Comparison of Easydo Activator, ultrasonic and needle irrigation techniques on sealer penetration and smear layer removal in vitro.

The effects of Easydo Activator (EA), a new sonic irrigation system, on sealer penetration at the root apex were compared to needle irrigation (NI) and passive ultrasonic irrigation (PUI) in this study. Forty-two single-rooted teeth were prepared and randomly divided into three groups (n = 14): group 1: NI; group 2: PUI; and group 3: EA. A solution of 3% sodium hypochlorite (NaOCl) was used for irrigation. Nine teeth in each group were filled with AH Plus sealer mixed with CY5 fluorescent dye and a single gutta-percha cone. The sealer penetration area, maximum penetration depth and percentage of sealer penetration at 5 mm and 1 mm from the apex were analyzed by confocal laser scanning microscopy (CLSM). The remaining 5 teeth in each group were subjected to test smear layer scores by scanning electron microscopy (SEM). The CLSM evaluation showed that increases in the area, depth and percentage of sealer penetration were detected at 1 and 5 mm from the root apex in the PUI group compared with the NI group, and greater increases were observed in the EA group (P < 0.05). The SEM experiment showed that the lowest scores for the smear layer and debris removal were achieved by the EA group when compared with the PUI and NI groups (P < 0.05). In conclusion, EA was superior to PUI and NI regarding sealer penetration at the root apex during endodontic treatment, and it could provide a new technical idea for clinical root canal therapy.

Cross-cultural adaptation and psychometric properties of the Mainland Chinese version of the manchester orofacial pain disability scale (MOPDS) among college students.

BACKGROUND: Orofacial pain (OFP) is a highly prevalent disorder in mainland China that predisposes to an associated physical and psychological disability. There is lack of a good properties mainland Chinese version of instrument to examine OFP. This study aims to cross-cultural adaptation and evaluate psychometrics properties of the Manchester Orofacial Pain Disability Scale (MOPDS) in mainland Chinese Mandarin context. METHODS: Translation and cross-cultural adaption of the mainland Chinese version MOPDS were conducted following accepted guidelines of self-report measures. Chinese college students (N = 1039) completed the mainland Chinese version of the MOPDS for item analysis, reliability and validity tests, and measurement invariance analysis, and after a one-month interval, around 10% of the sample (n = 110) were invited to retest. To conduct the CFA and measurement invariance analysis, Mplus 8.4 was used. IBM SPSS Statistics 26 software were used for all additional studies. RESULTS: We found that the mainland Chinese version of MOPDS contains 25 items, divided into two categories: physical disability and psychological disability. The scale demonstrated excellent internal reliability, test-retest reliability, and validity. The measurement invariance results proved that the scale could be applied to people of different gender, age, and health consultation status. CONCLUSIONS: The results demonstrated the mainland Chinese version of MOPDS has good psychometric properties and can be used to measure the level of physical and psychological disability of Chinese OFP peoples.

Tooth ultrastructure changes induced by a nonsense mutation in the FAM83H gene: insights into the diversity of amelogenesis imperfecta.

OBJECTIVES: The current research on single-nucleotide polymorphism (SNP) mutation sites at different positions of the FAM83H gene and their phenotypic changes leading to amelogenesis imperfecta (AI) is inconsistent. We identified a previously reported heterozygous nonsense mutation c.1192C>T (p.Q398*) in the FAM83H gene and conducted a comprehensive analysis of the dental ultrastructure and chemical composition changes induced by this mutation. Additionally, we predicted the protein feature affected by this mutation site. The aim was to further deepen our understanding of the diversity of AI caused by different mutation sites in the FAM83H gene. METHODS: Whole-exome sequencing (WES) and Sanger sequencing were used to confirm the mutation sites. Physical features of the patient's teeth were investigated using various methods including cone beam computer tomography (CBCT), scanning electron microscopy (SEM), contact profilometry (roughness measurement), and a nanomechanical tester (nanoindentation measurement). The protein features of wild-type and mutant FAM83H were predicted using bioinformatics methods. RESULTS: One previously discovered FAM83H heterozygous nonsense mutation c.1192C>T (p.Q398*) was detected in the patient. SEM revealed inconsistent dentinal tubules, and EDS showed that calcium and phosphorus were lower in the patient's dentin but higher in the enamel compared to the control tooth. Roughness measurements showed that AI patients' teeth had rougher occlusal surfaces than those of the control tooth. Nanoindentation measurements showed that the enamel and dentin hardness values of the AI patients' teeth were both significantly reduced compared to those of the control tooth. Compared to the wild-type FAM83H protein, the mutant FAM83H protein shows alterations in stability, hydrophobicity, secondary structure, and tertiary structure. These changes could underlie functional differences and AI phenotype variations caused by this mutation site. CONCLUSIONS: This study expands the understanding of the effects of FAM83H mutations on tooth structure. CLINICAL RELEVANCE: Our study enhances our understanding of the genetic basis of AI and may contribute to improved diagnostics and personalized treatment strategies for patients with FAM83H-related AI.

Factorial structure and measurement invariance of the Chinese version of the Oral Health Impact Profile-14 among clinical populations and non-clinical populations: an evidence for public oral investigations.

OBJECTIVE: Oral health-related quality of life (OHRQoL) is a multidimensional concept that is commonly used to examine the impact of oral health status on quality of life. The purpose of this study was to examine the optimal factor model of the Chinese version of the Oral Health Impact Profile (OHIP-14) questionnaire in clinical populations, measurement invariance across clinical status and gender cohorts. This would ensure equal validity of the Chinese version of OHIP-14 in different populations and further support public oral investigations. METHODS: The Chinese version of OHIP-14 was used to investigate 490 dental patients and 919 college students. Confirmatory factor analysis (CFA), item analysis and reliability, measurement invariance, and the t-test were used for data analyses. RESULTS: We found that the 7-factor structure had the best-fit index in the sample (CFI = 0.970, TLI = 0.952; SRMR = 0.029, RMSEA = 0.052(0.040,0.063)). The reliability of the scales was satisfactory (Cronbach's α = 0.942). The error variance invariance fitted the data adequately in measurement invariance, indicating that measurement invariance is acceptable both across the clinical and non-clinical populations (∆CFI=-0.017, ∆RMSEA = 0.010) and across genders in the clinical population (∆CFI = 0.000, ∆RMSEA=-0.003). T-test for scores showed that the clinical populations scored significantly higher than the non-clinical populations, as did the overall score (t = 7.046, p < 0.001, d = 0.396), in terms of functional limitation (t = 2.178, p = 0.030, d = 0.125), physical pain (t = 7.880, p < 0.001,d = 0.436), psychological discomfort (t = 8.993, p < 0.001, d = 0.514), physical disability (t = 6.343, p < 0.001, d = 0.358), psychological disability (t = 5.592, p < 0.001, d = 0.315), social disability (t = 5.301, p < 0.001,d = 0.304), social handicap (t = 4.452, p < 0.001, d = 0.253), and that in the non-clinical populations, females scored significantly higher than males, as did in terms of physical pain (t = 3.055, p = 0.002, d = 0.280), psychological discomfort (t = 2.478, p = 0.014, d = 0.222), and psychological disability (t = 2.067, p = 0.039, d = 0.188). CONCLUSION: This study found that the Chinese version of OHIP-14 has measurement invariance between the clinical and non-clinical populations and across genders in the clinical populations, and can be widely used in OHRQoL assessment for public oral investigations.

Comparison of EASYDO ACTIVATOR, passive ultrasonic, and needle irrigation techniques on the treatment of apical periodontitis: a study in rats.

OBJECTIVES: To evaluate the long-term therapeutic effect of EASYDO ACTIVATOR, passive ultrasonic irrigation, and needle irrigation in experimental apical periodontitis in rats. MATERIALS AND METHODS: Sprague-Dawley male rats were used to produce periapical lesions. The pulp chambers of the bilaterally first mandibular molars were exposed and left open for 21 days. The rats were divided into four groups according to different irrigation protocols. Seven days after irrigation, the mandibles were removed for micro-CT, histological, and immunohistochemical analysis. Serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were assessed by enzyme-linked immunosorbent assays (ELISA). Statistical data were analyzed by one-way analysis of variance (ANOVA) with LSD tests. RESULTS: The passive ultrasonic irrigation and EASYDO ACTIVATOR groups had the smallest apical lesions compared to the other groups (P < 0.05), while the needle irrigation group had smaller lesions than the control group (P < 0.05). The EASYDO ACTIVATOR group had less inflammation infiltration compared to the control and needle irrigation groups (P < 0.05). The control and needle irrigation groups had more TNF-α expression compared to the passive ultrasonic irrigation and EASYDO ACTIVATOR groups (P < 0.05). The lowest IL-6 expression was observed in the EASYDO ACTIVATOR group. The EASYDO ACTIVATOR group had the lowest serum level of TNF-α than other groups (P < 0.05). IL-6 expression was significantly lower in the EASYDO ACTIVATOR group in comparison with the control and needle irrigation groups (P < 0.05). CONCLUSIONS: EASYDO ACTIVATOR can significantly reduce the apical lesions and decrease the inflammatory response around the periapical area. CLINICAL RELEVANCE: EASYDO ACTIVATOR is recommended for clinical application.

Comparison of sealer penetration of sonic activation versus conventional needle irrigation: a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Most existing studies comparing the efficiency of sonic irrigation (SI) and conventional needle irrigation (CNI) in increasing the penetration of sealers into dentine tubules are controversial; and this study aimed to determine whether the use of SI can lead to greater sealing ability than CNI, during the root canal treatment. METHODS: The EMBASE, PubMed, and Cochrane Library databases were used to find confocal laser scanning microscopy studies evaluating percentage and maximum depth of sealer penetration following the use of SI or CNI in mature permanent teeth until October 2022. The critical estimative checklist of randomized controlled trials of the standardized Joanna Briggs Institute was adopted to independently score the quality of each study. The random-effect model for meta-analysis was used to analyse for each canal segment (apical, middle, coronal). The results are shown in the forest plots as weighted mean differences (WMDs) with 95% confidence intervals (95% CIs). RESULTS: Ninety-seven articles were included in the preliminary screening, and nine of them were included in this study. Eight studies were included in the meta-analysis.The meta-analysis exhibited great increases in the coronal (WMD: 8.09, 95% CI 2.78-13.40/WMD: 165.32, 95% CI 128.85-201.80), and middle segments (WMD: 8.81, 95% CI 5.76-11.87/WMD: 132.98, 95% CI 68.71-197.25) for the percentage and maximum depth of sealer penetration, respectively. The percentage of sealer penetration in the apical thirds region was nonsignificant (WMD: 4.73, 95% CI - 2.34-11.80). However, the maximum depth of sealer penetration in the apical thirds region was significant (WMD: 121.46, 95% CI 86.55-156.38). Chi-squared analysis revealed heterogeneity scores of 0.0-70.0% and 44.0-90.0% for the percentage and maximum depth of sealer penetration, respectively. DISCUSSION: This review verified that SI significantly improves tubular dentin sealer penetration in most areas of the root canal; thus, SI may lead to better filling efficiency and anti-reinfection effects than CNI during and after the root canal therapy. Nevertheless, a large heterogeneity in the current data comparing the irrigation efficiency of SI versus CNI in the apical third of the root canal was found, implying the necessity to standardize root canal irrigation procedures and obtain more accurate results in this area. TRIAL REGISTRATION: INPLASY database (INPLASY202270116).

Role of NOD2 and hepcidin in inflammatory periapical periodontitis.

The immunological response occurring during periapical inflammation includes expression of nucleotide binding oligomerization domain containing 2 and hepcidin. Nucleotide binding oligomerization domain containing 2 deficiency increases infiltration of inflammatory cells close to alveolar bone. Hepcidin has an important role in iron metabolism affecting bone metabolism.We investigated the role of nucleotide binding oligomerization domain containing 2 and hepcidin in inflammatory periapical periodontitis. Periapical periodontitis was induced in rats and confirmed by micro-computed tomography. Nucleotide binding oligomerization domain 2 and hepcidin were evaluated through immunohistochemistry. Bioinformatics analysis was undertaken usingthe Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. Micro-computer tomography revealed alveolar bone resorption in the periapical region and furcation area of mandibular molars in rats of the periapical periodontitis group. Immunohistochemistry showed increased expressionof nucleotide binding oligomerization domain containing 2 and hepcidin around root apices in rats of the periapical periodontitis group. Bioinformatics analysis of differentially expressed genes in inflamed and non-inflamed tissues revealed enrichment in the NOD-like receptor signaling pathway. Our data suggest that nucleotide binding oligomization domain contain2 and hepcidin have important roles in periapical periodontitis severity because they can reduce alveolar bone loss.They could elicit new perspectives for development of novel strategies for periapical periodontitis treatment.

The Chinese version of the Oral Health Impact Profile-14 (OHIP-14) questionnaire among college students: factor structure and measurement invariance across genders.

BACKGROUND: The Oral Health-related Quality of Life (OHRQoL) is a multi-dimensional concept commonly used to examine the impact of health status on quality of life, and the Oral Health Impact Profile-14 (OHIP-14) questionnaire is a good self-assessment tool. This study was designed to investigate the factor structure of the OHIP-14 scale Chinese version, measurement invariance and latent mean differences across genders among college students. METHODS: The online survey was completed by 919 college students. This study used confirmatory factor analysis (CFA) to check the structural models of the OHIP-14 scale, The correlation of each item with the scale total score could test homogeneity, and Cronbach's alpha (Cronbach's α) could evaluate internal consistency. Multi-group CFA was used to explore whether the Chinese version of the OHIP-14 scale was used in male and female populations for measurement consistency. T-test compared scores between men and women. Regression analyses were used to evaluate the relationship between age, gender, education, subject, and the score on the OHIP-14 scale. RESULTS: We found that the 7-factor structure had the best fit index in the sample. According to Cronbach's α, the overall score of OHIP was 0.958, and Cronbach's α for 7 factors was: functional limitation was 0.800, physical pain was 0.854, psychological discomfort was 0.902, physical disability was 0.850, psychological disability was 0.768, social disability was 0.862, social handicap was 0.819 and the test-retest reliability interval was 0.723. Multi-group confirmatory factor analysis supported residual measurement invariance across gender. T-test for scores showed that females scored higher significantly than men as did the overall score, in terms of physical pain (p<0.001), physical disability (p<0.001), and psychological disability (p<0.001). CONCLUSIONS: This study found the OHIP-14 Chinese version to be a good tool for assessing the college students' OHRQoL in China, allowing people to conduct self-assessments.

Adiponectin regulates osteocytic MLO-Y4 cell apoptosis in a high-glucose environment through the AMPK/FoxO3a signaling pathway.

Clinical studies have shown that persistent hyperglycemia following oxidative stress is associated with the apoptosis of osteocytes in diabetics. Adiponectin (APN) can ameliorate oxidative stress, and its receptors have been identified in bone-forming cells. However, the relationship between APN and osteocyte apoptosis has not been fully elucidated. This study aimed to investigate whether APN could prevent osteocyte apoptosis and regulate reactive oxygen species (ROS) generation in a high-glucose environment. Hoechst staining and fluorescence microscopy were used to observe the apoptosis of osteocytic MLO-Y4 cells. Real-time quantitative polymerase chain reaction and Western blot analysis were used to detect the expression of Caspase 3, Caspase 8, and Bcl-2. ROS generation was investigated with an active oxygen kit and fluorescence microscopy. Furthermore, the expression of proteins in the AMPK/FoxO3A signaling pathway was also studied by Western blot analysis. In a high-glucose environment, APN promoted the proliferation of MLO-Y4 osteocytes and the expression of Bcl-2 but inhibited the expression of Caspase 3, Caspase 8, and ROS in a dose-dependent manner. APN promoted the activation of p-AMPK and p-AMPK/AMPK, which reached their highest levels at 10 min and returned to baseline at 30 min. The expression of p-FoxO3A/FoxO3A in both the cytoplasm and nucleus peaked at 15 min, and this expression was returned to baseline at 60 min. In summary, APN has an antiapoptotic effect and regulates ROS generation in MLO-Y4 osteocytes in a high-glucose environment. The AMPK/FoxO3A signaling pathway might be a key signaling pathway that participates in the effect of APN on regulating osteocyte apoptosis in diabetics.

miR-124-3p increases in high glucose induced osteocyte-derived exosomes and regulates galectin-3 expression: A possible mechanism in bone remodeling alteration in diabetic periodontitis.

The mechanisms underlying the two-way relationship between diabetes mellitus (DM) and periodontitis are unclear. We examined a possible effect of galectin-3 (Gal-3), a factor in DM and bone metabolism, on periodontitis with or without DM. Using enzyme-linked immunosorbent assay, we detected saliva Gal-3 in patients with periodontitis, with or without type 2 diabetes mellitus (T2DM). In animal models, we measured periodontal bone microarchitecture via micro computed tomography, and detected Gal-3, Runt-related transcription factor 2 (Runx2), and interleukin-6 (IL-6) expression in alveolar bone. Applying dual luciferase reporter assay, we explored the target binding of miR-124-3p and Gal-3. We examined osteocyte-derived exosomes with transmission electron microscopy and detected miR-124-3p, Gal-3, and IL-6 expression in exosomes. Saliva Gal-3 was increased in DM compared with controls but decreased in patients with moderate periodontitis and DM compared with those who had moderate periodontitis only. Alveolar bone mass was increased in DM and exacerbated in DM with periodontitis. Gal-3 and Runx2 were both increased in periodontitis and DM compared with controls, but decreased in DM with periodontitis compared with DM alone. MiR-124-3p targeted and inhibited Gal-3 expression in vitro. Osteocytes secreted exosomes carrying miR-124-3p, Gal-3, and IL-6, which were influenced by high glucose. These findings indicate that osteocyte-derived exosomes carrying miR-124-3p may regulate Gal-3 expression of osteoblasts, especially under high-glucose conditions, suggesting a possible mechanism for DM-related alveolar bone pathologies.

Endoplasmic reticulum stress remodels alveolar bone formation after tooth extraction.

Bone healing in tooth extraction sockets occurs in a complex environment containing saliva and many microorganisms and is affected by many factors. Endoplasmic reticulum (ER) stress affects bone metabolism, but the role of ER stress in bone healing after tooth extraction remains unclear. We utilized a rat tooth extraction model, in which we promoted wound healing by using salubrinal to regulate the ER stress response. Western blot analysis showed increased expression of p-eIF2α/eIF2α, Runx2 and alkaline phosphatase (ALP) in bone tissue, and histological assays showed irregularly arranged and new bone with more collagen fibres 14 days after tooth extraction and after modulating the degree of ER stress. Micro-CT showed that modulating ER stress to an appropriate degree increases bone filling in regards to the density in the bottom and the surrounding bone wall of the tooth extraction wounds. Transmission electron microscopy showed rough ER expansion and newly formed collagen fibrils in osteoblasts after modulating ER stress to an appropriate degree. We also used different concentrations of salubrinal to evaluate the resistance to tunicamycin-induced ER stress in an osteogenic induction environment. Salubrinal restored the tunicamycin-induced decrease in the viability of primary calvarial osteoblasts and increased the expression of Runx2 and ALP, and decreased p-eIF2α/eIF2α in a dose-dependent manner. Taken together, the results demonstrate that ER stress occurred after tooth extraction, and regulating the degree of ER stress can promote bone healing in tooth extraction sockets, providing clinical evidence for bone healing.

Selective ß2-adrenoreceptor signaling regulates osteoclastogenesis via modulating RANKL production and neuropeptides expression in osteocytic MLO-Y4 cells.

The ß2-adrenergic receptor (ß2-AR) signaling on bone cells is the major contributor in the effect of the sympathetic nervous system on bone turnover. However, it remains unclear whether receptor activator of nuclear factor κ-Β ligand (RANKL) modulation and neuropeptides expression in osteocytes are responsible for the mechanism. This study used ß2-AR stimulation to investigate cell cycle and proliferation, the gene and protein expression of RANKL, and osteoprotegerin (OPG), as well as neuropeptides regulation in osteocytic MLO-Y4 cells. Clenbuterol (CLE; a ß2-AR agonist) slightly promoted the growth of MLO-Y4 cells in a concentration-dependent effect but had no effect on the proliferation index. And the concentration of 10-8 M showed a significant increase in the S-phase fraction on day 3 in comparison with the control. Additionally, CLE-promoted osteoclast formation and bone resorption in osteocytic MLO-Y4 cell-RAW264.7 cell cocultures. RANKL expression level and the ratio of RANKL to OPG in MLO-Y4 cells were enhanced in CLE treatment but were rescued by blocking ß2-AR signaling. However, neuropeptide Y and α-calcitonin gene-related peptide, two neurogenic markers, were inhibited in CLE treatment of MLO-Y4 cells, which was reversed by a ß2-AR blocker. The results indicate that osteocytic ß2-AR plays an important role in the regulation of RANKL/OPG and neuropeptides expression, and ß2-AR signaling in osteocytes can be used as a new valuable target for osteoclast-related pathologic disease.

Serum and salivary ferritin and Hepcidin levels in patients with chronic periodontitis and type 2 diabetes mellitus.

BACKGROUND: Iron disorder and abnormal expression of hepcidin play important roles in many diseases, but it is still unclear in chronic periodontitis (CP) and type 2 diabetes mellitus (T2DM). We aimed to assess ferritin and hepcidin levels in serum and saliva of CP patients with or without T2DM. METHODS: Serum and unstimulated whole saliva samples were collected from 88 participants, who were categorized into 4 groups based on the presence or absence of CP or T2DM. Demographics and general health parameters were recorded. Full-mouth clinical periodontal parameters including probing pocket depth, clinical attachment loss, bleeding index, and plaque index were recorded. Chemiluminescence microparticle immunoassay and enzyme-linked immunosorbent assay were used to detect ferritin and hepcidin concentrations, respectively, in serum and saliva. RESULTS: Serum ferritin and hepcidin levels in the CP and CP with T2DM groups were higher than in the control group (P < 0.05). Serum hepcidin and serum ferritin are linear correlated (P < 0.001). Serum hepcidin/ferritin values in the CP with T2DM group were significantly lower than those in the T2DM and control groups. Moreover, salivary ferritin levels in the CP and T2DM groups were higher than those in the control group (P < 0.05). There was positively correlation between salivary ferritin and serum ferritin (P = 0.017). Hepcidin concentrations were relatively low in saliva. CONCLUSIONS: These results suggest that iron overload and hepcidin inadequacy existed in CP with T2DM patients. Salivary ferritin might provide a reference for body iron load. TRIAL REGISTRATION: ChiCTR-ROC-17012780.

[Remineralization of demineralized dentin induced by bioactive glass NovaMin].

OBJECTIVE: To explore the remineralization effect of bioactive glass NovaMin on demineralized dentin specimens, and to study the physical and chemical properties of formed structure at dentin surface.
 Methods: One mm-thickness coronal dentin slices were soaked in ethylene diamine tetraacetic acid (EDTA) for 48 h to prepare the completely demineralized dentin specimens and they were divided into 2 groups: an artificial saliva group (control group) and a NovaMin powder group. The specimens were treated with artificial saliva or NovaMin powder for 2 min (2 times every day), and the interval was 8 hours. Then, the specimens were soaked in the remineralization solution. After 7 days, the scanning electron microscope (SEM), energy dispersive X-ray (EDX), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray diffraction (XRD) were used to detect dentin morphology, the physical and chemical properties of the formed structure at dentin surface.
 Results: The results of SEM showed that a defined surface layer in the NovaMin powder group could be observed in the SEM imaging at the 7th day, which completely occluded dentinal tubules; the EDX, ATR-FTIR and XRD analysis found that the mineralized layer formed at dentin surface was mainly composed of calcium and phosphate elements, which was similar to the hydroxyapatite-like crystal. However, there were no materials formed at the dentin surface in the control group, and the dentinal tubules were still open.
 Conclusion: NovaMin can remineralize the demineralized dentin specimens and occlude the dentinal tubules in hydroxyapatite-like crystal structure.

Insulin-like growth factor-1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA-binding protein LARP6 expression.

This study explored the mechanism underlying the stimulation of collagen synthesis and osteoblastic differentiation by insulin-like growth factor 1 (IGF1) in primary mouse osteoblasts. Primary mouse calvarial osteoblasts were cultured and treated with various doses of IGF1 before transfection with siRNA targeting the collagen type I alpha 2 (Col1a2) or La ribonucleoprotein domain family member 6 (Larp6) genes. Alkaline phosphatase (ALP) activity, osteocalcin staining, alizarin red quantification and the expression level of runt-related transcription factor 2 (RUNX2) were performed to assess the differentiation of pre-osteoblasts. Based on Western blot analysis, IGF1 up-regulated COL1A2 protein expression in the primary osteoblasts in a dose- and time-dependent manner. In addition, Col1a2 interference inhibited the differentiation and mineralization of osteoblasts. IGF1 also stimulated the differentiation of mouse primary osteoblasts and increased LARP6 expression during osteogenic differentiation. RNA-Immunoprecipitation (IP) indicated that LARP6 could bind to Col1a2 mRNA after IGF1 stimulation. However, transfection of Larp6-specific siRNA significantly reduced collagen and ALP secretion, mineralization and inhibited the expression of osteocalcin and RUNX2, indicating that Larp6 interference inhibited the differentiation ability of primary mouse calvarial osteoblasts, and these effects could not be reversed by IGF1. Thus, IGF1 could promote COL1A2 expression and osteoblast differentiation in primary mouse calvarial pre-osteoblasts by increasing LARP6 expression via a posttranscriptional mechanism.

Insulin receptor substrate-1 time-dependently regulates bone formation by controlling collagen Iα2 expression via miR-342.

Insulin promotes bone formation via a well-studied canonical signaling pathway. An adapter in this pathway, insulin-receptor substrate (IRS)-1, has been implicated in the diabetic osteopathy provoked by impaired insulin signaling. To further investigate IRS-1's role in the bone metabolism, we generated Irs-1-deficient Irs-1smla/smla mice. These null mice developed a spontaneous mutation that led to an increase in trabecular thickness (Tb.Th) in 12-mo-old, but not in 2-mo-old mice. Analyses of the bone marrow stromal cells (BMSCs) from these mice revealed their differential expression of osteogenesis-related genes and miRNAs. The expression of miR-342, predicted and then proven to target the gene encoding collagen type Iα2 (COL1A2), was reduced in BMSCs derived from Irs-1-null mice. COL1A2 expression was then shown to be age dependent in osteoblasts and BMSCs derived from Irs-1smla/smla mice. After the induction of osteogenesis in BMSCs, miR-342 expression correlated inversely with that of Col1a2 Further, Col1a2-specific small interfering RNA (siRNA) reduced alkaline phosphatase (ALP) activity and inhibited BMSC differentiation into osteocyte-like cells, both in wild-type (WT) and Irs-1smla/smla mice. Conversely, in Irs-1smla/smla osteocytes overexpressing COL1A2, ALP-positive staining was stronger than in WT osteocytes. In summary, we uncovered a temporal regulation of BMSC differentiation/bone formation, controlled via Irs-1/miR-342 mediated regulation of Col1a2 expression.-Guo, Y., Tang, C.-Y., Man, X.-F., Tang, H.-N., Tang, J., Wang, F., Zhou, C.-L., Tan, S.-W., Feng, Y.-Z., Zhou, H.-D. Insulin receptor substrate-1 time-dependently regulates bone formation by controlling collagen Iα2 expression via miR-342.

The dentin tubule occlusion effects of desensitizing agents and the stability against acids and brushing challenges.

PURPOSE: To investigate the effects of desensitizing agents on dentin tubule occlusion, acid and tooth brushing challenge, and microhardness change of human dentin. METHODS: Partially demineralized dentin slabs were divided into four groups (n = 30): (1) Control, (2) Non-desensitizing toothpaste, (3) Pro-Argin toothpaste, (4) CPP-ACP paste. The specimens were treated with these dentifrices for 2 minutes/day and soaked in artificial saliva (AS) for 24-hour remineralization. Then the dentin discs were divided into three subgroups for removal resistance tests: acid challenge, mechanical brushing challenge and blank control. Changes in dentin morphology were observed using scanning electron microscopy (SEM). Vickers microhardness measurements were performed at baseline and after 24-hour remineralization for all groups. RESULTS: A surface layer and intra-tubular crystals were observed in SEM imaging of Pro-Argin toothpaste and CPP-ACP paste groups, which occluded most of the dentin tubules for those specimens. But the dentin tubules were opened after the acid challenge again. Moreover, the dentin microhardness showed a slight increase after 24-hour AS immersion. The percentage microhardness gain (PMG) values of these two groups were 5.4% and 5.1% respectively, which were significantly higher than the other groups (P < 0.05).

Dental health and treatment needs in people with leprosy in China.

OBJECTIVES: To evaluate the dental health status and treatment needs of people affected by leprosy in China, and provide a basis for the development of national or regional dental health programmes to cover the treatment needs of this population. DESIGN: A cross-sectional study with 613 former leprosy patients was carried out in six leprosy villages, in three provinces in China (Nanjing, Taixing and Jiangyan in Jiangsu Province, Hanzhong and Shangluo in Shanxi Province and Yongzhou in Hunan Province). A questionnaire about demographic and clinical data was used. The World Health Organization's (WHO) basic methods were used to determine the tooth-based treatment needs. Periodontal status was determined by using the Community Periodontal Index of Treatment Needs (CPITN). In addition, prosthetic normative needs were assessed. RESULTS: Among the 613 people affected by leprosy, there were 472 people (77%) who had never visited a dentist and 172 people (28.1%) had never brushed their teeth; 302 (49.3%) brushed their teeth once a day. However, there were 267 people (43.6%) who thought their dental health was at an average level and 108 (17.6%) thought they had good dental health. 55.6% of the subjects required dental fillings, 32.7% required pulp care and restoration, and 71.1% required extraction. On CPITN, 23.2% of the subjects scored 2, 28.6% scored three and 48.0% scored four, showing that these people required systematic periodontal treatment. In addition, 84.5% of the subjects needed normative prosthetic treatment. CONCLUSIONS: Most of the subjects with leprosy in this study lacked self-care knowledge on dental health, and especially self-awareness of dental conditions. Normative treatment needs of people affected by leprosy were very high. This result calls for improved oral health education and oral health care in people with leprosy. Oral health education might preferably be integrated into already existing leprosy rehabilitation programs.

The Gothenburg Trismus Questionnaire in China: Cross-cultural adaptation and measurement invariance.

OBJECTIVES: The Gothenburg Trismus Questionnaire (GTQ) is a comprehensive scale for screening and assessing trismus in head and neck (H&N) cancer and temporomandibular joint disorders (TMD) patients. This study aimed to translate and cross-culturally adapt the GTQ in China, and to test its measurement invariance. METHODS: This study comprised 278 H&N cancer, 245 TMD, and 507 control patients. Internal consistency and test-retest reliability were tested to assess the GTQ's reliability. The validity was evaluated through composite reliability (CR), average variance extracted (AVE), and correlation tests. Multi-group confirmatory factor analysis (CFA) was used to investigate the GTQ's measurement invariance across clinical status and gender. T tests were employed to compare score differences across clinical status and gender. RESULTS: The Chinese version of GTQ scale shows excellent internal consistency and test-retest reliability. The CR, AVE, and correlation values demonstrate the good validity of GTQ. The multi-group CFA supported configural invariance across clinical status but not metric invariance, while it supported strict invariance across gender. Additionally, t tests revealed that patients with H&N cancer and TMD scored higher than the control group, while males scored higher than females. CONCLUSIONS: The Chinese version of GTQ serves as an effective tool for screening and assessing trismus.

Prevotella, a dominant bacterium in young people with stage â ¢ periodontitis, related to the arachidonic acid metabolism pathway.

OBJECTS: As periodontitis progresses, the oral microbiome changes dynamically. The aim of this study is to evaluate the dominant bacteria of adults with stage III periodontitis and investigate potential pathways related to the dominant bacteria. MATERIALS AND METHODS: 16S rRNA sequencing was carried out to detect the differences in the oral microbiome between adult with stage Ⅰ and stage Ⅲ periodontitis and find the dominant bacteria in each group. The inhibitor of the predominant pathway for stage Ⅲ periodontitis was used to investigate the role of the dominant bacteria in periodontitis in vivo and in vitro. RESULTS: There was no significant difference in the α-diversity between the two groups. The results of ß-diversity showed that the samples were divided into different groups according to the stage of periodontitis. The dominant bacteria in youths with stage Ⅲ periodontitis was Prevotella and may be related to the arachidonic acid metabolism pathway. Administration of SKF-86002 suppressed the expression of inflammation mediators in vivo and vitro. CONCLUSIONS: Prevotella was the one dominant bacteria in young people with stage Ⅲ periodontitis and was related to the arachidonic acid metabolism pathway.