Investigation of the effects of Bipolar Radiofrequency Energy on the Structural Morphology of Dental Plaque.

OBJECTIVES: To investigate the effects of radiofrequency (RF) energy, applied through a power toothbrush, on the structural morphology of dental plaque and its bacteria components. Previous studies showed that a toothbrush powered by RF (ToothWave) effectively reduces extrinsic tooth stains, plaque, and calculus. However, the mechanism by which it reduces dental plaque deposits is not fully established. MATERIALS AND METHODS: Multispecies plaques at sampling time points of 24, 48, and 72 hours were treated with the application of RF using ToothWave with the toothbrush bristles 1 mm above the plaque surface. Groups that underwent the same protocol but without RF treatment served as paired controls. Confocal laser scanning microscope (CLSM) was used to determine cell viability at each time point. Plaque morphology and bacteria ultrastructure were viewed using scanning electron microscope (SEM) and transmission electron microscope (TEM), respectively. STATISTICAL ANALYSIS: Data were analyzed statistically using analysis of variance (ANOVA) and Bonferroni post-tests. RESULTS: At each time, RF treatment significantly (p < 0.05) reduced the viable cells in plaque and caused a substantial disruption of plaque morphology, while the untreated plaque had intact morphology. Cells in treated plaques showed disrupted cell walls, cytoplasmic material, huge vacuoles, and heterogeneity in electron density, while these organelles remained intact in untreated plaques. CONCLUSION: The application of RF via a power toothbrush can disrupt plaque morphology and kill bacteria. These effects were enhanced by the combined application of RF and toothpaste.

Changes in oral health during aging in a novel non-human primate model.

Oral health plays a significant role in the quality of life and overall well-being of the aging population. However, age-related changes in oral health are not well understood due to challenges with current animal models. In this study, we analyzed the oral health and microbiota of a short-lived non-human primate (i.e., marmoset), as a step towards establishing a surrogate for studying the changes that occur in oral health during human aging. We investigated the oral health of marmosets using cadaveric tissues in three different cohorts: young (aged ≤6 years), middle-aged, and older (>10 years) and assessed the gingival bacterial community using analyses of the V3-V4 variable region of 16S rRNA gene. The oldest cohort had a significantly higher number of dental caries, increased dental attrition/erosion, and deeper periodontal pocket depth scores. Oral microbiome analyses showed that older marmosets had a significantly greater abundance of Escherichia-Shigella and Propionibacterium, and a lower abundance of Agrobacterium/Rhizobium at the genus level. Alpha diversity of the microbiome between the three groups showed no significant differences; however, principal coordinate analysis and non-metric multidimensional scaling analysis revealed that samples from middle-aged and older marmosets were more closely clustered than the youngest cohort. In addition, linear discriminant analysis effect size (LEFSe) identified a higher abundance of Esherichia-Shigella as a potential pathogenic biomarker in older animals. Our findings confirm that changes in the oral microbiome are associated with a decline in oral health in aging marmosets. The current study suggests that the marmoset model recapitulates some of the changes in oral health associated with human aging and may provide opportunities for developing new preventive strategies or interventions which target these disease conditions.

Evaluation of an Artificial Mouth for Dental Caries Development.

This study validated a microbial caries model (artificial mouth) for dental caries development to determine the optimal time to create early caries suitable for evaluation of the efficacy of caries therapeutic agents. In all, 40 human enamel blocks were placed in an artificial mouth at 37 °C and 5% CO2 and were exposed to brain heart infusion broth inoculated with S. mutans in continuous circulation (0.3 mL/min). The culture medium was replaced three times daily. Samples were exposed to 10% sucrose for 3 min, 3 times daily to promote biofilm growth. Five samples were harvested from the chamber after 3, 4, 5, 6, 7, 14, 21, and 28 days. At the end of experiment, samples were assessed visually by ICDAS criteria, while lesion depth (LD) and mineral loss (ML) were measured using polarizing light microscopy and transverse microradiography. Data were analyzed by Pearson correlation, ANOVA, and Tukey comparison test (p < 0.05). Results showed significant and strong positive correlation (p < 0.01) between all variables and biofilm growth time. LD and ML profiles of 7-day lesions seem to be the most suitable for remineralization studies. In conclusion, using the evaluated artificial mouth, early-stage caries suitable for products' evaluation studies was produced within 7 days of exposure to microbial biofilm.

In vitro effect of an oral spray and mouthrinses on dual species cariogenic bacteria biofilm.

PURPOSE: To determine the efficacy of an oral spray and oral rinses to inhibit oral cariogenic dual species biofilm formation on hydroxyapatite (HA) discs. METHODS: The Streptococcus mutans (NCTC 10449, ATCC), Lactobacilli casei (NCIB 8820, ATCC) dual species biofilm formation and inhibition on HA disc was tested using five antimicrobial products, i.e., oral spray (Oral Shield), Mouthrinse (Listerine Ultra Clean, Listerine Cool Mint, Crest Pro-Health, ACT Restoring). An untreated group served as control. The established biofilm on the surface of each disc was treated or untreated with oral spray and mouthrinse for 2 minutes after 24 or 48 hours. The dual species biofilm formation and inhibition on HA discs was determined using the spread plate method and colonies were counted and expressed as colony forming units (CFU/mL). Further, the HA disc was subjected to confocal laser scanning microscope (CLSM) examination to determine the viability of cells using live-dead staining and a scanning electron microscope (SEM) to examine the effect on bacteria biofilm and morphology. The cytotoxic effect of test spray and mouthrinse was tested on OKF6/TERT-2 cells using the MTT method. RESULTS: At each time point, 24- or 48-hours, S. mutans and L. casei mixed biofilm on HA discs had a significantly (P> 0.001) fewer number of bacteria in the treated groups than the untreated one. The oral spray and mouthrinses had a detrimental effect on bacteria biofilm, morphology and cell wall, whereas no significant changes were observed in the untreated group. Cytotoxic assay revealed that the oral spray was safe for human oral keratinocyte cells. CLINICAL SIGNIFICANCE: The tested oral spray could offer potential to inhibit the cariogenic bacteria and protect the tooth enamel from cariogenic bacterial biofilm.

ß2-Adrenergic receptor agonist induced hepatic steatosis in mice: modeling nonalcoholic fatty liver disease in hyperadrenergic states.

Nonalcoholic fatty liver disease (NAFLD) is a spectrum of disorders ranging from hepatic steatosis [excessive accumulation of triglycerides (TG)] to nonalcoholic steatohepatitis, which can progress to cirrhosis and hepatocellular carcinoma. The molecular pathogenesis of steatosis and progression to more severe NAFLD remains unclear. Obesity and aging, two principal risk factors for NAFLD, are associated with a hyperadrenergic state. ß-Adrenergic responsiveness in liver increases in animal models of obesity and aging, and in both is linked to increased hepatic expression of ß2-adrenergic receptors (ß2-ARs). We previously showed that in aging rodents intracellular signaling from elevated hepatic levels of ß2-ARs may contribute to liver steatosis. In this study we demonstrate that injection of formoterol, a highly selective ß2-AR agonist, to mice acutely results in hepatic TG accumulation. Further, we have sought to define the intrahepatic mechanisms underlying ß2-AR mediated steatosis by investigating changes in hepatic expression and cellular localization of enzymes, transcription factors, and coactivators involved in processes of lipid accrual and disposition-and also functional aspects thereof-in livers of formoterol-treated animals. Our results suggest that ß2-AR activation by formoterol leads to increased hepatic TG synthesis and de novo lipogenesis, increased but incomplete ß-oxidation of fatty acids with accumulation of potentially toxic long-chain acylcarnitine intermediates, and reduced TG secretion-all previously invoked as contributors to fatty liver disease. Experiments are ongoing to determine whether sustained activation of hepatic ß2-AR signaling by formoterol might be utilized to model fatty liver changes occurring in hyperadrenergic states of obesity and aging, and thereby identify novel molecular targets for the prevention or treatment of NAFLD.NEW & NOTEWORTHY Results of our study suggest that ß2-adrenergic receptor (ß2-AR) activation by agonist formoterol leads to increased hepatic TG synthesis and de novo lipogenesis, incomplete ß-oxidation of fatty acids with accumulation of long-chain acylcarnitine intermediates, and reduced TG secretion. These findings may, for the first time, implicate a role for ß2-AR responsive dysregulation of hepatic lipid metabolism in the pathogenetic processes underlying NAFLD in hyperadrenergic states such as obesity and aging.

Biodegradation and Characterization of Streptomyces sp. (JMCACA3) from Acid Corroded Iron Plate.

From acid corroded iron plates five different types of actinobacteria were isolated. Among the five, JMCACA3 strain was selected for the present study. In ISP media, JMCACA3 strain showed well-developed aerial and substrate mycelia were observed. This strain showed good growth in 12 different carbon and 4 different nitrogen sources. The 16S rRNA sequence of phylogenetic analysis by neighbor-joining method identified the studied strain belongs to Streptomyces sp. The biodegradation activity of the strain analyzed by UV and FTIR analysis, which revealed that the various concentrations of Benzimidazole inhibitor with JMCACA3 culture showed slightly varied results. For weight loss method, mild steel coupons incubated with JMCACA3 culture, Benzimidazole inhibitor + JMCACA3 culture and mixed sample showed that JMCACA3 strain utilized the inhibitor as their energy source and the weight the coupons were slightly varied, evidenced by XRD spectra and showed Fe2O3 corrosion products. Our study concluded that the JMCACA3 strain, an iron-reducing actinobacteria which utilizes and converted the corrosion inhibitor Benzimidazole as their energy source. So, it is very urgent to develop more powerful corrosion inhibitor from green biocide or microbial-based biocide and their analog which incorporated into the pre-existing Benzimidazole to increase the corrosion inhibitor level against the biofilm of actinobacterial influenced corrosion.

In vitro evaluation of the effects of Ultrasound Tongue Scraper on bacteria and biofilm formation.

AIM: Oral malodor is a common condition caused by some Gram-negative oral bacteria, among which are the 3 red complex bacteria (RCB). The present study investigated the effectiveness of the Ultrasound Tongue Scraper (UTS) to disrupt the structural morphology of the bacteria and their biofilm. METHODS: While developing over 72 hours, multispecies biofilms of RCB (Porphromonas gingivalis, Tryponema denticola, Tannerella forsythia) were treated every 24 hours with 1.6-MHz ultrasound waves generated with UTS. An untreated group served as controls. Confocal laser scanning microscopy was used to determine the biofilm thickness, biomass and live : dead cell ratio at each time point (24, 48 and 72 hours). Biofilm morphology and bacteria ultrastructure were viewed using scanning/transmission electron microscopy, respectively. Data were analyzed using ANOVA and Tukey tests. RESULTS: At each time point, the 3 variables were significantly lower in treated samples than the untreated. Significant biofilm disruption was observed in treated samples at each time period while the untreated had intact biofilm morphology. Cells in treated samples showed disrupted cell wall, cytoplasmic material, huge vacuoles and heterogeneity in electron density, while these cell organelles remained intact in untreated samples. CONCLUSION: The UTS has an inhibitory effect on RCB and could be useful for oral malodor management.