Abstract Submandibular salivary gland inflammation has been suggested as one of the mechanisms underlying impaired salivary secretion associated with sleep deprivation (SD). However, whether the salivary inflammatory response occurs to the same extent in paradoxical sleep deprivation with or without sleep recovery remains unknown. Objective: This study evaluated the extent to which inflammation influences salivary impairments associated with paradoxical sleep deprivation with or without sleep recovery. Methodology: Male Wistar rats were randomly assigned into three groups as control, partial SD (PSD) with sleep recovery for four hours a day and total SD (TSD). Paradoxical SD was carried out for seven days in the SD groups, after which saliva, blood, and submandibular gland samples were taken. Levels of interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α), and nitrite were determined in saliva, serum, and the submandibular salivary gland. Leucocyte count and neutrophil-lymphocyte ratio were determined in all groups. One-way ANOVA and the Tukey's post hoc tests were used for data analysis. P-values < 0.05 were considered statistically significant. Results: Levels of TNF-α, IL-6, and nitrite in the submandibular salivary glands were significantly higher in the TSD groups (p=0.04,p<0.001, p=0.03, respectively) than in the control. Saliva level of TNF-α was higher in the PSD and TSD groups (p=0.003 and p=0.01 respectively) than in the control. Neutrophil-lymphocyte ratio was significantly higher in both PSD and TSD groups than in the control (p<0.01 for both). Conclusion: While total SD produced higher inflammatory response in the submandibular salivary gland, four-hour sleep recovery ameliorated this impact. This finding suggests that sleep recovery is crucial to improve inflammatory salivary gland dysfunction induced by sleep deprivation.

Abstract Guided bone regeneration (GBR) is an alternative treatment for craniofacial bone defects reconstruction through membrane barrier adaptation, such as demineralized dentin material membrane (DDMM). DDMM is used as a substitute for GBR material, which aligns with Green Economy principles, it has a good biological osteoinductive and osteoconductive effects, and its structure resembles bones. The balance of bone remodeling when experiencing craniofacial defects will be altered and allow changes to resorption activity, so the mechanisms of osteoclastogenesis and bone resorption are vital. Objective: this article aims to analyze the expression of TNF-α, RANKL, and osteoclast cells count after application of DDMM as GBR in mandibular bone defects. Methodology: this is an experimental study with a post-test only control group design, which began with the randomization of 120 rats into five groups: K(−), without membrane implantation; K(+), PPCM; P1, DDMM; P2, DDMM + bone graft; P3, PPCM + bone graft. The expression of TNF-α, RANKL, and osteoclast cells count were observed, followed by analysis using a one-way ANOVA and post hoc Tukey HSD comparison test. Results: there were significant differences in the expression of TNF-α, RANKL, and osteoclast cells count in all study groups (p=0.000). TNF-α showed a decreasing difference with the highest expression in the K(−) group on day 3 of 12.00±2.16. RANKL expression increased on day 14 and decreased on day 21 in all groups. The osteoclast cells count generally showed a critical period with the highest increase in the K(−) group on day 14 of 73.00±0.00. Conclusion: DDMM has the potential to be a superior membrane substitute compared to PPCM as GBR in alternative treatment for craniofacial bone defects reconstruction.

Abstract Objectives: Depth of invasion (DOI) in oral squamous cell carcinoma (OSCC) guides treatment and prognosis but lacks three-dimensional (3D) insight. Thus, this study aimed to investigate the feasibility and accuracy of Lugol's iodine-enhanced micro-computed tomography (CT) for the 3D measurement of DOI in OSCC samples. Methodology: In total, 50 in vitro OSCC samples from Nanjing Stomatological Hospital (July 2022 to January 2024) were subjected to micro-CT imaging with a slice thickness of 50 μm following 3% Lugol iodine staining for 12 h, followed by pathological examination and staining. The panoramic diagnostic scanner digitally measured pathological DOI. The micro-CT DOI was measured by evaluating the voxel value of the boundary of the tumor lesion and comparing it with the pathological examination results. Experienced physicians analyzed both measurements, and statistical analyses were performed to determine their correlation. Results: Lugol iodine-enhanced micro-CT imaging distinguishes various tissue structures, such as tumor tissue, epithelial tissue, muscle tissue, blood vessel structure, and other major tissue structures in 3D space. This imaging technique found and localized micro-tumor lesions (1.82×1.5×1 mm3) when in conjunction with pathological sections. Statistical analysis indicated a strong correlation between pathological DOI and micro-CT DOI (P<.001; r=0.986). During DOI measurement, Lugol iodine-enhanced micro-CT imaging effectively compensated for the loss of 3D space information in the pathological measurements, improving the accuracy of the DOI measurement. Conclusions: Lugol iodine-enhanced micro-CT improves OSCC DOI 3D measurements, enhances pathological staging accuracy, and aids treatment decisions and prognosis.

Abstract Pulpitis, which is often triggered by caries and trauma, is a significant clinical challenge due to its prevalence. This research aims to uncover potential metabolic biomarkers for pulpitis and map out the implicated metabolic pathways, thereby laying a foundation for enhanced diagnostic and preventive strategies. Methodology We analyzed pulp samples from 12 participants (six who had pulpitis and six who had healthy teeth) using serum metabolomics via ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry. Important biomarkers were pinpointed via multivariate analysis and orthogonal partial least squares discriminant analysis. Additionally, correlation and biomarker pathway enrichment analyses were conducted to explore the relations between differentially expressed biomarkers and their associated biological pathways. Specific metabolites of interest were further examined via alkaline phosphatase (ALP) staining, Alizarin Red staining, and RT-qPCR analysis. Results We identified 22 significant biomarkers (13 increased, nine decreased) related to 18 metabolic pathways in pulpitis cases. Key biomarkers included ascorbic acid, inosine, allopurinol riboside, and L-asparagine, in which ascorbic acid and inosine showed the most substantial downregulation and strongest association with pulpitis. Notably, aminoacyl-tRNA biosynthesis and retrograde endocannabinoid signaling pathways were closely linked with pulpitis. Ascorbic acid enhanced the osteogenic differentiation, calcium deposition, as well as the expression of osteogenic genes of human dental pulp stem cells (DPSCs).Conclusions: The identified biomarkers and metabolic pathways offer insights into the pathogenesis of pulpitis and have potential applications in developing preventive treatments.

Abstract The acquired enamel pellicle (AEP) consists of an organic, acellular, and bacteria-free film, formed in vivo as a result of biomolecules adsorption onto the tooth surface. It is composed of proteins, glycoproteins, lipids, phospholipids, and other macromolecules, such as carbohydrates. The AEP formation process is complex and can be divided into three stages: initiation, development, and maturation. The pellicle has two main layers: the globular and basal layers. The basal layer offers the most protection against demineralization, as the subsequent globular layer is weaker and less tenacious. The formation of the AEP can be influenced by various factors, such as the physicochemical properties of the teeth, location in the oral cavity, pathologies, and even the oral microbiota. With the advancement of “omics” techniques, it has been possible to observe the presence of acid-resistant proteins in the AEP, which allowed the development of the “acquired pellicle engineering” strategy. This strategy involves enriching and modifying the basal layer with acid-resistant proteins. Among these proteins, hemoglobin, statherin-derived peptide, and a protein derived from sugarcane stand out. The objective of this literature review is to provide a comprehensive overview of the AEP, detailing its composition, formation process, and protective functions. Additionally, the review aims to explore recent advances in the field of “acquired pellicle engineering,” highlighting the acid-resistant proteins of the AEP and their potential applications in dentistry. Finally, the review intends to highlight the clinical implications of these findings and how they may contribute to the development of new strategies for the prevention and treatment of dental pathologies according to published studies.

Abstract The nasopalatine duct cyst (NPDC) is the most common nonodontogenic jaw cyst, with a higher prevalence reported in males. Diagnosing jaw cysts remains challenging for general dentists due to their overlapping clinical and radiographic presentations. An updated case series and integrative literature review may assist in improving the accurate diagnosis of NPDC. Objective This study aimed to describe the clinicopathological and imaging characteristics of 63 NPDC cases and to review previously reported cases in the literature. Methodology An international, multicenter, retrospective NPDC case series was conducted. Demographic, radiographic, and histopathological data were collected from clinical records. Additionally, a PubMed/MEDLINE search was performed to identify articles on NPDC. Results A total of 63 NPDC cases were evaluated, with a mean patient age of 47 years and no significant sex predilection. Twenty-one cases were asymptomatic, while 34 presented with symptoms such as pain and swelling. Radiographically, NPDC appeared as a well-defined radiolucent lesion located between the upper central incisors, bordered by a radiopaque margin. The integrative literature review identified 67 studies, comprising 51 case reports, 12 retrospective studies, and four case series, totaling 1,003 reported NPDC cases. The clinicopathological and radiographic findings from the literature aligned with those in this case series. Conclusion The 63 cases analyzed in this study showed consistent findings across six international centers, with no sex predilection observed, contrasting with the male dominance reported in the literature. NPDC should be considered in the differential diagnosis of intraosseous lesions in the anterior maxilla. Accurate diagnosis requires a combination of radiographic and histopathological evaluations to prevent misdiagnosis and improper treatment.

Abstract Aim This cross-sectional study investigated the salivary proteomic profile associated with generalized gingivitis in pregnant women with obesity. Methodology Pregnant women in the third trimester (≥27 weeks of gestation) were divided into two groups based on bleeding on probing (BOP): G1 (BOP>50%; n=9) and G2 (BOP 0–30%; n=9). Collected unstimulated saliva samples were individually analyzed using nano liquid chromatography electron spray ionization tandem mass spectrometry. Identified proteins were classified according to gene ontology for biological processes, molecular functions, immune system involvement, and cellular components. Differential protein expression was determined using thresholds of p<0.05 for downregulation and 1-p>0.95 for up-regulation proteins. Results Of the 183 identified proteins, 100 were shared between groups, totaling 57 up-regulated and 27 downregulated proteins in G1. Key biological processes included antimicrobial humoral response and hydrogen peroxide catabolism, with proteins linked to immune function and endopeptidase regulation. Functional analysis showed that Lactotransferrin (5-fold increase in G1), Haptoglobin (4-fold), and Immunoglobulin J chain (3-fold) were up-regulated, whereas Statherin (5-fold) and Protein S100-A8 (4-fold) were downregulated in G1. Conclusions Pregnant women with obesity and generalized gingivitis exhibited a distinct salivary proteomic profile characterized by the up-regulation of immune-related proteins and downregulation of tissue-protective proteins. These findings suggest potential salivary biomarkers for detection and targeted management of periodontal inflammation in this high-risk population.

Abstract Aim To evaluate the clinical effectiveness of ozonated sunflower oil (Oz) as an adjunctive of non-surgical periodontal therapy in patients with type 2 diabetes mellitus (DM2), on fibroblast cell viability and migration and the effectiveness of Oz on a Candida albicans (C. albicans) culture. Methodology In total, 32 sites in 16 DM2 with moderate to advanced periodontal disease with periodontal pocket depths ≥5mm were selected. The treatments were divided into two groups: control, saline solution (SS) as an adjunctive of scaling and root planing (SRP+SS), and test, Oz as an adjunctive of SRP (SRP+Oz). Hematological [fasting glucose level (FGL) and hemoglobin A1c (HbA1c)] and microbiological samples were collected from the participants at baseline and three months after periodontal treatment and the microbiological samples were analyzed by PCR. C. albicans was previously tested by the agar diffusion test. The effect of Oz was tested on cell viability and fibroblast migration. Results The groups showed no statistically significant differences (paired t-test-p>0.05) regarding hematological parameters, FGL (median - baseline 171.41, 3 months 164mg/dL), and HbA1c (baseline 8%, 3 months 7.5%) (Kruskal-Wallis One-Way Nonparametric-p>0.05) after periodontal therapy. The groups showed statistical differences for periodontal parameters between baseline and three months (paired t-test-p<0.05). PCR analysis showed a reduction in the percentage of C. albicans in the SRP+Oz group after three months (McNemar’s test-p=0.002). Cell viability was lower in the high glucose Dulbecco’s modified Eagle’s medium (4500 mg/L) than in low glucose (1000 mg/L) (RM-ANOVA-p<0.0001). The wound healing test showed reduced fibroblast migration (one-way ANOVA with Dunnett’s post-test-p<0.01). Oz showed high C. albicans antifungal inhibition (Kruskal-Wallis test-p=0.0001). Conclusions SRP+Oz effectively reduced C. albicans in-vitro and in-vivo but showed no clinical improvements compared to the control. Cell viability and wound healing of fibroblasts showed no improvements.

Abstract Circadian rhythm disorders and NF-κB are closely linked and can exacerbate periodontitis. However, the mechanisms via which circadian rhythm-related genes influence periodontitis are not yet fully understood. Objective We investigated the effect of brain and muscle Arnt-like protein-1 (BMAL1) on the NF-κB pathway and downstream inflammatory factors on periodontitis. In this study, Bmal1 homozygous knockout and periodontitis mouse models were established. Methodology Bone marrow-derived macrophages (BMDMs) from Bmal1-/- mice were cultured and stimulated with lipopolysaccharides. Bone resorption was detected using micro-computed tomography and histological analyses. Gene and cytokine expression was assessed using quantitative reverse-transcription PCR and ELISA. The nuclear translocation of p65 was detected using immunofluorescence. Results Our findings indicate that Bmal1 knockout exacerbates periodontitis severity in mice by activating the NF-κB signaling pathway with increased nuclear translocation of p65 (p<0.05), as well as increased expression of Il-1b, Il-6, and Tnfα (p<0.01), along with decreased Nr1d1 expression (p<0.05) in BMDMs under inflammation. Conclusion The results highlight the protective role of Bmal1 in periodontitis and suggest its potential link to the circadian clock's influence on the disease.

Abstract Objective This study evaluated whether hypoglycemic drug metformin enhances the anti-cancer effects of cisplatin in YD-9 cells. Methodology YD-9 cells, derived from oral mucosal squamous cell carcinoma of oral mucosa, were used to assess the combined effects of metformin and cisplatin by means of MTT assay, live and dead cell staining, and colony formation assays to evaluate cell viability and proliferation. Reactive oxygen species level was measured using a Muse cell analyzer. Apoptosis, epithelial-mesenchymal transition, and related molecular pathways were analyzed by western blot. Wound healing assays and Transwell migration assays examined cell migration, whereas monophosphate-activated protein kinase inhibitor Compound C, was utilized to investigate the AMPK pathway. Results Sequential treatment of YD-9 cells with metformin and cisplatin resulted in decreased cell viability and proliferation, increased ROS levels, and elevated apoptosis compared with the individual drugs. Moreover, the treatment inhibited EMT, wound healing, and cell migration. These results correlated with increased AMPK phosphorylation, a key regulator of cellular energy homeostasis. Introduction of Compound C pre-treatment upregulated N-cadherin and α-smooth muscle actin along with enhanced cell migration. Conclusion This study found synergism in anti-cancer effects between metformin and cisplatin. Additionally, introduction of Compound C confirmed that EMT inhibition is AMPK dependent. These findings indicate the potential use of metformin as an adjunct drug in anti-cancer treatments, warranting further investigation.

Abstract Generalized Joint Hypermobility (GJH) is one of the pathophysiological contributing factors for the development of temporomandibular disorders (TMD). There are, however, several counterpoints on the potential relation between TMD and joint hypermobility, especially when considering the temporomandibular joint (TMJ), event known as TMJ hypertranslation. Additionally, there is no consensus regarding the clinical and imaging diagnostic criteria for such condition. Hence, this scoping review addresses the association between GJH, TMJ hypertranslation and TMD, highlighting the lack of consensus concerning TMJ hypertranslation diagnosis. Eligibility criteria included book sections, clinical trials, meta-analyses, multicenter studies, observational studies, and reviews published in English between 1964 and 2024. Bibliographic search was conducted on the PubMed, SciELO, LILACS and Science Direct databases using the following Medical Subjective Headings (MeSH) terms: “temporomandibular joint disorders,” “temporomandibular joint,” “joint instability” and “joint dislocations.” “TMJ hypermobility” and “TMJ subluxation,” non-indexed terms, were applied as individual searches in the same databases. Manual search was performed in selected works by cross-referencing the included studies and book sections. Additional search was conducted in the grey literature. All searches were performed from January to June 2024. After selection, 54 texts were included. While some studies suggest that joint hypermobility (generalized or TMJ specific) may be a risk factor for TMD, especially of the intra-articular type, others rule out this association. No consensus on the potential association between joint hypermobility and TMD was achieved due to the diverse methodologies used to define TMJ hypertranslation diagnosis. More robust and controlled studies are needed to establish a diagnostic criteria and, consequently, understanding of its potential repercussions on masticatory structures, as well as management and prevention of the clinical manifestations.

Abstract Inflammation, oxidative damage, and adenosine triphosphate (ATP) depletion play a role in the pathogenesis of cisplatin (CIS)-induced oral mucositis. Objective: The purpose of this research is to examine the impact of ATP against potential oral mucositis development in cisplatin-treated rats. Methodology All rats were randomly assigned to four groups, namely healthy control group (HG), ATP group (ATPG), Cisplatin group (CISG), and ATP + Cisplatin group (ATCS). Firstly, ATP 4 mg/kg was administered via intraperitoneal injection (IP) to both ATPG and ATCS groups. The same volume of normal saline was injected into HG and CISG groups. After 1 h, cisplatin 5 mg/kg was administered via IP to CISG and ATCS groups. The drugs were taken 1x1 for 7 d. Later, tongue tissues were collected from all groups. Biochemical, macroscopic, and histopathological examinations were performed on all tissues. Results: ATP inhibited cisplatin-induced oxidative damage and pro-inflammatory cytokines levels in tongue tissue. In the CIS group, a significant number of distinct sulcus formations were found in the apex and corpus, as well as a few ulcer foci in the corpus, significant papilla loss, and bleeding. Meanwhile, in the ATP group, a similar appearance to healthy tissue was observed. Histopathologically, it was determined that in cisplatin-aggravated tongue tissue damage, filiform papillae decreased when ATP was administered, and the arrangement and structures of the epithelium, blood capillaries, muscle groups, and adipose cell groups were normal. Conclusions: Oral mucositis caused by cisplatin is alleviated by ATP. These findings may be useful for developing new therapeutic approaches to prevent or treat mucositis, a side effect so severe that can lead to treatment discontinuation.

Abstract This study evaluated the effect of Stemregen® nutritional supplement on inflammation and resorption in apical periodontitis using a rat model. Methodology Rats were divided in three groups: negative control (n=7), positive control (n=10), and Stemregen® (Stem) (n=10). Apical periodontitis was induced in the positive control and Stem groups, and all rats were sacrificed on the 30th day. Serum phosphorus (P), calcium (Ca), and alkaline phosphatase (ALP) were analyzed. Histopathological assessments measured osteoblastic and osteoclastic activity, inflammation, fibrosis, and abscess density. Immunohistochemical analyses evaluated RANKL, TRAP, and OPG levels. Results Results showed significantly lower osteoblastic activity in the negative control compared to Stem and positive control groups (p=0.005). Osteoclastic activity was higher in the positive control (p=0.032). Inflammation and abscess formation were reduced in the Stem group compared to the positive control (p<0.001). OPG levels were lower in the negative control compared to the other groups (p=0.005). Conclusion Stemregen® effectively reduced inflammation and bone destruction, suggesting potential benefits for apical periodontitis management, though further research is needed.

Abstract Objectives The oral cavity harbors a plethora of bacterial species. Dysbiosis of oral and gut microbiota is associated with several oral and systemic pathologies, such as cancer, obesity, diabetes, atherosclerosis and gastrointestinal diseases. Imbalance in the oral-gut microbial axis has been associated with head and neck squamous cell carcinoma (HNSCC). This study aims to analyze the bacterial profile of HNSCC across various taxonomic units, investigate molecular patterns associated with prevalent bacterial phylum in HNSCC, and compare the bacterial profile in HNSCC and gastrointestinal (GI) carcinoma using computational analysis. Methodology The microbe-host transcriptomic, proteomic, and epigenetic analyses of HNSCC and GI carcinomas were performed using The Cancer Microbiome Atlas (TCMA) database. The differential expression of the host’s mRNA transcripts and proteins associated with tumor microbiome were analyzed using The University of Alabama at Birmingham Cancer data analysis (UALCAN) and Clinical Proteomic Tumor Analysis Consortium (CPTAC) websites. Results A decrease in Actinobacteria and an enrichment of Flavobacteria at the class level, Neisseriales, Pasteurellales, and Campylobacterales at the order level, Pasteurellaceae, Flavobacteriaceae, Campylobacteraceae, and Peptoniphilaceae at the family level, and Hemophilus, Porphyromonas, and Leptotrichia at the genus level were observed in HNSCC compared to the normal mucosa. RICTOR protein, mRNA transcripts (HIST1H2BB, SCARNA11, TBC1D21 gene), and hsa-miR-200a-5p miRNA were significantly correlated with prevalent bacterial species in HNSCC. A major increase in Actinobacteria, Fusobacteria, and Spirochaetes was observed in HNSCC compared to GI carcinoma. Conclusion The oral-gut microbial dysbiosis, as reflected by the differential abundance of bacterial species in oral and GI carcinomas, suggests the implication of tumor microbiome and their genomic interactions with the host in carcinogenesis.