Near-infrared imaging in orthodontic intraoral scanners for early interproximal caries detection.

INTRODUCTION: Intraoral scanners commonly used in orthodontic offices now offer near-infrared imaging (NIRI) technology, advertised as a screening tool to identify interproximal caries. This study aimed to evaluate the reliability and validity of NIRI detection of interproximal carious lesions in a common intraoral scanner (iTero Element 5D; Align Technology, San Jose, Calif) with and without bitewing radiograph complement, compared with a microcomputed tomography (micro-CT) reference standard. METHODS: Extracted human posterior teeth (premolars and molars) were selected for early (noncavitated) interproximal carious lesions (n = 39) and sound control surfaces (n = 47). The teeth were scanned via micro-CT for evaluation by 2 blinded evaluators using consensus scoring. The teeth were mounted to simulate anatomic interproximal contacts and underwent a NIRI scan using iTero Element 5D and bitewing radiographs. Two trained, calibrated examiners independently evaluated (1) near-infrared images alone with clinical photograph, (2) bitewing radiograph alone with clinical photograph, and (3) near-infrared images with bitewing radiograph and clinical photograph in combination, after at least a 10-day washout period between each evaluation. RESULTS: Interrater reliability was highest for NIRI alone (k = 0.533) compared with bitewing radiograph alone (k = 0.176) or in combination (k = 0.256). NIRI alone showed high specificity (0.83-0.96) and moderate sensitivity (0.42-0.63) compared with a micro-CT reference standard. Dentin lesions were significantly more reliably detected than enamel lesions. CONCLUSIONS: After rigorous training and calibration, NIRI can be used with moderate reliability, high specificity, and moderate sensitivity to detect noncavitated interproximal carious lesions.

Microcomputed Tomography Mineral Density Profile as Reference Standard for Early Carious Lesion Activity Assessment.

Early caries diagnosis is crucial to treatment decisions in dentistry and requires identification of lesion activity: whether a carious lesion is active (progressively demineralizing) or arrested (progressively remineralizing). This study aimed to identify microtomographic (micro-CT) differences between active and arrested smooth surface enamel lesions, to quantify those micro-CT differences by creating thresholds for ex vivo caries activity assessment to serve as a future reference standard, and to validate those thresholds against the remaining sample. Extracted human permanent teeth (n = 59) were selected for sound surfaces and non-cavitated smooth surface carious lesions. Each surface was then examined for caries activity by calibrated individuals via visual-tactile examination using the International Caries Classification and Management System (ICCMS) activity criteria. Each tooth was scanned via micro-CT and the mineral density was plotted against lesion depth. The area under the curve (AUC) was calculated and represented the loss of density for the outermost 96 µm of enamel. AUC thresholds obtained from micro-CT were established to classify sound, remineralized, and demineralized surfaces against the gold standard examiner's lesion assessment of sound, inactive, and active lesions, respectively. The established AUC thresholds demonstrated moderate agreement with the assessment in identifying demineralized lesions (k = 0.45), with high sensitivity (0.73) and specificity (0.77). This study demonstrated quantifiable differences among demineralized lesions, remineralized lesions, and sound surfaces, which contributes to the establishment of micro-CT as a reference standard for caries activity that may be used to improve clinical and laboratorial dental caries evaluations.

Selenomonas sputigena acts as a pathobiont mediating spatial structure and biofilm virulence in early childhood caries.

Streptococcus mutans has been implicated as the primary pathogen in childhood caries (tooth decay). While the role of polymicrobial communities is appreciated, it remains unclear whether other microorganisms are active contributors or interact with pathogens. Here, we integrate multi-omics of supragingival biofilm (dental plaque) from 416 preschool-age children (208 males and 208 females) in a discovery-validation pipeline to identify disease-relevant inter-species interactions. Sixteen taxa associate with childhood caries in metagenomics-metatranscriptomics analyses. Using multiscale/computational imaging and virulence assays, we examine biofilm formation dynamics, spatial arrangement, and metabolic activity of Selenomonas sputigena, Prevotella salivae and Leptotrichia wadei, either individually or with S. mutans. We show that S. sputigena, a flagellated anaerobe with previously unknown role in supragingival biofilm, becomes trapped in streptococcal exoglucans, loses motility but actively proliferates to build a honeycomb-like multicellular-superstructure encapsulating S. mutans, enhancing acidogenesis. Rodent model experiments reveal an unrecognized ability of S. sputigena to colonize supragingival tooth surfaces. While incapable of causing caries on its own, when co-infected with S. mutans, S. sputigena causes extensive tooth enamel lesions and exacerbates disease severity in vivo. In summary, we discover a pathobiont cooperating with a known pathogen to build a unique spatial structure and heighten biofilm virulence in a prevalent human disease.

Dental caries and their microbiomes in children: what do we do now?

The oral cavity is an unique ecosystem formed by different structures, tissues, and a complex microbial community formed by hundreds of different species of bacteria, fungi, viruses, phages, and the candidate phyla radiation (CPR) group, all living in symbiosis with healthy individuals. In an opposite state, dental caries is a biofilm-mediated dysbiosis that involves changes in the core microbiome composition and function, which leads to the demineralization of tooth tissues due to the fermentation of dietary carbohydrates, producing acid by select oral bacteria. The cariogenic biofilm is typically characterized by bacterial species with the ability of adhering to the saliva-coated tooth surface, production of exopolysaccharides-rich matrix (which will limit the diffusion of acidic products of carbohydrate fermentation), and the ability of surviving in this acidic environment. Besides years of research and dental treatment, dental caries remains the most common chronic disease in children worldwide. This article aims to bring an insightful discussion about important questions that remain unanswered in the Cariology and Oral Microbiology fields, to move Science forward, characterize the interrelationships of these communities, and understand mechanistic functions between microorganisms and the host, therefore leading to translatable knowledge that benefits the provision of care to our pediatric patients.

BZINB Model-Based Pathway Analysis and Module Identification Facilitates Integration of Microbiome and Metabolome Data.

Integration of multi-omics data is a challenging but necessary step to advance our understanding of the biology underlying human health and disease processes. To date, investigations seeking to integrate multi-omics (e.g., microbiome and metabolome) employ simple correlation-based network analyses; however, these methods are not always well-suited for microbiome analyses because they do not accommodate the excess zeros typically present in these data. In this paper, we introduce a bivariate zero-inflated negative binomial (BZINB) model-based network and module analysis method that addresses this limitation and improves microbiome-metabolome correlation-based model fitting by accommodating excess zeros. We use real and simulated data based on a multi-omics study of childhood oral health (ZOE 2.0; investigating early childhood dental caries, ECC) and find that the accuracy of the BZINB model-based correlation method is superior compared to Spearman's rank and Pearson correlations in terms of approximating the underlying relationships between microbial taxa and metabolites. The new method, BZINB-iMMPath, facilitates the construction of metabolite-species and species-species correlation networks using BZINB and identifies modules of (i.e., correlated) species by combining BZINB and similarity-based clustering. Perturbations in correlation networks and modules can be efficiently tested between groups (i.e., healthy and diseased study participants). Upon application of the new method in the ZOE 2.0 study microbiome-metabolome data, we identify that several biologically-relevant correlations of ECC-associated microbial taxa with carbohydrate metabolites differ between healthy and dental caries-affected participants. In sum, we find that the BZINB model is a useful alternative to Spearman or Pearson correlations for estimating the underlying correlation of zero-inflated bivariate count data and thus is suitable for integrative analyses of multi-omics data such as those encountered in microbiome and metabolome studies.

BZINB model-based pathway analysis and module identification facilitates integration of microbiome and metabolome data.

Integration of multi-omics data is a challenging but necessary step to advance our understanding of the biology underlying human health and disease processes. To date, investigations seeking to integrate multi-omics (e.g., microbiome and metabolome) employ simple correlation-based network analyses; however, these methods are not always well-suited for microbiome analyses because they do not accommodate the excess zeros typically present in these data. In this paper, we introduce a bivariate zero-inflated negative binomial (BZINB) model-based network and module analysis method that addresses this limitation and improves microbiome-metabolome correlation-based model fitting by accommodating excess zeros. We use real and simulated data based on a multi-omics study of childhood oral health (ZOE 2.0; investigating early childhood dental disease, ECC) and find that the accuracy of the BZINB model-based correlation method is superior compared to Spearman’s rank and Pearson correlations in terms of approximating the underlying relationships between microbial taxa and metabolites. The new method, BZINB-iMMPath facilitates the construction of metabolite-species and species-species correlation networks using BZINB and identifies modules of (i.e., correlated) species by combining BZINB and similarity-based clustering. Perturbations in correlation networks and modules can be efficiently tested between groups (i.e., healthy and diseased study participants). Upon application of the new method in the ZOE 2.0 study microbiome-metabolome data, we identify that several biologically-relevant correlations of ECC-associated microbial taxa with carbohydrate metabolites differ between healthy and dental caries-affected participants. In sum, we find that the BZINB model is a useful alternative to Spearman or Pearson correlations for estimating the underlying correlation of zero-inflated bivariate count data and thus is suitable for integrative analyses of multi-omics data such as those encountered in microbiome and metabolome studies.

Human Tooth as a Fungal Niche: Candida albicans Traits in Dental Plaque Isolates.

Candida albicans, a fungus typically found in the mucosal niche, is frequently detected in biofilms formed on teeth (dental plaque) of toddlers with severe childhood caries, a global public health problem that causes rampant tooth decay. However, knowledge about fungal traits on the tooth surface remains limited. Here, we assess the phylogeny, phenotype, and interkingdom interactions of C. albicans isolated from plaque of diseased toddlers and compare their properties to reference strains, including 529L (mucosal isolate). C. albicans isolates exhibit broad phenotypic variations, but all display cariogenic traits, including high proteinase activity, acidogenicity, and acid tolerance. Unexpectedly, we find distinctive variations in filamentous growth, ranging from hyphal defective to hyperfilamentous. We then investigate the ability of tooth isolates to form interkingdom biofilms with Streptococcus mutans (cariogenic partner) and Streptococcus gordonii (mucosal partner). The hyphal-defective isolate lacks cobinding with S. gordonii, but all C. albicans isolates develop robust biofilms with S. mutans irrespective of their filamentation state. Moreover, either type of C. albicans (hyphae defective or hyperfilamentous) enhances sucrose metabolism and biofilm acidogenicity, creating highly acidic environmental pH (<5.5). Notably, C. albicans isolates show altered transcriptomes associated with pH, adhesion, and cell wall composition (versus reference strains), further supporting niche-associated traits. Our data reveal that C. albicans displays distinctive adaptive mechanisms on the tooth surface and develops interactions with pathogenic bacteria while creating an acidogenic state regardless of fungal morphology, contrasting with interkingdom partnerships in mucosal infections. Human tooth may provide new insights into fungal colonization/adaptation, interkingdom biofilms, and contributions to disease pathogenesis. IMPORTANCE Severe early childhood caries is a widespread global public health problem causing extensive tooth decay and systemic complications. Candida albicans, a fungus typically found in mucosal surfaces, is frequently detected in dental plaque formed on teeth of diseased toddlers. However, the clinical traits of C. albicans isolated from tooth remain underexplored. Here, we find that C. albicans tooth isolates exhibit unique biological and transcriptomic traits. Notably, interkingdom biofilms with S. mutans can be formed irrespective of their filamentation state. Furthermore, tooth isolates commonly share dental caries-promoting functions, including acidogenesis, proteolytic activity, and enhanced sugar metabolism, while displaying increased expression of pH-responsive and adhesion genes. Our findings reveal that C. albicans colonizing human teeth displays distinctive adaptive mechanisms to mediate interkingdom interactions associated with a disease-causing state on a mineralized surface, providing new insights into Candida pathobiology and its role in a costly pediatric disease.

Oral biofilm dysbiosis during experimental periodontitis.

OBJECTIVES: We have previously characterized the main osteoimmunological events that occur during ligature periodontitis. This study aims to determine the polymicrobial community shifts that occur during disease development. METHODS: Periodontitis was induced in C57BL/6 mice using the ligature-induced periodontitis model. Healthy oral mucosa swabs and ligatures were collected every 3 days from 0 to 18 days post-ligature placement. Biofilm samples were evaluated by 16SrRNA gene sequencing (Illumina MiSeq) and QIIME. Time-course changes were determined by relative abundance, diversity, and rank analyses (PERMANOVA, Bonferroni-adjusted). RESULTS: Microbial differences between health and periodontal inflammation were observed at all phylogenic levels. An evident microbial community shift occurred in 25 genera during the advancement of "gingivitis" (3-6 days) to periodontitis (9-18 days). From day 0 to 18, dramatic changes were identified in Streptococcus levels, with an overall decrease (54.04%-0.02%) as well an overall increase of Enterococcus and Lactobacillus (23.7%-73.1% and 10.1%-70.2%, respectively). Alpha-diversity decreased to its lowest at 3 days, followed by an increase in diversity as disease advancement. Beta-diversity increased after ligature placement, indicating that bone loss develops in response to a greater microbial variability (p = 0.001). Levels of facultative and strict anaerobic bacteria augmented over the course of disease progression, with a total of eight species significantly different during the 18-day period. CONCLUSION: The data supports that murine gingival inflammation and alveolar bone loss develop in response to microbiome shifts. Bacterial diversity increased during progression to bone loss. These findings further support the utilization of the periodontitis ligature model for microbial shift analysis under different experimental conditions.

Is Fluorescence Technology a Promising Tool for Detecting Infected Dentin in Deep Carious Lesions?

The purpose of this study was to determine if the degree of fluorescence detected by fluorescence-aided caries excavation (FACE) correlates with dentin bacterial microbiome diversity, as assessed by 16S rRNA gene amplicon sequencing, and with traditional tactile dentin caries assessment. Unidentified human teeth were obtained from a dental facility. The included teeth had a carious lesion two-thirds into the dentin, verified by radiography, and were red-fluorescing (RF) using FACE technology (SIROInspect; Sirona, Bensheim, Germany). Two independent examiners performed visual/tactile assessment of the lesions. RF sites were sampled with a sterile spoon excavator and dentin characteristics were evaluated. Once RF dentin was removed, a second sample of pink-fluorescing (PF) dentin was obtained. After excavation with a sterile round bur to nonfluorescing (NF) dentin, a third sample was collected with a slow-speed round bur. The samples were processed at the UNC (University of North Carolina at Chapel Hill) Microbiome Core Facility. Out of 134 extracted teeth collected, 21 fit the inclusion criteria, yielding 61 dentin samples. RF samples had the highest number of observed operational taxonomic units (n = 154), followed by PF (n = 109) and NF (n = 100). RF carious dentin was primarily "soft," and NF dentin was assessed as "hard" 100% of the time by both examiners (rank correlation χ2: p < 0.001). However, approximately one-third of the tactile assessments of hard dentin still displayed some fluorescence, either pink or red. We concluded that the sampled fluorescing (RF and PF) and NF carious dentin layers displayed diverse bacterial taxa, and tactile assessments of soft, leathery, and hard corresponded with RF, PF, and NF.

Influence of sonic application on surface roughness of bulk-fill resin composites.

PURPOSE: To evaluate the influence of sonic application on the surface roughness of bulk-fill resin composite restorations. METHODS: 80 intact bovine incisors had their incisal thirds removed, their buccal surfaces flattened, and standard preparations mimicking Class II preparations performed on their buccal surfaces. Specimens were then randomly assigned for restoration with the bulk-fill resin composites Tetric EvoCeram Bulk Fill, SureFil SDR flow+, and SonicFill 2, with sonic application for 15 seconds and 30 seconds as well as no sonic application. Filtek Supreme Ultra applied without sonic application served as control. Sonic application was accomplished with the KAVO SONICflex handpiece. Surface roughness was measured using a 3D scanner and data were analyzed using the Kruskal-Wallis and Mann-Whitney tests at a significance level of 0.05. RESULTS: No statistical differences were found between groups treated with and without sonic application. When materials were compared, Tetric EvoCeram Bulk Fill showed the lowest surface roughness in the 15-second sonic application subgroup; and SureFil SDR flow+ showed the highest surface roughness when applied without sonic application and in the 30-second sonic application subgroups. CLINICAL SIGNIFICANCE: Clinicians should expect a similar performance of bulk-fill resin composites in terms of surface roughness when applied with or without sonic activation. Precaution should be taken during the finishing/ polishing procedures of SureFil SDR flow+ resin composite.

The oral bacterial microbiome of occlusal surfaces in children and its association with diet and caries.

Dental caries is the most prevalent disease in humans globally. Efforts to control it have been invigorated by an increasing knowledge of the oral microbiome composition. This study aimed to evaluate the bacterial diversity in occlusal biofilms and its relationship with clinical surface diagnosis and dietary habits. Anamneses were recorded from thirteen 12-year-old children. Biofilm samples collected from occlusal surfaces of 46 permanent second molars were analyzed by 16S rRNA amplicon sequencing combined with the BLASTN-based search algorithm for species identification. The overall mean decayed, missing and filled surfaces modified index [DMFSm Index, including active white spot lesions (AWSL)] value was 8.77±7.47. Biofilm communities were highly polymicrobial collectively, representing 10 bacterial phyla, 25 classes, 29 orders, 58 families, 107 genera, 723 species. Streptococcus sp_Oral_Taxon_065, Corynebacterium matruchotii, Actinomyces viscosus, Actinomyces sp_Oral_Taxon_175, Actinomyces sp_Oral_Taxon_178, Actinomyces sp_Oral_Taxon_877, Prevotella nigrescens, Dialister micraerophilus, Eubacterium_XI G 1 infirmum were more abundant among surfaces with AWSL, and Streptococcus gordonii, Streptococcus sp._Oral_Taxon_058, Enterobacter sp._str._638 Streptococcus australis, Yersinia mollaretii, Enterobacter cloacae, Streptococcus sp._Oral_Taxon_71, Streptococcus sp._Oral_Taxon_F11, Centipeda sp._Oral_Taxon_D18 were more abundant among sound surfaces. Streptococcus mutans was detected on all surfaces in all patients, while Streptococcus sobrinus was detected only in three patients (mean relative abundances 7.1% and 0.6%, respectively). Neither species differentiated healthy from diseased sites. Diets of nine of the subjects were scored as high in fermentable carbohydrates (≧2X/day between meals). A direct association between relative abundances of bacteria and carbohydrate consumption was observed among 18 species. High consumption of fermentable carbohydrates and sound surfaces were associated with a reduction in bacterial diversity. PCoA plots displayed differences in bacterial community profiles between sound and diseased surfaces. Our study showed that, in addition to mutans streptococci, other species may be associated with the initiation of dental caries on occlusal surfaces, and that biofilm diversity of tooth surfaces is influenced by carbohydrate consumption and a surface's health status.

Métodos de detecção de cárie / Methods of caries detection

A Odontologia contemporânea não deve restringir-se a identificar cavidades, mas, também, despertar para um diagnóstico precoce das lesões cariosas iniciais. Os mais diversos e modernos métodos de detecção de cárie vêm sendo desenvolvidos com objetivo de tornar mais preciso o diagnóstico de cárie dental. Os métodos de detecção mais utilizados são os convencionais, porém a associação dos métodos proporciona melhores resultados ao exame clínico tradicional isolado. Este estudo teve como propósito a revisão de literatura sobre os métodos de detecção da doença cárie disponíveis atualmente, baseados nos métodos radiográficos, fluorescência a laser e corrente elétrica.

The contemporary Odontology cannot be restricted to just identify cavities, but must be interested in an early diagnosis of caries lesions. Varied and modern methods of caries detection are being developed, aiming to turn the diagnosis of dental caries more accurate. The conventional methods are most used for detection, however, the association of methods offer better results to the clinician. The purpose of this paper was to do a literature review and to compare the methods of caries detection, based on radiographic methods, laser fluorescence and electric chain.

Contribuição ao estudo dos fatores de risco para cárie em crianças infectadas pelo Hiv: hábitos de contatos salivares / Contribution to the risk factors study for dental caries in hiv infected children: contact salivary habits

O objetivo do trabalho foi avaliar a relação entre oshábitos de contatos salivares (CHS)de responsáveis por crianças infectadas pelo HIV (HIV+) e por crianças sem evidências de imunossupressão (HIV-) com a experiência de cárie. Após consentimento assinado pelos responsáveis, a cavidade bucal de 63 crianças HIV+ (33 do sexo feminino) e de 58 crianças HIV- (21 do sexo feminino), com idade entre 3 e 12 anos, foram examinadas utilizando espelho bucal, sonda exploradora e gaze sob luz ambiente. Os responsáveis foram questionados sobre a presença de algum HCS, como: usar a mesma colher, limpar a chupeta na boca, beijar a boca da criança e assoprar a colher para esfriar alimentos. Para análise dos resultados foram utilizados os testes Qui-quadrado e Kruskal-Wallis. O ceod e CPOD médios foram 8,5(± 13,0) e 3,7(±7,2) para i grupo de crianças HIV+ e, 5,6(±6,5) e 2,1(±3,4) para o grupo HIV-, respectivamente. O grau de parentesco mais frequente foi de pais em ambos os grupos. 28 (44,4 por cento) responsáveis por crianças infectada pelo HIV e 69 por cento das crianças sem evidências de imunossupressão relataram a presença de algum HCS (p=0,007). O mais frequente foi assoprar a colher para esfriar alimento em ambos os grupos. Pode-se concluir que embora a frequência de HCS fosse maior no grupo de crianças HIV-, o mesmo não foi observado em relação à experiência de cárie, mostrando não haver relação entre a experiência nestes dois grupos e os HCS