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.
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.
Copyright (c) 2026 https://creativecommons.org/licenses/by/4.0
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
Monocytes and Macrophage-derived mediators influence the behavior of squamous cell carcinoma cell lines
Immune cells play diverse roles in cancer development. Myeloid cells are key drivers of tumor-escape mechanisms as they suppress immune responses, facilitate metastasis, and contribute to therapy resistance. In particular, macrophages can be polarized into an inflammatory M1 (anti-tumor) or anti-inflammatory M2 (pro-tumor) phenotype. M2 macrophages are associated with tumor progression, as they secrete factors that promote tumor angiogenesis, suppress T-cell activity, and correlate with poor clinical outcomes in squamous cell carcinoma (SCC). Given this context, this study aims to demonstrate the biological effects of monocytes and both M1 and M2 macrophages in squamous cell carcinoma. Our data indicate higher CD163 immunoreactivity in biopsies from SCC patients. Furthermore, we found that a conditioned medium (CM) containing bioactive compound generated by M2 macrophages enhances the proliferation and invasion of the SCC-25 cell line in vitro. Surprisingly, CM derived from blood CD14+ monocytes increased SCC-25 proliferation at the same rate of M2 macrophages-CM. M1 macrophages conditioned medium significantly enhanced the motility and decreased proliferation in Detroit 562 cells. The analysis of tumor-associated transcripts showed that both M1 and M2 conditioned medium induced high levels of EPCAM mRNA and significantly decreased the expression of MYC, an epithelial-to-mesenchymal transition marker, in SCC cell lines. Detroit cells exposed to conditioned medium from monocytes and macrophage also showed elevated SOX2 mRNA levels. The findings suggest that monocytes and macrophage mediators exert distinct biological effects on SCC cell lines.
Copyright (c) 2025 Graziela Perri, Raíssa Gabrieli Candido, Luiz Henrique Camargo Soares, Rafael Carneiro Ortiz, Izabel de Camargo, Maria Renata Sales Nogueira, Edgard José Franco Merllo Júnior, Ana Lucia Coelho, Edwin M Posadas, Cory Hogaboam, Karen A Cavassani, Ana Paula Campanelli https://creativecommons.org/licenses/by/4.0
Three-dimensional soft tissue comparison in non-growing patients treated for skeletal Class II Division I malocclusion: mandibular advancement surgery vs. Herbst Appliance
Objective This study used three-dimensional (3D) stereophotogrammetry to compare changes in facial soft tissues of patients with skeletal class II malocclusion with mandibular retrognathia treated with orthognathic surgery (OS) or Herbst appliance (HA). Methodology This retrospective study included 15 adults treated with bilateral sagittal split osteotomy without genioplasty (OS) and 16 adults treated with HA. The patients were treated successfully, with class I occlusal relationships with normal overjet and overbite. Three-dimensional stereophotogrammetric records acquired in habitual occlusion from before and after fixed orthodontic treatment were analyzed. Lateral cephalometric radiographs were used to evaluate skeletal parameters and sagittal oropharyngeal airway length. Results In HA-group, mandibular corpus length, anterior facial height, posterior facial height, Bº, vertical angle, and mentolabial angle increased; lower facial width, mandibular angle, mandibular convexity angle, ANBº, and Aº decreased. In OS-group, mandibular length, mandibular corpus length, and facial convexity angle increased significantly compared with those in HA-group; posterior facial height, Bº, vertical angle, and mentolabial angle also increased, but mandibular angle and ANBº decreased significantly. Only a significant difference in coordinate changes was observed for the pogonion in the sagittal direction. According to the cephalometric analysis, SNBº and Pg-NA perpendicular measurements increased significantly in the OS group compared with the HA group. In both groups, sagittal oropharyngeal airway length increased significantly post-treatment, with no significant intergroup difference. Conclusion In patients treated with OS, skeletal advancement resulted in greater increases in mandibular and corpus lengths, along with forward positioning of the pogonion. Based on 3D soft tissue and cephalometric comparisons, the Herbst appliance seems to be a viable non-surgical alternative for young adults with moderate Class II malocclusion. Both treatment modalities also contributed to significant improvements in sagittal oropharyngeal airway length.
Copyright (c) 2025 Gökhan Çoban, Taner Öztürk, Gökhan Türker, Ebru Topcuoğlu Hashimli, Yusuf Nuri Kaba https://creativecommons.org/licenses/by/4.0
Vagus nerve stimulation alleviates alveolar bone loss and inflammation in ligature-induced periodontitis rat model
Objective Vagus nerve stimulation (VNS) can inhibit inflammation in various diseases by activating the cholinergic anti-inflammatory pathway. However, whether VNS could attenuate periodontitis by activating α7-nicotinic acetylcholine receptor (α7-nAChR) remains unknown. Methodology Ligature induction was utilized to establish the periodontitis rat model. Periodontal indices like bleeding, tooth mobility, and probing depth, were measured. Bone mineral density, trabecular thickness, and length of the cement-enamel junction to the alveolar bone crest were analyzed using micro-CT. Immunohistochemistry assessed bone morphology and inflammatory levels. Inflammatory cytokines were detected using enzyme-linked immunosorbent assay and quantitative reverse transcription polymerase chain reaction. For the in vitro inflammation model, RAW264.7 cells were stimulated with lipopolysaccharides and acetylcholine to study inflammatory responses. Results VNS significantly improved periodontal health and reduced alveolar bone loss in periodontitis rats. VNS alleviated inflammation by suppressing pro-inflammatory cytokines, including IL-1β, IL-6, and IL-18, enhanced bone formation, and activated the cholinergic anti-inflammatory pathway, as evidenced by increased α7-nAChR expression. Additionally, acetylcholine activation of α7-nAChR in RAW264.7 cells inhibited pro-inflammatory responses and promoted anti-inflammatory responses. Conclusion These findings suggest that VNS can effectively reduce inflammation and improve periodontal outcomes in periodontitis.
Copyright (c) 2025 Honghui Xie, Ping Huang, Aihong Tan, Zhijian Su, Ying Fu https://creativecommons.org/licenses/by/4.0
Effect of wavelength and radiant emittance on light transmittance and temperature increase in dental resin-based composites
Objectives: To evaluate the effect of wavelength and radiant emittance on light transmittance and heat generation through dental resin-based composites. Methodology: Light-emitting diodes on the blue, green, and red wavelength spectra were assembled and characterized using a spectrometer (MARC Resin Calibrator, BlueLight Analytics). Voltage (V) and amperage (A) from each LED was set up to emit a 500-, 1000-, 2000-, 3000-, or 4000-mW/cm2 radiant emittance. A self-cured resin-based composite model was fabricated in two pastes, one with benzoyl peroxide and another with ethyl 4-dimethylaminobenzoate. The two pastes were mixed and placed into a mold (ø=10 mm, 2 mm thick) until completely hardened. A power analysis was conducted to determine the sample size to provide a power of at least 0.8 and α=0.05. Light transmittance through the 2-mm thick composite-based sample was evaluated using a spectrometer (n=10). Heat generation (oC) in the resin-based composite sample induced by the exposure to the tested wavelength spectra and radiant emittances were recorded using an infrared camera (FLIR ONE PRO, FLIR Systems) (n=10). Statistical analyses were performed using analysis of variance and the Tukey's test for multiple comparisons. Results: Light transmission systematically increased based on radiant and wavelength emittances (p<0.0001). Heat generation was directly proportional to radiant emittance but indirectly proportional to wavelength emittance (p<.0.0001). Conclusions: Despite its limitations, this study found that increasing wavelength emittance seems to configure a great alternative to increase light transmittance through resin-based composite restorations while reducing heat generation.
Copyright (c) 2025 Sharanya Singh, Mateus Garcia Rocha, Shannon M Roulet, Jean-François Roulet, Mario Alexandre Coelho Sinhoreti, Dayane Oliveira https://creativecommons.org/licenses/by/4.0
Single local injection of 3S-HMGB1 enhances early bone healing and titanium implant osseointegration in type 2 diabetic mice
Diabetes significantly impairs bone healing via several mechanisms, including sustained inflammation, oxidative stress, and poor osteogenic responses. Fully reduced isoform of High Mobility Group Box 1 with three serine substitutions (3S-HMGB1) has shown regenerative properties while being resistant to oxidative degradation. Objective: This study evaluated the therapeutic potential of a single local injection of 3S-HMGB1 in early osseointegration under diabetic conditions in mice. Methodology: A total of 48 male 129/Sv mice (24 non-diabetic [ND], 24 diabetic [D]) received titanium implants following maxillary first molar extraction. ND and D mice (n:12) were injected with either saline (control) or 3S-HMGB1 (0.75 mg/kg) into the fresh extraction socket. Osseointegration was evaluated at 7 and 21 days post-implantation using microCT, histology (bone-to-implant contact [BIC] and birefringence), and immunohistochemistry for Runx2 and CXCR4. Results: ND controls exhibited early osteogenic activity, with a predominance of Runx2-positive cells at 7 days and successful osseointegration by 21 days. In contrast, D controls showed reduced numbers of Runx2-positive cells and markedly lower BV/TV, indicating compromised bone healing at 21 days. Treatment with 3S-HMGB1 resulted in significantly increased bone volume at the implant site in D animals (55.6±7.20% vs. 44.6±6.23%) and restored BIC from 44.9±9.32% (D controls) to 61.78±11.31% (D 3S-HMGB1), near ND levels (65.16±7.64%). Both ND and D groups treated with 3S-HMGB1 presented enhanced collagen organization. No significant differences were found in CXCR4 between groups; however, in D animals, a distinct peri-implant staining pattern suggested impaired recruitment despite preserved stem cell niches in the bone marrow. Conclusions: Collectively, our findings indicate that a single injection of redox-stable 3S-HMGB1 may represent a promising regenerative strategy to mitigate early implant failure in diabetes. Future studies should explore sustained delivery approaches to enhance long-term outcomes.
Copyright (c) 2025 Bhuvana Lakkasetter Chandrashekar, Alexandra Arteaga, Evelin Rios, Jimena Mora, Gustavo Pompermaier Garlet, Danieli C. Rodrigues, Claudia Cristina Biguetti https://creativecommons.org/licenses/by/4.0
Calcium carbonate in dentistry: a bibliometric review of emerging applications and trends
Calcium carbonate (CaCO₃) has garnered increasing attention in dental research due to its versatile bioactive properties and expanding applications in preventive, restorative, and regenerative therapies. Objective This study aims to comprehensively assess the bibliometric features of articles evaluating the utilization of calcium carbonate (CaCO₃) in dentistry by conducting a bibliographic search on the Web of Science databases until March 2025. Methodology The following data were collected: number and density of citations; authorship; year, journal of publication, and impact factor; study design and theme; keywords; institution and country of origin. VOSviewer software was used to generate collaborative network maps for authors and keywords. Results A total of 91 highly cited articles were identified, with citation counts ranging from 123 to zero. Most articles (74%) were published after 2010, with the highest prevalence in Asia (44%), especially China (10%). The most frequent study design was in vitro (55%), primarily focused on restorative dentistry (29%) and cariology (23%). The most common keywords were “Calcium Carbonate” and “Hydroxyapatite”. DeVizio W. was the most prolific author, with four publications. Conclusions Bibliometric analysis highlights a growing interest in the application of calcium carbonate in dentistry, with a progressive increase in scientific output over the years. The findings underscore the global distribution of research and emphasize the relevance of this biomaterial in various dental specialties. This study reinforces several key points for research groups worldwide engaged in the development of innovative dental materials, providing valuable direction for future investigations, which remain limited in scope, especially regarding clinical applications and long-term outcomes.
Copyright (c) 2025 Andressa da Silva Barboza, Samira Schons de Oliveira, Adriana Poli Castilho Dugaich, Maurício Malheiros Badaró, Ana Paula Varela Brown Martins, Rafael Guerra Lund, Sheila Cristina Stolf, Juliana Silva Ribeiro de Andrade https://creativecommons.org/licenses/by/4.0
A randomized prospective clinical study evaluating the effectiveness of the Beautibond Xtreme adhesive system using different bonding techniques in class I and II restorations: one-year results
Objective This randomized, parallel-group clinical trial aimed to evaluate the one-year clinical performance of the Beautibond Xtreme adhesive system (Shofu Inc., Kyoto, Japan) applied with different bonding strategies in Class I and Class II posterior restorations. Methodology A total of 22 patients (14 female and 8 male, aged ≥18 years) requiring restorative treatment provided 152 teeth with Class I or Class II carious lesions or defective restorations. Restorations were randomly assigned to six groups, including two control groups (Class I and II using the total-etch technique) and four test groups (Class I and II using either the self-etch technique or selective enamel etching). All restorations were performed using Beautibond Xtreme adhesive combined with Beautifill LS composite resin. Clinical performance was assessed at baseline, six months, and one year using modified United States Public Health Service (USPHS) criteria, including anatomic form, marginal adaptation, marginal discoloration, color match, surface texture, secondary caries, postoperative sensitivity, and retention. Randomization was performed with a computer-generated sequence, and two calibrated, blinded examiners (Kappa = 0.84) conducted all evaluations. Results No significant changes were observed in anatomic form, color match, surface texture, secondary caries, postoperative sensitivity, or retention over time in any group (p>0.05). However, restorations performed using the self-etch technique showed significant deterioration in marginal adaptation and marginal discoloration from baseline to six months and one year (p<0.05). Significant differences were also observed when comparing these restorations to the total-etch and selective enamel etching groups (p<0.05). Conclusion The Beautibond Xtreme adhesive system demonstrated better clinical performance when applied with total-etch or selective enamel etching techniques compared to the self-etch mode for both Class I and Class II restorations after one year of follow-up.
Copyright (c) 2025 Walter Raucci Neto, Ana Flávia Simões Barbosa, Adrielle Fracaroli Baltazar, Larissa Fernanda Pereira, Yara Teresinha Corrêa Silva Sousa, Danielle Cristine Furtado Messias, Taylane Soffener Berlanga de Araujo https://creativecommons.org/licenses/by/4.0
PPARα modulation of macrophage polarization and inflammatory signaling in mimic periodontitis
Objective This study investigates the role of peroxisome proliferator-activated receptor alpha (PPARα) in regulating macrophage polarization and inflammatory signaling under stimulation by periodontal pathogens. Methodology THP-1-derived macrophages were stimulated with Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) in the presence or absence of PPARα agonists fenofibrate and WY14643, or the antagonist GW6471. Protein expression levels of TNF-α, IL-10, and phosphorylated NF-κB were assessed by Western blot. Immunofluorescence staining was used to evaluate IL-10, NF-κB, and CD36 expression. Flow cytometry quantified changes in macrophage polarization markers, including CD14+CD86+ (M1) and CD68+CD206+/CD163+ (M2) populations. THP-1 cells transfected with a secreted embryonic alkaline phosphatase (SEAP) reporter plasmid were treated with Pg-LPS (1 μg/mL) ± fenofibrate (50 μM) to assess NF-κB/AP-1 activity. PPARα reporter cells were treated with increasing concentrations of GW590735 or WY14643 and exposed to TNF-α, LPS, or GW6471+LPS to evaluate PPARα transcriptional activity. Results PPARα activation by fenofibrate reduced TNF-α expression in Pg-LPS-stimulated macrophages and attenuated NF-κB signaling via both TLR2 and TLR4 pathways. Fenofibrate significantly increased IL-10 and CD36 expression, inhibited Pg-LPS-induced NF-κB nuclear translocation, and promoted a phenotypic shift from pro-inflammatory M1 to anti-inflammatory M2 macrophages. Moreover, inflammatory stimuli such as TNF-α and LPS suppressed PPARα activity, which could be restored by potent PPARα agonists. Conclusion These findings suggest that PPARα activation modulates macrophage polarization and suppresses inflammatory signaling in response to periodontal bacterial antigens.
Copyright (c) 2025 Arthur Hu, Yvette Y Chen https://creativecommons.org/licenses/by/4.0
Association between single nucleotides polymorphisms in the DR2D, ANKK1, COMT, and 5-HTT genes with temporomandibular disorder
Background Temporomandibular disorders (TMDs) comprise a heterogeneous group of musculoskeletal conditions affecting the temporomandibular joint, masticatory muscles, and associated structures. Growing evidence highlights the role of genetic predisposition as a significant contributor to TMD. Aim This study aimed to investigate genetic aspects involved in TMDs etiology. Methodology A cross-sectional study was conducted with 249 adolescents, of whom 149 were affected by TMD. Genomic DNA was extracted from buccal cells, and single nucleotide polymorphisms (SNPs) in DRD2 (rs6275 and rs6276), ANKK1 (rs1800497), COMT (rs6269 and rs4818), and 5-HTT genes (rs3813034 and 1042173) were analyzed. Allelic and genotypic distribution, haplotype, and diplotype analysis were performed using PLINK software version 1.06. Multifactor dimensionality reduction (MDR) was applied to identify SNP-SNP interactions and generate an interaction graph. Results In total, three possible single-locus allele combinations were obtained for haplotype and diplotype analyses (rs6275|rs6276 in DRD2, rs6269|rs4818 in COMT and rs3813034|rs1042173 in 5-HTT), but no associations with TMD were observed (p>0.05). However, MDR analysis for gene-gene interactions revealed a synergistic relationship between rs6275 (DRD2), rs6269 (COMT), and rs1042173 (5-HTT) that predisposes to TMD (p=0.050). Conclusion MDR analysis suggests a possible synergistic interaction among SNPs in the DRD2, COMT, and 5-HTT genes that may contribute to TMD susceptibility in adolescents.
Copyright (c) 2025 Otavio Augusto Pozza, Fernanda Mara de Paiva Bertoli, Franciele Topolski, Paula Porto Spada, Marilisa Carneiro Leão Gabardo, Juliana Feltrin de Souza, Erika Calvano Küchler, João Armando Brancher https://creativecommons.org/licenses/by/4.0
Panax notoginseng saponins alleviate alveolar bone loss and reprogram macrophages in diabetic periodontitis rats
Diabetic periodontitis (DP) features accelerated alveolar bone destruction driven by impaired immune regulation and compromised bone metabolism. Objective This study investigated the therapeutic potential of Panax notoginseng saponins (PNS) against alveolar bone loss in a rat DP model and elucidated its mechanisms of action. Methodology Male Sprague-Dawley rats were allocated to four groups: control, periodontitis control (CP), diabetic periodontitis (DP), and DP + PNS (80 mg/kg/day). Diabetes was induced by streptozotocin injection, followed by ligature-induced periodontitis at the maxillary first molar. After 4 weeks of PNS treatment, alveolar bone samples were analyzed by micro-CT, histomorphometry, immunohistochemistry, and immunofluorescence. Results Micro-CT and H&E analyses revealed severe alveolar bone resorption in DP rats, whereas PNS treatment substantially mitigated these destructive changes. TRAP staining demonstrated that PNS significantly suppressed osteoclast formation and activity. Immunohistochemistry detected upregulated expression of OCN in PNS-treated groups, indicating enhanced osteogenic differentiation. Immunofluorescence analysis showed that PNS promoted a phenotypic shift in macrophages, reducing pro-inflammatory M1 polarization (iNOS+) while increasing anti-inflammatory M2 populations (Arg-1+). This shift correlated with decreased interleukin-6 (IL-6) and elevated interleukin-10 (IL-10) levels. Conclusions PNS attenuates alveolar bone loss in diabetic periodontitis by inhibiting osteoclastogenesis, stimulating osteogenic activity, and modulating macrophage polarization toward an anti-inflammatory M2 phenotype. These actions collectively reduce inflammation and promote tissue regeneration, highlighting PNS as a promising candidate for managing DP-related bone destruction.
Copyright (c) 2025 Xue-wei Jiang, Jun Zhu, Wen Cheng, De-sheng Fan, Lei Zhen https://creativecommons.org/licenses/by/4.0
Comprehensive analysis of transcriptome and pathway interactions in periodontitis
Periodontitis involves complex immune, epithelial, and metabolic disruptions. High-throughput transcriptomic approaches can reveal key regulatory networks underpinning disease progression. Objective To investigate the transcriptomic profile and dysregulated molecular pathways associated with severe periodontitis. Methodology Gingival tissues from patients with severe periodontitis (n=11) and periodontally healthy controls (n=11) were compared using RNA sequencing in this cross-sectional study. Differentially expressed genes (DEGs) were identified and analyzed using Ingenuity Pathway Analysis (IPA). Selected DEGs were validated at the protein level via immunohistochemistry (IHC). Results A total of 909 DEGs were upregulated and 742 were downregulated in periodontitis versus healthy tissues. Highly upregulated genes included MT-RNR1, MTRNR2L12, pseudogenes, long non-coding RNAs, and immunoglobulins. Downregulated genes included ADGRG7, C6orf15, members of enzyme families, keratin family members, loricrin (LOR), and immune modulators, such as CD207 (Langerin) and DEFB4A. IPA predicted the Wnt/β-catenin pathway as the most upregulated and the IL-17 signaling pathway as the most suppressed canonical pathway in periodontitis. The expression of the level of protein of LOR, Wnt10b, JUN, and FOS was confirmed by IHC. Conclusion Dysregulation in key canonical signaling pathways and the altered expression of genes critical to cell chemotaxis, innate immunity, and epithelial barrier integrity seem to play a pivotal role in the pathogenesis of periodontitis.
Copyright (c) 2025 Bruno César de Vasconcelos Gurgel, Nathalia Vilela, Kaio Henrique Soares, Ikramuddin Aukhil, Poliana Mendes Duarte, Luiz Eduardo Rodrigues Juliasse https://creativecommons.org/licenses/by/4.0
The impact of surface pre-reacted glass-ionomer nanoparticles on ion dynamics and the physio-mechanical properties of denture base resin
Dentures containing nanoparticles must maintain appropriate ion dynamics and physio-mechanical properties. Objective this study evaluated the effects of incorporating 5wt% and 10wt% surface pre-reacted glass-ionomer S-PRG nanoparticles into a polymethylmethacrylate (PMMA) resin on multiple-ion exchange rates following recharge with an S-PRG solution, flexural strength, surface roughness, and hardness. Methodology In total, 72 disc-shaped and 32 rectangular specimens were fabricated from PMMA resin containing 0, 5, and 10% S-PRG nanoparticles, and 20% S-PRG microparticles by weight. In total, six-disc specimens per group were tested for surface roughness and hardness using a contact profilometer and the Vickers hardness test, respectively. The remaining discs were immersed in 11 ml of deionized water for 24 hours, followed by an analysis of their aluminum, boron, sodium, silicate, strontium, and fluoride concentrations. They were then recharged with a 10% S-PRG solution for 24 hours, alternating with deionized water for five cycles, before their ion levels were reanalyzed. Rectangular specimens underwent flexural strength testing via the three-point bending method. Data were analyzed using two-way repeated-measures analysis of variance with a significance level of p<0.05. Results PMMA resin modified with S-PRG nanoparticles released all ions except aluminum. The 10wt% nanoparticle group showed significantly higher ion release than the 5wt% nanoparticle and control groups. While the initial ion release from the 10wt% nanoparticle was comparable to the 20wt% microparticle, recharge resulted in slower ion release from nanoparticles. Incorporating S-PRG nanoparticles altered the surface characteristics and flexural strength of the resin (which remained within international standards). Conclusion Incorporating 5 and 10wt% S-PRG nanoparticles into PMMA resin facilitated the release of boron, silicate, strontium, sodium, and fluoride ions, but not of aluminum ones following recharge with an S-PRG solution and maintained compliance with standards. The 10wt% nanoparticles achieved a balance between enhanced ion release and the physio-mechanical properties required for denture base resins.
Copyright (c) 2025 Kwanchanok Ratanakupt, Toshiyuki Nakatsuka, Kritirat Kiatsirirote https://creativecommons.org/licenses/by/4.0