O Journal of Applied Oral Science é uma continuação da Revista da Faculdade de Odontologia de Bauru. O Journal of Applied Oral Science opera na modalidade de publicação contínua eletrônica, formato que permite a publicação de volumes anuais sem especificações de fascículos. A Revista da Faculdade de Odontologia de Bauru foi publicada no período de 1993-2001, sendo continuada, em 2002, pelo Journal of Applied Oral Science, volume 11, número 1, mantendo a numeração de sua antecessora. O Journal of Applied Oral Science tem como objetivo principal publicar resultados de pesquisas originais, e a convite do editor, revisões nos campos da Odontologia, Fonoaudiologia e áreas correlatas. A abreviatura de seu título é J. Appl. Oral Sci., que deve ser usada em bibliografias, notas de rodapé e referências.
Universidade de São Paulo. Faculdade de Odontologia de Bauru
Todo o conteúdo do periódico, exceto onde está identificado, está licenciado sob uma Licença Creative Commons do tipo atribuição CC-BY.
Effects of highly leukotoxic Aggregatibacter actinomycetemcomitans associated with ligature-induced periodontitis on oral and gut tissues in male rats
Objective To evaluate the effects of gastric administration of the highly leukotoxic Aggregatibacter actinomycetemcomitans JP2 strain (Aa JP2), with or without ligature-induced periodontitis, on periodontal and small intestinal tissues in a rat model. Methodology Forty-eight-week-old male Wistar rats were divided into four groups (n=10/group): Control (CG), Periodontitis (PG), Periodontitis and Aa JP2 (PAaG), and Aa JP2 only (AaG). Ligatures were placed in PG and PAaG on day one and maintained for 42 days. After six weeks of twice-weekly gastric Aa JP2 inoculation, mandibles and small intestines were analyzed using stereomicroscopy, micro-computed tomography (CT), and histopathology to assess alveolar bone loss, trabecular architecture, inflammatory infiltrate, vascular congestion, goblet cell density, and villus morphology. Statistical analyses included the Kruskal–Wallis, Mann–Whitney, and Chi-square tests for group comparisons (p<0.05), as well as Spearman’s correlation test. Results PG and PAaG exhibited significantly greater bone loss compared to CG and AaG (p<0.05). Micro-CT analysis revealed reduced trabecular thickness in AaG (342.4±23.1µm) compared to PG (378.2±28.1µm) and PAaG (385.5±45.1µm) (p<0.05). PAaG and AaG presented higher inflammatory infiltrate scores (>751 inflammatory cells) compared to CG and/or PG (p<0.05). Elevated vascular congestion and/or goblet cell hyperplasia in the jejunum and ileum were observed in PAaG and AaG compared to CG and PG (p<0.05). Villus height (duodenum/ileum) and villus width (jejunum/ileum) differed significantly among groups (p<0.0001). Significant correlations were identified: the alveolar bone volume/total volume ratio was positively associated with duodenal vascular congestion (rho=0.738; p=0.037); the periodontal inflammatory infiltrate was positively associated with jejunal goblet cell counts (rho=0.704; p=0.007); alveolar bone loss was positively associated with both duodenal and jejunal villus height (rho≥0.762; p<0.05); and bone volume was negatively associated with both jejunal vascular congestion (rho=-0.696; p=0.003) and jejunal goblet cell counts (rho=-0.617; p=0.011). Conclusions Gastric inoculation with Aa JP2 exacerbates ligature-induced periodontitis and independently induces intestinal morphological changes.
Copyright (c) 2026 https://creativecommons.org/licenses/by/4.0
Effects of live and heat-killed Bifidobacterium lactis in rats with induced periodontitis
Background Probiotics and postbiotics have emerged as promising adjunctive therapies in managing periodontal disease. Bifidobacterium animalis subsp. lactis HN019 has shown antimicrobial and immunomodulatory effects in both experimental and clinical settings when administered orally. However, the systemic impact of this strain, independent of local oral effects, remains unclear. Objective To evaluate the systemic effects of live and heat-killed B. animalis subsp. lactis HN019 in a rat model of ligature-induced periodontitis, excluding direct contact with the oral cavity. Methodology A total of 32 Wistar rats were randomly assigned to four groups (n=8): control (C), periodontitis only (EP), periodontitis + probiotic (PRO), and periodontitis + postbiotic (POS). Periodontitis was induced by placing a cotton ligature around the cervical region of the lower right first molar, inserted into the gingival sulcus. Treatments were administered via oral gavage for 30 days before and 15 days after periodontitis induction. Alveolar bone loss and periodontal parameters were assessed using micro-computed tomography (microCT) and histomorphometric analysis. Results MicroCT revealed that ligature effectively induced periodontitis, reducing BV/TV and Tb.N and increasing Tb.Sp and Po.Tot. Probiotic and postbiotic treatments did not improve outcomes. Bone loss was lowest in the control group, with no differences between EP, PRO, and POS. Conclusion Systemic administration of Bifidobacterium animalis subsp. lactis HN019 or its derived postbiotic resulted in no significant improvements in periodontal outcomes in this experimental model. Further investigations using integrative approaches are needed to better characterize the systemic effects of probiotics and postbiotics.
Hydrogel microarchitecture as a regulatory cue for in vitro odontogenic differentiation
Objective This study introduces an innovative, cost-effective, and easily reproducible strategy for engineering three-dimensional bioprinted GelMA-based scaffolds, designed with ordered macroporous and tubular architectures, and integrated microfluidic channels to advance structural and functional performance. Their geometric features were specifically designed to investigate how microarchitectural cues influence the mineralizing cell differentiation of human dental pulp cells (HDPCs). Methodology The scaffolds were fabricated via an indirect bioprinting process using resin molds, resulting in cylindrical structures with distinct grid or honeycomb surface architectures. Biomaterials were characterized for morphology, surface topography, porosity, pore diameter, and degradability. Biological performance was evaluated by culturing HDPCs for 21 days to assess viability, proliferation, and mineralizing differentiation (ANOVA/Tukey; α=0.05). Results Both scaffold designs exhibited interconnected porous networks, with the honeycomb configuration presenting significantly larger pores. HDPCs cultured on the scaffolds showed high viability and proliferation, with the honeycomb architecture promoting elevated ALP activity. However, the grid architecture more effectively influenced odontoblastic differentiation and mineralized matrix deposition. Conclusion Our findings highlight the impact of biomaterial architecture on cellular behavior and reveal the potential of this novel bioprinting approach for bioactive dentin regeneration in dental tissue engineering.
The effect of Stage I-II periodontitis on the mechanical, structural, and biological properties of leukocyte and platelet-rich fibrin
Objective This study aimed to compare the mechanical, biological, and structural properties of a leukocyte- and platelet-rich fibrin (L-PRF) membrane in periodontally healthy patients and those with periodontitis. A total of 30 individuals (17 men and 13 women) with a median age of 41 years were divided into two groups as periodontally healthy (control group) and as having periodontitis (test group). Methodology Blood samples were collected and centrifuged at 2700 rpm for 12 minutes. A tensile test was performed to determine the mechanical properties of L-PRF membranes. Elasticity-modulus, tensile strength, and stretch at rupture were calculated. Platelet and leucocyte counts and ratios were calculated. Fibril thickness and density were evaluated by scanning electron microscopy imaging. The comparisons between groups were analyzed by the Mann–Whitney U test, and a p<0.05 value was considered statistically significant. Results In the comparison of the control group versus the test group, pocket depth (3.55 (1.59-3.97) vs. 1.52 (1.47-3.53)), clinical attachment level (3.11 (1.77-4.23) vs. 0.06 (0.05-3.68)), plaque index (1.63 (0.05-1.75) vs. 0.04 (0.04-1.58)), gingival index (1.55 (0.04-1.63) vs. 0.05 (0.04-1.49)), and bleeding on probing (56.25 (3.7-70.11) vs. 3.57 (2.67-47.12)) were statistically significantly higher in the test group than in the control group (p= 0.009, p=0.008, p=0.008, p=0.009, p=0.009, respectively). Elasticity-modulus, tensile strength, and stretch at rupture values showed no statistically significantly differences between groups (p=0.745, p=0.754, p=0.602, respectively). Fibril thickness and density values showed no statistically significant differences between groups (p=1.000, p=0.753, respectively). Platelet count and platelet and leukocyte ratio values showed no statistically significant differences between groups (p=0.754, p=0.600, p=0.142, respectively). Leukocyte count was found to be statistically significantly higher in the control group than in the test group (p=0.028). Conclusion This study showed no effect of Stage I-II periodontitis on the mechanical, structural, and biological properties of L-PRF. Further studies with larger sample sizes are required to confirm these findings.
Bond strength in class V cavities: an in vitro study on beveling, universal adhesive strategy, and composite type
Cervical Class V lesions present restorative challenges due to their multifactorial etiology and complex bonding substrates. Objectives: This study evaluated the influence of marginal bevel, bonding using a universal adhesive (etch-and-rinse or self-etch), and the resin composite type (conventional or bulk-fill) on the bond strength of simulated Class V cavities. Methodology: A total of 80 dental fragments (7mm×7 mm×2 mm) were obtained from bovine incisors. Cylindrical cavities with enamel margins and internal dentin walls were prepared centrally in each specimen. Samples were randomly assigned to groups based on three factors: presence or absence of bevel, bonding strategy (etch-and-rinse or self-etch), and resin composite type (conventional or bulk-fill). Groups with bevel received a 1 mm enamel bevel along the preparation margins using a conical diamond bur. Adhesive and restorative procedures were performed following the manufacturer’s recommendations. Bond strength was assessed using a push-out test on a universal testing machine. Data were analyzed using three-way ANOVA and Tukey’s post-hoc test (α=5%). Results: Presence of a marginal bevel did not significantly influence bond strength. The etch-and-rinse bonding strategy resulted in significantly higher bond strength than the self-etch approach, and bulk-fill composites presented superior performance compared with conventional composites. Conclusions: Combining a universal adhesive applied with the etch-and-rinse strategy and a bulk-fill composite proved to be an effective approach for enhancing bond strength with enamel and dentin substrates. Beveling of enamel margins did not improve the bond strength of Class V restorations, suggesting that this procedure may be unnecessary when margins are in enamel. Avoiding beveling in such situations contributes to preserve sound tooth structure and supports a more conservative restorative approach.
Comparative analysis of connective tissue graft versus ozone-enriched platelet-rich fibrin in the treatment of Cairo class I gingival recessions
Gingival recession is characterized by the apical displacement of the marginal gingiva, exposing the root surface and causing aesthetic and functional issues such as dentin hypersensitivity and cervical lesions. Subepithelial connective tissue grafts (SCTG) are considered the gold standard for root coverage. Recently, ozone-enriched advanced platelet-rich fibrin (A-PRF) has emerged as a promising alternative due to its regenerative potential and biocompatibility. Objectives: To compare the clinical efficacy of ozone-enriched A-PRF and SCTG combined with the coronally advanced flap (CAF) technique to treat Cairo Class I gingival recessions. Methodology: This randomized, double-blind, controlled clinical trial included 22 healthy patients (mean age 32.23±11.86 years, range 20–60 years), each with bilateral Cairo Class I recessions. Sites were treated with either CAF + SCTG (control) or CAF + ozone-enriched A-PRF (test). Clinical parameters were assessed at baseline, 90, and 180 days. Data were analyzed using the Shapiro-Wilk, repeated measures ANOVA, Tukey, and Friedman tests (α=0.05) on GraphPad Prism 8.0®. This study adhered to the CONSORT guidelines for randomized clinical trials. Results: Complete root coverage (100%) was achieved in 13 of 22 sites (60%) in the SCTG group and in nine of 22 cases (43%) in the ozone-enriched A-PRF group. Both groups statistically and significantly improve over time in selected clinical parameters (p<0.05). The SCTG group showed significant gains in all evaluated outcomes, whereas the ozone-enriched A-PRF group showed significant improvements in clinical attachment level, recession height, recession width, and dentin hypersensitivity. SCTG more greatly increased gingival thickness (0.77±0.52 mm) and keratinized tissue width (0.81±0.95 mm) than ozone-enriched A-PRF (0.27±0.55 mm and 0.22±0.42 mm, respectively). Conclusions: Both treatment approaches effectively managed Cairo Class I gingival recessions. SCTG promoted superior soft tissue augmentation, whereas ozone-enriched A-PRF achieved comparable reductions in recession-related parameters. The limitations of this study include its short follow-up period and the absence of histological assessment. Clinical Trial Register: RBR-537bzqt
Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm in the oral cavity, characterized by high morbidity and mortality. Various proteins, including matrix metalloproteinases (MMPs), have been investigated as potential biomarkers for diagnosing and prognosing this disease. Objective: To evaluate the utility of MMP-2, MMP-9, and the tissue inhibitor of metalloproteinase-2 (TIMP-2) as biomarkers in diagnosing and prognosing OSCC. Methodology: This retrospective case-control study involved 60 cases and 30 controls from the University Hospital Fundación Santa Fe de Bogotá, part of the Colombian cohort of the InterChange/Headspace study. Saliva samples were collected using a 60-second mouth rinse with 10 mL of sterile saline solution, along with serum and oral tissue samples and clinical and sociodemographic data. MMP-9, MMP-2, and TIMP-2 concentrations in saliva were determined using ELISA, whereas tissue samples were analyzed by immunohistochemistry. MMP activity in saliva was quantified using a generic activity assay. Results: Salivary MMP-9 and MMP-2 concentrations were significantly higher in cases than in controls, whereas no significant differences were observed for TIMP-2. ROC curve analysis showed excellent discriminatory capacity for salivary MMP-9 (AUC=0.946; sensitivity=0.817; specificity=0.867), good performance for MMP-2 (AUC=0.708; sensitivity=0.717; specificity=0.633), and lower discriminatory ability for TIMP-2 (AUC=0.630; sensitivity = 0.617; specificity=0.500). Elevated MMP-9 concentrations were observed in 33% of the serological samples. The immunohistochemistry of MMP-2, MMP-9, and TIMP-2 showed differences between cases and controls. Conclusions: This study highlights salivary MMP-9 and MMP-2, together with MMP activity, as biomarkers of interest for OSCC that may contribute to the understanding of disease-related molecular changes detectable by liquid biopsy. While elevated salivary levels were observed in OSCC cases when compared to controls, these findings should be interpreted as exploratory. Elevated serological MMP-9 levels were observed in a small subset of cases. However, the limited sample size and the absence of a control group in this study preclude diagnostic interpretation. Larger, well-powered, and externally validated studies are required to determine the potential clinical utility of these biomarkers.
Squamous cell carcinoma of head and neck
Assessment of chitosan addition to at-home bleaching gel with and without calcium on the physical properties of tooth enamel
Dental bleaching is a widely used esthetic treatment, but concerns persist regarding its potential effects on enamel integrity. The incorporation of remineralizing and protective agents into bleaching gels, such as calcium and chitosan, has been proposed to mitigate these effects. Objective: This in vitro study evaluated the effects of adding 2% chitosan to at-home bleaching gels with and without calcium on enamel physical properties. Methodology: Bovine enamel specimens stained with black tea were assigned to five groups (n = 12): control (no bleaching), 7.5% hydrogen peroxide (7.5% HP), 7.5% HP + 2% chitosan (7.5% HP+ Chi), 7.5% HP + calcium (7.5% HP + Ca), and 7.5% HP + calcium + chitosan (7.5% HP + Ca + Chi). Bleaching was performed for 14 days (1 hour/day). Color (∆L*, ∆a*, ∆b*, ∆Eab, ∆E00, and ∆WID), surface roughness, and surface microhardness (SMH) were evaluated at baseline (T1) and after treatment (T2). Mineral content (Ca, P, and the Ca/P ratio) was assessed by energy-dispersive X-ray spectroscopy and enamel morphology by scanning electron microscopy. Statistical analyses included the Kruskal-Wallis and Dunn's tests, mixed models, and ANOVA. Results: Groups containing chitosan (7.5% HP + Chi and 7.5% HP + Ca + Chi) showed the lowest surface roughness and the highest microhardness values. For these groups, SMH showed no decrease after treatment (T2), remaining similar to the initial values (T1). Color changes were similar among all bleached groups. Calcium, phosphorus, and Ca/P levels were the same for all groups. Conclusion: Adding 2% chitosan to 7.5% hydrogen peroxide-based at-home bleaching gels appears to be a promising approach to protect enamel against changes in roughness, microhardness, and mineral content without compromising bleaching efficacy.
Integrated analysis identifies CCNA2 as a candidate diagnostic and prognostic biomarker in oral tongue squamous cell carcinoma
Background Oral tongue squamous cell carcinoma (OTSCC) is an aggressive malignancy with poor prognosis, necessitating reliable biomarkers. Methodology Genes with significantly higher expression in OTSCC tumor tissues compared to normal tongue tissues were identified via integrated transcriptomic analysis of seven GEO datasets. To assess their diagnostic and prognostic potential, these genes were further characterized using multi-omic and clinical data from the TCGA-OTSCC and CPTAC-OTSCC cohorts. Results A total of 1,117 genes were found to be upregulated in OTSCC tissues, among which only CCNA2 (Cyclin A2) was significantly associated with both reduced overall survival (OS) and disease-free survival (DFS) in the TCGA-OTSCC cohort (n=128), based on Cox proportional hazards regression and Kaplan–Meier analyses. CCNA2 showed moderate prognostic performance (AUC=0.63 for OS; AUC=0.65 for DFS) and was significantly upregulated in higher-grade tumors (p=0.01) and in deceased patients (p=0.03). No somatic mutations or promoter methylation alterations were observed in CCNA2 based on TCGA data. In CPTAC-OTSCC samples (n=18), CCNA2 protein expression was significantly higher in tumor tissues than in non-tumoral tissues (p <0.0001), with a positive correlation between mRNA and protein levels (r=0.56, p=0.01). Both mRNA and protein forms showed strong diagnostic performance (AUC=0.92 and AUC=0.82, respectively), consistent with observations across multiple tumor types. While CCNA2 protein levels showed prognostic relevance for OS (AUC=0.69, p=0.01), the mRNA-based prediction did not reach statistical significance (AUC=0.63, p=0.36). Functional enrichment analysis of CCNA2 co-expressed genes predicted involvement in cell cycle, mismatch repair, and DNA replication pathways. Additionally, protein–protein interaction analysis positioned CCNA2 as a central hub, suggesting its potential role in OTSCC pathogenesis. Conclusions These findings indicate that CCNA2 is a promising diagnostic and prognostic biomarker candidate in OTSCC. Given the small size of the CPTAC validation cohort, further studies in larger, independent OTSCC cohorts are warranted to confirm its clinical utility.
Evaluation of bleaching efficacy and cytotoxicity of an experimental hydrogel containing MnO-Doped Biosilicate® activated by violet LED light
Although hydrogen peroxide (HP) at 35% is effective in dental bleaching, the cytotoxicity associated with its use remains a significant concern. In this context, this study aimed to develop a bleaching hydrogel containing 6% HP, Biosilicate® (BioS) doped with manganese oxide (MnO_BioS), and irradiated with violet LED light (LED). Enamel/dentin discs were submitted to the following bleaching treatments (n=08): 35%HP (positive control), 6%HP gel containing BioS or MnO_BioS (0 and 10 wt%), with or without LED irradiation. The discs were adapted to artificial pulp chambers (APCs), with the enamel exposed for bleaching and the dentin facing the culture medium (DMEM). Bleaching was performed in three 30-minute sessions with 7-day intervals. After bleaching, the extracts (DMEM + bleaching gel components diffused through the discs) were collected and applied to the odontoblast-like MDPC-23 cells. Color change (ΔE00) and changes in the whiteness index (ΔWID) were determined before (T0) and after the last bleaching session (T2). Cell viability (MTT, %), HP diffusion (µg/mL), oxidative cell stress (OxS), and cell fluorescence (live/dead assay, by confocal microscopy) were assessed after the first session (T1). The data were analyzed using ANOVA and Tukey’s test (p=0.05). The addition of 6%HP_MnO_BioS_LED exceeded ΔWID compared to 35%HP, showing statistical differences from the other groups, while ΔE00 was statistically similar to 35%HP. 6%HP_MnO_BioS_LED demonstrated higher cell viability, lower HP diffusion, and reduced oxidative stress (OxS) compared to the other groups (p<0.05). 6%HP_MnO_BioS_LED increased bleaching potential and presented lower cytotoxicity compared to 35%HP.
Notch2 is involved in regulating apical tissue repair following severe intrusive luxation
This study aims to characterize apical repair dynamics in immature permanent teeth after severe intrusive luxation, and to explore Notch signaling pathway regulation in this process using a Sprague-Dawley rat model. Methodology In this study, a modified tool was utilized to establish an intrusive luxation model in SD rats. Tissue repair was assessed via micro-computed tomography (Micro-CT) and hematoxylin-eosin (H&E) staining. Differential gene expression analysis of stem cells from the apical papilla (SCAPs) was performed using the publicly available GEO dataset. Immunohistochemical detection of Notch2 receptor expression in the apical region of injured teeth was subsequently conducted, guided by bioinformatics screening. Finally, the Notch signaling pathway was pharmacologically inhibited using DAPT (γ-secretase inhibitor), and its impact on post-injury prognosis was evaluated. Results Histological analysis revealed prevalent pulp hypoplasia and apical tissue fibrosis after injury. Differential gene analysis suggested that the Notch signaling pathway, particularly Notch2, is involved in the regulation of angiogenesis-related pathways in SCAPs. Immunohistochemistry showed positive Notch2 expression in fibrotic apical regions. Notably, inhibition of Notch signaling significantly reduced aberrant fibrosis while enhancing vascular proliferation in the injury site. Conclusions Severe intrusive luxation is primarily characterized by aberrant fibrous differentiation in the apical region. The Notch signaling pathway may negatively regulate angiogenesis in the damaged apical area, suggesting that targeted inhibition of this pathway could promote tissue repair following embedded dental injuries.
Bioactive luting cements on the adhesive interface of fiber post and root dentin
Innovative self-etching dental cements have been considered a versatile material with recent formulations incorporating bioactive additives. Objective This study analyzed the role of these ingredients in systems used for luting intracanal posts. Methodology Bond strength (BS), water sorption (WS), water solubility (WSB), and in situ zymography (IZ) were assessed. Three cements were tested: RelyX U200 (RU, bioactive-free control), RelyX Luting Plus (RL, fluoride-containing glass-ionomer), and BeautiCem SA (BC, S-PRG-based). Bond strength was evaluated in bovine root dentin (n=10) at the cervical, middle, and apical thirds at 7 days and 6 months. WS/WSB were assessed using 10 cement disks (10×1 mm) cycled through deionized water immersion (m1), desiccation (m2), and a second immersion (m3). For IZ, the specimens (n=3) were sectioned and incubated with fluorescein-conjugated gelatin for 48h at 37°C at both storage times (7 days and 6 months) for analysis by Confocal Laser Scanning Microscopy. Fluorescence intensity was quantified using Image J. Data were analyzed using ANOVA and Tukey’s test (p<0.05). Results RU presented the highest BS and the lowest WS/WSB, while RL showed the opposite trend. BC demonstrated intermediate performance with no statistical differences from the other materials. For all cements, the cervical third and the 6-month period yielded the highest BS values. WS/WSB were greater for the bioactive cements (BC, RL). IZ results aligned with BS findings, revealing suppressed enzymatic activity over time. Based on these results, it can be concluded that RU exhibited superior mechanical stability despite lacking bioactivity. Conclusion Among bioactive cements, BC emerged as an interesting alternative, offering improved bond strength, water stability, and bioactive potential compared to RL.
Copyright (c) 2026 Tiago Carvalho dos Santos, Giovanna Speranza Zabeu, Ana Carolina Almeida Lima, Mylena Proença Costa, Marcela Rocha Carrilho, Linda Wang https://creativecommons.org/licenses/by/4.0
Tooth discoloration is a frequent concern among dental patients. While many professionals and patients choose veneers or other invasive solutions, Minimal Intervention Dentistry (MID) recommends non- and microinvasive approaches to preserve tooth structure. Objective To assess Brazilian dentists’ knowledge and practices on tooth discoloration and how education and professional profile may influence them. Methodology A cross-sectional observational study was conducted with 384 volunteer dentists via an online series of questions on causes, diagnosis, treatment (per tissue removal), and professional-patient relationship. Answers were scored (1=correct/affirmative, 0=incorrect/negative). Descriptive and multivariate regression analyses were performed (α=0.05). Results Most respondents were female, aged 20-49, from the Southeast region. Correct answer rates: 69.9% for causes, 69.4% for diagnosis, 48.8% for treatment, 83.6% for professional-patient relationship, 56.4% overall. Graduation from public universities and holding graduate training in Operative or Pediatric Dentistry or Orofacial Harmonization were associated with better outcomes. Conclusion Knowledge of non- and microinvasive approaches to tooth discoloration remains limited. Broader educational efforts are needed, especially among specific groups of professionals who showed lower performance.
Effects of bromelain, melatonin, and thymoquinone on neutrophil extracellular trap formation and macrophage polarization in experimental periodontitis
Periodontitis is a chronic inflammatory disease characterized by dysregulated immune responses leading to alveolar bone loss. Formation of neutrophil extracellular traps and macrophage polarization play critical roles in its pathogenesis, and natural agents with immunomodulatory properties are being investigated as potential adjunctive therapies. Objective This study aimed to compare the effects of systemically administered melatonin, bromelain, and thymoquinone on serum neutrophil extracellular trap formation and macrophage polarization in experimental periodontitis. Methodology Forty rats were assigned to five groups: control, experimental periodontitis, and experimental periodontitis treated with melatonin, bromelain, or thymoquinone. Periodontitis was induced by ligature placement for 14 days. After ligature removal, the treatment groups received daily oral doses of melatonin, bromelain, or thymoquinone for 14 consecutive days. Biochemical, histologic, histomorphometric, and immunohistochemical analyses were performed. Comparisons between groups were performed using one-way ANOVA. Neutrophil extracellular trap levels measured before and after treatment within the same group were analyzed using a paired t-test. Results Thymoquinone treatment significantly reduced alveolar bone loss compared to the bromelain and melatonin groups (p=0.00). The highest value was observed in the Periodontitis group and the lowest in the Thymoquinone group for neutrophil extracellular trap levels, with intergroup comparisons showing no statistically significant differences (p=0.597). All immunohistochemical analyses showed significant differences in all treatment groups compared to both the control and periodontitis groups (p=0.000). CD68(+) macrophages were most abundant in the periodontitis group and least in the Melatonin group (p=0.00). CD80 expression was highest in the Melatonin group and lowest in the Thymoquinone group (p=0.000), while CD206 expression was highest in the Bromelain group and lowest in the Thymoquinone group (p=0.00). Conclusion Thymoquinone exhibited comparatively more favorable patterns, and overall, all agents showed trends that may support their consideration as adjunctive approaches in periodontitis management.
DNA methyltransferases expressions in mice tongue exposed to waterpipe smoke
Objective Waterpipe smoking has gained widespread use nowadays, especially among young adults. As this type of smoke has been described to contain toxins and carcinogens, this study evaluated DNA methyltransferase (DNMT) 1 and 3b expression, and inflammation, on the dorsal surface, ventral surface and lateral border of Swiss mice’s tongues exposed to waterpipe smoke. Methodology Animals were divided into 6 groups (n=60): control, 7, 15, 30, 60, and 90 days of consecutive exposure to waterpipe smoke in a whole-body exposure system. After each period, tongues were analyzed using hematoxylin/eosin staining for inflammation status and immunohistochemistry for DNMT1 and DNMT3b. Results DNMT3b showed lower immunoexpression from 7 to 60 days; at 90 days, expression was similar to that of control or there was upregulation on the ventral surface compared with control. DNMT1 exhibited lower expression at all exposure times, with the ventral surface showing similar expression to that of control at 90 days. Waterpipe smoke was not able to induce acute or chronic inflammation in mice tongue. Conclusion Waterpipe smoke may result in a DNA hypomethylation pattern in initial exposure periods, contributing to activate proto-oncogenes and/or genomic instability. Over long periods, it may lead to a methylation pattern similar to that of control or even to hypermethylation, silencing tumor suppressor genes. These alterations in the genome due to hypo/hypermethylation contribute largely for the development of diseases such as oral cancer.
Squamous Cell Carcinoma of Head and Neck
Maqui berry cystatin inhibits cathepsin k activity and stimulates osteogenic differentiation of human dental pulp cells
Objectives Cystatins, endogenous inhibitors of cysteine proteases, regulate extracellular matrix degradation. Their plant-derived homologs (phytocystatins) include MaquiCPI-3, a recombinant protein obtained from Aristotelia chilensis (maqui berry). This study aimed to investigate the inhibitory effect of MaquiCPI-3 on human cathepsin K (CTSK) activity and its cytotoxicity and impact on the proliferation, migration, and osteogenic differentiation of human dental pulp cells (hDPCs). Methodology The inhibitory activity of MaquiCPI-3 against CTSK was measured using a spectrofluorometer with the fluorogenic substrate Z-Phe-Arg-AMC. The hDPCs from third molars were characterized by flow cytometry for mesenchymal (CD90, CD73, CD105) and hematopoietic (CD34, CD45) markers. The hDPCs, either exposed to MaquiCPI-3 or left untreated (control), were assessed for viability (MTT assay), proliferation (bromodeoxyuridine incorporation), chemotaxis (Transwell assay), mineralized nodule formation (Alizarin Red S staining), alkaline phosphatase activity (thymolphthalein release), and expression of mineralization-related genes (qPCR). Data were analyzed using one- or two-way ANOVA with appropriate post hoc tests or nonparametric alternatives (α=0.05). Results MaquiCPI-3 potently inhibited CTSK (Ki=1.72 nM, Ki,app=2.08 nM), showed no cytotoxicity, and significantly enhanced ALP activity, mineralized nodule formation, and expression of BMP-2 and osteocalcin, stimulating no hDPC proliferation or migration when compared with the control. Conclusions MaquiCPI-3 increased no cell proliferation or migration, its ability to inhibit CTSK activity and induce an osteogenic phenotype shows promising potential therapeutic strategies aimed at repairing and regenerating pulp and periapical tissues and controlling bone resorption.
Diversity of Streptococcus mutans serotypes isolated from dental plaque of Brazilian patients with a history of acute myocardial infarction
Background: Streptococcus mutans serotypes (c, e, f, k) are linked to dental caries, with less common serotypes (e, f, k) and collagen-binding genes (CBG: cnm and cbm) suggested to be associated with cardiovascular diseases. Objective: This study investigated the presence of S. mutans serotypes and collagen-binding genes (CBGs) in dental plaque and their possible association with acute myocardial infarction (AMI) in adults. Methodology: A total of 31 infarcted and 17 non-infarcted patients underwent oral and blood examinations. DNA from plaque samples was analyzed using PCR to identify S. mutans serotypes and cnm/cbm genes. S. mutans was detected in 22.6% (7/31) of infarcted patients and 11.8% (2/17) of non-infarcted patients. Results: Serotype c was the most prevalent in infarcted patients (57.1%, 4/7), followed by e (42.9%, 3/7) and k (14.3%, 1/7); serotype f was not detected. Only serotype c was found in non-infarcted patients. Less common serotypes (e, k) and co-occurring serotypes (c, k) were exclusive to infarcted patients. No CBGs were detected in any serotypes. No association was found between S. mutans and dental caries in either group. Patients with cholesterol levels <190 mg/dL and no S. mutans had a 47.4% lower risk of infarction. Conclusion: Although S. mutans, including less common serotypes, was more prevalent in infarcted patients, no significant association with AMI was observed. Further research is needed to elucidate the role of S. mutans in cardiovascular diseases.
SHED-exosomes functionalized micro-nano bioactive glass/PCL membrane regulates macrophage polarization and promotes osteogenic differentiation
Micro-nano bioactive glass (MNBG) has bone regenerative potential. However, the difficulty of MNBG molding restricts its clinical applications in bone regeneration. Exosomes carry proteins, lipids, and nucleic acids for communication between cells. In this study, we adhered human exfoliated deciduous teeth (SHED)-derived exosomes (SHED-Exos) to electrospun micro-nano bioactive glass/polycaprolactone (MNBG/PCL) membrane to control inflammation and promote bone regeneration. MNBG/PCL membrane showed biocompatibility and osteogenic ability. Next, we demonstrated that SHED-Exos internalized into macrophages attenuated LPS-induced expression of M1-macrophage markers iNOS, IL-6, and IL-1β and upregulated M2-macrophage marker IL-10 expression, indicating a switch from M1 to M2 macrophage phenotype. MNBG/PCL-loaded SHED-Exos membrane supported the survival and growth of mouse bone marrow stromal cells (mBMSCs). When M1 macrophages were co-cultured with mBMSCs on the membrane, the SHED-Exos-loaded membrane showed higher Runx2, Alp, and Ocn gene expression. Our findings indicate that SHED-Exos-functionalized MNBG/PCL membrane has anti-inflammatory and osteoinductive potential, suggesting its potential as a candidate material for bone regeneration.
Factors associated with orofacial myofunctional condition in patients with temporomandibular disorder and somatosensory tinnitus
Tinnitus is frequently reported in individuals with temporomandibular disorders (TMD), especially in the somatosensory subtype, which can be modulated by afferent input from orofacial and cervical structures. While the impact of orofacial dysfunctions on tinnitus has been explored, the potential reverse relationship remains understudied. Objective To investigate the association between somatosensory tinnitus and orofacial myofunctional condition in individuals with TMD. Methodology An observational, cross-sectional study was conducted with 47 adults aged 18 to 45 years diagnosed with TMD and self-reported tinnitus. Participants were assessed in a university-based outpatient setting. The primary outcome was the orofacial myofunctional condition, measured using the Orofacial Myofunctional Evaluation with Scores (OMES) protocol. Additional variables included tinnitus intensity and frequency (via acuphenometry), tinnitus-related distress (Tinnitus Handicap Inventory – THI), cervical disability (Neck Disability Index – NDI), craniofacial pain and disability (Craniofacial Pain and Disability Inventory – CF-PDI), and psychological symptoms (Hospital Anxiety and Depression Scale – HADS). Data were analyzed using Spearman correlation and multiple linear regression, and adopted statistical significance was set at p<0.05. Results OMES scores showed negative correlations with tinnitus intensity (r=−0.353), THI, NDI, CF-PDI, and age. In the final regression model, only tinnitus intensity remained a predictor of OMES score (β =−0.356; R2=0.127, Confidence Interval −0.20 to −0.02), indicating that greater tinnitus intensity was associated with poorer orofacial myofunctional condition. Conclusion These findings suggest that somatosensory tinnitus may negatively impact orofacial motor function in patients with TMD, underscoring the need for integrated and multidisciplinary assessment approaches.
Copyright (c) 2026 Gislaine Aparecida Folha, Nelma Ellen Zamberlan-Amorim, Carina Ferreira Pinheiro-Araujo, Luana Maria Ramos Mendes, Gesseany Joice Rodrigues dos Santos, Debora Bevilaqua-Grossi https://creativecommons.org/licenses/by/4.0
Genetic variants of EBI3, tumor Epstein-Barr virus, and human cytomegalovirus status in HPV-negative oral cancer
Oral squamous cell carcinoma (OSCC), a malignancy with high recurrence rate and poor survival, may be influenced by immune-related genetic variations and oncogenic viruses. Objective This study evaluated the clinical relevance of EBI3 polymorphisms, along with tumor Epstein–Barr virus (EBV) and Human Cytomegalovirus (HCMV) status, in prognostically adverse HPV-negative OSCCs. Methodology EBI3 (rs4740, rs4905, rs428253) genotyping was performed by qPCR in 95 HPV-negative OSCC patients and 108 age- and sex-matched controls. Tumor HPV, EBV, and HCMV status were assessed by qPCR. EBV viral load was calculated by exponential approach and a relative estimate of EBV copies per 105 cells. Associations with overall survival (OS) and recurrence-free survival (RFS) were evaluated by Kaplan-Meier and Cox regression analyses. Results EBV-positive tumors showed a significant association with increasing nodal stage (P=0.020). EBV viral load stratification (negative, low, high) presented a non-significant trend toward association with advanced tumor stage (P=0.060). Notably, EBI3 rs428253 predicted worse OS in EBV-positive patients, whereas rs4740 and rs4905 variants were associated with advanced tumor stage (P=0.024 and P=0.018). rs4740 and rs4905 variants were inversely associated with OSCC risk in dominant and overdominant models. Analysis detected HCMV in 7.4% of tumors but was not clinically relevant. Conclusions EBI3 genetic variants and EBV status may have prognostic relevance in HPV-negative OSCC. EBV may interact with the host genetics to influence nodal metastases and outcomes, suggesting a potential EBV-EBI3 axis, which warrants further investigation. Future precision oncology approaches may incorporate host and viral genetic markers to identify and stratify high-risk HPV-negative OSCC patients.
Copyright (c) 2026 Marko Simonovic, Ruzica Kozomara, Sasa Jovic, Gordana Velikic, Elizabeta Ristanovic, Nikoleta Djordjevski, Danijela Djuric-Petkovic, Srboljub Stosic, Gordana Supic https://creativecommons.org/licenses/by/4.0
Biofilm formation of mixed Candida albicans and methicillin-sensitive Staphylococcus aureus and surface properties of a 3D-printed denture base resin under different printing parameters
Objective To evaluate the formation of mixed-species biofilms of Candida albicans and methicillin-sensitive Staphylococcus aureus (MSSA) on the surface of a 3D-printed denture base resin, as well as its surface properties, under varying printing parameters. Methodology Discs (n=40 per group, 10×1.2 mm) of a denture base resin (priZma 3D Bio Denture) were fabricated using two 3D-printers—Liquid Crystal Display (LCD) and Digital Light Processing (DLP)—at three different angles (0°, 45°, or 90°). Surface roughness was measured using a digital profilometer and expressed as Ra (µm). For surface energy (SE) analysis, contact angles were measured using a tensiometer. Discs were incubated at 37 °C for 90 minutes and 48 hours to enable biofilm formation using C. albicans and MSSA inocula. Cell viability was assessed by colony-forming unit (CFU/mL) counts, and metabolic activity was evaluated using the XTT assay (absorbance). Microbial counts and XTT results were analyzed by three-way ANOVA (printer type, printing angle, incubation period). Surface roughness was analyzed by two-way ANOVA (printer type, printing angle), with Tukey’s test and a significance level of 0.05. Results For both CFU/mL and XTT assays, incubation period was the only significant factor (p<0.001 and p=0.006, respectively), while other factors and interactions were not statistically significant (p>0.05). Surface roughness was significantly influenced by printer type, printing angle, and their interaction (p=0.027). The LCD 0° and LCD 90° groups produced smoother surfaces compared with LCD 45° (p=0.002), which showed similar values to all DLP groups regardless of angle (p>0.05). The DLP printer did not show significant roughness variations across the tested angles (p>0.05). The LCD groups presented numerically lower SE values compared to the DLP groups. Conclusion The LCD system performs better than DLP in reducing surface roughness at 0° and 90°. Moreover, the analyzed factors did not significantly affect microbial adhesion or the formation of mixed-species biofilms.
Copyright (c) 2026 Larianne S. Moisés, Hamile Emanuella do Carmo Viotto, Sabrina Romão Gonçalves Coelho, Danny Omar Mendoza Marin, Raphael Freitas Souza, Ana Carolina Pero https://creativecommons.org/licenses/by/4.0
Antigenemia and oral lesions in paracoccidioidomycosis: is there a correlation?
Background Paracoccidioidomycosis is a systemic mycosis caused by the dimorphic fungus Paracoccidioides brasiliensis and related species, which are endemic to Latin America. Oral manifestations are highly relevant for diagnosis, representing the primary anatomical site biopsied for confirmation. Objective To investigate whether the virulence of P. brasiliensis influences the levels of circulating antigen in susceptible (B10.A) and resistant (A/Sn) murine models at multiple post-infection time points. The presence of oral lesions and their correlation with antigenemia were also assessed. Methodology One-week-old female B10.A and A/Sn mice were inoculated with the P. brasiliensis lineages Pb18 or Pb265, whereas controls received sterile saline. At four, eight, 12, and 16 post-inoculation weeks, sera were collected via sub-axillary plexus incision under anesthesia. Circulating antigen levels were quantified using competitive ELISA. Specific antibody titers were determined by indirect non-competitive ELISA and P. brasiliensis antigens. Intergroup comparisons were performed using two-way ANOVA, followed by multiple comparisons. Oral cavities were visually examined by three independent evaluators to find mulberry stomatitis. Prevalence was expressed as the percentage of affected animals. Results Significant differences in the circulating antigen levels were detected between susceptible mice infected with Pb18 and Pb265 and between susceptible and resistant mice infected with Pb18 16 weeks after inoculation. The highest levels of antibodies were found when both B10.A and A/Sn mice were infected with Pb18. During infection, no mice showed mulberry-like lesions in the oral cavity. Conclusion The virulence of the infecting P. brasiliensis strain, although not specifically linked to the gp43 antigen, plays a critical role in antigenemia. Despite absent oral lesions-an inherent limitation of this murine model-this study provides relevant insights into the relationship between fungal virulence and circulating antigen levels.
Copyright (c) 2026 Tatiana Teixeira de Miranda, Eva Burger https://creativecommons.org/licenses/by/4.0
Resin composites containing S-PRG fillers: effects on pH modulation of the surrounding medium, surface roughness, and gloss following erosive/abrasive challenge
Composite resins containing S-PRG fillers may degrade under pH fluctuations and abrasion, influenced by their composition, particle size, exposure time, and solution acidity. Objective: This study evaluated the ability of resin composites containing S-PRG fillers to modulate the pH of the surrounding medium, as well as their surface roughness and gloss following an erosive/abrasive challenge. Methodology: Resin disks (⌀4mm × 2mm) (n=15) were prepared using Filtek Z350 XT (control), Beautifil II, Beautifil II Enamel, and Beautifil II LS (the latter three containing S-PRG fillers). To assess buffering capacity, disks (n=5) were immersed in a demineralizing solution, and pH was measured at baseline and after 1, 3, 7, 14, and 21 days. Separate specimens (n=10) underwent daily erosion cycles using 0.3% citric acid, combined with abrasion via toothbrushing, conducted over a 5-day period to simulate an acidic diet and twice-daily oral hygiene. Surface roughness (Ra) and gloss (GU) were assessed at baseline, after polishing, and following the challenge; surface topography was analyzed by a scanning electron microscope (SEM). Data were analyzed using a generalized linear mixed model for repeated measures over time, in addition to Tukey–Kramer, Kruskal–Wallis, Dunn, Friedman, and Nemenyi tests (α=0.05). Results: All materials showed a significant increase in the pH of the surrounding medium over time (p<0.05). After 24 hours, Beautifil II exhibited a significantly higher pH than the others (p<0.05). Filtek Z350 XT demonstrated lower surface roughness than the other materials after the erosive/abrasive challenge (p<0.0001). Following the challenge, Beautifil II and Beautifil II Enamel presented higher gloss values (p<0.05). Conclusions: Resin composites containing S-PRG fillers slightly increased the pH of the surrounding medium over time. Surface roughness increased after the erosive/abrasive challenge, although the gloss of the S-PRG-containing composites also improved.
Copyright (c) 2025 Carlos Alberto Campos de Oliveira Neto, Mayara Zaghi Dal Picolo, Flávia Lucisano Botelho do Amaral, Carlos Rocha Gomes Torres, Kamila Rosamilia Kantovitz, Thomas Attin, Roberta Tarkany Basting https://creativecommons.org/licenses/by/4.0
Microstructures and mechanical characteristics of primary teeth affected by regional odontodysplasia
Regional odontodysplasia (ROD) is a rare, localized disorder of tooth hard tissues that can affect primary and permanent dentitions. However, its definitive etiology remains unclear and only a few studies have reported the detailed microstructures and mechanical characteristics of the affected teeth. Objective This study investigates the microstructural and mechanical characteristics of primary teeth affected by ROD. Methodology A total of two ROD-affected primary teeth from two different cases underwent clinical examinations. In addition to control samples of caries-free retained primary teeth, the affected samples were examined using micro-computed tomography (micro-CT), X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and nanoindentation analysis. Results Clinical findings revealed yellowish discoloration, rough surfaces, and root resorption on the affected teeth. Radiographs indicated hypocalcified enamel, widened pulp chambers, and delayed development of permanent teeth. Micro-CT showed affected teeth with thinner and uneven enamel, disordered dentin, and reduced mineral density. XRD analysis found reduced crystallinity. SEM and TEM analyses revealed hypoplastic and loosely packed enamel crystals, whereas dentin exhibited disorganized collagen fibrils and poorly mineralized crystals. EDS analysis showed a reduced calcium/phosphorus ratio and an increased magnesium/calcium ratio in the affected enamel. Nanoindentation tests found reduced hardness and elastic modulus in ROD-affected enamel compared with control teeth. Conclusion ROD-affected primary teeth display significant microstructural abnormalities and compromised mechanical properties, underscoring the need for early intervention and long-term monitoring to prevent complications.
Copyright (c) 2026 Wenxian Jiang, Minjian Shen, Jian Wang, Mengyao Bian, Haiyan Zheng, Zihuai Zhou, Ying Shi, Zhifang Wu https://creativecommons.org/licenses/by/4.0
Selective HDAC4 inhibition by SP1-PTD promotes odontoblast differentiation
Background Vital pulp therapy is limited by incomplete dentin regeneration and dose-limiting toxicities of current histone deacetylase (HDAC) inhibitors. Previous structural studies have identified critical determinants of HDAC4-silencing mediator for retinoid and thyroid hormone receptor (SMRT) protein interactions, providing a rationale for developing selective inhibition strategies. Objective This study evaluated SMRT peptide 1-protein transduction domain (SP1-PTD), which is a cell-penetrating peptide designed to selectively disrupt HDAC4–SMRT interaction based on structural insights, for promoting odontoblast differentiation with improved safety compared to pan-HDAC inhibitors. Methodology SP1-PTD comprises an SMRT-derived sequence fused to a PTD, enabling targeted inhibition without affecting HDAC catalytic activity. Effects on odontoblast differentiation were assessed in murine dental papilla cell lines and primary human dental pulp cells using gene expression analysis, functional mineralization assays, and mechanistic studies including chromatin immunoprecipitation and RUNX2 acetylation analysis. Cytotoxicity was directly compared with suberoylanilide hydroxamic acid (SAHA) and trichostatin A. Results SP1-PTD treatment significantly enhanced odontoblast differentiation with 15.9-fold increase in dentin sialophosphoprotein (Dspp) expression alongside upregulation of RUNX2, osteocalcin, and bone sialoprotein. Functional analysis revealed 1.8-fold increased mineralization capacity. Mechanistically, SP1-PTD increased RUNX2 protein acetylation and histone acetylation at the Dspp promoter, indicating derepression of RUNX2-mediated transcription. Importantly, SP1-PTD did not show cytotoxicity across a wide therapeutic range (0.1-20 μM) and promoted cell proliferation, contrasting sharply with dose-dependent toxicity of pan-HDAC inhibitors. Direct comparison revealed SP1-PTD induced 14-fold increase in Dspp expression while SAHA suppressed it despite comparable Runx2 induction. Conclusions SP1-PTD represents a first-in-class selective HDAC4 inhibitor that achieves robust pro-differentiation effects with an exceptional safety profile. By specifically targeting HDAC4–SMRT interactions, SP1-PTD overcomes limitations of conventional HDAC inhibitors and offers translational promise for dental regenerative medicine.
Copyright (c) 2025 Yeo-Kyeong Shin, Jung-Sun Moon, Su-Kyeong Son, Bin-Na Lee, Chungoo Park, Sun-Hun Kim, Young Chu Lee, Min-Seok Kim https://creativecommons.org/licenses/by/4.0
Modification of restorative glass ionomer cement with zinc oxide nanoparticles and calcium glycerophosphate microparticles: in vitro assessment of mechanical properties and antimicrobial activity
The incorporation of bioactive agents into resin-modified glass ionomer cement (RMGIC) is a promising strategy to improve its mechanical strength and biofilm control, especially for patients with active dental caries. Objective This study aimed to evaluate the effects of incorporating ZnONPs and CaGP into RMGIC on its mechanical and microbiological properties. Design Six groups were tested: 1) RMGIC (without CaGP/ZnONPs); 2) RMGIC-1.0%ZnONPs; 3) RMGIC-2.0%ZnONPs; 4) RMGIC-3.0%CaGP; 5) RMGIC-3.0%CaGP-1.0%ZnONPs; and 6) RMGIC-3.0%CaGP-2.0%ZnONPs. The compressive strength (CS), diametral tensile strength (DTS), and surface hardness (SH) were evaluated after 24 hours and 7 days. Antimicrobial and antibiofilm activity were evaluated using agar diffusion and biofilm metabolic activity (XTT) assays. Results After 24 hours, all the groups showed similar DTS values (p>0.05), except for RMGIC-3.0%CaGP-1.0%ZnONPs, which showed the highest DTS value (p<0.05). Comparing 24 hours and 7 days, the DTS values of RMGIC-3.0%CaGP-2.0%ZnONPs, RMGIC-3.0%CaGP, and RMGIC-3.0%CaGP-2.0%ZnONPs were similar (p=0.360). After 24 hours, the RMGIC group showed the CS highest value, followed by RMGIC-2.0%ZnONPs (p < 0.05). After 7 days, the RMGIC-3.0%CaGP-1.0%ZnONPs group exhibited the highest CS value, approximately 15% higher than RMGIC (p<0.05). The RMGIC-1.0%ZnONPs group exhibited significantly higher SH at 24 hours (p=0.621). At 7 days, the highest SH value was observed for the RMGIC-3.0%CaGP-1.0%ZnONPs group (p<0.05). Regarding antimicrobial and antibiofilm activity, including results from biofilm metabolism assays, the RMGIC-3.0%CaGP-1.0%ZnONPs group demonstrated the most effective antimicrobial and inhibitory effects (p<0.05). Conclusion This study demonstrated that adding ZnONPs and CaGP to RMGIC enhanced its mechanical and antimicrobial and antibiofilm properties, suggesting enhanced mechanical performance and improved protection against cariogenic biofilms—critical factors for successful restorative treatments. Therefore, the addition of ZnONPs and CaGP is a promising strategy to develop advanced restorative materials that improve clinical outcomes, especially for patients with active dental caries.
Copyright (c) 2025 Maria Fernanda Cavalcante Meira, Gabriela Leal Peres Fernandes, Andréa Simone Stucchi de Camargo, Leandro Piaggi Ravaro, Marylyn Setsuko Arai, Maria Fidela de Lima Navarro, Fernanda Lourenção Brighenti, Analú Barros de Oliveira, Marcelle Danelon https://creativecommons.org/licenses/by/4.0
Evaluation of the levels of Fractalkine (CX3CL1), TNF-α, and TGF-β in the gingival crevicular fluid/tissue of patients with gingival overgrowth: a cross-sectional observational study
Fractalkine (CX3CL1) is expressed by various cells, contributing to the pathogenesis of diseases such as diabetes mellitus, vascular pathologies, and rheumatoid arthritis via immunological mechanisms. The CX3CL1–CX3CR1 axis regulates cellular responses such as proliferation and collagen production, which are implicated in gingival overgrowth (GO). Objectives This study aimed to assess the levels of CX3CL1, tumor necrosis factor-α (TNF-α), and transforming growth factor-beta (TGF-β) in both gingival crevicular fluid (GCF) and gingival tissues among patients with biofilm- and amlodipine-induced GO. Additionally, the potential relationship between these biomarkers and clinical periodontal parameters was evaluated. Methods The study included 17 participants with biofilm-induced GO (Group I), 18 participants with amlodipine-induced GO (Group A), and 10 systemically healthy participants without GO (Control). CX3CL1, TNF-α, and TGF-β levels in GCF samples were assessed using enzyme-linked immunosorbent assay (ELISA). Moreover, mRNA expression levels of CX3CL1, TNF-α, and TGF-β in tissue samples were determined by quantitative real-time PCR (qPCR). Results The total GCF CX3CL1 level was significantly higher in Group I and Group A compared to controls. However, tissue CX3CL1 and TNF-α levels were significantly higher in Group I than Group A (p<0.05). In Group A, total GCF CX3CL1 levels showed a positive correlation with the gingival index (GI) (r=0.644), bleeding on probing (BOP) (r=0.622), and GCF volume (r=0.720). A significant positive correlation was observed between tissue CX3CL1 and TNF-α levels (r=0.762) (p<0.05). In Group I, a significant correlation was observed between total GCF CX3CL1 and TNF-α, TGF-β, and GCF volume levels, respectively (r=0.865, r=0.845, r=0.651). A positive correlation (p<0.05) was also found between tissue CX3CL1 and TNF-α and TGF-β levels, respectively (r=0.689, r=0.903). Conclusion CX3CL1 may have a potential role in the development of GO-associated tissue fibrosis and its inflammatory mechanisms.
Copyright (c) 2025 Nazife Hamurcu, Ahu Uraz Çörekci, Rahşan Ilikçi Sağkan, Serpil Cula, Deniz Özbay Çetiner https://creativecommons.org/licenses/by/4.0
Clinical performance of posterior resin composite restorations bonded with universal adhesive system using three different application modes: a 26-month randomized clinical trial
Objective This study aimed to evaluate the clinical performance of posterior resin composite restorations (Class I and II), bonded with a universal adhesive using three application modes over 26 months, comparing the adapted FDI and USPHS criteria. Methodology In total, 138 Class I and II cavities were restored using Scotchbond Universal adhesive (3M ESPE) in three application modes: etch-and-rinse (ER); selective enamel etch (SEE), and self-etch (SE). All cavities were restored with FiltekTM Supreme composite resin (3M ESPE). The restorations were evaluated by two trained and calibrated evaluators using FDI and adapted USPHS criteria at baseline (12.02±5.68 days) and after 26 (± 2.36) months. The obtained data were subjected to statistical analysis between the groups using the Friedman and Wilcoxon test for intra-group analysis and the McNemar test for comparisons between FDI and USPHS criteria (α=0.05). Results No statistically significant differences were observed between the ER, SEE, and SE groups after 26 months, except for marginal discoloration between the ER and SE groups (p≤0.05). Conclusion Class I and II resin composite restorations performed with Scotchbond Universal adhesive using ER, SEE, and SE modes showed acceptable clinical performance over a 26-month evaluation period. Moreover, the adapted FDI and adapted USPHS criteria yielded comparable results. Brazilian Clinical Trials Registry (Rebec) — RBR-9p3hdp
Copyright (c) 2025 Thaís Silva Mendonça, Andreia Assis Carvalho, Jéssica Karla Maia Zago, Terezinha de Jesus Esteves Barata, Érica Miranda Torres, Gersinei Carlos de Freitas, Lawrence Gonzaga Lopes https://creativecommons.org/licenses/by/4.0
Oral microbiome patterns of dental caries in Kazakhstani adolescents
Objective The oral microbiome is one of the most complex microbial ecosystems in the host. This study aimed to investigate and characterize the oral microbiome composition in Kazakhstani adolescents associated with dental caries. Methodology The study included 312 adolescents, with 241 individuals presenting with caries and 71 caries-free, aged 12-15 years. Dental caries assessment was performed using DMFT (Decayed, missed, filled teeth) index. Oral samples were collected, and 16S rRNA (16S ribosomal ribonucleic acid) gene sequencing targeting the V3-V4 hypervariable regions on an Illumina MiSeq platform was performed to profile the microbial communities. Functional metagenomic predictions were generated using PICRUSt2 v2.5.0, using the KEGG database for bacterial pathway abundance estimation. Data analysis was conducted using Python 3.9.16 and R 4.2.2. Results The alpha diversity was insignificant, while beta diversity analysis demonstrated clear distinctions by Bray-Curtis (F=2.5, p=0.003) and weighted UniFrac distances (F=4.4, p=0.002). The Neisseria and Prevotella genera, and Gammaproteobacteria class showed significant associations with dental caries (MaAsLin2 p≤0.05, LDA≥2), stronger predictive power (AUC=0.65, F1=0.83), and higher predicted functional activity through glutathione metabolism, RNA degradation, and unsaturated fatty acid metabolism pathways. Conclusions This study identified specific oral microbiome patterns associated with dental caries in Kazakhstani adolescents, revealing interactions between key bacterial taxa and metabolic pathways.
Copyright (c) 2025 Argul Issilbayeva, Zharkyn Jarmukhanov, Samat Kozhakhmetov, Yermekbayeva Bakytgul, Laura Chulenbayeva, Graciela Muniz-Terrera, Masae Furukawa, Hiroki Nikawa, Adil Supiyev, Almagul kushugulova, Anara Zhumadilova https://creativecommons.org/licenses/by/4.0
Development of liver disease caused by chronic periodontitis in rats
Periodontitis is an inflammatory disease characterized by producing alterations in periodontal tissues, but it has been associated with the manifestation of systemic alterations, related to those produced in liver tissue. This study aims to evaluate the progression of liver damage caused by ligation-induced periodontitis in rats with time intervals of ten to 80 days. Methodology Overall, 40 rats were divided into five groups: control (no ligatures), P10, P20, P40, and P80 (teeth with ligatures at intervals of ten, 20, 40, and 80 days) in which we verified liver disease caused by periodontitis. Oral parameters were evaluated: gingival bleeding index (GBI), probing pocket depth (PPD), dental mobility (TM), myeloperoxidase activity (MPO), alveolar bone height (ABH). Liver parameters were evaluated: liver weight, histopathological scores for steatosis, inflammation, and necrosis in the liver; glutathione (GSH) and malondialdehyde (MDA). Serum parameters were also evaluated: concentrations in liver tissues, blood levels of albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), glucose, total cholesterol, and total proteins. Results The results showed that the hepatic steatosis score gradually increased (P<0.05) in rats with induced periodontitis for up to 20 days (P20); and the rats with ligatures for 40 (P40) and 80 days (P80) had stable scores compared to the P20, without any further worsening, similarly occurred with GSH, MDA, and total cholesterol. Conclusions This study indicated that liver alterations caused by ligature-induced periodontitis are progressive in early stages (0-20 days) and reach a plateau in later stages (40-80 days).
Any Carolina Cardoso Guimarães Vasconcelos
Copyright (c) 2025 Karen Neisman Rodriguez Ayala, Vinícius da Silva Caetano, Any Carolina Cardoso Guimarães Vasconcelos, Maria Vitoria Pereira de Sousa, Nikaely Brandão Barbosa, Luanna Maria Soares Mesquita, Paulo Roberto Carneiro Gomes, André Luis dos Reis Barbosa, Hélio Mateus Silva Nascimento , Daniel Fernando Pereira Vasconcelos https://creativecommons.org/licenses/by/4.0