Glyburide inhibits the bone resorption induced by traumatic occlusion in rats.

OBJECTIVE: To examine whether glyburide inhibits bone destruction caused by traumatic occlusion in a rat occlusal trauma model. BACKGROUND: Excessive mechanical stress, such as traumatic occlusion, induces expression of IL-1ß and may be involved in bone resorption. NLRP3 inflammasomes have been linked to IL-1ß expression, but it is currently unclear whether glyburide, the inhibiter of NLRP3 inflammasome, suppresses occlusal trauma in rats. METHODS: Male SD rats aged 7 weeks were used. In the trauma group, the occlusal surface of the maxillary first right molar was raised by attaching a metal wire to apply occlusal trauma to the mandibular first right molar. In the trauma + glyburide group, the NLRP3 inhibitor glyburide was administered orally every 24 hours from 1 day before induction of occlusal trauma. Rats were euthanized after 5 or 10 days, and the maxillary first molars were harvested with the adjacent tissues for histopathological investigation. Immunohistochemical expression of IL-1ß, NLRP3, and RANKL was also assessed. RESULTS: On day 5, bone resorption was significantly greater in the trauma group compared with the control group or the trauma + glyburide group, and there were significantly higher numbers of osteoclasts and cells positive for IL-1ß, NLRP3, and RANKL in the trauma group. CONCLUSION: In this study, glyburide inhibits bone resorption by traumatic occlusion in rats. It suggests that the NLRP3/IL-1ß pathway might be associated with bone resorption induced by traumatic occlusion.

Toll-like receptor 2 activation primes and upregulates osteoclastogenesis via lox-1.

BACKGROUND: Lectin-like oxidized low-density-lipoprotein receptor 1 (Lox-1) is the receptor for oxidized low-density lipoprotein (oxLDL), a mediator in dyslipidemia. Toll-like receptor (TLR)-2 and - 4 are receptors of lipopolysaccharide (LPS) from Porphyromonas gingivalis, a major pathogen of chronic periodontitis. Although some reports have demonstrated that periodontitis has an adverse effect on dyslipidemia, little is clear that the mechanism is explained the effects of dyslipidemia on osteoclastogenesis. We have hypothesized that osteoclast oxLDL has directly effect on osteoclasts (OCs), and therefore alveolar bone loss on periodontitis may be increased by dyslipidemia. The present study aimed to elucidate the effect of Lox-1 on osteoclastogenesis associated with TLRs in vitro. METHODS: Mouse bone marrow cells (BMCs) were stimulated with macrophage colony-stimulating factor into bone marrow macrophages (BMMs). The cells were also stimulated with synthetic ligands for TLR2 (Pam3CSK4) or TLR4 (Lipid A), with or without receptor activator of nuclear factor kappa-B ligand (RANKL), and assessed for osteoclastogenesis by tartrate-resistant acid phosphatase (TRAP) staining, immunostaining, western blotting, flow activated cell sorting (FACS) analysis, real-time polymerase chain reaction (PCR), and reverse transcription PCR. RESULTS: Lox-1 expression was significantly upregulated by Pam3CSK4 and Lipid A in BMCs (p < 0.05), but not in BMMs. FACS analysis identified that Pam3CSK4 upregulated RANK and Lox-1 expression in BMCs. TRAP-positive cells were not increased by stimulation with Pam3CSK4 alone, but were increased by stimulation with combination combined Pam3CSK and oxLDL. Expression of both Lox-1 and myeloid differentiation factor 88 (MyD88), an essential adaptor protein in the TLR signaling pathway, were suppressed by inhibitors of TLR2, TLR4 and mitogen-activated protein kinase (MAPK). CONCLUSIONS: This study supports that osteoclastogenesis is promoted under the coexistence of oxLDL by TLR2-induced upregulation of Lox-1 in BMCs. This indicates that periodontitis could worsen with progression of dyslipidemia.

Efficacy of an Electric Toothbrush With Monitor in Dental Plaque Removal: A Crossover Randomized Controlled Trial.

BACKGROUND AND PURPOSE: Plaque control is very important in the treatment of periodontitis. However, plaque is difficult to remove because one cannot see one's own oral cavity. The purpose of this study was to verify the plaque removal effect of a prototype device that has a built-in image sensor in the head of an electric toothbrush, enabling the user to brush while checking the condition of the tooth surface on a monitor in real time and to assess their sense of use. MATERIALS AND METHODS: The subjects were 10 fifth-year students from the Graduate School of Dental Science, Fukuoka Dental College, Fukuoka, Japan. The subjects were divided into those who used electric toothbrushes while having the condition of the tooth surface checked with a monitor (monitor group) and those without a monitor (non-monitor group). O'Leary plaque control records before and after brushing and the brushing time were measured, and questionnaires were given to the subjects after brushing. Scaling and professional tooth cleaning were performed after completing the questionnaire. One week later, subjects were switched to the opposite group and had the same measurements and questionnaires. The Wilcoxon signed-rank test was used to compare both groups before and after the examination at a 5% significance level. RESULTS: The monitor group had significantly better plaque removal than the non-monitor group. In addition, the monitor group spent significantly more time brushing than the control group. CONCLUSION: Brushing while monitoring oral conditions in real time using an electric toothbrush with a built-in image sensor showed that significantly better plaque removal can be achieved with a longer brushing time.

Development of a School-Based Online Periodontal Education Programme for Adolescents.

INTRODUCTION AND AIMS: This study aimed to investigate the effects of a developed school-based online health education programme with a periodontal examination results sheet for high school students on their subjective oral symptoms, knowledge and attitudes regarding oral health, and oral health behaviours. METHODS: The participants were first- and second-year students aged 15 to 17 years (n = 847) at a high school in Japan. The students underwent a periodontal examination and were divided into periodontal condition (PC) and nonperiodontal condition (non-PC) groups. The students participated in the online oral health education programme, which included a periodontal examination results sheet after the examination. The data for identifying the effect of the programme were collected via questionnaire surveys at the periodontal examination (baseline), after 3 months, and after 1 year, and they were compared between baseline and 3 months later and between baseline and 1 year later by the chi-square test. Logistic regression analysis was used to determine the associations between the measured variables related to oral health at 3 months or 1 year and the presence/absence of periodontal conditions after adjustment for sex and variables at baseline. RESULTS: A total of 628 students (74.1%) participated in this study. The percentages of individuals with knowledge of how many teeth they had, knowledge of periodontal disease, and experience with toothbrushing instruction increased significantly after 1 year in both groups (p < .05). The awareness of gingival swelling and bleeding and the use of fluoride toothpaste at 3 months were positively associated with the presence of periodontal conditions. CONCLUSIONS: This study showed that an online oral health education programme contributed to improving oral health knowledge and behaviours among high school students and that the awareness of periodontal conditions according to the periodontal examination results sheet might improve the awareness of gingival swelling and bleeding at 3 months.

Current status and factors of periodontal disease among Japanese high school students: a cross-sectional study.

INTRODUCTION: This study aimed to investigate the prevalence of periodontal disease and the factors of the disease among high school students. METHOD: The participants were all students aged 15-18 years (n = 1202) at a high school in Japan. The data on oral health perceptions and behaviours were collected by a questionnaire survey. The prevalence of periodontal disease among them was investigated with the partial community periodontal index (PCPI). A logistic regression analysis was used to identify the factors associated with the PCPI. RESULTS: A total of 1069 students (88.9%) participated in this study. The prevalence of gingival bleeding, calculus, pocket depth of 4-5 mm, and pocket depth of 6 mm or more were 44.2%, 42.2%, 11.4%, and 1.6%, respectively. Approximately one-third of the students had a fear of dental treatment, and only 28.4% used dental floss. The results of logistic regression analysis, adjusted for sex and school year, showed that not visiting dentists regularly, not using dental floss, brushing teeth for less than 5 min, fear of dental treatment, and drinking sports drinks frequently were positively associated with periodontal conditions. CONCLUSION: This study identified a high prevalence of periodontal disease among Japanese high school students aged 15-18 years and its risk factors, such as poor oral health behaviours and fear of dental treatment.

A histopathologic study of the controlling role of T cells on experimental periodontitis in rats.

BACKGROUND/PURPOSE: The onset and progression of periodontitis involve bacterial infection and the immune response. T cells function in the immune response and reportedly induce bone resorption in inflammatory bone loss. However, the exact role of T cells in periodontal destruction remains unclear. Using our experimental model of periodontitis, we aimed to investigate the influence of T cells on periodontal destruction. MATERIALS AND METHODS: Male athymic nude (Nu) and euthymic wild-type (WT) rats were divided into the immunized (I-Nu and I-WT), non-immunized (nI-Nu and nI-WT). The immunized groups were immunized intraperitoneally with lipopolysaccharide (LPS). The non-immunized groups received phosphate-buffered saline (PBS). Nothing was administered to the non-treated groups. LPS was applied to the right palatal gingival sulcus in the immunized and non-immunized groups daily for 20 days. Loss of attachment, numbers of inflammatory cells and osteoclasts, and levels of alveolar bone were investigated histopathologically and histometrically. Osteoclasts were stained with tartrate-resistant acid phosphatase. The numbers of IL-4-positive cells were evaluated immunohistologically. RESULTS: Loss of attachment, numbers of inflammatory cells, levels of alveolar bone, and the number of osteoclasts were significantly increased in the nI-WT group compared with the nI-Nu group. However, the parameters were significantly increased in the I-Nu group compared with the I-WT group. The number of IL-4-positive cells was greater in the I-WT group than in the I-Nu group. CONCLUSION: T cells promote inflammation in non-immunized animals; however, they regulate these processes in immunized animals.

Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor.

Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors.

RANKL pretreatment plays an important role in the differentiation of pit-forming osteoclasts induced by TNF-α on murine bone marrow macrophages.

BACKGROUND: Osteoclasts differentiated from bone marrow macrophages (BMMs) induced by TNF-α alone do not have resorbing activity. When BMMs are stimulated with receptor activator of NF-κB ligand (RANKL) before TNF-α stimulation, pit-forming osteoclasts are differentiated. However, the details of the effect of RANKL pretreatment on the pit-forming osteoclast differentiation by TNF-α have not been established. The aim of this study is to examine the condition of RANKL pretreatment for differentiation of pit-forming osteoclasts induced by TNF-α. Murine BMMs were stimulated with various concentrations of RANKL for 24h in the presence of M-CSF, then the medium was changed and TNF-α was added. Osteoclasts and pits formation were examined. Osteoprotegerin (OPG), decoy receptor of RANKL, was added to the culture to examine the necessity of co-existing RANKL with TNF-α on the formation of pit-forming osteoclasts. To investigate the influence of RANKL of sufficient concentration as pretreatment for pit-forming osteoclast formation by TNF-α, dose- and time-dependent changes of osteoclast formation were checked. RESULTS: The pit formation by osteoclasts in response to TNF-α required 10ng/mL RANKL pretreatment. Stimulation with this concentration of RANKL led to the differentiation of mature osteoclasts in the 72h culture. The pit formation was not inhibited by the OPG. CONCLUSION: These results suggested that the concentration of RANKL pretreatment, which also alone can differentiate BMMs into osteoclasts, may be important in the differentiation of pit-forming osteoclasts by TNF-α. In addition, the effects of TNF-α after RANKL treatment might be independent of RANKL.