DLX6-AS1 regulates odonto/osteogenic differentiation in dental pulp cells under the control of BMP9 via the miR-128-3p/MAPK14 axis: A laboratory investigation.

AIM: The regenerative capacity of dental pulp relies on the odonto/osteogenic differentiation of dental pulp cells (DPCs), but dynamic microenvironmental changes hinder the process. Bone morphogenetic protein 9 (BMP9) promotes differentiation of DPCs towards an odonto/osteogenic lineage, forming dentinal-like tissue. However, the molecular mechanism underlying its action remains unclear. This study investigates the role of DLX6 antisense RNA 1 (DLX6-AS1) in odonto/osteogenic differentiation induced by BMP9. METHODOLOGY: Custom RT2 profiler PCR array, quantitative Real-Time PCR (qRT-PCR) and western blots were used to investigate the expression pattern of DLX6-AS1 and its potential signal axis. Osteogenic ability was evaluated using alkaline phosphatase and alizarin red S staining. Interactions between lncRNA and miRNA, as well as miRNA and mRNA, were predicted through bioinformatic assays, which were subsequently validated via RNA immunoprecipitation and dual luciferase reporter assays. Student's t-test or one-way ANOVA with post hoc Tukey HSD tests were employed for data analysis, with a p-value of less than .05 considered statistically significant. RESULTS: DLX6-AS1 was upregulated upon BMP9 overexpression in DPCs, thereby promoting odonto/osteogenic differentiation. Additionally, miR-128-3p participated in BMP9-induced odonto/osteogenic differentiation by interacting with the downstream signal MAPK14. Modifying the expression of miR-128-3p and transfecting pcMAPK14/siMAPK14 had a rescue impact on odonto/osteogenic differentiation downstream of DLX6-AS1. Lastly, miR-128-3p directly interacted with both MAPK14 and DLX6-AS1. CONCLUSIONS: DLX6-AS1 could regulate the odonto/osteogenic differentiation of DPCs under the control of BMP9 through the miR-128-3p/MAPK14 axis.

NLRP10 promotes AGEs-induced NLRP1 and NLRP3 inflammasome activation via ROS/MAPK/NF-κB signaling in human periodontal ligament cells.

Diabetes mellitus (DM), characterized by production and accumulation of advanced glycation end products (AGEs), induces and promotes chronic inflammation in tissues, including periodontal tissue. Increasing amount of epidemiological and experimental evidence demonstrated that more extensive inflammatory reaction and bone resorption occurred in periodontal tissues in diabetic patients with periodontitis, which is speculated to be related to NLRP3 inflammasome. NLRP10 is the only NOD-like receptor protein lacking leucine-rich repeats, suggesting that NLRP10 may be a regulatory protein. The aim of this study was to investigate the regulatory role of NLRP10 on NLRP1 and NLRP3 inflammasome in human periodontal ligament cells (HPDLCs) under AGEs treatment. Expression of NLRP10 in HPDLCs stimulated with 100 ug/mL AGEs for 24 h was observed. Detection of TRIM31 is conducted, and in TRIM31-overexpressed HPDLCs, the interaction between NLRP10 with TRIM31 as well as NLRP10 with ubiquitination were explored by immunoprecipitation. Under AGEs stimulation, the activation of reactive oxidative stress (ROS) and inflammatory signaling pathway (NF-κB, MAPK pathway) was detected by biomedical microscope and western blot (WB), respectively. After stimulation with AGEs for 24 h with or without silencing NLRP10, inflammatory cytokines (IL-6 and IL-1ß), NF-κB, MAPK pathway, ROS, and components of inflammasome were assessed. In HPDLCs, we found AGEs induced NLRP10 and inhibited TRIM31. TRIM31 overexpression significantly enhanced interaction between TRIM31 and NLRP10, then induced proteasomal degradation of NLRP10. Moreover, under AGEs stimulation, NLRP10 positively regulates NLRP1, NLRP3 inflammasomes by activating NF-κB, MAPK pathway, and increasing ROS, finally promoting the expression of inflammatory cytokines. Together, we, for the first time, confirmed that NLRP10 could promote inflammatory response induced by AGEs in HPDLCs via activation of NF-κB, and MAPK pathway and increasing ROS.

Micro-computed tomography analysis of calcium hydroxide delivery efficacy in C-shaped canal system of mandibular second molars.

BACKGROUND: Calcium hydroxide [Ca(OH)2] is widely accepted as a biocompatible interappointment intracanal medicament. This study aimed to analyze the efficacy of Ca(OH)2 placement into the C-shaped canal system of mandibular second molars using the syringe method with and without lentulo spiral utilizing micro-computed tomography (micro-CT). METHODS: Twenty-four extracted mandibular second molars were instrumented and classified into C-shaped floors (n = 12) and non-C-shaped floors (n = 12). Both groups were placed with Ca(OH)2 using the syringe system, then all teeth were scanned and cleaned, and placed with Ca(OH)2 again but with the syringe system followed by lentulo spiral and rescanned. The specimens were scanned using micro-CT to analyze the volume, volume percentage, uncontacted surface area, and uncontacted surface area percentage of Ca(OH)2 with the two delivery methods in the entire canal and at the apical 4 mm of the canal. Mann-Whitney test and Wilcoxon signed-rank test were used to determine the statistical differences among the groups. RESULTS: Syringe administration used in conjunction with lentulo spiral presented lower uncontacted surface area, a lower percentage of uncontacted surface area, larger volume, and a higher percentage of volume than syringe without lentulo spiral (P < 0.05). There was no significant difference between the C-shaped floor group and the non-C-shaped floor group (P > 0.05) in the Ca(OH)2 uncontacted surface area, volume, and percentages at different regions of canals and among different delivery techniques groups. CONCLUSIONS: The lentulo spiral and syringe technique combination can increase the volume and contacted surface area of Ca(OH)2 in the C-shaped canal system of mandibular second molars.

3D-printed coloured tooth model for inlay preparation in pre-clinical dental education.

INTRODUCTION: Accurate inlay preparation is extremely important in pre-clinical training. However, there is a lack of tools to guide students to efficiently practise inlay preparation. Therefore, a 3D-printed coloured tooth model for inlay preparation was designed to guide beginners to practise inlay preparation by themselves according to different colour prompts. This study aimed to evaluate the benefits of using a 3D-printed coloured tooth model in the pre-clinical training on inlay preparation. MATERIALS AND METHODS: Twenty-eight students in their fourth-year undergraduate dental program participated in this study. The participants were randomly assigned to two groups for the inlay preparation. Group 1 prepared a plain tooth model for the first and fourth attempts and a 3D-printed coloured tooth model for the second and third attempts (n = 14). Group 2 prepared four plain tooth models (n = 14). The first and fourth tooth models prepared by both groups were scored using an evaluation system (Fair Grade 2000, NISSIN). Next, questionnaires answered by students were used to evaluate the benefits of using a 3D-printed coloured tooth model and self-evaluate hands-on ability using a grading system (1 = strongly agree, 2 = agree, 3 = neutral, 4 = disagree, and 5 = strongly disagree). The scores were evaluated statistically using the Mann-Whitney U test, and the given grades are displayed as percentages and mean values. RESULTS: There was an overall increase in the clinical confidence of all students after repeated attempts to prepare an inlay; however, students from group 1, who had used the 3D-printed coloured tooth model, had more positive experiences and comments. The 3D-printed coloured tooth model for inlay preparation has been widely praised by participants. Comparing the average score of the first and fourth preparations, the average score of group 1 increased by 12% (Ø 54.46 ± 8.33, Ø 61.11 ± 7.13, p = .090), while that of group 2 increased by 0.72% (Ø 56.39 ± 9.59, Ø 56.80 ± 8.46, p = .925). CONCLUSION: Students favoured the use of the 3D-printed coloured tooth model, and this improved the average score for inlay preparation. The 3D-printed coloured tooth model for inlay preparation is expected to play an important role in dental education in the future.

Immortalized cell lines derived from dental/odontogenic tissue.

Stem cells derived from dental/odontogenic tissue have the property of multiple differentiation and are prospective in tooth regenerative medicine and cellular and molecular studies. However, in the face of cellular senescence soon in vitro, the proliferation ability of the cells is limited, so studies are hindered to some extent. Fortunately, immortalization strategies are expected to solve the above issues. Cellular immortalization is that cells are immortalized by introducing oncogenes, human telomerase reverse transcriptase genes (hTERT), or miscellaneous immortalization genes to get unlimited proliferation. At present, a variety of immortalized stem cells from dental/odontogenic tissue has been successfully generated, such as dental pulp stem cells (DPSCs), periodontal ligament cells (PDLs), stem cells from human exfoliated deciduous teeth (SHEDs), dental papilla cells (DPCs), and tooth germ mesenchymal cells (TGMCs). This review summarized establishment and applications of immortalized stem cells from dental/odontogenic tissues and then discussed the advantages and challenges of immortalization.

The role of MIF in periodontitis: A potential pathogenic driver, biomarker, and therapeutic target.

OBJECTIVE: Periodontitis is an inflammatory disease that involves an imbalance in the oral microbiota, activation of inflammatory and immune responses, and alveolar bone destruction. Macrophage migration inhibitory factor (MIF) is a versatile cytokine involved in several pathological reactions, including inflammatory processes and bone destruction, both of which are characteristics of periodontitis. While the roles of MIF in cancer and other immune diseases have been extensively characterized, its role in periodontitis remains inconclusive. RESULTS: In this review, we describe a comprehensive analysis of the potential roles of MIF in periodontitis from the perspective of immune response and bone regulation at the cellular and molecular levels. Moreover, we discuss its potential reliability as a novel diagnostic and therapeutic target for periodontitis. CONCLUSION: This review can aid dental researchers and clinicians in understanding the current state of MIF-related pathogenesis, diagnosis, and treatment of periodontitis.

The expression of cyclic GMP-AMP synthase in human apical periodontitis: A laboratory investigation.

AIM: As a key DNA sensor, cyclic GMP-AMP synthase (cGAS) has emerged as a major mediator of innate immunity and inflammation. Human apical periodontitis has yet to be studied for the presence of cGAS. This investigation was conducted to determine if cGAS is involved in the pathological process of human apical periodontitis. METHODOLOGY: Sixty four human periapical lesions, comprising 20 periapical granulomas and 44 radicular cysts, were employed in this investigation. Healthy gingiva (n = 6), dental pulp (n = 3), and apical papilla (n = 3) were used as control samples. The expression of cGAS in the periapical tissues was discovered using immunohistochemical staining. mRNA-Sequencing and qRT-PCR were utilized to determine the differentially expressed genes (DEGs) associated with DNA-sensing signalling in periapical lesions compared to the healthy control. Immunofluorescence labelling was used to identify the co-expression of cGAS, interleukin-1ß, and interleukin-18. RESULTS: A significantly greater expression level of cGAS was discovered in the periapical lesions, with no significant difference between radicular cysts and periapical granulomas. mRNA-Sequencing analysis and qRT-PCR identified differentially expressed mRNA, such as cGAS and its downstream DEGs, between periapical lesions and healthy control tissues. Immunofluorescence labelling further revealed that cGAS, interleukin-1, and interleukin-18 were co-localized. CONCLUSIONS: These observations imply that along with the synthesis of interleukin-1 and interleukin-18, cGAS may be involved in the aetiology of apical periodontitis.

An application framework of 3D assessment image registration accuracy and untouched surface area in canal instrumentation laboratory research with micro-computed tomography.

OBJECTIVES: The purpose of this study was to develop a customized framework for evaluating the registration accuracy of four registration techniques and measuring the untouched surface area of canal instrumentation by visually inspecting and calculating the overlapping area of the surfaces. METHODS: Twenty-one mandibular incisors were scanned by micro-computed tomography before and after instrumentation. Elastix registration, surface registration, manual registration, and DataViewer registration techniques were used to align the pre- and post-operative datasets. The customized MeVisLab framework was created to investigate the registration accuracy by visual inspection and calculating overlapping areas. The canal surfaces were imported into the same framework to measure the untouched surface area and the consistence test was validated. The correlation between registration accuracy and untouched surface area was analyzed. RESULTS: There is a statistically significant difference between manual registration and automatic registration (P < 0.05). There is no statistical difference between the two untouched surface measure methods (P > 0.05). The partial correlation coefficients for the untouched surface area and registration accuracy were 0.45 (P < 0.05). CONCLUSIONS: This application framework based on free customizable software, allows a new method to measure registration accuracy and untouched surface area in an efficient and sensitive way. The application of a precise registration method would improve the quality of micro-CT canal instrumentation studies. CLINICAL RELEVANCE: This study developed a customized framework based on free software for evaluating the registration accuracy of different registration techniques and measuring the untouched surface area of canal instrumentation could help researchers to improve the quality of micro-CT studies of canal instrumentation.

Microcomputed tomography analysis of the radicular residual dentin thickness in mandibular second molars after virtual fiber post placement: Identification of danger zones.

STATEMENT OF PROBLEM: Mandibular second molars have variable root, pulp chamber floor, and radicular groove morphologies, potentially affecting residual dentin thickness and post placement suitability. However, an identification of the danger zones is lacking. PURPOSE: The purpose of this in vitro study was to investigate the residual dentin thickness in the danger zone of mandibular second molars after virtual fiber post placement by using a simulation method based on microcomputed tomography (µCT). MATERIAL AND METHODS: A total of 84 extracted mandibular second molars were scanned using µCT and classified according to root morphology (separate or fused) and pulp chamber floor morphologies (C-shaped, non-C-shaped, or no pulp chamber floor). Fused-root mandibular second molars were further classified based on the radicular groove type (V-, U-, or Ω-shaped). All specimens were accessed, instrumented, and rescanned with µCT. Two types of commercial fiber posts were also scanned. Clinical fiber post placement was simulated in all prepared canals by using a multifunctional software program. The minimum residual dentin thickness of each root canal was measured and analyzed by using nonparametric tests to identify the danger zone. Perforation rates were calculated and recorded. RESULTS: Larger fiber posts decreased minimum residual dentin thickness (P<.05) and increased perforation rate. In regard to mandibular second molars with separate roots, the distal root canal exhibited a significantly higher minimum residual dentin thickness than the mesiobuccal and mesiolingual root canals (P<.05). However, no significant difference in minimum residual dentin thickness was found between the different canals in fused-root mandibular second molars with C-shaped pulp chamber floors (P<.05). Fused-root mandibular second molars with Ω-shaped radicular grooves had a lower minimum residual dentin thickness than those with V-shaped radicular grooves (P<.05) and demonstrated the highest perforation rate. CONCLUSIONS: The morphologies of the root, pulp chamber floor, and radicular groove were correlated with residual dentin thickness distribution in mandibular second molars after fiber post placement. A comprehensive understanding of mandibular second molar morphology is essential for determining the suitability of post-and-core crown restorations after endodontic treatment.

Emerging roles of lncRNAs in the pathogenesis, diagnosis, and treatment of trigeminal neuralgia.

Trigeminal neuralgia (TN) is one of the most common neuropathic pain disorders and is often combined with other comorbidities if managed inadequately. However, the present understanding of its pathogenesis at the molecular level remains lacking. Long noncoding RNAs (lncRNAs) play crucial roles in neuropathic pain, and many studies have reported that specific lncRNAs are related to TN. This review summarizes the current understanding of lncRNAs in the pathogenesis, diagnosis, and treatment of TN. Recent studies have shown that the lncRNAs uc.48+, Gm14461, MRAK009713 and NONRATT021972 are potential candidate loci for the diagnosis and treatment of TN. The current diagnostic system could be enhanced and improved by a workflow for selecting transcriptomic biomarkers and the development of lncRNA-based molecular diagnostic systems for TN. The discovery of lncRNAs potentially impacts drug selection for TN; however, the current supporting evidence is limited to preclinical studies. Additional studies are needed to further test the diagnostic and therapeutic value of lncRNAs in TN.

Inflammation down regulates stromal cell-derived factor 1α in the early phase of pulpitis.

The key to prevent pulp necrosis in the early stage of pulpitis is to promote tissue repair, which begins with cell migration. Stromal cell-derived factor 1α (SDF-1α) has been proven to promote cell migration. Related research has so far concentrated on the biological effects of SDF-1α while its expression in pulpitis is still unclear. We investigated the effect of inflammation on SDF-1α in dental pulp and the underlying regulatory mechanisms. First, rat pulpitis models were established by exposing pulp. SDF-1α was decreased on the 3rd day but increased on the 7th day. Next, lipopolysaccharide from Porphyromonas gingivalis (Pg.LPS) was applied to dental pulp cells (DPCs). Within 24 h, SDF-1α decreased, but after 48 h, it steadily increased. Similarly, SDF-1α expression in human chronic pulpitis tissues was also increased. To investigate the effect of altered SDF-1α on DPC migration, cell supernatants collected following Pg.LPS treatment were utilized to stimulate DPCs, and the number of migrated cells was correlated with changes in SDF-1α secretion. Finally, we explored the regulatory mechanisms of SDF-1α down-regulation in the early phase of pulpitis. Within 24 h, JNK/c-Jun pathway was activated in DPC inflammation. When JNK pathway was suppressed, SDF-1α rose. Furthermore, tumor necrosis factor receptor 2 (TNFR2) and apoptosis signal-regulated kinase-interacting protein 1 (AIP1) were up-regulated. Knockdown of them abolished Pg.LPS-induced activation of JNK and c-Jun(Ser63) and significantly enhanced SDF-1α. Our findings indicated that in the early phase of pulpitis, inflammation suppressed SDF-1α by up-regulating TNFR2 and AIP1, which activated JNK/c-Jun(Ser63) pathway.

Wnt4 regulates bone metabolism through IKK-NF-κB and ROCK signaling under occlusal traumatic periodontitis.

BACKGROUND AND OBJECTIVE: Occlusal trauma is one of the most important local contributing factors of periodontitis. It has been reported that Wnt4, a noncanonical Wnt ligand, can inhibit osteoclast formation and inflammation and promote bone formation in vivo. However, the prospects of Wnt4 application in occlusal trauma and periodontitis have not yet been described. This study aimed to investigate the function and the corresponding mechanism of Wnt4 to regulate bone metabolism in occlusal trauma and periodontitis. MATERIAL AND METHODS: Osteogenic-induced MC3T3-E1 cells were treated with or without Porphyromonas gingivalis lipopolysaccharide (Pg. LPS) under cyclic uniaxial compressive stress. After treatment with mouse recombinant protein Wnt4 (rWnt4), the expression of osteogenic markers and activation of the IKK-NF-κB signaling pathway were evaluated in vitro. To investigate whether Wnt4 can promote osteogenesis via the ROCK signaling pathway, the expression of RhoA was evaluated in vitro. Finally, we evaluated the change in bone quantity and the activation of the IKK-NF-κB and ROCK signaling in mice with occlusal trauma and periodontitis to demonstrate the therapeutic efficacy of rWnt4 injection. RESULTS: Stimulation of traumatic force and Pg. LPS stimulation suppressed the expression of osteoblast markers, but their expression was rescued after rWnt4 treatment in vitro. In addition, the inhibition of the ROCK signaling pathway induced by force loading was reversed when rWnt4 was applied in vitro. Micro-CT, H&E, and TRAP staining of the mandibles showed increased bone loss in the occlusal trauma-aggravated periodontitis group, whereas it was rescued after rWnt4 injection. The expression levels of IκBα and p65 were upregulated in occlusal trauma and periodontitis-bearing mice, whereas the expression levels of Runx2 and RhoA were downregulated. After rWnt4 injection, remarkably upregulation of Runx2 and RhoA expression was observed in occlusal trauma and periodontitis- bearing mice. CONCLUSION: Wnt4 not only inhibits IKK-NF-κB signaling but also activates ROCK signaling to inhibit osteoclast formation and promote bone regeneration in occlusal trauma and periodontitis-bearing mice.

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.

Innate immune sensing of nucleic acid in endodontic infection.

Innate immunity is the first line of defence and fights against microorganisms. Nucleic acids are important pathogen-associated molecular patterns to be recognized in innate immunity. There are three types of nucleic acid sensors, including endosomal sensors (NA-sensing TLRs), cytosolic DNA sensors (cGAS and AIM2) and cytosolic RNA sensors (RLG-I, MDA5 and LGP2). Recent studies have shown that nucleic acid sensors are expressed differently in a variety of dental pulp cells and mediate inflammation through complex pathways. Nucleic acid sensing may play a vital role in the development of endodontic infection. This review aims to summarize and analyse the possible roles of various nucleic acid sensors in endodontic infection. It may help better understand the pathogenesis of these diseases and find new strategies for prevention and treatment.

Micro-computed tomography analysis of root canal morphology and thickness of crown and root of mandibular incisors in Chinese population.

OBJECTIVES: The aim of this study was to investigate the root canal morphology and the thickness of crown and root of mandibular incisors in a Chinese subpopulation by micro-computed tomography (micro-CT). METHODS: In total, 208 mandibular incisors were scanned using micro-CT. The anatomical features of the canals (canal configuration, apical constriction, foramen-to-apex distance, accessory canal vertical distribution, and canal geometrical parameters) and the thickness of the crown and root 2/3 were evaluated. RESULTS: Three canal categories, labeled as Single (77.88%), Merged (15.87%), and Separated (6.25%), were summarized. The most frequent constriction type in main foramina was single constriction (42.53%). Wide and narrow diameters in a single main foramen were 0.37 ± 0.14 mm and 0.26 ± 0.07 mm, respectively. The distance from the anatomical foramen to the physiological foramen and the anatomical apex was 0.49 ± 0.20 mm and 0.36 ± 0.28 mm, respectively. During the virtual root-end resection, 97.12% of roots underwent successful resection at the 2-mm level, with the foramina visible on the resection surface. During 2-D cross-sectional analyses, the shape parameters of the root and canal showed significant positive correlation (P < 0.05). The thickest and thinnest walls were the incisal and proximal walls (P < 0.05) of the crown, respectively. The buccal walls were significantly thicker than the lingual walls on the crown (P < 0.05), whereas the lingual walls were thicker on the root 2/3 (P < 0.05). CONCLUSIONS: This study provides detailed information about the root canal morphology and thickness of the crown and root of mandibular incisors in a Chinese population. An understanding of morphology can benefit endodontic treatment. CLINICAL RELEVANCE: Adequate knowledge of the crown, root, and canal morphology, as well as the thickness distribution of the crown and root, is essential for successful root canal therapy and prevention of complications.

Shaping outcome of ProTaper NEXT for root canal preparation in mandibular incisors: a micro-CT study.

BACKGROUND: Relatively high incidence of single canals with oval or round shape were observed in human mandibular incisors. In order to investigate the influence of the root canal morphology on root canal preparation, the shaping outcome of ProTaper NEXT in oval and round canals of mandibular incisors were evaluated by using micro-computed tomography (micro-CT) analysis. METHODS: This experiment was approved by the School Medical Ethics Committee. The sample size calculation was conducted using G*Power software. Intact mandibular incisors with a single canal were selected. Oval canals (2 < aspect ratio (AR) ≤ 4) and round canals (AR ≤ 2) were pair-matched according to canal length, and assigned to two experimental groups (13 per group). ProTaper NEXT was used for root canal preparation for both groups. Untouched canal wall (UCW), root canal morphological parameters and three-dimensional (3D) dentin thickness were evaluated in the entire root canal and each canal third after micro-CT scanning. STATISTICAL ANALYSIS: Data were collected and analyzed with Mann-Whitney test and Friedman test using SPSS statistics software 25 (P < 0.05). Additionally, correlations of UCW area with canal morphological parameters were also investigated. RESULTS: After root canal preparation, no significant difference was observed in all analyzed parameters in the apical third between oval and round canal groups (P > 0.05). In the coronal two thirds of the canal, the post-operative structure model index (SMI), form factor and roundness were significantly increased, while the AR was significantly decreased in both groups (P < 0.05). In addition, in the coronal two thirds, significantly more UCW and higher UCWΔ% was observed in oval canal group (P < 0.05). Furthermore, UCW correlated very strongly to canal major diameter (0.924) and initial volume (0.938), and strongly to canal form factor (- 0.724), minor diameter (0.799) and canal area (0.882). Proximal dentin wall was associated with significantly thinner pre-operative dentin thickness and higher amount of dentin removal after root canal preparation in both oval and round canal groups. CONCLUSIONS: (1) Both types of canals were more conical after root canal preparation, but oval root canals tend to leave more UCW area than round canals in the coronal two thirds of mandibular incisors, which contributes to the challenge for endodontic infection control. (2) Root canal preparation usually results in excessive dentin removal in the proximal dentin wall comparing with buccal and lingual walls in both types of canals of mandibular incisors.

The involvement of semaphorin 7A in tumorigenic and immunoinflammatory regulation.

Semaphorins, a large group of highly conserved proteins, consist of eight subfamilies that are widely expressed in vertebrates, invertebrates, and viruses and exist in membrane-bound or secreted forms. First described as axon guidance cues during neurogenesis, semaphorins also perform physiological functions in other organ systems, such as bone homeostasis, immune response, and tumor progression. Semaphorin 7A (SEMA7A), also known as CDw108, is an immune semaphorin that modulates diverse immunoinflammatory processes, including immune cell interactions, inflammatory infiltration, and cytokine production. In addition, SEMA7A regulates the proliferation, migration, invasion, lymph formation, and angiogenesis of multiple types of tumor cells, and these effects are mediated by the interaction of SEMA7A with two specific receptors, PLXNC1 and integrins. Thus, SEMA7A is intimately related to the pathogenesis of multiple autoimmune and inflammation-related diseases and tumors. This review focuses on the role of SEMA7A in the pathogenesis of autoimmune disorders, inflammatory diseases, and tumors, as well as the underlying mechanisms. Furthermore, strategies targeting SEMA7A as a potential predictive, diagnostic, and therapeutic agent for these diseases are also addressed.

Materials for retrograde filling in root canal therapy.

BACKGROUND: Root canal therapy is a sequence of treatments involving root canal cleaning, shaping, decontamination, and obturation. It is conventionally performed through a hole drilled into the crown of the affected tooth, namely orthograde root canal therapy. When it fails, retrograde filling, which seals the root canal from the root apex, is a good alternative. Many materials are used for retrograde filling. Since none meets all the criteria an ideal material should possess, selecting the most efficacious material is of utmost importance. This is an update of a Cochrane Review first published in 2016. OBJECTIVES: To determine the effects of different materials used for retrograde filling in children and adults for whom retrograde filling is necessary in order to save the tooth. SEARCH METHODS: An Information Specialist searched five bibliographic databases up to 21 April 2021 and used additional search methods to identify published, unpublished, and ongoing studies. We also searched four databases in the Chinese language. SELECTION CRITERIA: We selected randomised controlled trials (RCTs) that compared different retrograde filling materials, with the reported success rate that was assessed by clinical or radiological methods for which the follow-up period was at least 12 months. DATA COLLECTION AND ANALYSIS: Records were screened in duplicate by independent screeners. Two review authors extracted data independently and in duplicate. Original trial authors were contacted for any missing information. Two review authors independently assessed the risk of bias of the included studies. We followed Cochrane's statistical guidelines and assessed the certainty of the evidence using GRADE. MAIN RESULTS: We included eight studies, all at high risk of bias, involving 1399 participants with 1471 teeth, published between 1995 and 2019, and six comparisons of retrograde filling materials. - Mineral trioxide aggregate (MTA) versus intermediate restorative material (IRM): there may be little to no effect of MTA compared to IRM on success rate at one year, but the evidence is very uncertain (risk ratio (RR) 1.09, 95% confidence interval (CI) 0.97 to 1.22; I2 = 0%; 2 studies; 222 teeth; very low-certainty evidence). - MTA versus super ethoxybenzoic acid (Super-EBA): there may be little to no effect of MTA compared to Super-EBA on success rate at one year, but the evidence is very uncertain (RR 1.03, 95% CI 0.96 to 1.10; 1 study; 192 teeth; very low-certainty evidence). - Super-EBA versus IRM: the evidence is very uncertain about the effect of Super-EBA compared with IRM on success rate at 1 year, with results indicating Super-EBA may reduce or have no effect on success rate (RR 0.90, 95% CI 0.80 to 1.01; 1 study; 194 teeth; very low-certainty evidence). - Dentine-bonded resin composite versus glass ionomer cement: compared to glass ionomer cement, dentine-bonded resin composite may increase the success rate of the treatment at 1 year, but the evidence is very uncertain (RR 2.39, 95% CI 1.60 to 3.59; 1 study; 122 teeth; very low-certainty evidence). Same result was obtained when considering the root as unit of analysis at one year (RR 1.59, 95% CI 1.20 to 2.09; 1 study; 127 roots; very low-certainty evidence). - Glass ionomer cement versus amalgam: the evidence is very uncertain about the effect of glass ionomer cement compared with amalgam on success rate at one year, with results indicating glass ionomer cement may reduce or have no effect on success rate (RR 0.98, 95% CI 0.86 to 1.12; 1 study; 105 teeth; very low-certainty evidence). - MTA versus root repair material (RRM): there may be little to no effect of MTA compared to RRM on success rate at one year, but the evidence is very uncertain (RR 1.00, 95% CI 0.94 to 1.07; I2 = 0%; 2 studies; 278 teeth; very low-certainty evidence). Adverse events were not assessed by any of the included studies. AUTHORS' CONCLUSIONS: Based on the present limited evidence, there is insufficient evidence to draw any conclusion as to the benefits of any one material over another for retrograde filling in root canal therapy. We conclude that more high-quality RCTs are required.

BMP9 is a potential therapeutic agent for use in oral and maxillofacial bone tissue engineering.

Oral and maxillofacial surgery is often challenging due to defective bone healing owing to the microbial environment of the oral cavity, the additional involvement of teeth and esthetic concerns. Insufficient bone volume as a consequence of aging and some oral and maxillofacial surgical procedures, such as tumor resection of the jaw, may further impact facial esthetics and cause the failure of certain procedures, such as oral and maxillofacial implantation. Bone morphogenetic protein (BMP) 9 (BMP9) is one of the most effective BMPs to induce the osteogenic differentiation of different stem cells. A large cross-talk network that includes the BMP9, Wnt/ß, Hedgehog, EGF, TGF-ß and Notch signaling pathways finely regulates osteogenesis induced by BMP9. Epigenetic control during BMP9-induced osteogenesis is mainly dependent on histone deacetylases (HDACs), microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), which adds another layer of complexity. As a result, all these factors work together to orchestrate the molecular and cellular events underlying BMP9-related tissue engineering. In this review, we summarize our current understanding of the SMAD-dependent and SMAD-independent BMP9 pathways, with a particular focus on cross-talk and cross-regulation between BMP9 and other major signaling pathways in BMP9-induced osteogenesis. Furthermore, recently discovered epigenetic regulation of BMP9 pathways and the molecular and cellular basis of the application of BMP9 in tissue engineering in current oral and maxillofacial surgery and other orthopedic-related clinical settings are also discussed.

The emerging roles of semaphorin4D/CD100 in immunological diseases.

In vertebrates, the semaphorin family of proteins is composed of 21 members that are divided into five subfamilies, i.e. classes 3 to 7. Semaphorins play crucial roles in regulating multiple biological processes, such as neural remodeling, tissue regeneration, cancer progression, and, especially, in immunological regulation. Semaphorin 4D (SEMA4D), also known as CD100, is an important member of the semaphorin family and was first characterized as a lymphocyte-specific marker. SEMA4D has diverse effects on immunologic processes, including immune cell proliferation, differentiation, activation, and migration, through binding to its specific membrane receptors CD72, PLXNB1, and PLXNB2. Furthermore, SEMA4D and its underlying signaling have been increasingly linked with several immunological diseases. This review focuses on the significant immunoregulatory role of SEMA4D and the associated underlying mechanisms, as well as the potential application of SEMA4D as a diagnostic marker and therapeutic target for the treatment of immunological diseases.