Association between more complex special care needs and overweight status and adolescents' difficulty with dental caries.

OBJECTIVE: Little is known about dental caries experience in adolescents with overweight and complex special health care needs (SHCNs). METHODS: Adolescent data (10-17 years) from the 2016-2020 National Survey of Children's Health (n = 91,196) was analyzed. The sample was grouped into the following: more complex SHCN and overweight, more complex SHCN without overweight, less complex SHCN and overweight, less complex SHCN without overweight, no SHCN but with overweight, and neither SHCN nor overweight. A multivariable-adjusted logistic regression model was conducted. RESULTS: Adolescents with more complex SHCNs with (OR: 1.82, 95% CI: 1.44-2.30, p < 0.001) or without overweight (OR: 1.51, 95% CI: 1.30-1.76, p < 0.001) were at higher odds of experiencing dental caries compared to healthy adolescents. No significant associations were observed between adolescents with less complex or no SHCN regardless of the overweight status with healthy adolescents. CONCLUSIONS: Adolescents with more complex SHCNs, irrespective of overweight status, experienced a higher caries severity than adolescents with no SHCNs or overweight.

A national sample of developmentally disabled adolescents with obesity and their utilization of preventive dental care services.

Background: Previous literature indicates that adolescents with developmental disabilities and obesity may have more oral health complications than healthy adolescents. However, dental care utilization among adolescents with developmental disabilities (DDs) and obesity is unclear. We investigated the differences in the utilization of preventive dental services between this high-risk group of adolescents and those with no DDs or obesity. Methods: Parent-reported data of adolescents 10-17 years (n = 68,942) from the 2016 to 2019 National Survey of Children's Health was used. In addition to descriptive and bivariate statistics, we ran three multiple logistic regression models guided by Andersen's Behavioral Model of Health Services Use, predicting the use of dental cleanings, fluoride treatments, and dental sealants. Results: Among adolescents with DDs and obesity, dental cleanings, fluoride treatments, and dental sealant utilization prevalence were 76%, 48%, and 21%, respectively. In comparison, adolescents with no DDs or obesity had a prevalence of 83%, 50%, and 19%, respectively. Multiple logistic regression analysis showed that adolescents with DDs and obesity did not significantly differ in their receipt of dental cleanings (p = .07), fluoride treatments (p = .55), and dental sealants (p = .23) compared to those with neither DDs nor obesity. Adolescents with DDs but no obesity were 22% and 30% more likely to receive fluoride treatments (p < .0001) and dental sealants (p < .0001), respectively. Conclusions: Fewer than half of adolescents with DDs and obesity utilized fluoride treatments, and less than one quarter utilized dental sealants but used all three preventive services at the same rate as those with no DDs or obesity. Implications: This study identified no differences in preventive dental care services utilization in adolescents with developmental disabilities (DDs) and obesity compared to those without DDs and obesity. However, the utilization of preventive dental services in this population is influenced by the federal poverty level and family background.

Correlation between Streptococcus mutans levels in dental plaque and saliva of children.

PURPOSE: This study was designed to compare the levels of Streptococcus mutans (S. mutans) in saliva with those in occlusal plaque on posterior teeth at different stages of dentition, and to explore the correlation with caries experience to determine the most suitable source of S. mutans for research. METHODS: Samples of saliva and occlusal plaque were collected from 83 patients (aged 3-17 years) over three months. S. mutans levels were determined by culture-based selective plating, morphological identification, and S.mutans-specific monoclonal antibody labeling. RESULTS: The mean age of the participants was 8.8 (±3.7) years, and 74.7% of them were Hispanic. Mean caries experience for children with primary, mixed, and permanent dentition was 5.2 (±4.7), 4.0 (±3.3), and 0.8 (±1.3), respectively. Children with primary and mixed dentition had a higher caries experience than children with permanent dentition (P < 0.01), despite having similar S. mutans levels and total bacteria. A positive correlation was observed between S. mutans levels in plaque and those in saliva, but not between S. mutans levels and caries experience. It was noteworthy that plaque samples harbored higher S. mutans levels (>105 CFU/mL) than saliva samples. CONCLUSION: Both plaque and saliva samples are useful sources for S. mutans isolation. S. mutans levels from both sources were not significantly correlated with caries experience, but occlusal plaque had greater sensitivity for quantification of high S. mutans levels.

Tooth-Specific Streptococcus mutans Distribution and Associated Microbiome.

Dental caries is multifactorial and polymicrobial in nature and remains one of the most common oral diseases. While caries research has focused on Streptococcus mutans as the main etiological pathogen, its impact at the tooth level is not fully understood. In this cross-sectional study, the levels and distribution of S. mutans in the posterior teeth at different dentition stages were investigated along with the corresponding tooth-specific microbiome. Occlusal plaque samples of 87 individual posterior teeth were collected from thirty children in three dentition stages (primary, mixed, and permanent). The S. mutans levels in the occlusal plaque of individual posterior teeth were quantified with qPCR, and those with preferential colonization were selected for tooth-specific microbiome analysis using 16S rRNA sequencing. Results: Quantification of S. mutans levels in the occlusal plaque confirmed the preferential colonization on the first primary and permanent molars. These teeth were selected for further tooth-specific microbiome sequencing, as they also displayed high caries experience. There were significant differences in the relative abundance of the four most abundant genera: Neisseria, Streptococcus, Rothia, and Veillonella. Furthermore, the tooth-level caries experience was correlated with a reduction in the microbiome diversity. Analyzing the different tooth-associated microbial communities, distinct tooth-specific core microbiomes were identified. Conclusions: Our findings suggest that caries susceptibility at the tooth level, depending on tooth type and dentition stage, is influenced by individual species as well as plaque community.

Effect of Cigarette and E-Cigarette Smoke Condensates on Candida albicans Biofilm Formation and Gene Expression.

Smoking triggers environmental changes in the oral cavity and increases the risk of mucosal infections caused by Candida albicans such as oral candidiasis. While cigarette smoke has a significant impact on C. albicans, how e-cigarettes affect this oral pathogen is less clear. Here, we investigated the effect of cigarette and e-cigarette smoke condensates (CSC and ECSC) on C. albicans growth, biofilm formation, and gene expression. Whereas pure nicotine (N) at the minimum inhibitory concentration (MIC, 4 mg/mL) prevented C. albicans growth, enhanced biofilm formation was observed at 0.1 mg/mL. In contrast, at this nicotine sub-MIC (0.1 mg/mL) concentration, CSC and ECSC had no significant effect on C. albicans biofilm formation. Additionally, N, CSC, and ECSC increased the expression of HWP1 and SAP2 genes. The ECSC group exhibited elevated expression levels of the EAP1 and ALS3 genes, compared to the nicotine-free ECSC (-) control. Moreover, our in vitro study illustrated that the antifungal drugs, fluconazole and amphotericin B, alleviated the effect of nicotine on C. albicans gene expression. Overall, the results of the study indicated nicotine from different sources may affect the pathogenic characteristics of C. albicans, including hyphal growth, biofilm formation, and particularly the expression of virulence-related genes.

Oral Microbiome: Streptococcus mutans/Caries Concordant-Discordant Children.

Dental caries remains the most common chronic disease in children, and the respective etiology is not fully understood. Though Streptococcus mutans is an important factor in the initiation and progression of caries, its presence is not always associated with the disease. The existence of caries discordant populations, in which S. mutans counts do not correlate with caries experience, poses a challenging problem. This study explored the possible correlation of S. mutans and other microorganism levels on caries-associated ecology of caries-concordant and discordant populations. A total of forty-seven children were analyzed in this study and stratified into four clinical groups based on their S. mutans levels in saliva (HS/LS: High/low S. mutans) and caries experience. Streptococcus mutans levels were determined by culture-based selective plating. The salivary microbiome of caries concordant and discordant populations was investigated by 16S rRNA gene sequencing and downstream bioinformatics analysis. The salivary microbial communities significantly clustered based on S. mutans levels and independent of their caries experience. In addition to S. mutans levels, significant differences in the abundance of other species were observed between HS and LS groups. Interestingly, disease-associated species such as Veillonella dispar, Streptococcus spp., and Prevotella spp. were significantly increased in HS groups and may contribute, in combination with S. mutans, to the caries progression. Furthermore, health-associated species exhibited higher abundance in the LS groups, such as Veillonella rogosae, Haemophilus sp., and Alloprevotella spp. but their possible contribution to the caries process remains to be elucidated. This study provides evidence that S. mutans may play a role in shaping the salivary microbial community. Our results highlight that future caries research should consider additional species as health/disease microbial markers in conjunction with S. mutans to improve diagnosis and caries management of the caries-discordant population.

The impact of fixed orthodontic appliances and clear aligners on the oral microbiome and the association with clinical parameters: A longitudinal comparative study.

INTRODUCTION: Orthodontic treatment interferes with oral hygiene and promotes plaque retention, which leads to gingival inflammation and enamel demineralization. Although removable clear aligners (CAs) are designed to improve oral hygiene compared with fixed appliances (FAs), comprehensive studies comparing their respective effects on the oral microbiome are limited. This longitudinal study investigated the microbial changes during orthodontic treatment with FA and CA in correlation with clinical parameters. METHODS: Clinical parameters and supragingival plaque were collected from 12 study participants for the FA or CA treatment groups at baseline and at least twice at the 1, 3, 6, and 12-month follow-up appointments. The plaque was also harvested from the aligner tray for the CA group. Microbiome composition was determined via 16S rRNA gene sequencing, compared between groups, and correlated with clinical parameters. RESULTS: Plaque (PI) and gingival indexes (GI) increased significantly in the FA but not the CA group. Beta but not alpha diversities of the microbial communities were distinct between the 2 treatment groups, even though genus-level differences were not significant except for Leptotrichia. The CA tray harbors a unique plaque community. Elevated PI and GI in the FA group correlated with a higher abundance of disease-related genera. CONCLUSIONS: Orthodontic treatments trigger appliance-related plaque community shifts from baseline, and the CA tray environment attracts distinct microbial communities. In comparison with FA, the use of CA resulted in better oral health index outcomes, which is reflected by the corresponding PI and GI-associated oral microbial communities.

Oral colonization of Candida albicans and Streptococcus mutans in children with or without fixed orthodontic appliances: A pilot study.

BACKGROUND/PURPOSE: Adolescents undergoing fixed orthodontic therapy have an increased risk of oral diseases due to additional plaque accumulation sites. However, the effect of fixed orthodontics appliances (FOAs) on the colonization of Candida albicans (Ca) and Streptococcus mutans (Sm), two synergistic oral pathogens, is largely unknown and was, therefore, the primary objective of this pilot investigation. MATERIAL AND METHODS: Sixteen children aged 10-15 years were enrolled, nine in the FOA and seven in the control groups. Saliva and occlusal plaque were collected, and the Ca and Sm levels were quantified with a quantitative real-time polymerase chain reaction (qPCR) assay. RESULTS: A trend of higher Ca levels was observed in the saliva and occlusal plaque of the FOA group, while the control group contained higher levels of Sm. Furthermore, for Sm levels, a positive correlation between saliva and occlusal plaque was shown in both the FOA and control groups; in contrast, Ca levels were negatively correlated between these samples only in the FOA group. Between Ca and Sm, a positive correlation was observed in saliva and occlusal plaque in the control group; however, this relationship was disrupted in the FOA group. CONCLUSION: Our preliminary study demonstrated that the presence of FOAs disturbs the colonization of Ca and Sm within the oral cavity. This perturbation might increase orthodontic patients' risk for Ca- and Sm-related diseases.

Pilot study on selective antimicrobial effect of a halitosis mouthrinse: monospecies and saliva-derived microbiome in an in vitro model system.

BACKGROUND: Halitosis refers to malodor emanating from the oral cavity. Several mouthrinses with halitosis-reduction exist on the market, but their effect on the oral microbiome is largely unknown. In this study, we used an efficient in vitro model system to investigate a test mouthrinse's impact on the oral microbiome. METHODS: Single halitosis-associated species and other common oral microorganism cultures were exposed to the test mouthrinse over time, and their viability was determined by culture-based selective plating. Next, the saliva-derived microbiome from healthy and halitosis-associated individuals was cultured in the presence of the test mouthrinse over time using the previously developed in vitro model system. The microbiome composition was assessed with 16S rRNA gene sequencing and downstream bioinformatics analyses. RESULTS: The test mouthrinse displayed antimicrobial activity against known anaerobic bacterial species producing halitosis-related compounds such as Fusobacterium nucleatum, F. periodonticum, and Prevotella intermedia but not against other common oral microorganisms. In the multispecies, saliva-derived cultures, mouthrinse exposure decreased the relative abundance of the Fusobacterium and Prevotella genera while not affecting overall diversity. CONCLUSIONS: The test mouthrinse had promising anti-halitosis characteristics at the microbiome level, as demonstrated by the reduction in the relative abundance of halitosis-associated taxa while maintaining microbial diversity.

Omics and interspecies interaction.

Interspecies interactions are key determinants in biofilm behavior, ecology, and architecture. The cellular responses of microorganisms to each other at transcriptional, proteomic, and metabolomic levels ultimately determine the characteristics of biofilm and the corresponding implications for health and disease. Advances in omics technologies have revolutionized our understanding of microbial community composition and their activities as a whole. Large-scale analyses of the complex interaction between the many microbial species residing within a biofilm, however, are currently still hampered by technical and bioinformatics challenges. Thus, studies of interspecies interactions have largely focused on the transcriptional and proteomic changes that occur during the contact of a few prominent species, such as Porphyromonas gingivalis, Streptococcus mutans, Candida albicans, and a few others, with selected partner species. Expansion of available tools is necessary to grow the revealing, albeit limited, insight these studies have provided into a profound understanding of the nature of individual microbial responses to the presence of others. This will allow us to answer important questions including: Which intermicrobial interactions orchestrate the myriad of cooperative, synergistic, antagonistic, manipulative, and other types of relationships and activities in the complex biofilm environment, and what are the implications for oral health and disease?

Deep metagenomics examines the oral microbiome during dental caries, revealing novel taxa and co-occurrences with host molecules.

Dental caries, the most common chronic infectious disease worldwide, has a complex etiology involving the interplay of microbial and host factors that are not completely understood. In this study, the oral microbiome and 38 host cytokines and chemokines were analyzed across 23 children with caries and 24 children with healthy dentition. De novo assembly of metagenomic sequencing obtained 527 metagenome-assembled genomes (MAGs), representing 150 bacterial species. Forty-two of these species had no genomes in public repositories, thereby representing novel taxa. These new genomes greatly expanded the known pangenomes of many oral clades, including the enigmatic Saccharibacteria clades G3 and G6, which had distinct functional repertoires compared to other oral Saccharibacteria. Saccharibacteria are understood to be obligate epibionts, which are dependent on host bacteria. These data suggest that the various Saccharibacteria clades may rely on their hosts for highly distinct metabolic requirements, which would have significant evolutionary and ecological implications. Across the study group, Rothia, Neisseria, and Haemophilus spp. were associated with good dental health, whereas Prevotella spp., Streptococcus mutans, and Human herpesvirus 4 (Epstein-Barr virus [EBV]) were more prevalent in children with caries. Finally, 10 of the host immunological markers were significantly elevated in the caries group, and co-occurrence analysis provided an atlas of potential relationships between microbes and host immunological molecules. Overall, this study illustrated the oral microbiome at an unprecedented resolution and contributed several leads for further study that will increase the understanding of caries pathogenesis and guide therapeutic development.

Effectiveness of the GumChucks flossing system compared to string floss for interdental plaque removal in children: a randomized clinical trial.

Flossing, an important oral hygiene skill, is technique-sensitive and challenging for children with developing manual dexterity. GumChucks is a novel flossing device designed to assist children with proper flossing technique. The aim of this study was to assess the efficacy of the GumChucks flossing device compared to string floss (SF). We conducted a randomized trial with 40 children aged 4-15 years at the UCLA Children's Dental Center from January- April 2017. Participants were randomly assigned to either GumChucks or SF. Interdental plaque score (IPS) and gingival index (GI) were recorded at baseline and 4-week post-usage. Flossing speed and interdental plaque reduction were also determined immediately after first use. In addition, questionnaires were completed by children, parents and dentists. Overall, children flossed significantly faster (p < 0.001) and achieved greater IPS reduction after first use (47.0% vs. 26.8%) with GumChucks compared to SF. After 4-week post-usage, children ages 10-15 in the GumChucks group demonstrated significantly greater improvement in GI and IPS from baseline (p < 0.01) and greater efficacy in interdental plaque removal compared to the SF group (p < 0.01). Children ages 4-9 flossed more effectively (p < 0.01) with GumChucks after first use, but no significant IPS and GI improvement after 4-week post-usage. Children preferred GumChucks (92.5%) over SF, with a similar positive attitude reported by parents and dentists. GumChucks is an effective alternative interdental plaque removal aid that allows children to floss with greater speed and efficacy, with recommended parental supervision for children under age 10.

Identification of the Bacterial Biosynthetic Gene Clusters of the Oral Microbiome Illuminates the Unexplored Social Language of Bacteria during Health and Disease.

Small molecules are the primary communication media of the microbial world. Recent bioinformatic studies, exploring the biosynthetic gene clusters (BGCs) which produce many small molecules, have highlighted the incredible biochemical potential of the signaling molecules encoded by the human microbiome. Thus far, most research efforts have focused on understanding the social language of the gut microbiome, leaving crucial signaling molecules produced by oral bacteria and their connection to health versus disease in need of investigation. In this study, a total of 4,915 BGCs were identified across 461 genomes representing a broad taxonomic diversity of oral bacteria. Sequence similarity networking provided a putative product class for more than 100 unclassified novel BGCs. The newly identified BGCs were cross-referenced against 254 metagenomes and metatranscriptomes derived from individuals either with good oral health or with dental caries or periodontitis. This analysis revealed 2,473 BGCs, which were differentially represented across the oral microbiomes associated with health versus disease. Coabundance network analysis identified numerous inverse correlations between BGCs and specific oral taxa. These correlations were present in healthy individuals but greatly reduced in individuals with dental caries, which may suggest a defect in colonization resistance. Finally, corroborating mass spectrometry identified several compounds with homology to products of the predicted BGC classes. Together, these findings greatly expand the number of known biosynthetic pathways present in the oral microbiome and provide an atlas for experimental characterization of these abundant, yet poorly understood, molecules and socio-chemical relationships, which impact the development of caries and periodontitis, two of the world's most common chronic diseases.IMPORTANCE The healthy oral microbiome is symbiotic with the human host, importantly providing colonization resistance against potential pathogens. Dental caries and periodontitis are two of the world's most common and costly chronic infectious diseases and are caused by a localized dysbiosis of the oral microbiome. Bacterially produced small molecules, often encoded by BGCs, are the primary communication media of bacterial communities and play a crucial, yet largely unknown, role in the transition from health to dysbiosis. This study provides a comprehensive mapping of the BGC repertoire of the human oral microbiome and identifies major differences in health compared to disease. Furthermore, BGC representation and expression is linked to the abundance of particular oral bacterial taxa in health versus dental caries and periodontitis. Overall, this study provides a significant insight into the chemical communication network of the healthy oral microbiome and how it devolves in the case of two prominent diseases.

Development of a quantitative assay amenable for high-throughput screening to target the type II secretion system for new treatments against plant-pathogenic bacteria.

Plant-pathogenic bacteria are the causative agents of diseases in important agricultural crops and ornamental plants. The severe economic burden of these diseases requires seeking new approaches for their control, particularly because phytopathogenic bacteria are often resistant to available treatments. The type II secretion (T2S) system is a key virulence factor used by major groups of phytopathogenic bacteria. The T2S machinery transports many hydrolytic enzymes responsible for degradation of the plant cell wall, thus enabling successful colonization and dissemination of the bacteria in the plant host. The genetic inactivation of the T2S system leads to loss of virulence, which strongly suggests that targeting the T2S could enable new treatments against plant-pathogenic bacteria. Accordingly, we have designed and optimized an assay to identify small-molecule inhibitors of the T2S system. This assay uses a double parametric output: measurement of bacterial growth and the enzymatic activity of cellulase, which is secreted via the T2S pathway in our model organism Dickeya dadantii. The assay was evaluated by screening natural extracts, culture filtrates isolated from rhizosphere bacteria, and a collection of pharmaceutically active compounds in LOPAC(1280). The calculated Z' values of 0.63, 0.63, and 0.58, respectively, strongly suggest that the assay is applicable for a high-throughput screening platform.