Teaching Radiographic Caries Detection and Treatment Planning: A Seminar Using an Audience Response System.

This study presents a seminar model for teaching radiographic caries detection and treatment planning at the Faculty of Dentistry, University of Oslo. The seminar is based partly on an audience response system (ARS) and uses patient cases to focus on caries risk assessment and treatment planning. This paper describes the seminar design, implementation, learning outcomes, and observational study of variability in caries registrations and students' attitudes to use of ARS. Dental and dental hygiene students participate in two seminar modules. Module 1 aims to develop and increase individual student skills in radiographic caries lesion detection, scoring, and differential diagnosis. Students perform trial registrations on bitewings using an ARS with anonymous live polling, and scorings are discussed in plenum. Students then perform individual registrations on 12 bitewing pairs. Using digital scoring, students detect and grade caries lesions on all approximal and occlusal tooth surfaces. After the session, students use the ARS to repeat scorings on selected tooth surfaces, and results are again discussed in plenum. Module 2 involves group exercises on 4 patient cases that are later presented with plenary discussions. In total, 1,624 caries registrations performed by 150 students attending the seminar between 2016 and 2018 were assessed for variability between students. As expected, variations in caries registrations were observed between students, mostly related to restored surfaces or tooth surfaces that were otherwise difficult to register. In 2022, 63 dental and dental hygiene students attending the seminar answered a questionnaire about use of ARS. The responses were scored using a five-point Likert scale. Overall, no significant difference in satisfaction with the ARS-based module was observed between dental and dental hygiene students (χ2 test, p > 0.05). The majority of the students were positive toward the use of ARS (94%), but some disagreed on the role of ARS in usefulness for understanding the seminar content (3.2%), and in increasing their confidence in radiographic registration of caries (3.2%). The ARS-based module provides a positive learning environment that ensures student anonymity, interactivity, and engagement, and combined with the other seminar module gives students basic skills in caries detection and treatment planning.

Phylogenetic group- and species-specific oligonucleotide probes for single-cell detection of lactic acid bacteria in oral biofilms.

BACKGROUND: The purpose of this study was to design and evaluate fluorescent in situ hybridization (FISH) probes for the single-cell detection and enumeration of lactic acid bacteria, in particular organisms belonging to the major phylogenetic groups and species of oral lactobacilli and to Abiotrophia/Granulicatella. RESULTS: As lactobacilli are known for notorious resistance to probe penetration, probe-specific assay protocols were experimentally developed to provide maximum cell wall permeability, probe accessibility, hybridization stringency, and fluorescence intensity. The new assays were then applied in a pilot study to three biofilm samples harvested from variably demineralized bovine enamel discs that had been carried in situ for 10 days by different volunteers. Best probe penetration and fluorescent labeling of reference strains were obtained after combined lysozyme and achromopeptidase treatment followed by exposure to lipase. Hybridization stringency had to be established strictly for each probe. Thereafter all probes showed the expected specificity with reference strains and labeled the anticipated morphotypes in dental plaques. Applied to in situ grown biofilms the set of probes detected only Lactobacillus fermentum and bacteria of the Lactobacillus casei group. The most cariogenic biofilm contained two orders of magnitude higher L. fermentum cell numbers than the other biofilms. Abiotrophia/Granulicatella and streptococci from the mitis group were found in all samples at high levels, whereas Streptococcus mutans was detected in only one sample in very low numbers. CONCLUSIONS: Application of these new group- and species-specific FISH probes to oral biofilm-forming lactic acid bacteria will allow a clearer understanding of the supragingival biome, its spatial architecture and of structure-function relationships implicated during plaque homeostasis and caries development. The probes should prove of value far beyond the field of oral microbiology, as many of them detect non-oral species and phylogenetic groups of importance in a variety of medical conditions and the food industry.

In vitro quantitative light-induced fluorescence to measure changes in enamel mineralization.

A sensitive, quantitative method for investigating changes in enamel mineralization of specimens subjected to in vitro or in situ experimentation is presented. The fluorescence-detecting instrument integrates a Xenon arc light source and an object positioning stage, which makes it particularly suitable for the nondestructive assessment of demineralized or remineralized enamel. We demonstrate the ability of in vitro quantitative light-induced fluorescence (QLF) to quantify changes in mineralization of bovine enamel discs that had been exposed in vitro to a demineralizing gel (n=36) or biofilm-mediated demineralization challenges (n=10), or were carried in situ by three volunteers during a 10-day experiment (n=12). Further experiments show the technique's value for monitoring the extent of remineralization in 36 specimens exposed in vitro to oral multispecies biofilms and document the repeatability of in vitro QLF measurements (n=10) under standardized assay conditions. The validity of the method is illustrated by comparison with transversal microradiography (TMR), the invasive current gold standard for assessing experimental changes in enamel mineralization. Ten discs with 22 measurement areas for comparison demonstrated a positive correlation between TMR and QLF (r=0.82). Filling a technological gap, this QLF system is a promising tool to assay in vitro nondestructively localized changes in mineralization of enamel specimens.

Effects of mouthrinses with triclosan, zinc ions, copolymer, and sodium lauryl sulphate combined with fluoride on acid formation by dental plaque in vivo.

Bacteriological tests demonstrated a slight synergistic effect of triclosan and sodium lauryl sulphate (SLS) on the growth of Streptococcus mutans NCTC 10449 and Streptococcus sanguis ATCC 10556 in vitro. A single mouthrinse with SLS (17.4 mM) or SLS plus triclosan (3.5 mM) significantly decreased the number of salivary mutans streptococci in a group of 12 subjects up to 90 min after rinsing. The effect on plaque pH of a mouthrinse with either 12.0 mM NaF, NaF plus 10.0 mM zinc acetate, NaF plus 17.4 mM SLS, or NaF plus SLS plus 3.5 mM triclosan with or without the addition of zinc ions or 0.65% w/v of a polyvinylmethyl ether/maleic acid copolymer was investigated. The plaque pH responses to a 10% w/v sucrose mouthrinse were measured in 2-day-old plaque with microtouch pH electrodes in six groups of 10 subjects 90 min after a single mouthrinse with test solution. There was no significant difference in plaque pH between the various mouthrinses. In conclusion, triclosan enhanced the growth-inhibitory activity of SLS against oral streptococci in vitro but not against salivary mutans streptococci in vivo. Neither triclosan incorporated into a mouthrinse containing SLS plus fluoride, nor the addition of zinc ions or copolymer affected acid formation by dental plaque in vivo.

Application of the Zürich biofilm model to problems of cariology.

The term biofilm is increasingly replacing 'plaque' in the literature, but concepts and existing paradigms are changing much more slowly. There is little doubt that biofilm research will lead to more realistic perception and interpretation of the physiology and pathogenicity of microorganisms colonizing plaques in the oral cavity. There is clear evidence that the genotypic and phenotypic expression profiles of biofilm and planktonic bacteria are different. Several techniques are available today to study multispecies biofilms of oral bacteria, each having its particular advantages and weaknesses. We describe a biofilm model developed in Zürich and demonstrate a number of applications with direct or indirect impact on prophylactic dentistry: spatial arrangement and associative behavior of various species in biofilms; multiplex fluorescent in situ hybridization analysis of oral bacteria in biofilms; use of the biofilm model to predict in vivo efficacy of antimicrobials reliably; mass transport in biofilms; de- and remineralization of enamel exposed to biofilms in vitro. The potential of biofilm experimentation in oral biology has certainly not yet been fully exploited and dozens of possible interesting applications could be investigated. The overall physiological parameters of multispecies biofilms can be measured quite accurately, but it is still impossible to assess in toto the multitude of interactions taking place in such complex systems. What can and should be done is to test hypotheses stemming from experiments with planktonic cells in monospecies cultures. In particular, it will be interesting to investigate the relevance to biofilm composition and metabolism of specific gene products by using appropriate bacterial mutants.

Quantitative detection of Porphyromonas gingivalis fimA genotypes in dental plaque.

We developed quantitative fimA genotype assays and applied them in a pilot study investigating the fimbrial genotype distribution of Porphyromonas gingivalis in European subjects with or without chronic periodontitis. P. gingivalis was found in 71% and 9% of the samples from patients and healthy subjects, respectively. Enumeration of total P. gingivalis cell numbers by polymerase chain reaction and immunofluorescence showed excellent correspondence (r = 0.964). 73% of positive samples contained multiple fimA genotypes, but generally one genotype predominated by one to three orders of magnitude. Genotype II predominated in 60% of the samples. Genotype IV occurred with similar prevalence (73%) as genotype II but predominated in only 20% of the samples. Genotypes I, III and V were of much lower prevalence and cell densities of the latter two remained sparse. Our results suggest marked differences among the fimA genotypes' ability to colonize host sites with high cell numbers.