OBJECTIVE: This study aimed to investigate the mechanisms underlying impaired oral wound healing in aged individuals, with a focus on the roles of neutrophil extracellular traps (NETs) and NLRP3 inflammasome activation....OBJECTIVE: This study aimed to investigate the mechanisms underlying impaired oral wound healing in aged individuals, with a focus on the roles of neutrophil extracellular traps (NETs) and NLRP3 inflammasome activation. METHODS: Palatal wound healing was compared between young and aged mice. We assessed the change of neutrophil infiltration, NETs formation, and NLRP3 inflammasome activation in mucosa using immunohistochemistry, RNA sequencing, and flow cytometry. In vitro co-culture assays evaluated NETs' effects on fibroblast function and NLRP3 inflammasome activation. The therapeutic potential of NETs-NLRP3 clearance was tested in vivo with DNase1 and MCC950. To validate clinical relevance, human single-cell RNA sequencing data were further analyzed. RESULTS: Aged mice exhibited delayed oral wound healing, characterized by increased neutrophil recruitment and a heightened propensity for fibrin-induced NETs formation. NETs activated the NLRP3 inflammasome in mucosal fibroblasts, suppressing their proliferation and migration. Notably, targeting NETs-NLRP3 inflammasome reduced inflammation and accelerated wound closure in the aged group. Corresponding human data further revealed increased NETs-associated neutrophil subsets and enriched NLRP3 pathways in aged oral mucosa. CONCLUSION: Our findings demonstrate that enhanced NETs formation and subsequent NLRP3 inflammasome impair wound healing in aged oral mucosa. Targeting NETs-NLRP3 thus represents a promising therapeutic strategy to improve mucosal repair.
OBJECTIVES: Users of Swedish nicotine pouches and traditional oral tobacco-derived pouches were compared to non-users for clinical tissue responses and oral health status. In addition, the habits associated with snus use...OBJECTIVES: Users of Swedish nicotine pouches and traditional oral tobacco-derived pouches were compared to non-users for clinical tissue responses and oral health status. In addition, the habits associated with snus use were also evaluated. MATERIAL AND METHODS: In this cross-sectional study, participants aged 18-30 were recruited from nine dental clinics: 126 nicotine pouch users, 63 oral tobacco-derived pouch users, and 83 non-users. Clinical examinations assessed mucosal changes and dental status, including erythema, ulceration, oedema, hyperkeratosis, etching, bone defects, and gingival recession. Participants completed a questionnaire on snus use, oral hygiene, and education. Tissue responses were graded and intergroup differences analysed. The study followed the STROBE checklist. RESULTS: Oral lesions were identified in 79% of nicotine pouch users and 89% of oral tobacco-derived pouch users. Nicotine pouch users more frequently exhibited erythema and reported more oral symptoms, while hyperkeratosis predominated among oral tobacco-derived pouch users. A non-significant trend suggested mint-flavoured nicotine pouches may be associated with more pronounced lesions. No increased risk of caries or periodontal disease was observed for ≤ 6 years of use. CONCLUSIONS: Nicotine pouches appear to induce broader mucosal responses than tobacco-containing snus. Further research is needed to clarify local and systemic effects.
BACKGROUND: This study aimed to examine multidimensional factors associated with self-reported Temporomandibular Disorders (TMD) symptom burden, assessed using the Fonseca Anamnestic Index (FAI), in young adults with mya...BACKGROUND: This study aimed to examine multidimensional factors associated with self-reported Temporomandibular Disorders (TMD) symptom burden, assessed using the Fonseca Anamnestic Index (FAI), in young adults with myalgia defined according to the Diagnostic Criteria (DC) for TMD. METHODS: This cross-sectional study included 104 young adults with myalgia diagnosed according to the DC/TMD criteria. TMD symptom burden was assessed using the FAI. Clinical, psychosocial, and ultrasonographic variables were evaluated. Backward multiple linear regression was used to identify variables associated with higher FAI scores. RESULTS: The final model explained 53.0% of the variance in FAI scores (adjusted R = 0.530, p < 0.001). Higher activity-related pain, greater neck disability, elevated anxiety levels, and a higher number of parafunctional activities were positively associated with higher FAI scores, whereas greater right masseter muscle thickness was inversely associated with FAI scores (p < 0.05). CONCLUSIONS: Self-reported TMD symptom burden among young adults with DC/TMD-defined myalgia was associated with functional pain, cumulative parafunctional behaviors, neck disability, anxiety, and masseter muscle thickness. These findings support a multidimensional perspective in the assessment of TMD-related symptoms in individuals with myalgia and highlight the need for longitudinal studies using diagnostically grounded designs.
AIM: To characterize the cellular composition and molecular microenvironment of smoking-related periodontitis using single-cell RNA sequencing. MATERIALS AND METHODS: Single-cell RNA sequencing was performed on periodont...AIM: To characterize the cellular composition and molecular microenvironment of smoking-related periodontitis using single-cell RNA sequencing. MATERIALS AND METHODS: Single-cell RNA sequencing was performed on periodontal tissues from healthy controls (H, n = 2), patients with periodontitis (PD, n = 2), and patients with smoking-related periodontitis (SPD, n = 3). Immunofluorescence validated key gene expression and specific cell subtypes involved in SPD. RESULTS: In periodontal tissues, gingival epithelial cells were the cell type most affected by smoking. Compared with the PD group, the SPD group showed increased CXCL16 secretion and features of cellular senescence. A subset of CXCR6+ regulatory T cells (CXCR6+ Tregs) was identified in SPD and potentially recruited by epithelial cells via the CXCL16-CXCR6 axis. Quantitative Set Analysis for Gene Expression indicated a potential regulatory role of CXCR6+ Tregs in T helper 17 (Th17) differentiation, while intercellular communication and pseudotime analyses suggested that CXCR6+ Tregs may amplify inflammatory responses by activating Th17 cells through the CD2-CD58 interaction. CONCLUSIONS: In SPD, gingival epithelial cells exhibit senescence with increased chemokine secretion. These senescent cells may recruit CXCR6+ Tregs via CXCL16 release, potentially regulating Th17 cell differentiation. These findings provide a molecular basis for understanding SPD pathogenesis.
BACKGROUND: This study assessed the feasibility of remote halitosis diagnosis by evaluating VSC stability over 7 days and their association with periodontal parameters. METHODS: One hundred adult patients attending a per...BACKGROUND: This study assessed the feasibility of remote halitosis diagnosis by evaluating VSC stability over 7 days and their association with periodontal parameters. METHODS: One hundred adult patients attending a periodontal examination were included. Oral gas samples were collected using plastic syringes and analyzed by gas chromatography (OralChroma) at baseline (D0) and after 7 days of storage at room temperature (D7). Hydrogen sulfide (HS), methyl mercaptan (CHSH), and dimethyl sulfide ((CH)S) were quantified. Periodontal parameters were recorded. Correlations and agreement analyses were performed. RESULTS: At D0, 64% and 31% of patients were classified as having halitosis using thresholds of > 120 ppb and > 300 ppb, respectively. Mean VSC concentrations were 10.2 (33.1) ppb for HS, 208.3 (443.2) ppb for CHSH, and 79 (166) ppb for (CH)S, and increased after 7 days of storage for all compounds except HS. Only (CH)S showed a significant correlation between D0 and D7 measurements. Agreement between halitosis diagnoses at D0 and D7 was poor for both thresholds (Cohen's kappa < 0.2). Associations between VSC levels and periodontal parameters were weak. CONCLUSION: Delayed analysis of oral gas samples significantly altered VSC measurements limiting their reliability for remote halitosis diagnosis. Periodontal parameters were not strongly associated with VSC levels.
OBJECTIVE: To evaluate intraoral-ultrasound (ioUS) features of benign and malignant oral lesions. METHODS: In this prospective cross-sectional study, patients with oral soft tissue lesions underwent ioUS followed by biop...OBJECTIVE: To evaluate intraoral-ultrasound (ioUS) features of benign and malignant oral lesions. METHODS: In this prospective cross-sectional study, patients with oral soft tissue lesions underwent ioUS followed by biopsy within 2 weeks. Sonographic features: size, echogenicity, cystic areas, calcifications, margins, and vascularity were compared with histopathology. Associations between sonographic features and malignancy were explored using univariate and multivariable analyses. ROC analysis was used descriptively to explore discriminatory ability. RESULTS: Fifty-one patients with 52 lesions (24 females, 27 males; mean age 60.1 ± 16.7 years, range 18-90) were recruited. Following exclusions, the study comprised 23 histologically confirmed malignant cases (squamous cell carcinoma, SCC) and 23 benign cases. Multivariate analysis identified maximal diameter (> 12 mm; sensitivity 0.76 [95% CI: 0.53-0.90], specificity 0.77 [95% CI: 0.57-0.90]), ill-defined margins (p < 0.001), and increased vascularity (p = 0.002) as being associated with SCC in this cohort. Combinations of these features showed high apparent discrimination (AUC 0.92 [95% CI: 0.84-1.00]), but likely reflect optimistic bias given the small sample size and lack of validation. CONCLUSIONS: This pilot study describes ioUS features in benign and malignant oral lesions and identifies sonographic features potentially associated with malignancy. These findings are exploratory and hypothesis-generating and require validation in larger, more diverse cohorts before any clinical application.
OBJECTIVE: This study investigated the timing of sequestrum formation and detection, and associated factors with medication-related osteonecrosis of the jaw (MRONJ) among patients who had undergone conservative managemen...OBJECTIVE: This study investigated the timing of sequestrum formation and detection, and associated factors with medication-related osteonecrosis of the jaw (MRONJ) among patients who had undergone conservative management. METHODS: This study retrospectively analyzed data from 214 patients with MRONJ who underwent conservative management after invasive dental procedures between January 2014 and July 2025. The time to sequestrum detection was measured from the dental procedure to sequestrum development. Demographic and clinical variables, including primary disease, comorbidities, medication type and duration, and medication cessation, were investigated. RESULTS: Among 101 included patients, the mean time to sequestrum formation and detection was 6.82 ± 5.44 months. Patients with osteoporosis (6.31 ± 5.31 months) developed sequestra significantly earlier than those with malignancy (9.99 ± 5.35 months) (p = 0.018). The time to detection was longer in patients with comorbidities compared with those without comorbidities (8.21 ± 5.30 vs. 5.98 ± 5.40, p = 0.045). Notably, there was a significant difference depending on whether medication was discontinued (5.61 ± 4.93 vs. 9.82 ± 5.56 months, p < 0.001). Multivariate analysis identified medication cessation as the only independent factor affecting sequestrum detection time (p = 0.002). CONCLUSIONS: Sequestra typically formed and were detected at approximately 6-7 months, and continuation of medication after invasive dental treatment was associated with delayed sequestrum development.
OBJECTIVES: To determine whether pretreatment radiomics can predict medication-related osteonecrosis of the jaw (MRONJ). METHODS: Patients with mandibular MRONJ at Hiroshima University Hospital (2009-2024) who underwent...OBJECTIVES: To determine whether pretreatment radiomics can predict medication-related osteonecrosis of the jaw (MRONJ). METHODS: Patients with mandibular MRONJ at Hiroshima University Hospital (2009-2024) who underwent MDCT before initiating bone resorption inhibitors were retrospectively identified. Records from 2009 to 2020 were also reviewed to identify patients who received these agents, had pretreatment MDCT, and remained free of MRONJ for at least 2 years. Two types of three-dimensional segmentations were performed: one including cancellous bone alone and the other including cancellous bone plus the surrounding cortical bone. Radiomic features were extracted and quantified. Least absolute shrinkage and selection operator (LASSO) regression was employed to select effective informative features. Machine learning models-Random Forest (RF), Support Vector Machine (SVM), and Multilayer Perceptron (MLP)-were trained and evaluated using sensitivity, specificity, and area under the curve (AUC). RESULTS: This study included 94 patients who underwent MDCT imaging at our institution before receiving bone resorption inhibitors. The cohort included 44 patients with mandibular MRONJ and 50 without. In testing for the cancellous bone-only group, RF, SVM, and MLP showed sensitivities of 0.568, 0.568, and 0.659; specificities of 0.720, 0.680, and 0.760; and AUCs of 0.741, 0.706, and 0.802, respectively. Using testing datasets from the group including both cancellous and mandibular border cortical bones, the results were: sensitivity: 0.818, 0.796, and 0.886; specificity: 0.843, 0.824, and 0.886; AUC: 0.931, 0.917, and 0.921, respectively. CONCLUSIONS: Radiomics analysis of pretreatment MDCT images may accurately predict MRONJ risk before exposure to bone resorption inhibitors.
BACKGROUND: R-Loops are three-stranded DNA/RNA hybrids implicated in immune responses. Their role in mesenchymal stem cell differentiation, particularly in dental pulp regeneration under inflammation, remains unclear. ME...BACKGROUND: R-Loops are three-stranded DNA/RNA hybrids implicated in immune responses. Their role in mesenchymal stem cell differentiation, particularly in dental pulp regeneration under inflammation, remains unclear. METHODS: Rat pulpo-dentinal complex (PDC) injury models and LPS-stimulated human dental pulp stem cells (hDPSCs) were used. R-Loops and cGAS-STING were evaluated via immunofluorescence, immunohistochemistry, RNA-seq, RT-qPCR, ALP, and Alizarin Red S staining. Interventions included RNase H1 overexpression (to degrade R-Loops) and STING inhibitor H151. RESULTS: Inflammation induced R-Loops accumulation and cGAS-STING activation in vivo and in vitro, correlating with impaired odontogenic differentiation. RNase H1 or H151 treatment downregulated inflammatory cytokines (IFN β, IL 6, TNF α) and restored mineralization capacity of hDPSCs. CONCLUSION: R-Loops promote inflammation and suppress odontogenesis via cGAS-STING signaling. Targeting this axis alleviates inflammation and enhances regenerative potential, offering a therapeutic strategy for pulpitis.
BACKGROUND: Combustible cigarette smoking is a well-established risk factor for poor oral health, but the implications of e-cigarette use and dual use remain uncertain. Distinguishing the effects of vaping from the linge...BACKGROUND: Combustible cigarette smoking is a well-established risk factor for poor oral health, but the implications of e-cigarette use and dual use remain uncertain. Distinguishing the effects of vaping from the lingering consequences of prior smoking is a persistent challenge in the literature. OBJECTIVES: To assess how distinct nicotine use profiles relate to oral health-related quality of life, self-reported oral health indicators and self-rated dental health among adults in England, while differentiating e-cigarette users by smoking history. METHODS: We conducted a cross-sectional analysis of 6027 adults (aged ≥ 16 years) from the nationally representative 2021 Adult Oral Health Survey for England. Participants were classified into five current smoking/vaping categories (never users, former smokers, exclusive cigarette smokers, exclusive e-cigarette users, dual users) and, separately, by combined vaping-smoking history. Outcomes included any oral health difficulty and the number of impacts derived from the 14-item Oral Health Impact Profile (OHIP-14), as well as loose teeth, gum disease history, dry mouth and self-rated dental health. Binary outcomes (any oral health difficulty, loose teeth, gum disease history) were analysed using logistic regression, and ordinal outcomes (number of OHIP-14 impacts, dry mouth, self-rated dental health) using ordered logistic regression, with adjustment for sociodemographic, behavioural and oral hygiene covariates. RESULTS: Compared with never users, exclusive cigarette smokers had the highest adjusted odds of reporting any oral health difficulty (OR = 1.85, 95% CI: 1.51-2.27), loose teeth (OR = 3.18, 95% CI: 2.47-4.09), gum disease history (OR = 1.94, 95% CI: 1.52-2.49) and poorer self-rated dental health (OR = 2.88, 95% CI: 2.36-3.52). Dual users showed similarly elevated odds for loose teeth (OR = 3.11, 95% CI: 1.95-4.97), the highest odds for gum disease history (OR = 3.21, 95% CI: 2.06-4.99) and elevated odds for any oral health difficulty (OR = 1.55, 95% CI: 1.04-2.30) and poorer self-rated dental health (OR = 2.45, 95% CI: 1.68-3.58). Exclusive e-cigarette users also showed elevated odds across outcomes, though generally of smaller magnitude. Estimates for the very small subgroup of never-smoking current e-cigarette users did not show significant excess odds across outcomes, but were imprecise and should be regarded as exploratory. CONCLUSION: In this cross-sectional population-based analysis, combustible cigarette smoking and dual use were associated with poorer self-reported oral health outcomes. Findings related to e-cigarette use require cautious interpretation because of prior smoking history, the sparse never-smoker vaping subgroup and the absence of detailed exposure data (e.g., smoking intensity, pack-years, time since cessation, vaping duration, device type and nicotine concentration). Smoking cessation should remain central to preventive dentistry.
BACKGROUND: The optimal management of early-stage oral squamous cell carcinoma (OSCC) with clinically node-negative neck remains controversial. A reliable biomarker to stratify occult metastasis risk for elective neck di...BACKGROUND: The optimal management of early-stage oral squamous cell carcinoma (OSCC) with clinically node-negative neck remains controversial. A reliable biomarker to stratify occult metastasis risk for elective neck dissection (END) is lacking. METHODS: This retrospective cohort included 231 early-stage cN0 OSCC patients. Tumor budding (TB) was graded by ITBCC criteria. Survival was analyzed using Kaplan-Meier and Cox regression; subgroup and additive interaction were tested. RESULTS: High-grade TB was found in 55.4% (128/231) and associated with poorer 5-year OS (80.4% vs. 93.0%, p = 0.0059) and DFS (66.2% vs. 80.6%, p = 0.019). END (67.5%, 156/231) independently improved OS (HR = 0.28, p < 0.001) and DFS (HR = 0.35, p = 0.003). In high-grade TB patients, END markedly raised 5-year OS (88.1% vs. 60.0%, p < 0.001) and DFS (76.2% vs. 39.8%, p < 0.001), with significant additive interaction (RERI = 7.12). No OS benefit was seen in low-TB patients. CONCLUSION: TB effectively stratifies END benefit. END is recommended for high-grade TB early stage cN OSCC, while watch-and-wait is suitable for low-grade TB patients.