625 nm Light Irradiation Prevented MC3T3-E1 Cells from Accumulation of Misfolded Proteins via ROS and ATP Production.

Osteoblasts must acquire a considerable capacity for folding unfolded and misfolded proteins (MPs) to produce large amounts of extracellular matrix proteins and maintain bone homeostasis. MP accumulation contributes to cellular apoptosis and bone disorders. Photobiomodulation therapy has been used to treat bone diseases, but the effects of decreasing MPs with photobiomodulation remain unclear. In this study, we explored the efficacy of 625 nm light-emitting diode irradiation (LEDI) to reduce MPs in tunicamycin (TM) induced-MC3T3-E1 cells. Binding immunoglobulin protein (BiP), an adenosine triphosphate (ATP)-dependent chaperone, is used to evaluate the capacity of folding MPs. The results revealed that pretreatment with 625 nm LEDI (Pre-IR) induced reactive oxygen species (ROS) production, leading to the increased chaperone BiP through the inositol-requiring enzyme 1 (IRE1)/X-box binding protein 1s (XBP-1s) pathway, and then restoration of collagen type I (COL-I) and osteopontin (OPN) expression relieving cell apoptosis. Furthermore, the translocation of BiP into the endoplasmic reticulum (ER) lumen might be followed by a high level of ATP production. Taken together, these results suggest that Pre-IR could be beneficial to prevent MP accumulation through ROS and ATP in TM-induced MC3T3-E1cells.

Detection of Abnormal Changes on the Dorsal Tongue Surface Using Deep Learning.

Background and Objective: The tongue mucosa often changes due to various local and systemic diseases or conditions. This study aimed to investigate whether deep learning can help detect abnormal regions on the dorsal tongue surface in patients and healthy adults. Materials and Methods: The study collected 175 clinical photographic images of the dorsal tongue surface, which were divided into 7782 cropped images classified into normal, abnormal, and non-tongue regions and trained using the VGG16 deep learning model. The 80 photographic images of the entire dorsal tongue surface were used for the segmentation of abnormal regions using point mapping segmentation. Results: The F1-scores of the abnormal and normal classes were 0.960 (precision: 0.935, recall: 0.986) and 0.968 (precision: 0.987, recall: 0.950), respectively, in the prediction of the VGG16 model. As a result of evaluation using point mapping segmentation, the average F1-scores were 0.727 (precision: 0.717, recall: 0.737) and 0.645 (precision: 0.650, recall: 0.641), the average intersection of union was 0.695 and 0.590, and the average precision was 0.940 and 0.890, respectively, for abnormal and normal classes. Conclusions: The deep learning algorithm used in this study can accurately determine abnormal areas on the dorsal tongue surface, which can assist in diagnosing specific diseases or conditions of the tongue mucosa.

Anti-Inflammatory and Mineralization Effects of Bromelain on Lipopolysaccharide-Induced Inflammation of Human Dental Pulp Cells.

Background and Objectives: Bromelain is a mixture of protease obtained from pineapple fruits or stems. Even though the biological mechanism of action of bromelain has not been completely understood, it is well known that bromelain possesses anticancer, anti-inflammatory and immunomodulatory effects. This study investigated the anti-inflammatory effects of bromelain on lipopolysaccharide (LPS)-induced human dental pulp cells (hDPCs). Materials and Methods: Cell viability after bromelain treatment was measured using WST-1 assay. We exposed hDPCs to 5 µg/mL of LPS with 2.5 or 5 µg/mL of bromelain. We performed reverse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay to detect interleukin-1ß, interleukin-6, and interleukin-8 levels. Western blots were used to detect intercellular adhesion molecules-1 (ICAM-1) and vascular cell adhesion molecules-1 (VCAM-1) levels. Immunofluorescence staining and Western blots were used to determine bromelain's anti-inflammatory mechanism. We also performed alkaline phosphatase and Alizarin red staining to verify mineralization nodule formation. Results: Bromelain at 2.5, 5, 10, or 20 µg/mL did not affect the viability of hDPCs significantly. LPS increased interleukin-1ß, interleukin-6, interleukin-8, ICAM-1 and VCAM-1 expression in hDPCs. Bromelain significantly decreased interleukin-1ß, interleukin-6, interleukin-8, ICAM-1, and VCAM-1 levels in hDPCs, which were stimulated by LPS. Bromelain treatment significantly reduced p65 phosphorylation in the cytoplasm and the nucleus. It also significantly decreased phosphorylation levels of extracellular signal-related kinases (ERK) and p38 mitogen-activated protein kinases (p38). Bromelain also promoted ALP activity and mineralized nodule formation. Conclusions: Bromelain inhibits the expression of inflammatory cytokines in LPS-stimulated hDPCs. The inhibitory effect of bromelain on inflammatory mediators is related to decreased NF-κB and the MAPK pathway. Therefore, bromelain might have the potential to be used for regenerative endodontics, including vital pulp therapy.

Anti-inflammatory effects of zinc in PMA-treated human gingival fibroblast cells.

OBJECTIVES: Abnormal cellular immune response has been considered to be responsible for oral lesions in recurrent aphthous stomatitis. Zinc has been known to be an essential nutrient metal that is necessary for a broad range of biological activities including antioxidant, immune mediator, and anti-inflammatory drugs in oral mucosal disease. The objective of this study was to investigate the effects of zinc in a phorbol-12-myristate-13-acetate (PMA)-treated inflammatory model on human gingival fibroblast cells (hGFs). STUDY DESIGN: Cells were pre-treated with zinc chloride, followed by PMA in hGFs. The effects were assessed on cell viability, cyclooxygenease-1,2(COX-1,2) protein expression, PGE2 release, ROS production and cytokine release, Results: The effects were assessed on cell viability, COX1/2 protein expression, PGE2 release, ROS production, cytokine release. The results showed that, in the presence of PMA, zinc treatment leads to reduce the production of ROS, which results in decrease of COX-2 expression and PGE2 release. CONCLUSIONS: Thus, we suggest that zinc treatment leads to the mitigation of oral inflammation and may prove to be an alternative treatment for recurrent aphthous stomatitis.

Photodynamic therapy (PDT) resistance by PARP1 regulation on PDT-induced apoptosis with autophagy in head and neck cancer cells.

BACKGROUND: Photodynamic therapy (PDT) is an anticancer treatment that generates excessive reactive oxygen species after photosensitizer treatments following specific wavelength irradiation. In another reports, PDT was regulated with autophagic cell death and apoptotic cell death. However, the mechanism of PDT resistance in PDT-stimulated cell death is unclear. In this study, we determined PDT resistance by autophagy and apoptosis in HP-PDT-treated oral cancer cells. MATERIALS & METHODS: Cells were treated hematoporphyrin and then irradiation with or without inhibitor. Cell lysates were checked protein expression with specific antibody. PDT resistance cells were generated with PDT repeated treatments. RESULTS: In HP-PDT, PDT induced autophagy through mTOR, ATG5, and LC3 in dose-dependent manners. Also, PDT at high dose induced apoptosis through caspase activation and PARP-1. Moreover, PARP-1 inhibitor protected cells against HP-PDT-induced cell death, but not by caspase inhibitor. At low dose of HP, autophagy inhibitor partially protected from HP-PDT-induced cell death. In autophagy phases, at low doses, HP-PDT regulated autophagic cell death through the inhibition of LC3II. Although autophagy inhibitor did not alter cell death directly, autophagy has associated with HP-PDT-induced apoptotic cell death by PARP-1 regulation. CONCLUSION: Taken together, HP-PDT induces apoptotic cell death with autophagy in oral cancer cells. PDT resistance is related to autophagy by PARP-1 regulation in oral cancer cells.

Association of Serum Vitamin D Level with Temporomandibular Disorder Incidence: A Retrospective, Multi-Center Cohort Study Using Six Hospital Databases.

The relationship between serum vitamin D levels and temporomandibular disorders (TMDs) remains unclear. Therefore, this study aimed to investigate the association between serum 25-hydroxyvitamin D (25[OH]D) levels and TMD incidence using large-scale health data. Clinical data from the electronic health records of six secondary or tertiary hospitals in Korea were used to evaluate the relationship between serum 25(OH)D levels and TMD incidence. The data were converted to the Observational Medical Outcomes Partnership Common Data Model. A cohort study was designed using the Cox proportional hazards model to examine the hazard ratio (HR) of TMD development after propensity score matching (PSM). An aggregate meta-analysis of the HR was subsequently performed. After 1:4 PSM, a target group with deficient 25(OH)D levels (<20 ng/mL) (N = 34,560) and comparator group with non-deficient 25(OH)D levels (≥20 ng/mL) (N = 47,359) were pooled from six hospital databases. HR meta-analysis demonstrated a significant association between deficient 25(OH)D levels and TMD incidence (pooled HR: 1.50; 95% confidence interval: 1.07-2.12). In conclusion, deficient 25(OH)D levels were found to be associated with an increased TMD risk. Therefore, vitamin D deficiency is a potential risk factor for TMD development.

Effect of passive jaw opening on the electromyographic activity of the temporalis, masseter, digastric, and infrahyoid muscles in healthy adults.

OBJECTIVE: To investigate the surface electromyography (EMG) activity of the temporalis, masseter, digastric, and infrahyoid muscles during passive jaw opening in healthy adults. METHODS: The EMG activity of the masseter, temporalis, digastric anterior belly, and infrahyoid muscles on the right side was recorded during the four jaw-opening tasks: active opening to 20 mm (AO20); active opening to 40 mm (AO40); passive opening to 40 mm with a rubber mouth prop on the right posterior teeth (POR40); and passive opening to 40 mm with a mouth prop on the left posterior teeth (POL40). RESULTS: The EMG amplitude of the digastric anterior belly and infrahyoid muscles in either POL40 or POR40 was significantly less than that in AO20 or AO40, respectively. CONCLUSION: Passive jaw opening reduces the EMG activity of the digastric and infrahyoid muscles significantly and could help reduce the load on these muscles during prolonged mouth-opening conditions.

Can the subject reliably reproduce maximum voluntary contraction of temporalis and masseter muscles in surface EMG?

OBJECTIVE: To evaluate the repeatability of the surface EMG variables of myoelectric signals from the masseter and temporalis muscles according to three methods to induce maximum voluntary contraction (MVC) in healthy adults. METHODS: Thirty healthy young subjects performed the following three MVC tasks three times each in three sessions on the same day without replacing surface electrodes: clenching the teeth (MVCIC) and biting down on two cotton rolls bilaterally with the posterior teeth (MVCBP) or first molars (MVCB6). RESULTS: The intra-class correlation coefficient (ICC) of the amplitudes during the three MVC tasks ranged from 65 to 79%. The ICCs of the spectral variables ranged from 78 to 86%. The ICCs of the asymmetry index of the masseter ranged from 77 to 86%, and those of the activity index ranged from 68 to 90%. CONCLUSION: The surface EMG measurements according to the three MVC methods exhibited good to excellent reproducibility.

Antibacterial Effect of Zirconia Nanoparticles on Polyethyl Methacrylate Resin for Provisional Crowns.

Objective: Mid-to-long term use of provisional crowns in the oral cavity is associated with bacterial adhesion and biofilm formation, thus necessitating provisional crowns exhibiting antibacterial properties to prevent the occurrence of gingivitis and periodontal disease. This study aimed to evaluate the antibacterial effect of zirconia nanoparticle-containing polyethyl methacrylate (PEMA) resin for provisional restorations. Methods: Zirconia nanoparticles were added to the monomer of PEMA resin for provisional restorations, and the mixture was stirred for 2 h using a magnetic stirrer. Disk-shaped specimens were prepared by mixing the polymer with the nanoparticle solution. The control group contained pure PEMA resin samples, and the experimental groups Group Z2, Group Z4, and Group Z8 included PEMA resin specimens containing 2, 4, and 8% w/v zirconia nanoparticles, respectively. After analyzing the sample surfaces, the antibacterial effect of the specimens was evaluated using Streptococcus mutans. Statistical analysis was performed using Tukey's test and Mann-Whitney U-test, according to the normality result in the Shapiro-Wilk test. Results: FE-SEM and EDX analyses revealed the successful addition of zirconia nanoparticles. Results showed no significant difference in the measured values for surface roughness and contact angle between the experimental and control groups; however, adhesion and biofilm thickness of S. mutans were significantly decreased in Group Z2, Group Z4, and Group Z8 compared to the control group (P < 0.05, P < 0.01, P < 0.01, respectively). Conclusion: The addition of zirconia nanoparticles can lower the incidence of adhesion and biofilm thickness of S. mutans on PEMA resin used for provisional crowns. Thus, incorporating zirconia nanoparticles in repair materials for provisional crowns and PEMA resin can produce an antibacterial effect and prevent gingivitis, periodontal disease, and dental caries.

Surface electromyography for evaluating patients with oromandibular dystonia.

OBJECTIVE: To investigate myoelectric signals of dystonic activities in oromandibular dystonia (OMD) subjects using surface electromyography (EMG). METHODS: Twelve OMD subjects were included in this study. Resting myoelectric activities of the superficial masseter, anterior temporalis, and anterior belly of the digastric muscle on both sides were monitored, and dystonic muscle contractions were recorded using surface EMG. Myoelectric signal amplitude, the type of muscle contraction, and contraction rate for phasic activities were evaluated. RESULTS: Surface EMG revealed that eight subjects had dystonic muscle activities in the phasic contraction pattern, three subjects had a tonic contraction pattern, and one subject had a mixed pattern. Synchronous contraction of dystonic muscles was frequently observed. Many of the monitored muscles showed high resting amplitudes. CONCLUSION: Surface EMG detects abnormal muscle activities related to oromandibular dystonia. Surface EMG can serve as an objective method for diagnosing oromandibular dystonia.

Diagnostic accuracy and reliability of panoramic temporomandibular joint (TMJ) radiography to detect bony lesions in patients with TMJ osteoarthritis.

BACKGROUND/PURPOSE: The value of the temporomandibular joint (TMJ) projections of panoramic radiography for diagnosing TMJ osteoarthritis is not completely elucidated. This study aimed to assess the diagnostic accuracy and reliability of panoramic TMJ radiography to detect bony lesions in patients with TMJ osteoarthritis. MATERIALS AND METHODS: This study included 55 TMJs of 44 subjects who were diagnosed with TMJ osteoarthritis. They underwent panoramic radiography (PanRad), lateral (LatTMJ) and frontal (FrnTMJ) projection panoramic TMJ radiography, and cone-beam computed tomography (CBCT). All images were examined by two observers for flattening, erosion, and osteophytes on the condylar head and articular eminence of the TMJ. RESULTS: For detecting flattening lesions on the mandibular condyle, the sensitivities of PanRad, LatTMJ, and FrnTMJ were less than 67% and the combination of LatTMJ and FrnTMJ (ComTMJ) had the highest sensitivity for both observers (67.6% and 79.7%, respectively). For erosion lesions, the sensitivity of ComTMJ for observer 1 was the highest, at 84.3%, whereas the specificity of ComTMJ was the lowest, at 37.5%. The sensitivities of all four methods for observer 2 were less than 54% and the specificities ranged from 75.0% to 100%. The overall diagnostic accuracy was highest for ComTMJ (64.3%), followed by LatTMJ (59.5%). The intraobserver reliability was good for one observer and excellent for the other, and the interobserver reliability was fair or moderate. CONCLUSION: Panoramic TMJ radiography demonstrated limited diagnostic accuracy and acceptable reliability in detecting bony lesions of the TMJ, although it was better than conventional panoramic radiography.

Characterization of a submandibular gland sialolith: micromorphology, crystalline structure, and chemical compositions.

OBJECTIVE: The aim of this study was to understand the mechanism of mineralization and growth of a sialolith by analyzing its micromorphology, crystalline structure, and chemical compositions. STUDY DESIGN: A sialolith was removed along with submandibular salivary gland from a patient. After cross-cutting and polishing the sialolith, its morphology, chemical compositions, crystalline structure, and chemical states were analyzed by using optical camera, scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffractometer, Fourier transform infrared spectrophotometer, and transmittance electron microscopy. RESULTS: The sialolith had a core composed of organic material, such as lipid compounds, and the surrounding mineralized shell structure mostly consisted of hydroxyapatite. In the transition zone between the organic core and mineralized shell layers, inorganic layers were arranged alternately with organic layers. Congregated globular structures were calcified with hydroxyapatite and whitlockite crystallites. Analysis of crystalline structures and chemical compositions suggested that calcium phosphate minerals containing magnesium, such as whitlockite, were developed in the initial stage and gradually transformed into crystallites composed of hydroxyapatite during the growth of crystallites. CONCLUSIONS: Sialolith with an organic core grew as inorganic materials were deposited and calcified in alternate layers. The mineralization process might include the initial whitlockite development and successive transformation into more stable hydroxyapatite.

Structures of PPARγ complexed with lobeglitazone and pioglitazone reveal key determinants for the recognition of antidiabetic drugs.

Peroxisome proliferator-activator receptor (PPAR) γ is a nuclear hormone receptor that regulates glucose homeostasis, lipid metabolism, and adipocyte function. PPARγ is a target for thiazolidinedione (TZD) class of drugs which are widely used for the treatment of type 2 diabetes. Recently, lobeglitazone was developed as a highly effective TZD with reduced side effects by Chong Kun Dang Pharmaceuticals. To identify the structural determinants for the high potency of lobeglitazone as a PPARγ agonist, we determined the crystal structures of the PPARγ ligand binding domain (LBD) in complex with lobeglitazone and pioglitazone at 1.7 and 1.8 Å resolutions, respectively. Comparison of ligand-bound PPARγ structures revealed that the binding modes of TZDs are well conserved. The TZD head group forms hydrogen bonds with the polar residues in the AF-2 pocket and helix 12, stabilizing the active conformation of the LBD. The unique p-methoxyphenoxy group of lobeglitazone makes additional hydrophobic contacts with the Ω-pocket. Docking analysis using the structures of TZD-bound PPARγ suggested that lobeglitazone displays 12 times higher affinity to PPARγ compared to rosiglitazone and pioglitazone. This structural difference correlates with the enhanced affinity and the low effective dose of lobeglitazone compared to the other TZDs.

Repeatability of measurements of surface electromyographic variables during maximum voluntary contraction of temporalis and masseter muscles in normal adults.

This study evaluated the intraexaminer repeatability of measurements of surface electromyography (EMG) variables and functional indices of the myoelectric signals from the masseter and temporalis muscles bilaterally in 15 healthy men. The test was repeated on two different days without templates. The resting muscle activity was recorded once, and two kinds of maximum voluntary contraction (MVC) tasks were performed and recorded three times. The two MVC tasks involved clenching the teeth and biting down on two cotton rolls bilaterally with the posterior teeth. The intraclass correlation coefficient (ICC) of amplitude was >88% and that of frequency was >95% during the two MVC tasks but not under resting conditions. The ICC of the asymmetry and activity indices during the two MVC tasks was >76%. A Bland-Altman analysis revealed no significant difference in amplitude or frequency or in the two indices between the two days during the MVC tasks. In conclusion, the measurements of surface EMG variables and the indices obtained according to the study protocol were highly repeatable in healthy men. Additional studies using templates and intraexaminer measurement errors are warranted in both men and women for complete validation of these findings.

Expression of cancer stem cell marker during 4-nitroquinoline 1-oxide-induced rat tongue carcinogenesis.

One of the theories regarding oral carcinogenesis is that the tumor growth is initiated from cancer stem cells (CSCs) that self-renew and give rise to differentiated tumor cells, like stem cells do in normal tissues. The most common methods of CSC identification are based on CSC marker expression in carcinogenesis. This study examined the expression of CD133 and CD44, the most commonly used CSC biomarkers in oral squamous cell sarcoma (SCC), with the goal of identifying molecular biomarkers whose expression is associated with the multistep oral carcinogenesis. The expression of CD133, CD44, proliferating cell nuclear antigen (PCNA), and Cytokeratin (CK) was examined by Western blot analysis and confirmed by immunohistochemistry in a 4-nitroquinoline 1-oxide-induced rat tongue carcinogenesis model. Also, the expression of aldehyde dehydrogenase 1 (ALDH1), OCT-4 and Nanog were investigated for alteration of cancer cell stemness by Western blot. Along with the progress of multistep carcinogenesis, there were slight increases of CD133 and CD44 expression in the dysplasia group compared with normal rats. However, CD133 protein level was significantly overexpressed in SCC. The expression of PCNA and CK were low in normal group, but sequentially increased in SCC. ALDH1, Nanog and OCT-4 expression were significantly increased according to SCC grade during carcinogenesis. The findings indicate that CD133 is useful in identifying oral CSCs, which suggests that CD133 may serve as a predictor to identify CSCs with a high risk of oral cancer development.

Anti-inflammatory effects of zinc in PMA-treated human gingival fibroblast cells

OBJECTIVES: Abnormal cellular immune response has been considered to be responsible for oral lesions in recurrent aphthous stomatitis. Zinc has been known to be an essential nutrient metal that is necessary for a broad range of biological activities including antioxidant, immune mediator, and anti-inflammatory drugs in oral mucosal disease. The objective of this study was to investigate the effects of zinc in a phorbol-12-myristate-13-acetate (PMA)-treated inflammatory model on human gingival fibroblast cells (hGFs).Study DESIGN: Cells were pre-treated with zinc chloride, followed by PMA in hGFs. The effects were assessed on cell viability, cyclooxygenease-1,2(COX-1/2) protein expression, PGE2 release, ROS production and cytokine release, RESULTS: The effects were assessed on cell viability, COX1/2 protein expression, PGE2 release, ROS production, cytokine release. The results showed that, in the presence of PMA, zinc treatment leads to reduce the production of ROS, which results in decrease of COX-2 expression and PGE2 release. CONCLUSIONS: Thus, we suggest that zinc treatment leads to the mitigation of oral inflammation and may prove to be an alternative treatment for recurrent aphthous stomatitis