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Techniques for continuously monitoring the formation of subgingival biofilm, in relation to the determination of species and their accumulation over time in gingivitis and periodontitis, are limited. In recent years, advancements in the field of optical spectroscopic techniques have provided an alternative for analyzing three-dimensional microbiological structures, replacing the traditional destructive or biofilm staining techniques. In this work, we have demonstrated that the use of confocal Raman spectroscopy coupled with multivariate analysis provides an approach to spatially differentiate bacteria in an in vitro model simulating a subgingival dual-species biofilm. The present study establishes a workflow to evaluate and differentiate bacterial species in a dual-species in vitro biofilm model, using confocal Raman microscopy (CRM). Biofilm models of Actinomyces denticolens and Streptococcus oralis were cultured using the "Zürich in vitro model" and were analyzed using CRM. Cluster analysis was used to spatially differentiate and map the biofilm model over a specified area. To confirm the clustering of species in the cultured biofilm, confocal laser scanning microscopy (CLSM) was coupled with fluorescent in vitro hybridization (FISH). Additionally, dense bacteria interface area (DBIA) samples, as an imitation of the clusters in a biofilm, were used to test the developed multivariate differentiation model. This confirmed model was successfully used to differentiate species in a dual-species biofilm and is comparable to morphology. The results show that the developed workflow was able to identify main clusters of bacteria based on spectral "fingerprint region" information from CRM. Using this workflow, we have demonstrated that CRM can spatially analyze two-species in vitro biofilms, therefore providing an alternative technique to map oral multi-species biofilm models., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kriem, Wright, Ccahuana-Vasquez and Rupp.)

Purpose: To compare clinical outcomes and oral fluid biomarkers in gingivitis subjects using an electric toothbrush/irrigator combination (test) or a manual toothbrush alone (control) over 8 weeks., Materials and Methods: Subjects were randomly assigned to two groups of n = 30. In both groups, toothbrushing was performed twice daily at home and no additional interdental cleaning aids were allowed. Plaque Index (PLI), Gingival Index (GI), whole saliva (WS), and gingival crevicular fluid (GCF) samples were collected at weeks 2, 4, and 8., Results: Subjects' mean age was 23 years and 52% were female. Overall baseline means were 1.31 for PLI, 1.07 for GI, and 34.9 for number of bleeding sites. At every follow-up visit, both groups differed statistically significantly (p < 0.001) from baseline for all clinical parameters. The test group demonstrated statistically significantly (p < 0.001) greater reductions in GI vs the control group by 18% at week 2, 17% at week 4 and 24% at week 8. The test group also demonstrated statistically significantly (p < 0.002) greater reductions in the number of bleeding sites vs the control group by 33% at week 2, 34% at week 4 and 43% at week 8. Between-group comparisons for both WS and GCF revealed numerical trends for decreased levels of interleukin (IL)-1β in GCF after 4 and 8 weeks, but these were not statistically significant., Conclusion: In subjects using the electric toothbrush/irrigator combination, increased clinical improvements may be found accompanied by similarly improved trends for oral fluid biomarkers such as IL-1β.

The study of oral disease progression, in relation to the accumulation of subgingival biofilm in gingivitis and periodontitis is limited, due to either the ability to monitor plaque in vitro. When compared, optical spectroscopic techniques offer advantages over traditional destructive or biofilm staining approaches, making it a suitable alternative for the analysis and continued development of three-dimensional structures. In this work, we have developed a confocal Raman spectroscopy analysis approach towards in vitro subgingival plaque models. The main objective of this study was to develop a method for differentiating multiple oral subgingival bacterial species in planktonic and biofilm conditions, using confocal Raman microscopy. Five common subgingival bacteria (Fusobacterium nucleatum, Streptococcus mutans, Veillonella dispar, Actinomyces naeslundii and Prevotella nigrescens) were used and differentiated using a 2-way orthogonal Partial Least Square with Discriminant Analysis (O2PLS-DA) for the collected spectral data. In addition to planktonic growth, mono-species biofilms cultured using the 'Zürich Model' were also analyzed. The developed method was successfully used to predict planktonic and mono-species biofilm species in a cross validation setup. The results show differences in the presence and absence of chemical bands within the Raman spectra. The O2PLS-DA model was able to successfully predict 100% of all tested planktonic samples and 90% of all mono-species biofilm samples. Using this approach we have shown that Confocal Raman microscopy can analyse and predict the identity of planktonic and mono-species biofilm species, thus enabling its potential as a technique to map oral multi-species biofilm models., Competing Interests: There are no conflicts of interest declared for this paper beyond the affiliations of the coauthors with P&G, with Renzo Alberto Ccahuana-Vasquez being a recognized expert in Oral Care and Kevin Wright an R&D microbiologist. There is no IP associated with this publication for any of the partners and no technologies pertaining to products either in development or on the market have been used in this work. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: Many adolescents have poor plaque control and sub-optimal toothbrushing behavior. Therefore, we compared the efficacy of an interactive power toothbrush (IPT) to a manual toothbrush (MT) for reducing dental plaque and improving toothbrushing compliance., Methods: In this randomized, parallel single-blind clinical study, adolescents brushed twice daily with either a MT (Oral-B® Indicator soft manual toothbrush) or an IPT (Oral-B® ProfessionalCare 6000 with Bluetooth). Subjects brushed for 2 min, plus an additional 10 s for each 'Focus Care Area'. At screening and Week 2, afternoon pre-brushing plaque was assessed via the Turesky Modification of the Quigley-Hein Plaque Index (TMQHPI), and supervised brushing duration was measured., Results: Sixty subjects were randomized; 98% completed. At Week 2, the mean reduction in whole mouth plaque relative to baseline was 34% (p < 0.001) for the IPT versus 1.7% (p = 0.231) for the MT. For Focus Care Areas, the IPT yielded a 38.1% mean TMQHPI reduction (p < 0.001) versus 6.2% for the MT (p < 0.001). Mean brushing time versus baseline increased 34 s in the IPT group (p < 0.001) while remaining flat in the MT group (p = 1.0)., Conclusions: Over 2 weeks, adolescents using an IPT experienced superior plaque reduction and increased overall brushing time versus those using a MT., Trial Registration: This trial was retrospectively registered ( ISRCTN10112852 ) on the 18th, June 2018.

Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Müeller-Hinton agar. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates.