NOX1/2 activation in human gingival fibroblasts by Fusobacterium nucleatum facilitates attachment of Porphyromonas gingivalis.

Periodontal diseases are infectious polymicrobial inflammatory diseases that lead to destruction of the periodontal ligament, gingiva, and alveolar bone. Sequential colonization of a broad range of bacteria, including Fusobacterium nucleatum and Porphyromonas gingivalis, is an important phenomenon in this disease model. F. nucleatum is a facultative anaerobic species thought to be a key mediator of dental plaque maturation due to its extensive coaggregation with other oral bacteria, while P. gingivalis is an obligate anaerobic species that induces gingival inflammation by secreting various virulence factors. The formation of a bacterial complex by these two species is central to the pathogenesis of periodontal disease. Reactive oxygen species (ROS) are produced during bacterial infections and are involved in intracellular signaling. However, the impact of oral bacteria-induced ROS on the ecology of F. nucleatum and P. gingivalis has yet to be clarified. In the present study, we investigated ROS production induced in primary human oral cells by F. nucleatum and P. gingivalis and its effect on the formation of their bacterial complexes and further host cell apoptosis. We found that in primary human gingival fibroblasts (GFs), two NADPH oxidase isoforms, NOX1 and NOX2, were activated in response to F. nucleatum infection but not P. gingivalis infection. Accordingly, increased NADPH oxidase activity and production of superoxide anion were observed in GFs after F. nucleatum infection, but not after P. gingivalis infection. Interestingly, in NOX1, NOX2, or NOX1/NOX2 knockdown cells, the number of P. gingivalis decreased when the cells were coinfected with F. nucleatum. A similar pattern of host cell apoptosis was observed. This implies that F. nucleatum contributes to attachment of P. gingivalis by triggering activation of NADPH oxidase in host cells, which may provide an environment more favorable to strict anaerobic bacteria and have a subsequent effect on apoptosis of host cells.

Activation of sonic hedgehog signaling by a Smoothened agonist restores congenital defects in mouse models of endocrine-cerebro-osteodysplasia syndrome.

BACKGROUND: Endocrine-cerebro-osteodysplasia (ECO) syndrome is a genetic disorder associated with congenital defects of the endocrine, cerebral, and skeletal systems in humans. ECO syndrome is caused by mutations of the intestinal cell kinase (ICK) gene, which encodes a mitogen-activated protein (MAP) kinase-related kinase that plays a critical role in controlling the length of primary cilia. Lack of ICK function disrupts transduction of sonic hedgehog (SHH) signaling, which is important for development and homeostasis in humans and mice. Craniofacial structure abnormalities, such as cleft palate, are one of the most common defects observed in ECO syndrome patients, but the role of ICK in palatal development has not been studied. METHODS: Using Ick-mutant mice, we investigated the mechanisms by which ICK function loss causes cleft palate and examined pharmacological rescue of the congenital defects. FINDINGS: SHH signaling was compromised with abnormally elongated primary cilia in the developing palate of Ick-mutant mice. Cell proliferation was significantly decreased, resulting in failure of palatal outgrowth, although palatal adhesion and fusion occurred normally. We thus attempted to rescue the congenital palatal defects of Ick mutants by pharmacological activation of SHH signaling. Treatment of Ick-mutant mice with an agonist for Smoothened (SAG) rescued several congenital defects, including cleft palate. INTERPRETATIONS: The recovery of congenital defects by pharmacological intervention in the mouse models for ECO syndrome highlights prenatal SHH signaling modulation as a potential therapeutic measure to overcome congenital defects of ciliopathies.

Thickness of posterior palatal masticatory mucosa: the use of computerized tomography.

BACKGROUND: Periodontal plastic surgery is used to fulfill the esthetic and functional demands of patients. The palatal masticatory mucosa is the main donor site for connective tissue, and the thickness of the graft tissue obtained is an important factor for the success of this technique. The aim of this study was to measure the thickness of masticatory mucosa in the posterior palatal area using computerized tomography (CT). METHODS: The thickness measurements were performed on the images of 100 adult subjects who underwent CT on the maxilla for implant surgery. Twenty-four standard measurement points were defined in the hard palate according to the gingival margin and the middle palatal suture. The radiographic measurements were used after calibration. The data were analyzed to determine the differences in the mucosal thickness according to gender, age, tooth position, and depth of the palatal vault. RESULTS: The overall mean thickness of the palatal masticatory mucosa was 3.83 +/- 0.58 mm (range: 2.29 to 6.25 mm). Females had significantly thinner mean masticatory mucosa (3.66 +/- 0.52 mm) than males (3.95 +/- 0.60 mm) (P <0.0001). The thickness of the palatal masticatory mucosa increased with age. The mean thickness according to tooth site was 3.46 mm (maxillary canine), 3.66 mm (first premolar), 3.81 mm (second premolar), 3.13 mm (first molar), 3.31 mm (the base of the interproximal papilla of the first and second molars), and 3.39 mm (second molar). There was an overall increase in the thickness of the palatal masticatory mucosa as the distance from the gingival margin to the middle palatine suture increased, with the exception of the Ca-d (a point at 12 mm from the gingival margin of the canine) region. There was no significant difference in the thickness of the palatal masticatory mucosa between the groups with high or low palatal vaults. CONCLUSIONS: The palatal masticatory mucosa thickness increased from the canine to premolar region but decreased at the first molar region and increased again in the second molar region, with the thinnest area at the first molar region and the thickest at the second premolar region. The canine to premolar region seems to be the most appropriate donor site that contains a uniformly thick mucosa. CT can be considered an alternative method for the measurement of palatal soft tissue thickness.

Electrocatalytic Conversion of Carbon Dioxide and Nitrate Ions to Urea by a Titania-Nafion Composite Electrode.

CO2 and nitrate ions were successfully converted to urea by a TiO2 -Nafion nanocomposite electrode under ambient conditions. The composite electrode was constructed by dropcasting the mixture of P25 titania and Nafion solution on an indium-doped tin oxide (ITO) electrode. When the electrode was electrolyzed in CO2 -saturated 0.1 m KNO3 (pH 4.5) solution at -0.98 V versus Ag/AgCl, urea was formed with a Faradaic efficiency of 40 %. The other reduced products obtained were NH3 , CO, and H2 .

Differential Matrix Metalloprotease (MMP) Expression Profiles Found in Aged Gingiva.

The periodontium undergoes age-related cellular and clinical changes, but the involved genes are not yet known. Here, we investigated age-related genetic changes in gingiva at the transcriptomic level. Genes that were differentially expressed between young and old human gingiva were identified by RNA sequencing and verified by real-time PCR. A total of 1939 mRNA transcripts showed significantly differential expression between young and old gingival tissues. Matrix metalloprotease (MMP) regulation was the top pathway involved in gingival aging. MMP3, MMP9, MMP12, and MMP13 were upregulated in old gingival tissues, concomitantly with interleukin-1 beta (IL1B) expression. In vitro experiments using human gingival fibroblasts (hGFs) showed that MMP12 was upregulated in old hGFs compared to young hGFs. Moreover, the MMP3, MMP9 and IL1B levels were more highly stimulated by infection with the oral bacterium, Fusobacterium nucleatum, in old hGFs compared to young hGFs. Collectively, these findings suggest that, in gingiva, the upregulation of MMP12 may be a molecular hallmark of natural aging, while the upregulations of MMP3, MMM9, and IL1B may indicate externally (e.g., infection)-induced aging. These findings contribute to our understanding of the molecular targets involved in gingival aging.