeneration. The inhibition of miR-143-5p upregulates the proangiogenic potential of hDPSCs under hypoxic conditions by directly targeting HIF-1α and RORA.

Aim: Epigenetic regulation of the key inflammatory genes plays a crucial role in controlling monocyte/macrophage-mediated local and systemic responses to bacterial challenges. However, it has not been addressed in apical periodontitis (AP). We aimed to explore the methylation pattern of the TNF-α gene promoter and its association with the inflammatory phenotype of peripheral blood monocytes from individuals with AP and controls.

Methods: A cross-sectional study was conducted, including otherwise healthy individuals with AP (n = 25) and controls (n = 29). Monocytes were isolated from the volunteer's blood samples using a Ficoll gradient followed by negative immunoselection. RNA and DNA were extracted. The DNA methylation profiles of the TNF-α gene promoter region were analyzed using bisulfite sequencing PCR. The mRNA expression levels of DNA methyltransferases 3a (DNMT3a) and Ten Eleven Translocation enzymes 1(TET1) were assessed by qPCR. A fraction of primary monocytes was also cultured for 24 h, and the supernatant was collected to measure cytokine levels through a Luminex assay. Generalized structural equation models (GSEM) evaluated the association between AP, DNA methylation, and TNF-α protein expression controlled for potential covariates. Models included the effect of the methylation of TNF-α gene promoter as a mediator of the association between AP and TNF-α protein expression levels.

Results: Monocytes from AP individuals exhibited a heightened secretion of TNF-α and IL-1β and hypomethylation of the TNF gene promoter (p < .05). AP diagnosis was associated with the TNF-α gene promoter´s hypomethylated profile and enhanced pro-inflammatory cytokine levels, while lower methylation of the gene promoter region and -163 CpG single site mediated TNF-α overexpression (p < .05).

Conclusions: DNA hypomethylation at the TNF-α gene mediates a proinflammatory phenotype in monocytes from AP patients, supporting a role in the systemic response.

Aim: High-quality, prospective clinical studies are needed to increase evidence for guided endodontics. This study aims to assess the clinical outcome of guided endodontics for treatment of teeth presenting with pulp canal obliteration (PCO) in comparison with freehand treatment.

Methodology: This trial was registered in the ISRCTN.com registry (ISRCTN75277265) and designed as a controlled clinical trial: Single arm trial, prospective, nonrandomized, single-centre study (ethical approval number S64630). Inclusion criteria were; tooth presenting with PCO and symptoms and/or signs of apical periodontitis (AP). An external control group was selected from clinical records of patients presenting the same criteria but treated freehanded. Guided root canal treatments were performed by the same operator on all patients. Freehanded treatments were performed by a specialist in endodontics under microscope with pre-operative CBCT available. Primary outcome for both groups was evaluated as: canal found, canal not found, or perforation. As secondary outcome, the qualitative accuracy of the drill path was assessed as: optimal precision, acceptable precision or technical failure. Patients were followed up yearly. Descriptive statistics on the study patient's demographics and healing outcome were performed and specific statistical analysis was performed on each outcome variable.

Results: A total of 133 teeth were included (n = 60 guided, n = 73 freehanded) from 128 patients (n = 59 guided, n = 69 freehanded). The primary outcome for the guided group was: 59 teeth canals found and 1 tooth canal not found. No perforations were recorded. In the freehanded group, the root canal was successfully found in 59 teeth, seven were not found, and seven had a perforation. An analysis of all data showed that guided endodontics presented statistically significant better outcome than freehand treatment (p < .05).

Conclusion: Guided endodontics showed a statistically significant better outcome than freehanded treatment resulting in less technical failures. However, it is a complex procedure which should be carried out by an experienced endodontist with the aid of a dental microscope.

Aim: Despite the high success rate of endodontic microsurgery (EMS), it is difficult to suggest EMS as a general treatment option considering the difficulty of the procedure. A surgical guide has been proposed to overcome this problem. This study aimed to evaluate the stability of the surgical guide of a new design concept, as well as the accuracy of root resection, and to introduce the manufacturing method of the newly designed surgical guide.

Methodology: The experiment was conducted on 59 roots (9 in the maxillary and 50 in the mandibular region) of adult human cadavers. The surgical guide was designed using CAD/CAM design software based on cone-beam computed tomography (CBCT) images and optical scan files. Unlike conventional surgical guides, the surgical guide proposed herein was designed to act as a tooth-bone-supported removable appliance. Two different types of guides were prepared: the osteotomy guide (O guide) for separation of the cortical bone above the root tip with a trephine bur with an outer diameter of 6 mm and the root resection guide (R guide) for resection of the root tip with a trephine bur with an outer diameter of 4 mm. For stability evaluation, the guides were pressed at five predetermined locations after installation and checked for the presence of any movement. For accuracy evaluation, the length at which the root tip was cut was measured and examined by overlapping the preoperative and postoperative CBCT images.

Results: Of the 15 R guides, 14 were stably installed without mobility. For the R guide group, the root tip was resected with an average of 3.2 mm, showing better results than the no-guide group with an average of 4.0 mm.

Conclusions: The newly designed surgical guide of this study can be applied more stably, enabling root resection to be performed more accurately and simply according to the preoperative plan than when performed without a guide.