Liensinine alleviates mouse intestinal injury induced by sepsis through inhibition of oxidative stress, inflammation, and cell apoptosis.

Sepsis is a clinical syndrome triggered by an imbalanced host response to pathogens that can lead to multiple organ dysfunction. The immune response and barrier function of the gut play an important role in the pathogenesis and progression of sepsis. This study aimed to explore the potential role of natural alkaloid Liensinine in the treatment of intestinal injury caused by sepsis and its possible molecular mechanism. In this study, a mouse model of sepsis was established by injecting LPS to explore the protective effect of Liensinine on intestinal injury in sepsis. The results showed that Liensinine could reduce the intestinal damage caused by LPS and increase the number of goblet cells. Furthermore, it decreased the release of inflammatory cytokines by inhibiting NF-kB phosphorylation and NLRP3 inflammasome synthesis. Liensinine also reduced the oxidative stress and ROS accumulation caused by LPS, and played an anti-oxidative stress role by regulating the Nrf2/keap1 signaling pathway. In addition, Liensinine alleviated the inhibition of intestinal autophagy caused by LPS by inhibiting the PI3K/Akt/mTOR pathway. And then it reduced the excessive apoptosis of intestinal cells. This study provides valuable insights for sepsis prevention and treatment, offering a potential therapeutic candidate to protect against intestinal injury and regulate the inflammatory response in sepsis.

Ferroptosis and tumor immunity: In perspective of the major cell components in the tumor microenvironment.

Ferroptosis is an iron-dependent form of cell death driven by lipid peroxidation, which is morphologically, biochemically, and genetically distinct from apoptosis, necrosis, and autophagy. Mounting studies on the essential role of ferroptosis have been published in the progression of solid tumors, metastasis, therapy, and therapy resistance. Studies showed that ferroptosis is a "double-edged sword" in tumor immunity, which means it may have both tumor-antagonizing and tumor-promoting functions. The tumor microenvironment (TME) comprises not only tumor cells but also surrounding immune cells, stromal cells, as well as noncellular components such as the extracellular matrix (ECM), cytokines, growth factors, and extracellular vesicles (EVs). In the complex and diverse condition in TME where tumor cells grow, changes in each constituent may impact tumor destiny differently. Recently, several studies have revealed the interaction between ferroptosis and different constituents in TME. Both tumor cells and nontumor cells have a dual role in tumor immunity and influence tumor progression through ferroptosis. Herein, this review aims at summarizing the role of ferroptosis in tumor immunity based on TME, focusing on the mechanisms of the interaction between the ferroptosis and the different constituents in TME, illuminating how ferroptosis plays its role in promoting or antagonizing tumors by acting with varying components in TME and proposing several questions in immunomodulatory effects of ferroptosis and ferroptosis-associated immunotherapy.

Crosstalk between ferroptosis and macrophages: potential value for targeted treatment in diseases.

Ferroptosis is a newly identified form of programmed cell death that is connected to iron-dependent lipid peroxidization. It involves a variety of physiological processes involving iron metabolism, lipid metabolism, oxidative stress, and biosynthesis of nicotinamide adenine dinucleotide phosphate, glutathione, and coenzyme Q10. So far, it has been discovered to contribute to the pathological process of many diseases, such as myocardial infarction, acute kidney injury, atherosclerosis, and so on. Macrophages are innate immune system cells that regulate metabolism, phagocytize pathogens and dead cells, mediate inflammatory reactions, promote tissue repair, etc. Emerging evidence shows strong associations between macrophages and ferroptosis, which can provide us with a deeper comprehension of the pathological process of diseases and new targets for the treatments. In this review, we summarized the crosstalk between macrophages and ferroptosis and anatomized the application of this association in disease treatments, both non-neoplastic and neoplastic diseases. In addition, we have also addressed problems that remain to be investigated, in the hope of inspiring novel therapeutic strategies for diseases.

Diagnosis of Odontogenic Maxillary Sinusitis by Cone-beam Computed Tomography: A Critical Review.

INTRODUCTION: This review aims to provide an up-to-date overview of the current applications of cone-beam computed tomography (CBCT) and other imaging modalities in diagnosing odontogenic maxillary sinusitis (OMS). Furthermore, the clinical operation procedures of radiography modalities in OMS diagnosis were summarized, with the goal of assisting clinicians in improving OMS diagnostic accuracy in clinical practice. METHOD: A comprehensive review of researches that discussed the applications of radiography modalities in the diagnosis of OMS was conducted. Pertinent information was evaluated and organized for this review. RESULTS: Cone-beam computed tomography (CBCT) offers a superior, high-resolution, and three-dimensional view of the maxillary tooth-bone-sinus complex compared to conventional radiography modalities. It enables a better understanding and classification of the spatial relationships between root apices/periapical lesions and maxillary sinus. The use of CBCT, combined with the advantages of other radiography modalities and proper image interpretation, is indispensable to enhance OMS diagnostic accuracy and mitigate the risk of missed or misdiagnosis. CONCLUSION: The significance of CBCT in the diagnosis of oral and maxillofacial conditions has garnered widespread recognition. It provides highly precise diagnostic information and classification basis for OMS. The clinical operation procedures for imaging examination are essential in ensuring the consistency and reliability of the diagnosis.

Disruption of the intestinal barrier by avermectin in carp involves oxidative stress and apoptosis and leads to intestinal inflammation.

Avermectin (AVM) is a widely used insecticide. Due to its sensitive toxicity to aquatic organisms, the toxicology of AVM on fish intestines remains unclear. Here, we established a 96 h AVM acute toxicity model to explore the effects of AVM on the intestinal tract of carp. The 96 h LC50 of carps exposed to AVM was 24.04 µg/L, 12.02 µg/L was selected as the high-dose group and 3.005 µg/L was selected as the low-dose group. After 96 h of exposure, intestinal tissues were collected and subsequently analyzed for histopathology, the activities of antioxidant oxidases (CAT, SOD, GSH-Px), and the expression of mRNA associated with oxidative stress, inflammation, and apoptosis. Our study showed that AVM exposure caused intestinal damage in carp, decreased the expression of the tight junction protein gene, activated oxidative stress, induced apoptosis, and induced intestinal inflammation in carp. Therefore, we demonstrated that AVM exposure compromised the integrity of the intestinal barrier in carp, activated oxidative stress, induced endogenous apoptosis, and induced intestinal inflammatory responses. These results indicate that AVM, as a drug-sensitive to aquatic organisms, has a much more complex toxic effect on the fish intestinal tract, which provides a new perspective for studying the toxicology of AVM on the fish intestinal tract.

Ameliorative effects of silybin against avermectin-triggered carp spleen mitochondrial dysfunction and apoptosis through inhibition of PERK-ATF4-CHOP signaling pathway.

The aim of this study was to investigate the splenic tissue damage of environmental biological drug avermectin to freshwater cultured carp and to evaluate the effect of silybin on the splenic tissue damage of carp induced by avermectin. A total of 60 carp were divided into 4 groups with 15 carp in each group, including the control group fed with basic diet, experimental group fed with basal diet and exposed to avermectin (avermectin group), experimental group fed with basal diet supplement silybin (silybin group), and experimental group fed with basal diet supplement silybin and exposed to avermectin (silybin + avermectin group). The whole test period lasted for 30 days, and spleen tissue was collected for analysis. In this study, H&E staining, mitochondrial purification and membrane potential detection, ATP detection, DHE staining, biochemical tests, qPCR, immunohistochemistry, and apoptosis staining were used to evaluate the biological processes of spleen tissue injury, mitochondrial function, oxidative stress, apoptosis, and endoplasmic reticulum stress. The results show that silybin protected carp splenic tissue damage caused by chronic avermectin exposure, decreased mitochondrial membrane potential, decreased ATP content, ROS accumulation, oxidative stress, apoptosis, and endoplasmic reticulum stress. Silybin may ameliorate the splenic tissue damage of cultured freshwater carp caused by environmental biopesticide avermectin by alleviating mitochondrial dysfunction and inhibiting PERK-ATF4-CHOP-driven mitochondrial apoptosis. Adding silybin into the diet becomes a feasible strategy to resist the pollution of avermectin and provides a theoretical basis for creating a good living environment for freshwater carp.

A bibliometric comparison of undergraduate and postgraduate endodontic education publications: The topics, trends, and challenges.

OBJECTIVES: This study employs bibliometric analysis to compare knowledge units and main topics in undergraduate and postgraduate endodontic education, aiming to identify similarities, differences, and connections. The insights gained are expected to inform the future of two-stage education to enhance continuity, highlighting evolving trends, challenges, and development directions. METHODS: Citation data were retrieved from the Web of Science Core Collections (WOSCC) database and non-WOSCC databases with two separate search formulas. VOSviewer and CiteSpace were used to analyze the distribution of research by publication years, citation-sources, co-authorship network of authors and countries, and clusters of keywords. RESULTS: The focus on undergraduate education preceded postgraduate education by nearly a decade. The United Kingdom has emerged as the most prominent contributor to endodontic literature at both levels, with the International Endodontic Journal representing the most voluminous and cited resource in this domain. Dummer is recognized as the most prolific author in undergraduate endodontic education, while Gulabivala spearheads the most extensive cluster of postgraduate education. Keywords clustering analysis reveals that undergraduate education places greater emphasis on fundamental knowledge, while postgraduate education concentrates more on clinical practice. Descriptive analyses from non-WOSCC databases align with the topics and findings from WOSCC-based bibliometric analysis. CONCLUSION: This bibliometric analysis revealed the emphasis on fundamental knowledge and teaching techniques at the undergraduate level versus advanced clinical knowledge and techniques at the postgraduate level, which originated from different learning aims and contexts. Updating the curriculum to meet the latest practices and innovations is crucial for aligning learning objectives with current and future needs, and the connection between the two levels remains a central challenge in endodontic education.

Liensinine pretreatment reduces inflammation, oxidative stress, apoptosis, and autophagy to alleviate sepsis acute kidney injury.

Liensinine is mainly derived from alkaloids extracted and isolated from lotus seeds (Nelumbo nucifera Gaertn). It possesses anti-inflammatory, and antioxidant, according to contemporary pharmacological investigations. However, the effects and therapeutic mechanisms of liensinine on acute kidney injury (AKI) models of sepsis are unclear. To gain insight into these mechanisms, we established a sepsis kidney injury model by LPS injection of mice treated with liensinine, and stimulation of HK-2 with LPS in vitro and treated with liensinine and inhibitors of p38 MAPK, JNK MAPK. We first found that liensinine significantly reduced kidney injury in sepsis mice, while suppressing excessive inflammatory responses, restoring renal oxidative stress-related biomarkers, reducing increased apoptosis in TUNEL-positive cells and excessive autophagy, and that this process was accompanied by an increase in JNK/ p38-ATF 2 axis. In vitro experiments further demonstrated that lensinine reduced the expression of KIM-1, NGAL, inhibited pro- and anti-inflammatory secretion disorders, regulated the activation of the JNK/p38-ATF 2 axis, and reduced the accumulation of ROS, as well as the reduction of apoptotic cells detected by flow cytometry, and that this process played the same role as that of p38 MAPK, JNK MAPK inhibitors. We speculate that liensinine and p38 MAPK, JNK MAPK inhibitors may act on the same targets and could be involved in the mechanism of alleviating sepsis kidney injury in part through modulation of the JNK/p38-ATF 2 axis. Our study demonstrates that lensinine is a potential drug and thus provides a potential avenue for the treatment of AKI.

Clinical Effect Evaluation of Concentrated Growth Factor in Endodontic Microsurgery: A Cross-Sectional Study.

INTRODUCTION: Concentrated growth factor (CGF) is the third-generation platelet concentrate product. This study aimed to evaluate whether the use of CGF during endodontic microsurgery had a positive influence on surgical outcomes. METHODS: Fifty-four patients who underwent endodontic microsurgery from January 2017 to November 2021 were enrolled. They were assigned to the CGF and the control groups according to whether CGF was used during the surgery and followed up at 6, 12, and 18 months after surgery. Preoperative classification of the cases and follow-up radiographic outcomes were based on Kim's classification and Molven's criteria, respectively, and evaluated by 2 calibrated endodontists. The Student t test and χ2 test were used to assess the baseline of 2 groups. Rank sum test was used to determine whether CGF had an impact on the surgical outcome. RESULTS: Thirty-one patients (41 periapical lesion sites) were included in the CGF group, and 23 patients (26 periapical lesion sites) were included in the control group. The overall success rate of endodontic microsurgery was greater than 90%. The baseline of the 2 groups had no difference (P < .05). In the CGF group, the success rate was always 100% in 3 follow-ups, whereas the success rate was 84.2%, 92.8%, and 90%, respectively, in the control group. The success rate between the CGF group and the control group was statistically significant in all 3 follow-up points (P < .05). CONCLUSIONS: The application of CGF during endodontic microsurgery might have a positive influence on surgical outcomes, thus, its prognosis. However, higher-grade evidence is needed to demonstrate its role.

Orthodontically induced external apical root resorption considerations of root-filled teeth vs vital pulp teeth: a systematic review and meta-analysis.

INTRODUCTION: The purpose of this systematic review was to research the difference between root-filled teeth (RFT) and vital pulp teeth (VPT) in orthodontically induced external apical root resorption (EARR) and to offer suggestions for clinicians on therapeutic sequence and timing when considering combined treatment of endodontic and orthodontic. MATERIALS AND METHODS: An electronic search of published studies was conducted before November 2022 in PubMed, Web of Science and other databases. Eligibility criteria were based on the Population, Intervention, Comparison, Outcome, and Study design (PICOS) framework. RevMan 5.3 software was used for statistical analysis. Single-factor meta-regression analysis was used to explore the sources of literature heterogeneity, and a random effects model was used for analysis. RESULTS: This meta-analysis comprised 8 studies with 10 sets of data. As there was significant heterogeneity among the studies, we employed a random effects model. The funnel plot of the random effects model exhibited a symmetrical distribution, indicating no publication bias among the included studies. The EARR rate of RFT was significantly lower than that of VPT. CONCLUSIONS: In the context of concurrent endodontic and orthodontic treatment, priority should be given to endodontic therapy, as it serves as the foundation for subsequent orthodontic procedures. The optimal timing for orthodontic tooth movement post-root canal therapy is contingent upon factors such as the extent of periapical lesion resolution and the degree of dental trauma sustained. A comprehensive clinical assessment is essential in guiding the selection of the most suitable approach for achieving optimal treatment outcomes.

Liensinine attenuates inflammation and oxidative stress in spleen tissue in an LPS-induced mouse sepsis model. / 莲心碱可减轻LPSè¯±å¯¼çš„å°é¼ è„“æ¯’ç—‡æ¨¡åž‹ä¸­è„¾è„ç»„ç»‡ç‚Žç—‡å’Œæ°§åŒ–åº”æ¿€ååº”.

Sepsis is a complex syndrome caused by multiple pathogens and involves multiple organ failure, particularly spleen dysfunction. In 2017, the worldwide incidence was 48.9 million sepsis cases and 11 million sepsis-related deaths were reported (Rudd et al., 2020). Inflammation, oxidative stress, and apoptosis are the most common pathologies seen in sepsis. Liensinine (LIE) is a bisbenzylisoquinoline-type alkaloid extracted from the seed embryo of Nelumbo nucifera. Lotus seed hearts have high content of LIE which mainly has antihypertensive and antiarrhythmic pharmacological effects. It can exert anti-carcinogenic activity by regulating cell, inflammation, and apoptosis signaling pathways (Manogaran et al., 2019). However, its protective effect from sepsis-induced spleen damage is unknown. In this research, we established a mouse sepsis model induced by lipopolysaccharide (LPS) and investigated the protective effects of LIE on sepsis spleen injury in terms of inflammatory response, oxidative stress, and apoptosis.

PD-1/PD-L1 pathway: A double-edged sword in periodontitis.

Periodontitis is a disease caused by infection and immunological imbalance, which often leads to the destruction of periodontal tissue. Programmed death protein 1 (PD-1) and its ligand: programmed death ligand 1 (PD-L1) are important "immune checkpoint" proteins that have a negative regulatory effect on T cells and are targets of immunotherapy. Studies have shown that the expression of PD-1 and PD-L1 in patients with periodontitis is higher than that in healthy individuals. The keystone pathogen Porphyromonas gingivalis (P. gingivalis) is believed to be the main factor driving the upregulation of PD-1/PD-L1. High expression of PD-1/PD-L1 can inhibit the inflammatory response and reduce the destruction of periodontal supporting tissues, but conversely, it can promote the "immune escape" of P. gingivalis, thus magnifying infections. In addition, the PD-1/PD-L1 pathway is also associated with various diseases, such as cancer and Alzheimer's disease. In this review, we discuss the influence and mechanism of the PD-1/PD-L1 pathway as a "double-edged sword" affecting the occurrence and development of periodontitis, as well as its function in periodontitis-related systemic disorders. The PD-1/PD-L1 pathway could be a new avenue for periodontal and its related systemic disorders therapy.

A GntR family transcription factor in Porphyromonas gingivalis regulates bacterial growth, acylpeptidyl oligopeptidase, and gingipains activity.

Porphyromonas gingivalis is a keystone pathogen for periodontitis. The function of the GntR family transcription factor is poorly studied in P. gingivalis. Numerous processes govern bacterial growth. The survival and pathogenicity of P. gingivalis depend heavily on its capacity to acquire amino acids as nutritional sources. In this investigation, a GntR transcription factor, pg1007, was identified in P. gingivalis, the deletion of which significantly inhibited bacterial growth. The mutant strain also exhibited an increased extracellular activity of gingipains and acylpeptidyl oligopeptidase (AOP). Global gene expression profiling revealed that the expression levels of 59 genes were significantly altered in the Δpg1007 mutant, with an upregulation in gene expression for AOP, ABC transporters, and some membrane proteins. In addition, His-PG1007 protein was purified as a recombinant protein from Escherichia coli, and the conserved DNA sequence bound by it was determined using electrophoretic mobility shift assays and DNase I footprinting assays. Consequently, this study demonstrated that pg1007 is a crucial transcription factor in P. gingivalis and regulates the bacterial growth and activity of gingipains and AOP. These findings may enhance our understanding of the regulation of bacterial proliferation and protease activity in P. gingivalis.

Macrophage M1 polarization mediated via the IL-6/STAT3 pathway contributes to apical periodontitis induced by Porphyromonas gingivalis.

OBJECTIVE: To investigate the involvement of IL-6/STAT3 signaling pathway activation in macrophage polarization and bone destruction related to apical periodontitis (AP) stimulated by Porphyromonas gingivalis. METHODOLOGY: Macrophage polarization, IL-6/STAT3 expression, and the presence of P. gingivalis were detected in human AP tissues via RT-qPCR, western blotting, and immunohistochemistry staining. Murine bone marrow derived macrophages were isolated and cultured with P. gingivalis W83 in vitro, and levels of macrophage IL-6 expression, STAT3 phosphorylation, and macrophage polarization with or without the selective STAT3 phosphorylation inhibitor Stattic (5 µM) were detected via ELISA, western blotting, RT-qPCR, and flow cytometry, respectively. P. gingivalis-induced murine AP models were constructed, and bone destruction and macrophage polarization in the apical region were evaluated. Transwell co-culture systems were used to investigate the effects of macrophages infected with P. gingivalis on osteogenesis and osteoclastogenesis. RESULTS: P. gingivalis was detected in human AP tissues that highly expressed IL-6/STAT3, and the M1 subtype of macrophages was more abundant in these tissues. P. gingivalis infection induced IL-6 expression, STAT3 phosphorylation, and M1 polarization of macrophages, while 5 µM of Stattic partially abolished these activation effects. Systemic STAT3 blockade via oral administration of Stattic at a dose of 25 mg kg-1 alleviated murine periapical bone resorption and apical infiltration of M1 macrophages induced by P. gingivalis infection in vivo. Furthermore, macrophages infected with P. gingivalis promoted bone destruction via secretion of IL-6, TNF-α, and RANKL, which hinder pre-osteoblast expression of Runx2 and accelerate pre-osteoclast expression of NFAT2. CONCLUSIONS: The activation of IL-6/STAT3 signaling pathway is involved in mediating macrophages M1 polarization in the P. gingivalis induced apical inflammatory context and may also be intimately involved in the bone loss caused by P. gingivalis infection, directing the M1 macrophage infiltration during the progression of AP.

Ferroptosis interaction with inflammatory microenvironments: Mechanism, biology, and treatment.

Ferroptosis is a newly discovered form of regulated cell death. Ferroptosis is an iron-dependent lipid peroxidation reaction of cell membrane lipids, and it is closely related to the occurrence and development of many inflammatory diseases, such as ischemia-reperfusion injury, nonalcoholic steatohepatitis, and tumors. Although the precise role of ferroptosis in these inflammatory diseases is still unclear, recent evidence indicates that the association between ferroptosis and inflammatory diseases is related to the interaction of ferroptosis and inflammatory microenvironments. In inflammatory microenvironments, ferroptosis can be regulated by metabolic changes or the secretion of related substances between microorganisms and host cells or between host cells. At the same time, ferroptotic cells can also recruit immune cells by releasing injury-related molecular patterns, which in turn induces the generation of inflammatory microenvironments. Molecular crosstalk between ferroptosis and other cell death types also exists in inflammatory microenvironments. In addition, the interaction of ferroptosis and the tumor microenvironment is also correlated with tumor growth. This article reviews the main metabolic processes of ferroptosis, describes the interaction mechanism between ferroptosis and inflammatory microenvironments, and summarizes the role of ferroptosis in the treatment of diseases.

Macrophages: A communication network linking Porphyromonas gingivalis infection and associated systemic diseases.

Porphyromonas gingivalis (P. gingivalis) is a Gram-negative anaerobic pathogen that is involved in the pathogenesis of periodontitis and systemic diseases. P. gingivalis has recently been detected in rheumatoid arthritis (RA), cardiovascular disease, and tumors, as well as Alzheimer's disease (AD), and the presence of P. gingivalis in these diseases are correlated with poor prognosis. Macrophages are major innate immune cells which modulate immune responses against pathogens, however, multiple bacteria have evolved abilities to evade or even subvert the macrophages' immune response, in which subsequently promote the diseases' initiation and progression. P. gingivalis as a keystone pathogen of periodontitis has received increasing attention for the onset and development of systemic diseases. P. gingivalis induces macrophage polarization and inflammasome activation. It also causes immune response evasion which plays important roles in promoting inflammatory diseases, autoimmune diseases, and tumor development. In this review, we summarize recent discoveries on the interaction of P. gingivalis and macrophages in relevant disease development and progression, such as periodontitis, atherosclerosis, RA, AD, and cancers, aiming to provide an in-depth mechanistic understanding of this interaction and potential therapeutic strategies.

Three-dimensional analysis of coronal root canal morphology of 136 permanent mandibular first molars by micro-computed tomography.

BACKGROUND/PURPOSE: Minimally invasive endodontic approach become a research hotspot and may prevent the fracture of endodontically-treated teeth. This research aims to measure the coronal root canal morphology of permanent mandibular first molars in 3D and propose a new minimally invasive endodontic approach based on this measurement. MATERIALS AND METHODS: Data of 136 permanent mandibular first molars were involved and reconstructed in 3D models with canals. Then, the morphology characteristics of the coronal root canal were measured. RESULTS: Overall, the distribution of root canal orifices was more centralized than other landmarks. The landmarks were located more mesiobuccally to the center of the occlusal plane of molars. Specifically, the measurements of the maximum curvature of coronal root canals in the axial direction were: in 3-canals 2-rooted teeth, the average angles of curvatures were 23°,25°,11° for mesiobuccal (MB), mesiolingual (ML) and distobuccal (DB) canals, respectively; in 4-canals 2-rooted teeth were 23°,25°,12°,16°for MB, ML, DB, and distolingual (DL) canals, respectively; in 4-canals 3-rooted teeth were 25°,27°,17°,39° for MB, ML, DB, and DL canals, respectively. The degrees of coronal root canal curvatures in the horizontal direction were: in 3-canals teeth, the average angles of curvatures were -1°,47°,-2° for MB, ML and DB canals, respectively; in 4-canals 2-rooted teeth were -4°,41°,-25°,48° for MB, ML, DB, and DL canals, respectively; in 4-canals 3-rooted teeth were -3°,33°,-43°,79° for MB, ML, DB, and DL canals, respectively. CONCLUSION: The results of this study are similar to those previously obtained using CBCT and can help us design endodontic approaches.

Potential relationship between clinical symptoms and the root canal microbiomes of root filled teeth based on the next-generation sequencing.

AIM: To determine the microorganism in root canal systems of root filled teeth with periapical disease and their relationship with clinical symptoms using next-generation sequencing. METHODOLOGY: The roots of 10 extracted teeth were collected from 10 patients who presented with post-treatment apical periodontitis (PTAP; six with symptoms and four without symptoms). Each root was divided horizontally into two parts (apical and coronal segments) and cryo-pulverized. Microbial communities were detected using 16S rDNA hypervariable V3-V4 region. The diversity, principal coordinate analysis and linear discriminant analysis effect size were performed in the symptomatic and asymptomatic groups (apical and coronal parts respectively). A Mann-Whitney test and an analysis of similarities were applied for intergroup analysis, at a significance level of 5%. RESULTS: A total of 23 phyla, 257 genera and 425 species were detected. Firmicutes was the most abundant phylum in all samples. Three phyla (Fusobacteria, Synergistetes and unidentified_Bacteria) and seven genera (Fusobacterium, Porphyromonas, Phocaeicola, Olsenella, Campylobacter, Tannerella and Fretibacterium) were significantly more abundant in the symptomatic patients (p < .05), whereas asymptomatic patients had more Sphingomonas. The species more significantly abundant in the symptomatic samples were Porphyromonas gingivalis, Phocaeicola abscessus, Campylobacter showae, Tannerella forsythia and Olsenella uli (p < .05). CONCLUSIONS: A greater microbial diversity was observed in root filled teeth with PTAP compared to earlier reports. Several genera and species in root canal systems might be associated with clinical symptoms of PTAP.

Macrophage M1 polarization mediated via the IL-6/STAT3 pathway contributes to apical periodontitis induced by Porphyromonas gingivalis

Abstract Objective: To investigate the involvement of IL-6/STAT3 signaling pathway activation in macrophage polarization and bone destruction related to apical periodontitis (AP) stimulated by Porphyromonas gingivalis. Methodology: Macrophage polarization, IL-6/STAT3 expression, and the presence of P. gingivalis were detected in human AP tissues via RT-qPCR, western blotting, and immunohistochemistry staining. Murine bone marrow derived macrophages were isolated and cultured with P. gingivalis W83 in vitro, and levels of macrophage IL-6 expression, STAT3 phosphorylation, and macrophage polarization with or without the selective STAT3 phosphorylation inhibitor Stattic (5 μM) were detected via ELISA, western blotting, RT-qPCR, and flow cytometry, respectively. P. gingivalis-induced murine AP models were constructed, and bone destruction and macrophage polarization in the apical region were evaluated. Transwell co-culture systems were used to investigate the effects of macrophages infected with P. gingivalis on osteogenesis and osteoclastogenesis. Results: P. gingivalis was detected in human AP tissues that highly expressed IL-6/STAT3, and the M1 subtype of macrophages was more abundant in these tissues. P. gingivalis infection induced IL-6 expression, STAT3 phosphorylation, and M1 polarization of macrophages, while 5 μM of Stattic partially abolished these activation effects. Systemic STAT3 blockade via oral administration of Stattic at a dose of 25 mg kg-1 alleviated murine periapical bone resorption and apical infiltration of M1 macrophages induced by P. gingivalis infection in vivo. Furthermore, macrophages infected with P. gingivalis promoted bone destruction via secretion of IL-6, TNF-α, and RANKL, which hinder pre-osteoblast expression of Runx2 and accelerate pre-osteoclast expression of NFAT2. Conclusions: The activation of IL-6/STAT3 signaling pathway is involved in mediating macrophages M1 polarization in the P. gingivalis induced apical inflammatory context and may also be intimately involved in the bone loss caused by P. gingivalis infection, directing the M1 macrophage infiltration during the progression of AP.

Horizontal bone augmentation of the edentulous area with simultaneous endodontic microsurgery of the adjacent tooth: A digitally-driven multidisciplinary case report with a 1-year follow-up.

PURPOSE: To introduce a novel and efficient procedure to solve a multidisciplinary issue connected to implant-related surgery in areas near periapical lesions of adjacent teeth using single-stage combined surgery while exploring a new way to prevent retrograde peri-implantitis. MATERIALS AND METHODS: A 31-year-old woman diagnosed with a Kennedy III dentition defect in the maxillary right central incisor and posttreatment apical periodontitis in the maxillary right lateral incisor was treated using a multidisciplinary procedure. First, the preoperative data were collected from intraoral, extraoral facial and CBCT scans. Then, the aesthetic appearance of the anterior teeth was planned digitally and implant insertion was simulated. Next, virtual bone augmentation was carried out with reference to the simulated implant position, and according to the virtual augmentation, the templates for bone shell harvesting (also used for apical osteotomy and root tip resection during endodontic microsurgery) and bone shell grafting of the edentulous area were designed and fabricated. The templates for combined surgery (endodontic microsurgery and horizontal bone augmentation) consisted of one basal template and multiple interchangeable attachments via a plugin design to make guided endodontic microsurgery and digitally guided bone augmentation more efficient. Combined surgery was then carried out using the templates for guidance. During surgery, the apical inflammation affecting the maxillary right lateral incisor was first removed and its preserved apical bony window was prepared as an autogenous bone shell for bone augmentation of the maxillary right central incisor site. Guided bone regeneration of the edentulous area and guided tissue regeneration were then performed for the adjacent tooth. Six months after the combined surgery, digital guided implant surgery was carried out for the edentulous area. The final prosthesis was delivered in accordance with the preoperative aesthetic design and achieved using an implant-supported restoration for the maxillary right central incisor, full crown restoration for the maxillary right lateral incisor, and ceramic veneers for the maxillary left central and lateral incisors for space closure. RESULTS: The horizontal bone augmentation in the edentulous area and endodontic microsurgery on the neighbouring tooth were performed successfully in a single-stage surgical procedure; thus, augmentation of the resorbed alveolar bone and removal of infection in the adjacent site were achieved simultaneously. At the 1-year follow-up after combined surgery, the healing of the natural maxillary right lateral incisor and the area having undergone bone augmentation showed promising results with no postoperative complications. CONCLUSIONS: This novel digital workflow appears effective in addressing the problem of periapical lesions in retained teeth adjacent to the edentulous area that requires horizontal bone augmentation in one surgical procedure, providing an efficient way of resolving the problem using endodontics and implantology, and preventing retrograde peri-implantitis.