Objective To investigate the differences in proliferation and osteogenic differentiation capacities between rat dental socket stem cells (DSCs) and periodontal ligament mesenchymal stem cells (PDLSCs), thereby providing a experimental basis for elucidating the functional characteristics of these two distinct sources of mesenchymal stem cells. Methods This study was approved by the Institutional Animal Care and Use Committee. Twenty-four 6- to 8-week-old male Sprague-Dawley rats were divided into four groups (n = 6 per group) based on the time points of 0, 1, 3, and 7 days post-extraction by the random number table method. A rat alveolar socket model was established by extracting the left maxillary molars. Hematoxylin and eosin (H&E) staining was performed to observe the histomorphological changes in the alveolar sockets at the specified time points. Immunohistochemistry was used to detect the expression and distribution of the p75 neurotrophin receptor (P75NTR). Based on the staining results, an optimal time point was selected to harvest early healing tissues from the alveolar sockets. DSCs were cultured using a combination of tissue block and enzymatic digestion methods. Concurrently, PDLSCs were isolated from the extracted molars using identical culture protocols. The surface markers CD44, CD45, CD90, and P75NTR were analyzed via flow cytometry. Cell proliferation and clonogenic potential were compared using the Cell Counting Kit-8 (CCK-8) assay and colony formation assays, respectively. After adipogenic induction, Oil Red O staining was conducted to observe lipid droplet formation. Following osteogenic induction, alkaline phosphatase (ALP) activity and alizarin red S (ARS) staining were utilized to evaluate ALP expression and mineralized nodule formation, respectively. Furthermore, the mRNA and protein expression levels of osteogenesis-related factors, including ALP and Runt-related transcription factor 2 (RUNX2), were quantified to comprehensively assess the osteogenic differentiation potential of both cell types. Results H&E staining revealed that the alveolar sockets were filled with erythrocytes 0 and 1 days post-extraction. By day 3, significant fibroblastic connective tissue proliferation was observed within the sockets, and bone formation increased markedly by day 7. Immunohistochemical analysis showed sparse P75NTR-positive staining limited to the inner socket wall 0 and 1 days post-extraction. On day 3, P75NTR was distributed throughout the fibrous connective tissue of the socket, with slight staining on the bone walls. By day 7, the receptor was expressed in the fibrous connective tissue at the edges of the trabeculae. Consequently, day 3 post-extraction was selected as the optimal time point for DSC isolation. Both PDLSCs and DSCs exhibited similar morphologies in vitro, characterized by high expression of mesenchymal stem cell markers (CD90, CD44) and P75NTR, alongside low expression of the hematopoietic marker CD45. CCK-8 assays demonstrated that the proliferation capacity of the DSCs was significantly higher than that of the PDLSCs (P < 0.05), accompanied by a greater colony formation rate (P < 0.001). Both cell types possessed adipogenic differentiation potential, as evidenced by vacuolar structures in the cytoplasm. Regarding osteogenesis, the DSCs exhibited a larger area and higher density of ALP staining (P < 0.001). ARS staining indicated that both cells formed orange-red mineralized nodules; however, the DSCs produced significantly more calcified nodules (P < 0.0001). Additionally, the mRNA and protein expression levels of ALP and RUNX2 were notably elevated in the DSCs compared with the PDLSCs (all P < 0.05). Conclusion Rat DSCs can be stably cultured in vitro. Compared with PDLSCs, DSCs exhibited superior capabilities in terms of proliferation, colony formation, and osteogenic differentiation.
Objective To investigate and analyze the influencing factors and intervention strategy preferences of preoperative dental anxiety in patients undergoing oral implant surgery, and to provide a basis for the clinical formulation and implementation of targeted interventions. Methods This study was approved by the Hospital Medical Ethics Committee, and written informed consent was obtained from all participants. Using convenience sampling, a cross-sectional survey was conducted from January to October 2025 among 210 patients in the Department of Implantology of a tertiary dental hospital. All participants had been clinically diagnosed and were scheduled to undergo oral implant surgery on the day of the survey. The survey was conducted using a General Information Questionnaire, the Modified Dental Anxiety Scale (MDAS), and an Anxiety Cause Questionnaire. The General Information Questionnaire collected data on patients’ gender, age, presence of systemic diseases, dental treatment history, number of implants, whether bone augmentation was required. The MDAS was used to assess patients’ dental anxiety levels via MDAS scores, with a score ≥ 12 defined as dental anxiety. The Anxiety Cause Questionnaire covered the following items: fear of pain, concern that the instruments might cause permanent damage to their teeth, concern about the dentist’s technique and inability to achieve ideal results, fear of local anesthesia, concern about cross-infection, financial pressure, exposure to an unfamiliar environment and strangers, and previous unpleasant dental experiences. The MDAS scores of the items in the Anxiety Cause Questionnaire and General Information Questionnaire were analyzed using the Mann-Whitney U test or Kruskal-Wallis H test for univariate analysis, and factors with statistical significance in the univariate analysis were further analyzed using binary logistic regression for multivariate analysis. An Intervention Strategy Preference Questionnaire was used to survey patients’ preferences for the following interventions: understanding the surgical procedure, verbal reassurance from medical staff, distraction methods such as listening to music or using stress balls, staying in a physically and mentally relaxing environment, and others. Results The average preoperative MDAS score of patients undergoing oral implant surgery was 10.91 ± 3.40, and the incidence of dental anxiety (MDAS ≥ 12) was 34.8% (73/210). Univariate analysis showed that female patients, patients aged < 45 years, had significantly higher MDAS scores (P < 0.05). Patients who selected “fear of pain” (79.0%), “fear of local anesthesia” (21.0%), and “exposure to an unfamiliar environment and strangers” (4.8%) as causes of their anxiety had higher MDAS scores than those who did not select these items (P < 0.05). Multivariate logistic regression analysis showed that patient's age (OR = 0.420), fear of pain (OR = 4.234), and exposure to an unfamiliar environment and strangers (OR = 8.220) were independent influencing factors of preoperative dental anxiety in patients undergoing oral implant surgery (P < 0.05). The Intervention Strategy Preference Questionnaire showed that “verbal reassurance from medical staff” (45.2%), “distraction” (43.3%), and “understanding the surgical procedure” (39.5%) were the most preferred intervention strategies among patients. Conclusions Preoperative dental anxiety is common among some patients undergoing oral implant surgery, particularly young and middle-aged patients, those treated by less experienced physicians, and those experiencing anxiety due to a fear of pain or exposure to an unfamiliar environment and strangers. Clinical attention should focus on these patient populations, and systematic preoperative education and psychological counseling should be provided accordingly.
Objective To deeply explore the systemic management dilemmas affecting patients with chronic periodontitis in underdeveloped regions, providing practical pathways for achieving oral health equity. Methods This study has been reviewed and approved by the Medical Ethics Committee, and written informed consent was obtained from the participants. Based on a comprehensive design integrating literature review and theory, an interview guide was developed. Using purposive sampling, semi-structured interviews were conducted with 20 patients diagnosed with chronic periodontitis who were from underdeveloped regions and attended the Stomatological Hospital of Southern Medical University between November 2025 and February 2026. The sample size was determined by data saturation. Data analysis employed Colaizzi’s seven-step method. Results Through interview with 20 participants, three core themes were extracted, encompassing eight sub-themes: individual cognitive-behavioral dilemmas (cognitive limitations and passive coping, inefficient and superficial self-management, and economic and psychological barriers to seeking care); systemic and environmental structural obstacles (distrust in primary healthcare capabilities, and the limited buffering effect and constraints of medical insurance); and the need for health empowerment and support systems (authoritative knowledge and management skill guidance, aspirations for access to high-quality specialized medical resources, and sustained and reminder-based remote support). Conclusion Patients with chronic periodontitis in underdeveloped regions face a multidimensional dilemma composed of limited health literacy, poor adherence to health behaviors, weak primary care service capacity, uneven medical insurance coverage, and insufficient high-quality specialized medical resources. Patients’ health literacy should be advanced through innovative health education, primary care capacity should be strengthened for periodontal disease prevention and treatment, and it is recommended to prioritize the inclusion of initial periodontal therapy in outpatient coordination or chronic disease management, and a remote medical support system based on digital technology should be established.
Objective To explore the metabolic molecular mechanism of regulatory T cells(Tregs)resisting ferropto-sis under a hypoxic stress microenvironment,and to provide scientific support for finding intervention targets to improve the immune microenvironment of oral squamous cell carcinoma(OSCC).Methods This study was approved by the In-stitutional Medical Ethics Committee.Multiplex immunofluorescence was used to detect the distribution of Tregs and the localization of hypoxic regions in clinical pathological specimens from patients with OSCC tissues.Animal experi-ments were approved by the Laboratory Animal Ethics Committee.A tumor-bearing model was established in C57BL/6 mice.The submandibular,subscapular,inguinal,and popliteal lymph nodes were collected along with the spleen,and were homogenized to prepare single cell suspensions.C57BL/6 mouse naive CD4⁺ T cells were induced to differentiate into Tregs in vitro,and a 1%hypoxic culture model was established.Normoxia and hypoxia groups were established,and Treg viability under hypoxic conditions was assessed by flow cytometry.Transcriptomics and metabolomics were ap-plied to analyze differentially expressed genes,metabolic pathways,and changes in cholesterol metabolites.Western blot was performed to detect the expression of emopamil-binding protein(EBP),glutathione peroxidase 4(GPX4),ferrop-tosis suppressor protein 1(FSP1),and acyl-CoA synthetase long-chain family member 4(ACSL4).The safe concentra-tion of the EBP inhibitor TASIN-30 was determined by the CCK-8 assay.On this basis,four groups were established:normoxia control,hypoxia,hypoxia+TASIN-30,and hypoxia+TASIN-30+7-dehydrocholesterol(7-DHC).Treg viability in each group was measured by flow cytometry;intracellular Fe²⁺ content,lipid peroxidation level,and reactive oxygen spe-cies(ROS)accumulation were detected by cellular immunofluorescence and flow cytometry;the reduced/oxidized gluta-thione(GSH/GSSG)ratio was measured using a microplate reader;and mitochondrial morphology was observed by trans-mission electron microscopy.In vivo tumor-inhibition experiments in C3H/HeJ mice were performed to validate the in vivo effects of EBP inhibition,and clinical prognostic survival analysis based on the TCGA-HNSC cohort was performed to evaluate the clinical prognostic value of ferroptosis resistance in Tregs.Results Tregs were highly enriched in OSCC tissue,preferentially accumulated in hypoxic areas,and could maintain cell survival under hypoxia.The choles-terol metabolism pathway was upregulated in the hypoxic group,with increased EBP mRNA expression(P<0.001)and 7-DHC accumulation.Compared with the hypoxic group alone,the hypoxic+TASIN-30 group showed decreased GPX4 and FSP1 expression(P<0.001,P=0.003);increased ACSL4 expression(P<0.001);a decreased viable cell ratio(P<0.001);increased Fe2+,lipid peroxidation,and ROS levels(P<0.001);a decreased GSH/GSSG ratio(P<0.001);and mitochondrial shrinkage.Exogenous supplementation of 7-DHC reversed the effects of EBP inhibition.In vivo inhi-bition of EBP significantly inhibited tumor growth(P<0.001).Conclusion Tregs upregulate EBP and promote 7-DHC accumulation in hypoxic microenvironments,thereby inhibiting Treg ferroptosis and maintaining their survival and immunosuppressive function.Inhibition of EBP can weaken the resistance of Tregs to ferroptosis under hypoxia.
Objective To explore the influencing factors of incident caries at age 5 among caries-free 3-year-old children in Beijing, construct a machine learning prediction model based on oral health behavioral data, and provide a scientific basis and practical tool for early risk identification, stratified prevention and control, and home-kindergarten-medical collaborative intervention of incident dental caries in preschool children. Methods This study has been reviewed and approved by the Medical Ethics Committee, and written informed consent was obtained from the guardians of the study participants and their guardians. Children with complete baseline data at age 3 and follow-up data at age 5 from Beijing public oral health programs were included. Oral examinations were conducted according to World Health Organization diagnostic criteria, and questionnaires were completed by guardians. Risk factors were screened by Logistic regression. The total sample was randomly divided into a test set (20%) and a validation set (80%). Five-fold cross-validation was performed on the validation set for model training and tuning, and the final model performance was evaluated on the test set. Four machine learning models—decision tree, random forest, Extreme Gradient Boosting (XGBoost), and Light Gradient Boosting Machine (LightGBM)—were constructed. Model performance was evaluated using the area under the curve (AUC), accuracy, and specificity. Shapley Additive Explanations (SHAP) was used for model interpretation. Results The incidence of incident caries for 3 372 5-year-old children in Beijing was 50.96%. Multivariate logistic regression showed that parental caries, sleeping with milk, frequent feeding at night, no postprandial cleaning, high sugar intake, insufficient bedtime tooth brushing, and non-fluoride toothpaste use were significant risk factors (P < 0.05). Although all four machine learning models performed well, the XGBoost model performed the best (AUC = 0.806). Feature importance and SHAP analysis indicated that the frequency of bedtime tooth brushing, the frequency of candy consumption, and parental caries status were the top three predictors. Conclusion The XGBoost model based on oral health behaviors at age 3 can accurately predict the risk of incident caries in 5-year-old children in Beijing. Bedtime tooth brushing, sugar control, and parental oral health management are key points for caries prevention in preschool children.
