Project description:Apical periodontitis is an inflammatory disease of the periradicular tissues caused by the host's immune response to infection of the root canal system. MicroRNAs (miRNAs) have been shown to play an important role in the regulation of inflammation and the immune response; however, their role in the pathogenesis of endodontic periapical disease has not been explored. The purpose of this study was to examine the differential expression of miRNAs in diseased periapical tissues as compared with healthy controls.We first compared miRNA profiles in diseased periapical tissues collected from patients undergoing endodontic surgery with those of healthy pulps by using microarray analyses. The target genes of the differentially expressed miRNAs were identified by using miRWalk and PubMed. Selected miRNAs linked to inflammation and the immune response were then confirmed in a separate cohort of diseased and healthy tissues by using quantitative reverse transcription-polymerase chain reaction. Healthy pulps and periodontal ligaments were used as controls. Data were normalized to the level of SNORD 44, which served as an endogenous control.Of the 381 miRNAs identified by using microarray, 24 miRNAs were down-regulated in diseased periapical tissues compared with controls (n = 13) (P < .003). The down-regulation of 7 miRNAs was confirmed from 9 selected miRNAs by using quantitative real-time polymerase chain reaction (n = 19) (P < .05). Target genes of these miRNAs include key mediators in the immune and inflammatory response such as interleukin-6, matrix metalloproteinase-9, and transforming growth factor-?.These findings offer new insight into the pathogenesis of endodontic disease and have the potential to impact the development of new methods for prevention, diagnosis, and treatment of apical periodontitis.
Project description:In this study, we have investigated the association between the baseline gene expression profile in periapical granuloma and periapical wound healing after surgical endodontic treatment. Twenty-seven patients aged between 15 and 57 years underwent periapical surgery. The retrieved periapical tissue sample was used for mRNA expression analysis of COL1A1, VTN, ITGA5, IL-4, TNF, ANGPT, VEGFA, and CTGF. All patients were recalled after 6 and 12 months for periapical healing evaluation. Healing was then correlated with baseline gene expression. Healing was observed in 15 patients at the end of 6 months, which increased to 21 patients after 12 months. Six patients showed no healing even after 12 months. Analysis of baseline expression levels of the tested genes with healing status showed the mean relative expression of VTN, VEGFA, ANGPT, TNF, and CTGF to be significantly different (p < 0.05) between the healing group (6 and 12 months) (72.99%) and the non-healing (94.42%) group. Periapical Index scores 3-5 exhibited a positive correlation with ITGA-5 expression. Overexpression of ANGPT and a strong positive correlation between ITGA5 and PAI scores in the non-healing group of patients may suggest these genes to be a potential prognostic biomarker for periapical wound non-healing after surgical endodontic treatment.
Project description:ObjectivesThis study compared the clinical and radiological outcomes of regenerative endodontic procedures (REPs) using blood clots (BCs), platelet-rich plasma (PRP), and platelet-rich fibrin (PRF) through intraoral periapical radiography (IOPAR) and cone-beam computed tomography (CBCT).Materials and methodsForty-five single-rooted necrotic teeth with periapical pathology were randomly allocated to receive BC, PRP, or PRF as an individual scaffold. Outcomes were evaluated in 35 teeth in 23 patients with a follow-up period of 12-24 months through qualitative IOPAR scoring and quantitative CBCT measurements. Healing of periapical lesions and in immature teeth, changes in the apical foramen diameter (AFD), root wall thickness (RWT), and root length (RL) were assessed. A p value less than 0.05 was considered to indicate statistical significance.ResultsAll teeth were asymptomatic except 1 in the PRP group. Periapical lesion healing was seen in all except 2 teeth in the BC group and 3 in the PRP group. Both IOPAR and CBCT revealed no significant differences in bone healing or changes in AFD, RWT, and RL among the 3 groups. A positive pulp sensibility response to the cold test was seen in 2 teeth in the BC group, but none to the electric pulp test. Intracanal calcification (ICC) was evident in more teeth in the BC group than in the PRP and PRF groups, and was also significantly higher in immature teeth.ConclusionsOur results revealed that BC, PRP, and PRF have similar potential as scaffolds in REPs, and ICC may be a concern for long-term outcomes.
Project description:Periapical periodontitis results from pulpal infection leading to pulpal necrosis and resorption of periapical bone. The current treatment is root canal therapy, which attempts to eliminate infection and necrotic tissue. But, in some cases periapical inflammation doesn't resolve even after treatment. Resolvins belongs to a large family of specialized pro-resolving lipid mediators that actively resolves inflammation signaling via specific receptors. Resolvin D2 (RvD2), a metabolite of docosahexaenoic acid (DHA), was tested as an intracanal medicament in rats in vivo. Mechanism was evaluated in rat primary dental pulp cells (DPCs) in vitro. The results demonstrate that RvD2 reduces inflammatory cell infiltrate, periapical lesion size, and fosters pulp like tissue regeneration and healing of periapical lesion. RvD2 enhanced expression of its receptor, GPR18, dentin matrix acidic phosphoprotein 1 (DMP1) and mineralization in vivo and in vitro. Moreover, RvD2 induces phosphorylation of Stat3 transcription factor in dental pulp cells. We conclude that intracanal treatment with RvD2 resolves inflammation and promoting calcification around root apex and healing of periapical bone lesions. The data suggest that RvD2 induces active resolution of inflammation with pulp-like tissue regeneration after root canal infection and thus maybe suitable for treating periapical lesions.
Project description:Introduction: Pharmacotherapy with analgesics and non-steroidal anti-inflammatory drugs has been traditionally used to relief post-operative pain of endodontic treatments. However, due to the side effects reported for these drugs, some efforts have been made to decrease the post-operative pain of the endodontic treatments through laser irradiation. The present study aimed to evaluate the effects of low level laser therapy (LLLT) on the reduction of pain after root canal retreatment. Methods: In this clinical trial, 61 patients requiring endodontic retreatments in posterior teeth were selected. A single visit endodontic retreatment was performed. After biomechanical preparation, low level laser was irradiated to the buccal and lingual mucosa overlying the apices of the target tooth in the experimental group. In the control group patients received placebo laser to eliminate the probable psychological effects of laser. Laser irradiation was done with a single dose of 808 nm wavelength (Whitening Lase II- Laser DMC, Samsung, Korea) with 100 mW power, and dose of 70 J/cm2 for 80 seconds. Pain severity was recorded before, immediately after and 4, 8, 12, 24 and 48 hours after the treatment by visual analogue scale (VAS). The pain scores were statistically analyzed by chi-square test between 2 groups. The effects of different variables on the post-operative pain experience were also studied by means of Logistic regression. Results: Pain scores decreased significantly through time until 48 hours after treatment. No significant differences were observed between the 2 modalities regarding pain scores at any time. According to regression analysis, pain severity scores were lower in the laser-irradiated specimens than control groups (OR = 5.69); however, this difference was not statistically significant. Consumption of analgesics after the treatment had significant effect in decreasing post-operative pain experience (OR = 56) while factors of age, gender, laser irradiation, pre-treatment pain scores and education level did not. Conclusion: Low level laser irradiation had limited effects to decrease pain associated with the endodontic retreatments in the first and second molars; however, more studies are required to assess the effects of different parameters of low level laser in this regard.
Project description:The Regenerative Endodontic Procedure (REP) is a biologically based method in which a damaged pulp-dentin complex is replaced by a new vital tissue. This umbrella review aimed to critically assess the available systematic reviews (SRs) on REP. An electronic database search was conducted (PubMed-Medline, CENTRAL, Scielo, Web of Science, and LILACS) until December 2020. Studies were included if they were an SR on REP. The Risk of Bias (RoB) of SRs was analyzed using the Measurement Tool to Assess SRs criteria 2 (AMSTAR2). The primary outcome was the methodological quality in each specific section of REP protocols and outcomes. From 403 entries, 29 SRs were included. Regarding the methodological quality, ten studies were of critically low, three of low, fourteen of moderate, and two were rated as high quality. The quality of evidence produced by the available SRs was not favorable. Future high standard SRs and well-designed clinical trials are warranted to better elucidate the clinical protocols and outcomes of REP.
Project description:Background: Many challenges encounter the endodontist, especially when dealing with necrotic teeth with chronic periapical lesions. Postoperative pain may be induced following conventional endodontic therapy and total eradication of the bacteria is almost unachievable even with recently available techniques. In recent years, diode laser usage in the endodontic field has gained acceptance. Thus, this study aimed to investigate the ability of the diode laser (DL) to decrease postoperative pain and achieve root canal sterility. Methods: 56 patients with anterior teeth with chronic periapical lesions in upper anterior teeth were randomly divided into two groups (n = 28). All patients were treated with two visits of conventional root canal treatment with ProTaper Universal. The DL group: root canals were irradiated with 200 µm fiber optic at both visits; the control group (Endo): the DL fiber was placed in root canal with no activation. Bacterial samples were collected from all the cases at each step of the treatment. Pain levels were evaluated using a numerical rating scale preoperatively, and after 6, 12, 24, 48 hours and 7 days. Bacterial count was used to detect both aerobic and anaerobic bacterial load. Results: The qualitative pain scores revealed statistically significant lower pain levels in the DL group compared with the Endo group at all time intervals (P<0.001), except preoperatively where there was no significant difference. There was a statistically significant lower bacterial count for both aerobic and anaerobic bacteria in the DL group compared with the Endo group in both S3 samples (after laser application) and S4 samples (bacterial colonization) (P<0.001). Conclusion: The 980 nm diode laser may be a successful adjunct to conventional endodontic treatment of necrotic cases with chronic periapical lesions in terms of postoperative pain and root canal disinfection. Trail registration: PACTR201511001275414 (date: 23 rd September 2015).
Project description:AimThe aim of the study was to measure serum levels of molecular markers of inflammation in patients undergoing non-surgical root canal retreatment (Re-RCT) and periapical surgery (PS) for the treatment of apical periodontitis and to establish if such levels are influenced by the size of apical radiolucencies at baseline and by the treatment outcome.MethodologyA total of 115 participants were recruited (n = 50 Controls, n = 35 Re-RCT, n = 30 PS). Preoperative periapical radiographs and cone beam CT (CBCT) scans of teeth were taken. Blood was collected from treatment groups at baseline, 3-, 6-, and 12-month post-treatment and from controls at baseline and 12 months. Serum levels of IL-1β, IL-6, IL-8, TNF-α, Pentraxin 3, ICAM-1, VCAM-1, hs-CRP, FGF-23, MMP-2, MMP-8, MMP-9, C3 and ADMA were analysed using multiplex immunoassay and enzyme-linked immunosorbent assay. Different time points within the same group were compared using Wilcoxon signed-rank test, and differences between groups were analysed using the Mann-Whitney test. Non-linear association between different factors was assessed using Spearman's correlation.ResultsPreoperative serum levels of FGF-23, IL-1β, hs-CRP and ADMA were significantly higher in the diseased groups compared with controls (p < .001; p = .008; p < .001; p = .013, respectively). The preoperative size of the radiolucency was associated with increased levels of FGF-23, IL-1β and IL-6. At 3-months following treatment, IL-1β, IL-8, hs-CRP, C3, MMP-2 and MMP-9 levels increased compared with baseline in treatment groups. IL-1β and IL-8 further increased at 6 months, whereas FGF-23, hs-CRP, C3, MMP2 and MMP-9 decreased. One-year post-treatment, FGF-23, pentraxin-3 and ADMA were significantly reduced below baseline levels. At the 1-year review, CBCT revealed that 25.9% of treated cases completely healed, while 63% were healing, and 11.1% failed. Treatment outcome was found to be influenced by preoperative levels of ADMA and IL-8 levels at 6 months.ConclusionsBoth symptomatic and asymptomatic apical periodontitis (AP) can contribute to increased levels of molecular markers of inflammation. A further transient inflammatory markers rise after root canal retreatment and apical surgery were demonstrated. Successful endodontic treatment and periapical surgery result in a long-term reduction in inflammatory marker levels.
Project description:OBJECTIVES: To detect the expression levels of MLKL and p-MLKL, and explore its potential roles in inflammatory cell infiltration, angiogenesis, and bone destruction, in human and mouse periapical lesions. METHODS: Forty-six human periapical tissues, including periapical granulomas (PGs, n =26), radicular cysts (RCs, n =20), and eight healthy control tissues were collected. Samples were fixed and analyzed by HE staining, RNA-Seq, western blot, and immunohistochemical staining. A periapical lesion mouse model was induced by pulp exposure in the first lower molars of 15 C57BL/6J mice. After lesion induction, the mice were sacrificed on days 0, 21, and 35. Mandibles were harvested for microcomputed tomography scanning, histologic observation, immunohistochemistry, enzyme histochemistry, and double immunofluorescence analysis. Double immunofluorescence was utilized to assess the colocalization of phosphorylated MLKL (p-MLKL) with CD34, matrix metalloproteinase-9 (MMP-9), and Cathepsin K (CTSK) in human and mouse periapical lesions. RESULTS: RNA-Seq analysis showed that, in comparison with healthy gingiva tissues, MLKL was more significantly upregulated in periapical lesions (P-value < 0.05). Immunohistochemistry staining showed that, MLKL and p-MLKL were significantly overexpressed in the RC and PG groups compared with the control group (P-value < 0.05). However, the difference between the RC and PG groups was insignificant (P-value > 0.05). p-MLKL-positive cells were mainly lymphocytes, epithelial cells, and endothelial cells around the vascular wall. In mouse periapical lesions, the expression levels of p-MLKL were positively correlated with the bone defect area and tartrate-resistant acid phosphatase-positive (TRAP+) cell amounts (R2=0.4108, P-value < 0.05; R2=0.5668, P-value < 0.05, respectively). The double-labeling analysis showed that p-MLKL colocalized with CD34 and MMP-9 in human samples, and with CTSK adjacent to the bone in mouse periapical lesions. CONCLUSION: MLKL and p-MLKL were overexpressed in human periapical lesions. p-MLKL exhibited a close relationship with angiogenesis and alveolar bone resorption.
Project description:Dental caries, one of the most prevalent infectious diseases worldwide, affects approximately 80% of children and the majority of adults. Dental caries may result in endodontic disease, leading to dental pulp necrosis, periapical inflammation and bone resorption, severe pain, and tooth loss. Periapical inflammation may also increase inflammation in other parts of the body. Although many studies have attempted to develop therapies for this disease, there is still an urgent need for effective treatments. In this study, we applied a novel gene therapeutic approach using recombinant adeno-associated virus (AAV)-mediated RNAi knockdown of Cathepsin K (Ctsk) gene expression, to target osteoclasts and periapical bone resorption in a mouse model. We found that AAV-sh-Cathepsin K (AAV-sh-Ctsk) impaired osteoclast function in vivo and furthermore reduced bacterial infection-stimulated bone resorption by 88%. Reduced periapical lesion size was accompanied by decreases in mononuclear leukocyte infiltration and inflammatory cytokine expression. Our study shows that AAV-RNAi silencing of Cathepsin K in periapical tissues can significantly reduce endodontic disease development, bone destruction, and inflammation in the periapical lesion. This is the first demonstration that AAV-mediated RNAi knockdown gene therapy may significantly reduce the severity of endodontic disease.