OBJECTIVE:This study aimed to investigate the biological effects and underlying mechanisms of Astragalus polysaccharides (APS) on human dental pulp stem cells (hDPSCs) under inflammatory conditions. DESIGN:hDPSCs were isolated by enzymatic digestion. Cell viability was assessed using the CCK-8 assay. Cells were divided into three groups: negative control (NC), LPS, and LPS + APS groups. The expression of inflammatory cytokines was evaluated by qRT-PCR and ELISA. Protein expression levels of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), osteocalcin (OCN), and osteopontin (OPN) were detected by Western blot. ALP and Alizarin Red staining were used to assess mineralization capacity. Cell migration was evaluated using wound healing and Transwell assays. Transcriptome sequencing was performed to analyze gene expression profiles. RESULTS:APS at concentrations below 1000 μg/mL did not significantly affect cell viability. LPS significantly increased the expression of IL-6, TNF-α, and IL-1β, whereas APS treatment reduced their expression levels. LPS inhibited the expression of DSPP, DMP-1, OCN, and OPN, as well as mineralization capacity, while APS treatment reversed these effects. In addition, APS further enhanced cell migration. Transcriptome analysis indicated that differentially expressed genes were mainly enriched in the NF-κB signaling pathway. CONCLUSION:APS attenuates inflammation and promotes odontogenic/osteogenic differentiation and migration of hDPSCs, potentially through inhibition of the NF-κB signaling pathway.
OBJECTIVE:Lithium chloride (LiCl) induces cleft palate by disrupting palatal shelf elevation, but the underlying metabolic mechanisms remain unclear. This study aims to investigate these mechanisms using an integrated multi-omics approach. DESIGN:A LiCl-induced cleft palate mouse model was established by intraperitoneal injection of LiCl (0.4 mg/g/day) from embryonic day (E) 10.5 to E13.5. Palatal shelves were collected at E16.5 for targeted metabolomics using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and transcriptome sequencing. Integrated pathway analysis was performed to identify key pathways. Immunofluorescence, reverse transcription quantitative PCR (RT-qPCR), and Hydroxyproline assay validated findings in vivo and in human embryonic palatal mesenchymal (HEPM) cells. Further temporal analysis was conducted at E13.5, E14.0, and E14.5 to identify the key stage. RESULTS:In LiCl-induced cleft palate mice, multi-omics identified 24 differentially abundant metabolites and 595 differentially expressed genes, with arginine and proline metabolism as the most enriched pathway. Within this pathway, 4-Hydroxyproline was downregulated, proline was upregulated, accompanied by downregulation of P4HA3, a key enzyme for 4-Hydroxyproline synthesis. Immunofluorescence revealed increased β-catenin and decreased P4HA3 in cleft palate tissues, with partial colocalization. Temporal analysis showed these changes became significant at E14.0, coinciding with disrupted palatal elevation. In vitro, LiCl simultaneously upregulated β-catenin and downregulated P4HA3 in HEPM cells. CONCLUSION:LiCl-induced cleft palate in mice involves β-catenin-associated downregulation of P4HA3, which contributes to a reduction in 4-Hydroxyproline and impaired palatal elevation. These findings reveal a novel metabolic mechanism in cleft palate pathogenesis.
Purpose: To evaluate Portuguese parents' knowledge about ankyloglossia, examine how it may affect child development, and describe diagnostic and treatment practices of the condition. Methods: During March 2025, a 6-section multiple-choice questionnaire, based on an Australian study, assessed Portuguese parents' knowledge of ankyloglossia both in person at a university pediatric dental clinic and online. The data were analyzed with descriptive statistics, using Fisher's exact tests with Monte Carlo simulations. Results: This study collected 259 valid responses, predominantly from the central region of Portugal (76.1%). More than half of the participants (58.7%) were familiar with the term "ankyloglossia", primarily associating it with issues in speech (91.4%), chewing (70.8%) and breastfeeding (66.1%). The main source of information about ankyloglossia was healthcare professionals (77.9%). Approximately one-fifth of participants (20.1%) had a child diagnosed with ankyloglossia. Diagnoses were typically based on professional opinion, breastfeeding difficulties and tongue appearance, with physicians, dentists, speech therapists and lactation consultants being the primary diagnosticians. Surgical treatment, mainly laser frenectomy, was performed in 88.5% of cases, with significant involvement from dentists and speech therapists. Parents reported high satisfaction with treatment outcomes and the information provided. Conclusion: This study found a moderate level of awareness about ankyloglossia among Portuguese parents and caregivers. Clearer diagnosis and treatment protocols and further research are essential to improve care and enhance parental knowledge.
Purpose: To evaluate the effectiveness of positive pre-visit imagery in reducing dental anxiety (DA) in children undergoing procedures requiring local anesthesia. Methods: A triple-blind, randomized controlled clinical trial was conducted with 69 children aged 7 to 12 years of age at a university pediatric dental clinic in Brazil. Participants were randomly assigned to either an experimental group (positive dental images) or a control group (neutral nature images). Images were shown 2 minutes prior to receiving local anesthesia. DA was assessed before and after treatment using the Venham picture test (VPT). Statistical analyses included the Mann-Whitney U test, Wilcoxon signed-rank test and ordinal logistic regression to identify predictors of post-procedure anxiety. Results: No significant differences in VPT scores were found within or between groups before and after treatment (P>0.05). Baseline anxiety, age, sex and group allocation did not predict post-treatment anxiety. Conclusion: Positive pre-visit imagery was not effective in reducing DA when measured by the VPT.
Purpose: To analyze electronic health records from pediatric patients at a university hospital to evaluate the association between vitamin D deficiency and dental caries in hospitalized children from birth to 9 years of age. Methods: Electronic health records from 124,091 pediatric patients between the years 2011 and 2025 were analyzed. International Classification of Diseases-10 codes identified diagnoses of vitamin D deficiency and dental caries. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. Secondary analyses included stratification by race to account for potential demographic confounders. Results: Dental caries was diagnosed in 8% of children with vitamin D deficiency versus 2.6% of those without it. The unadjusted odds ratio for caries in vitamin D-deficient children was 3.17 (95% CI=2.20 to 4.54; P<0.0001). After adjustment for African American ethnicity, the association remained significant (OR=4.581; 95% CI=2.52 to 8.33; P<0.0001). Conclusions: Vitamin D deficiency was associated with increased odds of dental caries in the 0 to 9 years age group. Further research is needed to explore the influence of race/ethnicity and nutritional factors on pediatric caries risk.
Vitamin C is crucial for normal physiology and function. Severely insufficient levels of vitamin C can disrupt these processes, resulting in scurvy, a condition characterized by compromised collagen production and tissue degradation. The purpose of this report is to present a complex case of a pediatric patient with autism spectrum disorder exhibiting nonspecific oral lesions. Histopathological examination of the lesions revealed granulomatous inflammation, suggesting an initial diagnosis of orofacial granulomatosis, with a recommendation to rule out Crohn's disease. Further investigation confirmed the presence of vitamin C deficiency, with the oral lesions responding positively to vitamin C supplementation, indicating that the oral lesions were a manifestation of scurvy. In the United States, scurvy is rare and usually associated with underlying conditions and restrictive diets; if left untreated, severe complications affecting patients' life expectancy may occur. Understanding the clinical manifestations of vitamin C deficiency is crucial for timely diagnosis and effective management.
Purpose: To evaluate and compare the efficacy of physics forceps versus conventional forceps in primary mandibular second molar extractions among children aged 7 to 10 years of age. Methods: A randomized split-mouth trial was conducted with children requiring bilateral primary mandibular second molar extractions. Outcomes assessed included extraction time, tooth fracture, anxiety, and complications. Results: Fifty children participated. Physics forceps significantly reduced extraction time (mean difference=7.7 seconds, 95 percent confidence interval= -11.5 to -3.9; P<0.001) and anxiety reduction was greater (-1.94 versus -0.56; P<0.001). Tooth fractures were less frequent with physics forceps (12% versus 32%; P=0.05). No post-operative complications occurred. Conclusions: Physics forceps shortened extraction time, reduced anxiety and lowered fracture incidence, making them advantageous for pediatric dental extractions.
Purpose: To determine the prevalence of 3-rooted mandibular molars (3RMMs) in pediatric patients and evaluate associations between their occurrences across different teeth. Methods: A retrospective analysis of digital bitewing and panoramic radiographs from children 4 to 12 years of age was performed. Prevalence was calculated at both the patient and tooth levels. Associations examined included bilateral versus unilateral occurrence, sex and side differences, and correlations between adjacent primary molars as well as between primary and permanent molars. Results: Radiographs of 998 children were evaluated. Of 5,988 mandibular molars assessed, 52 exhibited an additional root (0.9%). At the patient level, 32 children (3.2%) had at least one 3RMM. Prevalence was 1.2% in primary first molars, 1.3% in primary second molars and 1.3% in permanent first molars. Bilateral involvement (43.8%) was significantly more frequent than in unilateral cases (P=0.035). A left-side predominance was noted for primary second molars (P=0.049), while no sex differences were observed. The presence of a primary 3RMM increased the likelihood of a permanent 3RMM (13.6% vs 1.3%; P=0.002). When the first primary molar was 3-rooted, the prevalence of a 3-rooted primary second molar was higher (25% vs 1% in children without a 3-rooted first primary molar, P<0.001), as was the prevalence of a 3-rooted permanent first molar (23.1% vs 1% in children without a 3-rooted first primary molar, P<0.001). Conclusions: Although rare, 3RMMs demonstrate consistent associations between adjacent and successive teeth, supporting a developmental link. Early radiographic recognition in primary molars may help anticipate similar anatomy in permanent molars and reduce the risk of procedural complications.
Purpose: To explore parents' perceptions, acceptance and concerns regarding the use of silver diammine fluordie (SDF) to manage early childhood caries (ECC). Methods: The authors conducted interviews with 39 parents whose children had participated in a randomized clinical trial using SDF to manage ECC. A mixed-methods design was employed: thematic analysis for open-ended responses and descriptive analysis for closed-ended questions. Results: Parental acceptance of SDF was strongly linked to trust in the dentist (100%), its painless application (97.4%), absence of anesthesia, needles or drilling (92.3%), and its ability to reduce pain (97.4%). Most parents appreciated the fast treatment (89.7%) and limited alternatives during the pandemic (89.7%). Nearly all participants (94.9%) reported they would choose SDF again, with higher acceptance for posterior teeth (87.2% vs. 69.2% for anterior teeth). Additionally, 74.4% preferred SDF over restorations. Thematic analysis highlighted 4 key areas: (1) acceptance of SDF, (2) pain and comfort, (3) reactions to color change and (4) concerns and uncertainties. Conclusions: Parents valued SDF as a safe, effective and non-invasive option for managing ECC, particularly given its ability to avoid drilling, needles and anesthesia. However, esthetic concerns about staining underscored the need for clear provider-parent communication and ongoing education about treatment limitations and the importance of follow-up applications for successful caries arrest.
Purpose: To investigate through a meta-analysis and systematic review the influence of the combined use of silver diammine fluoride (SDF) and potassium iodide (KI) on the bonding of adhesive materials to sound and carious dentin. Methods: The PubMed/MEDLINE, Scopus, Embase, LILACS, Web of Science databases and grey literature were searched. In vitro studies comparing the bond strength of adhesive systems (AD) or glass ionomer cements (GIC) to SDF+KI and SDF-treated dentin were included. Mean differences were estimated, separately by dentin condition (sound or carious dentin) and tooth type (primary or permanent), considering the adhesive material as a subgroup. Results: Of 1,689 potentially eligible studies, 45 were selected for full-text analysis and 14 were included. There was no significant difference in the bond strength to permanent carious dentin pre-treated with SDF or SDF+KI (P=0.17; I²=97%), regardless of the adhesive material (GIC: P=0.32; AD: P=0.25). Subgroup analyses for primary carious dentin pre-treated with SDF or SDF+KI, performed separately for GIC and AD, also showed no significant differences (GIC: P=0.13; AD: P=0.28). However, when pooling all datasets, a statistically significant difference was observed (P=0.005; I²=94%), indicating that SDF+KI improved bond strength. No difference in the bond strength of GIC to permanent sound dentin pre-treated with SDF and SDF+KI was found (P=0.80; I²=0%). Conclusion: Pooled in vitro data suggest that SDF + KI do not affect bond strength to permanent carious or sound dentin and may enhance bond strength in primary carious dentin.
Root fractures in adolescents present unique esthetic and prosthodontic challenges, especially when complicated by oblique or vertical fracture lines that preclude traditional root submergence. A 15-year-old female sustained multiple traumatic dental injuries, including a complex root fracture of the maxillary left central incisor, following a motor vehicle accident. Polytrauma management included pulpal therapy, splinting and interim prosthesis fabrication, but the incisor demonstrated both horizontal and vertical fracture components, contraindicating conventional root submergence therapy. Socket shield therapy (SST) ridge preservation enabled buccal bone retention while minimizing the patient's surgical burden, thereby facilitating immediate removable prosthesis fabrication and esthetic rehabilitation. The preserved shield will facilitate future implant placement during adulthood without requiring grafting. This case highlights SST as a conservative pediatric trauma management strategy, providing immediate esthetic reassurance for the adolescent patient and reducing trauma-related distress during a critical developmental stage.
OBJECTIVES:Our study has shown an enhanced effect on osteogenesis of periodontal ligament (PDL) cells upon treatment with osteoclasts' conditioned medium. However, characterization of osteoclasts' extracellular vesicles (OC-EVs), their osteogenic enhancement, and the underlying mechanism within PDL cells have been unexplored. DESIGNS:Before PDL cell treatment and staining with alkaline phosphatase, Alizarin Red, or von Kossa, OC-EVs were characterized by transmission electron microscopy, nanoparticle tracking analysis with their sizes demonstrated at 100-200 nm, and immunoblotting for expressions of OC-EVs' markers. Expressions of receptor activator of nuclear factor-kappa B (RANK) on OC-EVs' membrane and RANK ligand (RANKL) on PDL cells' membrane were determined by flow cytometry and immunofluorescence, respectively. Involvement of the RANK-RANKL and AKT signaling in OC-EVs-induced osteogenesis of PDL cells was studied by pretreatment with osteoprotegerin, RANKL, or MK-2206, an AKT inhibitor. RESULTS:RANK expression was found at 5.02% of total OC-EVs. Treatment of PDL cells with OC-EVs significantly increased osteogenic differentiation, mineralization, and gene expressions (P < 0.05). Pretreatment of osteoclasts with GW4869, a blocker of EVs' secretion, abolished the expression of RANK on OC-EVs and significantly reduced the enhanced osteogenesis of PDL cells (P < 0.05). Moreover, pretreatment with osteoprotegerin or RANKL before treatment with OC-EVs significantly diminished the enhanced osteogenesis (P < 0.05), without any inhibition on enhanced phosphorylated AKT expression by OC-EVs. Pretreatment with MK-2206 could significantly decrease differentiation and mineralization of OC-EVs-treated PDL cells (P < 0.05). CONCLUSION:OC-EVs can induce osteogenesis of PDL cells via RANK-RANKL signaling and AKT activation as parallel contributions.
Objective Cancer cells frequently exploit the Pentose Phosphate Pathway (PPP) to fuel antioxidant defenses and the synthesis of essential biomolecular precursors. It is hypothesized that sustained upregulation of glucose flux through the PPP is crucial for meeting the anabolic demands of cancer cells and mitigating oxidative stress. We investigated whether pharmacological inhibition of the rate-limiting PPP enzyme, glucose 6-phosphate dehydrogenase (G6PD), could decrease proliferation, disrupt cell-cell interactions, and impede migration in oral squamous cell carcinoma (OSCC). Design Non-tumorigenic keratinocytes (HaCaT), less invasive (CAL27) and highly invasive (SCC9) OSCC had the G6PD pathway inhibited using 6-aminonicotinamide (6-AN) and then submitted to proliferation, spheroid formation, and cell migration assays. Results Inhibition of G6PD significantly decreased cell proliferation, disrupted spheroid integrity, and reduced migratory capacity across both OSCC cell lines, while exhibiting only moderate cytotoxicity towards non-tumorigenic keratinocytes, yielding Selectivity Index (SI) values of 1.47 for CAL27 and 3.39 for highly invasive SCC9 cells. Conclusion Given that cancer cells often detach and migrate within challenging microenvironments, the ability of 6-AN to induce metabolic stress and impede both cell-cell interactions and migration highlights its potential as an adjuvant therapeutic strategy against OSCC progression.
OBJECTIVE:To investigate whether exosomes derived from human umbilical cord mesenchymal stem cells (UCMSCs) at two osteogenic induction stages (undifferentiated and late-stage) differentially affect periodontal ligament stem cells (PDLSCs), and to explore the potential molecular basis. DESIGN:UCMSCs and PDLSCs were isolated and cultured. Exosomes were harvested from undifferentiated UCMSCs (Exo-D0) and UCMSCs after 14 days of osteogenic induction (Exo-D14). PDLSCs were treated with both exosome types. Proliferation and migration were analyzed using EdU and scratch assays, the latter under serum-free conditions. Early osteogenic differentiation was assessed by alkaline phosphatase staining and quantitative reverse transcription PCR (qRT-PCR). Differentially expressed miRNAs were identified by high-throughput sequencing and further analyzed through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. RESULTS:Both exosome types promoted PDLSC migration. Exo-D0 enhanced early osteogenic differentiation, whereas Exo-D14 enhanced proliferation but reduced early osteogenic marker expression. Sequencing identified 21 differentially expressed miRNAs (13 upregulated, 8 downregulated). Bioinformatic prediction suggested that the putative target genes were enriched in Ras signal transduction, regulation of kinase activity, and focal adhesion, and further predicted significant enrichment in the MAPK, Ras, and PI3K-Akt signaling pathways, which are central to cell proliferation and osteogenic differentiation. CONCLUSIONS:Exosomes from undifferentiated and osteogenically induced UCMSCs exerted distinct effects on PDLSCs, potentially associated with differentially packaged miRNAs. These findings offer a basis for hypotheses about exosome-mediated mechanisms and support matching exosome sources to the intended therapeutic outcome as potential cell-free strategies for periodontal tissue regeneration and alveolar bone repair.
OBJECTIVE:To evaluate the effects of erythritol on functional cariogenic traits in separate single-species Streptococcus mutans and Lacticaseibacillus acidophilus models and to characterize relative species composition in a simplified dual-species biofilm. DESIGN:Functional microbial, biochemical, structural, molecular, and hard-tissue outcomes were evaluated in separate single-species models. Species-specific qPCR was used to quantify the relative abundance of S. mutans and L. acidophilus in a simplified dual-species biofilm. RESULTS:Erythritol reduced bacterial growth and viable colony formation in both species. Medium acidification was attenuated, with slower pH decline and lower lactic acid accumulation in erythritol-treated groups. Biofilm biomass and metabolic activity were lower at both 24 h and 48 h. In the dual-species biofilm, the relative abundance of S. mutans decreased, whereas that of L. acidophilus increased correspondingly. Microscopy showed reduced viable bacterial distribution, weaker dentin-surface adhesion, less compact biofilm architecture, and diminished extracellular matrix deposition. Separate single-species hard-tissue analyses demonstrated lower Ca2 + release, higher endpoint surface microhardness, lower surface hardness loss, and higher cross-sectional microhardness profiles. RT-qPCR showed reduced expression of selected virulence-related genes, particularly in S. mutans. Across most assays, the effects observed at 6% and 8% erythritol were similar. CONCLUSIONS:Erythritol attenuated multiple cariogenic traits in separate single-species S. mutans and L. acidophilus assays and altered relative species composition in a dual-species biofilm. The composition data do not establish reduced dual-species functional virulence or clinical carioprotection.
OBJECTIVE:To investigate dental plaque metabolic profile differences between preschoolers with Black Stain and Caries-free (BSCF) versus Severe Early Childhood Caries with Black Stain (SECCBS) to elucidate S-ECC pathogenesis. DESIGN:Dental plaque was collected from 59 preschoolers aged 3-6 (BSCF, n = 29; SECCBS, n = 30) and analyzed using ultra-high performance liquid chromatography-tandem mass spectrometry. Differential metabolites (DMs) were identified via partial least squares discriminant analysis. KEGG pathway analysis and Metorigin provided functional and source-tracking insights. Weighted Gene Co-expression Network Analysis (WGCNA) identified hub metabolites linked to clinical traits, while 10 machine learning models were developed for diagnostic prediction. RESULTS:Profiling identified 1069 metabolites, predominantly lipids/lipid-like molecules (24.60%) and organic acids/derivatives (11.51%). Eighty-five DMs were identified between groups; four were significantly downregulated ( Δ17-6-keto prostaglandin F1α, VLH, DPK, and N'-[1-(2-hydroxyphenyl)ethylidene]-3-methoxybenzohydrazide). L-glutamate was significantly upregulated in SECCBS, whereas L-(+)-citrulline was downregulated. KEGG analysis mapped DMs primarily to nucleotide, carbohydrate, amino acid, and lipid metabolism. Metorigin revealed 78.6% of metabolites were co-derived from diet, microbiota, and pharmaceuticals. WGCNA identified 31 modules, two positively correlating with SECCBS. The top-performing machine learning model achieved robust AUC performance for clinical stratification. CONCLUSIONS:Distinct plaque metabolomic profiles differentiate SECCBS from BSCF. Alterations in nucleotide metabolism, the pentose phosphate pathway, and specific amino acid pathways drive SECC progression. Furthermore, 2'-deoxyuridine and 6-phosphogluconic acid emerge as potential biomarkers for early SECC diagnosis, prevention, and targeted clinical intervention.
Objectives Phosphatase and Tensin Homolog (PTEN) is a key negative regulator of intracellular signalling pathways that govern cell growth, survival, differentiation, and inflammatory responses. Growing evidence suggests that PTEN plays a critical role in regulating the biological functions of dental mesenchymal stromal cells (MSCs) with stem-like properties, which are central to dental tissue regeneration and repair. This review focuses on the functional roles of PTEN in dental MSCs Design A literature search was conducted in PubMed and SCOPUS from inception through October 12, 2025. English-language original studies investigating PTEN-mediated regulation of dental MSC function in vitro models were included. Data were synthesised qualitatively due to heterogeneity in experimental designs. Results Nine studies met the inclusion criteria. The findings demonstrate that PTEN regulates key biological processes in dental MSCs, including dental pulp stem cells, stem cells derived from exfoliated deciduous teeth, periodontal ligament stem cells, and gingival fibroblast cells, primarily through the PI3K/AKT signalling axis. Other mechanisms, including DNA methylation, microRNAs, and circular RNAs, participate in PTEN modulation, which is associated with altered cell behaviours, influencing lineage commitment, inflammatory responses, apoptosis, and differentiation. Conclusions PTEN functions as a context-dependent regulator of the fate and homeostasis of dental MSCs. Precise modulation of PTEN signalling holds translational potential for modulating cell responses.
