Probiotics-containing beverages and dental caries: A systematic review and meta-analysis.

The aim was to evaluate the effect of probiotics-containing beverages on dental caries development by a systematic review and meta-analysis (PROSPERO CRD42021258010). PubMed, Embase, and Web of Science databases were assessed (up to October 2023) for randomized placebo-controlled trials that evaluated the consumption of probiotics-containing beverages having dental caries as endpoint. Risk of bias assessment (Cochrane Handbook) was performed for all studies. Risk ratio (RR), standardized mean difference (SMD) and 95% confidence interval (CI), were calculated for caries prevalence and caries increment, respectively. Meta-analysis was performed. Qualitative analysis of more than 2000 individuals aged from 1 to 84 years (9 studies) showed inconsistent effects of these probiotics-containing beverages. Synthesis of more than 1000 children and adolescents did not demonstrate a statistically significant difference in the proportion of individuals with cavitated lesions (RR 0.60 [95 % CI 0.34 to 1.08]; n = 2), but a significantly smaller increment of both cavitated and non-cavitated carious lesions (SMD -0.23 [95 % CI -0.39 to -0.08]; n = 4) was related to the consumption of probiotics-containing beverages. Certainty of evidence for caries prevalence and caries increment were moderate and low, respectively. Current evidence is not substantial to strengthen the impact of these beverages on caries development.

Consensus for teaching dental caries in the Portuguese Language at Brazilian dental schools.

The aim of this paper was to present a summary of the process of developing and preparing the final documents of the national consensus for teaching undergraduate Brazilian dental students the dental caries curriculum in the Portuguese language. The final document was developed in three steps: a) The ABENO and LAOHA cariology group invited experts from all five regions of Brazil to participate in the discussion. The theoretical support for crafting the first draft of the consensus was based on two publications: National Curriculum Guidelines of the Dentistry graduation in Brazil, Ministry of Education (2021) and the competences described in the European Core Curriculum for Cariology (ORCA-ADEE, 2011); b) The group of experts was divided into 5 working groups: G1-Domain, Main and Specific Competences, G2-Essential knowledge, G3-Life course perspective, G4-Social determinants and dental caries, G5- Glossary. The document was finalized by thoroughly reviewing the process using Delphi methodology; c) The 5-chapter document (one from each working group) was submitted to three open public consultations in 2022 (May-June, August, and October) using Google-forms. The suggestions (content/wording) were discussed within the group as: totally accepted, partially accepted, and rejected. A total of 192 suggestions were registered from 31 dental schools in all regions of Brazil. The number of suggestions received per Group were: 84, 28, 26, 24, 30 suggestions for G1, G2, G3, G4 and G5, respectively. The majority of suggestions were totally accepted by the group of experts (n = 172, 89.6%), 15 were partially accepted (7.8%), and 5 were rejected. Conclusion The final document could be considered to be the first national consensus for teaching the dental caries curriculum in Brazil.

Hub genes and pathways related to caries-free dental biofilm: clinical metatranscriptomic study.

OBJECTIVE: This study aimed to evaluate the microbial functional profile of biofilms related to caries-free (CF, n = 6) and caries-arrested (CI, n = 3) compared to caries-active (CA, n = 5) individuals. MATERIALS AND METHODS: A metatranscriptomic was performed in supragingival biofilm from different clinical conditions related to caries or health. Total RNA was extracted and cDNAs were obtained and sequenced (Illumina HiSeq3000). Trimmed data (SortMeRNA) were submitted to the SqueezeMeta pipeline in the co-assembly mode for functional analysis and further differential gene expression analysis (DESeq2) and weighted gene co-expression network analysis (WCGNA) to explore and identify gene modules related to these clinical conditions. RESULTS: A total of 5303 genes were found in the metatranscriptomic analysis. A co-expression network identified the most relevant modules strongly related to specific caries status. Correlation coefficients were calculated between the eigengene modules and the clinical conditions (CA, CI, and CF) discriminating multiple modules. CA and CI showed weak correlation coefficient strength across the modules, while the CF condition presented a very strong positive correlation coefficient (r = 0.9, p value = 4 × 10-9). Pearson's test was applied to further analyze the module membership and gene significance in CF conditions, and the most relevant were HSPA1s-K03283, Epr- K13277, and SLC1A-K05613. Gene Ontology (GO) shows important bioprocesses, such as two-component system, fructose and mannose metabolism, pentose and glucuronate interconversions, and flagellar assembly (p-adjust < 0.05). The ability to use different carbohydrates, integrate multiple signals, swarm, and bacteriocin production are significant metabolic advantages in the oral environment related to CF. CONCLUSIONS: A distinct functional health profile could be found in CF, where co-occurring genes can act in different pathways at the same time. Genes HSPA1s, Epr, and SLC1A may be appointed as potential biomarkers for caries-free biofilms. CLINICAL RELEVANCE: Potential biomarkers for caries-free biofilms could contribute to the knowledge of caries prevention and control.

Biofilm dysbiosis and caries activity: a surface or an individual issue?

OBJECTIVE: This study aimed to analyze the functional profile of supragingival biofilm from sound (CAs), active (CAa), and inactive (CAi) enamel caries lesions from caries-active individuals to provide insights into the diversity of biological processes regarding biofilm dysbiosis. METHODOLOGY: A metatranscriptome analysis was performed in biofilm samples collected from five caries-active individuals. Total RNA was extracted, and the microbial cDNAs were obtained and sequenced (Illumina HiSeq3000). Trimmed data were submitted to the SqueezeMeta pipeline in the co-assembly mode for functional analysis and further differential gene expression analysis (DESeq2). RESULTS: Bioinformatics analysis of mRNAs revealed a similar functional profile related to all analyzed conditions (CAa, CAi, and CAs). However, active and inactive surfaces share up-regulated genes (gtsA; qrtT; tqsA; pimB; EPHX1) related to virulence traits that were not overrepresented in sound surfaces. From a functional perspective, what matters most is the individual carious status rather than the surface condition. Therefore, pooling samples from various sites can be carried out using naturally developed oral biofilms but should preferably include carious surfaces. CONCLUSION: Metatranscriptome data from subjects with caries activity have shown that biofilms from sound, arrested, and active lesions are similar in composition and function.

Comparison of oral cavity protein abundance among caries-free and caries-affected individuals-a systematic review and meta-analysis.

Objective: Some salivary proteins seem to be differently abundant among caries-free (CF) and caries-affected (CA) individuals, but previous results are contradictory precluding that definitive conclusion be drawn. A pooled analysis of the available evidence may provide more robust data on identifying oral cavity protein patterns among CF and CA individuals. This systematic review and meta-analysis (PROSPERO CRD42021269079) aimed to compare the oral cavity protein abundance among caries-free and caries-affected individuals. Methods: This study was conducted following PRISMA guidelines. PubMed, Embase, and Web of Science databases were systematically assessed (up to February 2023) to retrieve clinical studies written in English, German, or in Latin-based languages that compared the oral cavity protein abundance among CF and CA individuals. Data extraction and methodological quality assessment (NIH guidelines) were independently performed by two investigators. Qualitative synthesis was performed from all included studies and meta-analysis was performed using a random-effects model with inverse variance for studies that reported the concentration of proteins or enzymatic activity. Standardized mean difference (SMD) with respective 95% confidence interval (CI) were calculated for each outcome. Results: A total of 90 studies (two cohort and 88 cross-sectional designs) of more than 6,000 participants were selected for data extraction, being the quality of evidence graded as "fair" for most of them. The oral cavity of CF individuals presented lower total protein concentration [SMD = 0.37 (95% CI: 0.07-0.68; 18 studies)], lower total antioxidant capacity [SMD = 1.29 (95% CI: 0.74-1.85); 17 studies], and lower carbonic anhydrase activity [SMD = 0.83 (95% CI: 0.58-1.09); three studies], whereas CA individuals presented lower carbonic anhydrase concentration [SMD = -0.66 (95% CI: -1.00 to -0.32); three studies], urease [SMD = -0.95 (IC 95%: -1.72 to -0.17); four studies], and arginine deiminase system [SMD = -2.07 (95% CI: -3.53 to -0.62); three studies] activities. Antimicrobial peptides, secretory immunoglobulin-A concentrations and alpha-amylase activity were similar among individuals. Conclusion: Differences on oral cavity protein abundance were observed among CF and CA individuals. These data indicate some protein patterns for the oral health and dental caries conditions. Even when statistically significant, some of the results were not very consistent. Cohort studies need to be conducted to validate these results.

Acid tolerance of Lactobacillus spp. on root carious lesions: A complex and multifaceted response.

Lactobacillus spp. are acidogenic and aciduric bacteria and are among the main cariogenic microorganisms associated with the carious process. OBJECTIVE: This study aimed to identify genes involved in the acid-tolerance of Lactobacillus spp. and potential functions attributed to these genes within the metatranscriptome of sound root surfaces and carious root surfaces. DESIGN: Genomic libraries were built from mRNA isolated from the biofilm samples (10 from sound root and 9 from carious root using Illumina HiSeq 2500). Reads generated by RNA-seq were mapped against 162 oral microbial genomes and genes potentially related to acid tolerance were manually extracted from the Lactobacillus spp. genomes using L. paracasei ATCC 344 as reference genome. The R package DESeq2 was used to calculate the level of differential gene expression between those two clinical conditions. RESULTS: Fifteen Lactobacillus spp. genomes were identified and a total of 653 acid tolerance genes were overexpressed in carious root surfaces. Multiple functions, as translation, ribosomal structure and biogenesis, transport of nucleotides and amino acids, are involved in Lactobacillus spp. acid tolerance. Species-specific functions also seem to be related to the fitness of Lactobacillus spp. in acidified environments such as that of the cariogenic biofilm associated with carious root lesions. CONCLUSIONS: The response of Lactobacillus spp. to an acidic environment is complex and multifaceted. This finding suggests several possible avenues for further research into the adaptive mechanisms of these bacteria.

Chapter 5.2: Oral Microbial Biofilms.

Bacteria, fungi, archaea, protozoa, viruses, and bacteriophages colonize the oral cavity and, in combination, they form the oral microbiome. The coexistence of different microorganisms and the microbial balance at each specific site are warranted by synergistic and antagonist interactions among members of the microbial communities. This microbiological balance suppresses the growth of potentially pathogenic microorganisms, generally keeping them at low abundance in the colonized sites. Microbial communities coexist in harmony with the host being compatible with a health condition. On the other hand, stressors exert selective pressure on the microbiota, promoting disruption in microbial homeostasis leading to dysbiosis. In this process, potentially pathogenic microorganisms become more abundant, resulting in microbial communities with altered properties and functions. Once the dysbiotic state has been reached, increased disease risk is expected. Biofilm is essential for caries development. The knowledge of the composition and metabolic interactions in the microbial community is fundamental for developing effective preventive and therapeutic measures. Studying both health and cariogenic conditions will bring an essential understanding of the disease process. Recent advances in omics approaches provide an unparalleled potential to reveal new insights about dental caries. This chapter will discuss a broader perspective on the etiology and pathogenesis of coronal dental caries from biofilm structure to microbial interactions.

Biofilm dysbiosis and caries activity: a surface or an individual issue?

Abstract Objective This study aimed to analyze the functional profile of supragingival biofilm from sound (CAs), active (CAa), and inactive (CAi) enamel caries lesions from caries-active individuals to provide insights into the diversity of biological processes regarding biofilm dysbiosis. Methodology A metatranscriptome analysis was performed in biofilm samples collected from five caries-active individuals. Total RNA was extracted, and the microbial cDNAs were obtained and sequenced (Illumina HiSeq3000). Trimmed data were submitted to the SqueezeMeta pipeline in the co-assembly mode for functional analysis and further differential gene expression analysis (DESeq2). Results Bioinformatics analysis of mRNAs revealed a similar functional profile related to all analyzed conditions (CAa, CAi, and CAs). However, active and inactive surfaces share up-regulated genes (gtsA; qrtT; tqsA; pimB; EPHX1) related to virulence traits that were not overrepresented in sound surfaces. From a functional perspective, what matters most is the individual carious status rather than the surface condition. Therefore, pooling samples from various sites can be carried out using naturally developed oral biofilms but should preferably include carious surfaces. Conclusion Metatranscriptome data from subjects with caries activity have shown that biofilms from sound, arrested, and active lesions are similar in composition and function.

Consensus for teaching dental caries in the Portuguese Language at Brazilian dental schools

Abstract The aim of this paper was to present a summary of the process of developing and preparing the final documents of the national consensus for teaching undergraduate Brazilian dental students the dental caries curriculum in the Portuguese language. The final document was developed in three steps: a) The ABENO and LAOHA cariology group invited experts from all five regions of Brazil to participate in the discussion. The theoretical support for crafting the first draft of the consensus was based on two publications: National Curriculum Guidelines of the Dentistry graduation in Brazil, Ministry of Education (2021) and the competences described in the European Core Curriculum for Cariology (ORCA-ADEE, 2011); b) The group of experts was divided into 5 working groups: G1-Domain, Main and Specific Competences, G2-Essential knowledge, G3-Life course perspective, G4-Social determinants and dental caries, G5- Glossary. The document was finalized by thoroughly reviewing the process using Delphi methodology; c) The 5-chapter document (one from each working group) was submitted to three open public consultations in 2022 (May-June, August, and October) using Google-forms. The suggestions (content/wording) were discussed within the group as: totally accepted, partially accepted, and rejected. A total of 192 suggestions were registered from 31 dental schools in all regions of Brazil. The number of suggestions received per Group were: 84, 28, 26, 24, 30 suggestions for G1, G2, G3, G4 and G5, respectively. The majority of suggestions were totally accepted by the group of experts (n = 172, 89.6%), 15 were partially accepted (7.8%), and 5 were rejected. Conclusion The final document could be considered to be the first national consensus for teaching the dental caries curriculum in Brazil.

Effect of Stevia rebaudiana Bertoni infusion on enamel demineralization and dental biofilm formation: an in-situ study / Efeito da infusão de Stevia rebaudiana Bertoni na desmineralização do esmalte e na formação do biofilme dental: um estudo in situ

Aim: Stevia rebaudiana Bertoni (Stevia) is a natural non--caloric sweetener that can modify the cariogenicity of biofilms. This study aimed to evaluate the effect of Ste-via infusion in microbial and biochemical composition of biofilms formed in the presence of sucrose and on enamel demineralization. Materials and Methods: In a cross-over design, eleven volunteers wore an intraoral palatal appliance containing 4 slabs of bovine enamel during 3 phases of 7 days each. Sucrose solution (20%) was dripped onto slabs 8 times/day and 0.9% sodium chloride (NaCl), 0.12% chlorhexidine (CHX), or 5% infusion of Stevia were dripped 2x/day. Biofilm formed on the slabs was collected and analyzed for counts of microorganisms (total bacteria, Lactobacilli, Candida spp., and Streptococcus mutans) biochemical composition in terms of soluble and insoluble extracellular polysaccharides and qualitative assessment by scanning electron microscopy. The per-centage of surface hardness loss (%SHL) was determined on enamel slabs taken baseline and post-biofilm Knoop surface hardness values. Results: The % SHL in the CHX treatment was statistically lower in comparison to NaCl (p < 0.05). No differences were found between Stevia and CHX and between Stevia and NaCl. No other difference was found among the experimental groups with respect to the other outcomes. Discussion: Under high carioge-nic conditions resembling frequent exposure to sucrose and absence of mechanical disruption, use of Stevia can neither modify the counts of cariogenic microorganisms nor the concentration of extracellular polysaccharides on in situ formed biofilms. This may have occurred due to the exposure of the biofilm to high sucrose concentration for all treatments and the condi-tion of the microorganism growth in situ, which may hinder the diffusion of substances through the thick biofilm. Conclusion: Biofilm exposed to a high cariogenic challenge and without mechanical disruption is not affected by an infusion of Stevia rebaudiana Bertoni.

Objetivo: A Stevia rebaudiana Bertoni (Stevia) é um adoçante natural não calórico que pode modificar a cariogenicidade de biofilmes. Este estudo teve como objetivo avaliar o efeito da infusão de Stevia na composição microbiana e bioquímica de biofilmes formados na presença de sacarose e na desmineralização do esmalte. Materiais e métodos: Em um desenho cruzado, onze voluntários usaram um aparelho intraoral palatino contendo 4 pla-cas de esmalte bovino durante 3 fases de 7 dias cada. A solução de sacarose (20%) foi gotejada em placas 8 vezes/dia e cloreto de sódio a 0,9% (NaCl), clorexidina a 0,12% (CHX) ou infusão a 5% de Stevia foram gotejados 2x/dia. O biofilme formado nas placas foi coletado e analisado para contagem da composição bioquímica de microrganismos (bactérias totais, Lactobacilos, Candida spp. e Streptococcus mutans) em termos de polissaca-rídeos extracelulares solúveis e insolúveis e avaliação qualitativa por microscopia eletrônica de varredura. A porcentagem de perda de dureza superficial (%SHL) nos blocos de esmalte foi determinada com base nos valores de dureza superficial Knoop tomadas no início e pós-biofilme. Resultados: O % SHL no tratamento CHX foi estatisticamente menor em comparação ao NaCl (p < 0,05). Não foram encontradas diferenças entre Stevia e CHX e entre Stevia e NaCl. Nenhuma outra diferença foi encontrada entre os grupos experimentais em re-lação aos outros resultados. Discussão: Sob condições cariogênicas elevadas que se assemelham a exposição frequente à sacarose e ausência de disrupção mecânica, o uso de Stevia não pode modificar as contagens de microrganismos cariogênicos nem a concentração de polissacarídeos extracelulares em biofilmes formados in situ. Isso pode ter ocorrido devido à exposição do biofilme à alta concentração de sacarose para todos os tratamentos e à condição de crescimento do microrganismo in situ, o que pode dificultar a difusão de substâncias através do biofilme espesso. Conclusão: O biofilme exposto a um alto desafio cariogênico e sem disruptucao mecânica não é afetado por uma infusão de Stevia rebaudiana Bertoni.

Dietary sugars modulate bacterial-fungal interactions in saliva and inter-kingdom biofilm formation on apatitic surface.

Bacteria and fungi can interact to form inter-kingdom biofilms in the oral cavity. Streptococcus mutans and Candida albicans are frequently detected in saliva and in dental biofilms associated with early childhood caries (tooth-decay), a prevalent oral disease induced by dietary sugars. However, how different sugars influence this bacterial-fungal interaction remains unclear. Here, we investigate whether specific sugars affect the inter-kingdom interaction in saliva and subsequent biofilm formation on tooth-mimetic surfaces. The microbes were incubated in saliva containing common dietary sugars (glucose and fructose, sucrose, starch, and combinations) and analyzed via fluorescence imaging and quantitative computational analyses. The bacterial and fungal cells in saliva were then transferred to hydroxyapatite discs (tooth mimic) to allow microbial binding and biofilm development. We found diverse bacterial-fungal aggregates which varied in size, structure, and spatial organization depending on the type of sugars. Sucrose and starch+sucrose induced the formation of large mixed-species aggregates characterized by bacterial clusters co-bound with fungal cells, whereas mostly single-cells were found in the absence of sugar or in the presence of glucose and fructose. Notably, both colonization and further growth on the apatitic surface were dependent on sugar-mediated aggregation, leading to biofilms with distinctive spatial organizations and 3D architectures. Starch+sucrose and sucrose-mediated aggregates developed into large and highly acidogenic biofilms with complex network of bacterial and fungal cells (yeast and hyphae) surrounded by an intricate matrix of extracellular glucans. In contrast, biofilms originated from glucose and fructose-mediated consortia (or without sugar) were sparsely distributed on the surface without structural integration, growing predominantly as individual species with reduced acidogenicity. These findings reveal the impact of dietary sugars on inter-kingdom interactions in saliva and how they mediate biofilm formation with distinctive structural organization and varying acidogenicity implicated with human tooth-decay.

Streptococcus mutans Gene Expression and Functional Profile in Root Caries: An RNA-Seq Study.

The literature is still scarce on studies describing Streptococcus mutans global gene expression under clinical conditions such as those found on complex biofilms from sound root surfaces (SRS) and carious root surfaces (RC). This study aimed to investigate the S. mutans gene expression and functional profile within the metatranscriptome of biofilms from SRS and from RC in an attempt to identify enriched functional signatures potentially associated with the healthy-to-disease transitioning process. Total RNA was extracted, and prepared libraries (SRS = 10 and RC = 9) were paired-end sequenced using the Illumina HiSeq2500. A read count assigned to each gene of the S. mutans UA159 strain was obtained. Differentially expressed genes (DEG) between SRS and RC were identified using the DESeq2 R package, and weighted gene co-expression network analysis (WGCNA) was performed to explore and identify functional modules related to SRS and RC. We found seventeen DEG between SRS and RC samples, with three overexpressed in RC and related to membrane protein, alanyl-tRNA synthetase, and GTP-binding protein, with the remaining ones overexpressed in SRS samples and related to hypothetical protein, transposon integrase, histidine kinase, putative transporter, bacteriocin immunity protein, response regulator, 6-phospho-beta-galactosidase, purine metabolism, and transcriptional regulator. Key-functional modules were identified for SRS and RC conditions based on WGCNA, being 139 hub genes found on SRS key-module and 17 genes on RC key-module. Functional analysis of S. mutans within the metatranscriptome of biofilms from sound root and from carious root revealed a similar pattern of gene expression, and only a few genes have been differentially expressed between biofilms from SRS and those from root carious lesions. However, S. mutans presented a greater functional abundance in the carious lesion samples. Some functional patterns related to sugar (starch, sucrose, fructose, mannose, and lactose) and heterofermentative metabolisms, to cell-wall biosynthesis, and to acid tolerance stress seem to be enriched on carious root surfaces, conferring ecological advantages to S. mutans. Altogether, the present data suggest that a functional signature may be associated with carious root lesions.

Antimicrobial potential of essential oils mouthrinses with and without alcohol: a randomized clinical trial

Aim: This study aimed to compare the microbiological potential and gustatory perception of essential oils (EO) mouthrinses containing and not containing alcohol. Methods: Twenty healthy adult volunteers rinsed with 10mL of the following test solutions: EO with alcohol, EO without alcohol, or a control solution (saline solution with mint essence). A washout period of at least seven days was adopted after a single-use protocol of the respective solution. All participants used all three tested substances. Antimicrobial potential was assessed by counting salivary total viable bacteria both before and after each rinse. Gustatory perception was evaluated using the Visual Analogue Scale (VAS). Multiple comparisons were performed with the Wilcoxon test, using Bonferroni correction. Results: Both EO solutions presented a higher antimicrobial potential in comparison to the control solution (p<0.017). However, no significant difference in antimicrobial potential was observed between EO containing or not containing alcohol (p=0.218). VAS of EO with alcohol (median: 2.7) was similar to control solution (median: 1.6) (p=0.287). A better gustatory perception was observed of the EO without alcohol (median 7.6) when compared to the control solution (p<0.0001). When EO groups were compared, EO without alcohol also demonstrated a significantly better gustatory perception (p=0.001). Conclusion: Mouthrinse containing EO without alcohol presented a better taste perception when compared to the EO with alcohol, but no difference was observed in the antimicrobial potential of both EO solutions after a single rinse protocol

Functionally Active Microbiome in Supragingival Biofilms in Health and Caries.

The oral microbiome is unique at inter and intra-individual levels at various sites due to physical and biological factors. This study aimed to compare the bacterial composition of supragingival biofilms collected from enamel sites with different caries activity, from active and inactive-caries subjects, and from caries-free (CF) subjects. Twenty-two individuals (aged between 13 and 76 years old; med = 23.5 years old) were allocated into 3 groups: caries-active (CA) (n = 10), caries-inactive (CI) (n = 6), and CF (n = 6). From the CA group, 3 sites were sampled: CA (active non-cavitated lesion), CI (inactive non-cavitated lesion), and sound enamel surface (S). From the subjects of the CI group, biofilm from a CI lesion was collected (INCL), while for the CF subjects, a pool of biofilm from sound enamel surfaces was sampled. The total RNA was extracted, and cDNA libraries were prepared and paired-end sequenced (Illumina HiSeq 3,000). Final dental biofilm samples analysed from CA was 16 (ANCL-CA = 6, INCL-CA = 4, S-CA = 6); from CI, 3 (INCL-CI = 3); and from CF, 6 (S-CF = 6) (some samples were lost by insufficient genetic material). Read sequences were processed and analysed using the Metagenomics RAST server. High-quality sequences (3,542,190) were clustered into operational taxonomic units (97% identity; SILVA SSU), representing 915 genera belonging to 29 phyla (higher abundant: Actinobacteria, Firmicutes, Bacteroidetes, and Fusobacteria). The presence of a core microbiome was observed (123 shared genera). The alpha diversity analysis showed less bacterial diversity in disease (S-CA) compared to health (S-CF). The dominant genera included Actinomyces, Corynebacterium, Capnocytophaga, Leptotrichia, Veillonella, Prevotella, Streptococcus, Eubacterium, and Neisseria. Veillonella and Leptotrichia were related with disease and Prevotella with health. Corynebacterium, Capnocytophaga, and Actinomyces clustered together presenting high abundance in health and disease. The Metric Multidimensional Scaling Ordination analysis shows that sites from active subjects (ANCL-CA, INCL-CA, and S-CA) are closer to each other than either INCL-CI subjects or S-CF subjects. In conclusion, supragingival bacterial communities presented intra-individual similarities, but inter-individual diversity and difference in bacterial composition reveal that the subject's caries activity status matters more than sites.

Management of dental caries lesions in Latin American and Caribbean countries.

Caries management at the lesion level is dependent on the lesion activity, the presence of a cavitation (either cleanable or non-cleanable), and lesion depth as evaluated via radiographic examination. A variety of non-invasive, micro-invasive, and minimally invasive treatment (with or without restoration) options are available for primary and permanent teeth. Non-invasive strategies include oral hygiene instructions, dietary counseling, and personal as well as professional use of fluoridated products that reduce demineralization and increase re-mineralization. Micro-invasive procedures include the use of occlusal resin sealants and resin infiltrants, while minimally invasive strategies comprise those related to selective removal of caries tissues and placement of restorations. Deep caries management includes indirect pulp capping, while exposed pulp may be treated using direct pulp capping and partial or complete pulpotomy. The aim of the present study was to review available evidence on recommended preventive and restorative strategies for caries lesions in Latin American/Caribbean countries, and subsequently develop evidence-based recommendations for treatment options that take into consideration material availability, emphasize ways to adapt available treatments to the local context, and suggest ways in which dentists and health systems can adopt these treatments.

Microbiome and oral squamous cell carcinoma: a possible interplay on iron metabolism and its impact on tumor microenvironment.

There is increasing evidence showing positive association between changes in oral microbiome and the occurrence of oral squamous cell carcinoma (OSCC). Alcohol- and nicotine-related products can induce microbial changes but are still unknown if these changes are related to cancerous lesion sites. In an attempt to understand how these changes can influence the OSCC development and maintenance, the aim of this study was to investigate the oral microbiome linked with OSCC as well as to identify functional signatures and associate them with healthy or precancerous and cancerous sites. Our group used data of oral microbiomes available in public repositories. The analysis included data of oral microbiomes from electronic cigarette users, alcohol consumers, and precancerous and OSCC samples. An R-based pipeline was used for taxonomic and functional prediction analysis. The Streptococcus spp. genus was the main class identified in the healthy group. Haemophilus spp. predominated in precancerous lesions. OSCC samples revealed a higher relative abundance compared with the other groups, represented by an increased proportion of Fusobacterium spp., Prevotella spp., Haemophilus spp., and Campylobacter spp. Venn diagram analysis showed 52 genera exclusive of OSCC samples. Both precancerous and OSCC samples seemed to present a specific associated functional pattern. They were menaquinone-dependent protoporphyrinogen oxidase pattern enhanced in the former and both 3',5'-cyclic-nucleotide phosphodiesterase (purine metabolism) and iron(III) transport system ATP-binding protein enhanced in the latter. We conclude that although precancerous and OSCC samples present some differences on microbial profile, both microbiomes act as "iron chelators-like" potentially contributing to tumor growth.

Interplay Among the Oral Microbiome, Oral Cavity Conditions, the Host Immune Response, Diabetes Mellitus, and Its Associated-Risk Factors-An Overview.

This comprehensive review of the literature aimed to investigate the interplay between the oral microbiome, oral cavity conditions, and host immune response in Diabetes mellitus (DM). Moreover, this review also aimed to investigate how DM related risk factors, such as advanced age, hyperglycemia, hyperlipidemia, obesity, hypertension and polycystic ovary syndrome (PCOS), act in promoting or modifying specific mechanisms that could potentially perpetuate both altered systemic and oral conditions. We found that poorly controlled glycemic index may exert a negative effect on the immune system of affected individuals, leading to a deficient immune response or to an exacerbation of the inflammatory response exacerbating DM-related complications. Hyperglycemia induces alterations in the oral microbiome since poor glycemic control is associated with increased levels and frequencies of periodontal pathogens in the subgingival biofilm of individuals with DM. A bidirectional relationship between periodontal diseases and DM has been suggested: DM patients may have an exaggerated inflammatory response, poor repair and bone resorption that aggravates periodontal disease whereas the increased levels of systemic pro-inflammatory mediators found in individuals affected with periodontal disease exacerbates insulin resistance. SARS-CoV-2 infection may represent an aggravating factor for individuals with DM. Individuals with DM tend to have low salivary flow and a high prevalence of xerostomia, but the association between prevalence/experience of dental caries and DM is still unclear. DM has also been associated to the development of lesions in the oral mucosa, especially potentially malignant ones and those associated with fungal infections. Obesity plays an important role in the induction and progression of DM. Co-affected obese and DM individuals tend to present worse oral health conditions. A decrease in HDL and, an increase in triglycerides bloodstream levels seem to be associated with an increase on the load of periodontopathogens on oral cavity. Moreover, DM may increase the likelihood of halitosis. Prevalence of impaired taste perception and impaired smell recognition tend to be greater in DM patients. An important interplay among oral cavity microbiome, DM, obesity and hypertension has been proposed as the reduction of nitrate into nitrite, in addition to contribute to lowering of blood pressure, reduces oxidative stress and increases insulin secretion, being these effects desirable for the control of obesity and DM. Women with PCOS tend to present a distinct oral microbial composition and an elevated systemic response to selective members of this microbial community, but the association between oral microbiome, PCOS are DM is still unknown. The results of the studies presented in this review suggest the interplay among the oral microbiome, oral cavity conditions, host immune response and DM and some of the DM associated risk factors exist. DM individuals need to be encouraged and motivated for an adequate oral health care. In addition, these results show the importance of adopting multidisciplinary management of DM and of strengthening physicians-dentists relationship focusing on both systemic and on oral cavity conditions of DM patients.

Management of dental caries lesions in Latin American and Caribbean countries

Abstract Caries management at the lesion level is dependent on the lesion activity, the presence of a cavitation (either cleanable or non-cleanable), and lesion depth as evaluated via radiographic examination. A variety of non-invasive, micro-invasive, and minimally invasive treatment (with or without restoration) options are available for primary and permanent teeth. Non-invasive strategies include oral hygiene instructions, dietary counseling, and personal as well as professional use of fluoridated products that reduce demineralization and increase re-mineralization. Micro-invasive procedures include the use of occlusal resin sealants and resin infiltrants, while minimally invasive strategies comprise those related to selective removal of caries tissues and placement of restorations. Deep caries management includes indirect pulp capping, while exposed pulp may be treated using direct pulp capping and partial or complete pulpotomy. The aim of the present study was to review available evidence on recommended preventive and restorative strategies for caries lesions in Latin American/Caribbean countries, and subsequently develop evidence-based recommendations for treatment options that take into consideration material availability, emphasize ways to adapt available treatments to the local context, and suggest ways in which dentists and health systems can adopt these treatments.

Dynamics of Mono- and Dual-Species Biofilm Formation and Interactions Between Paracoccidioides brasiliensis and Candida albicans.

The oral cavity is a highly diverse microbial environment in which microorganisms interact with each other, growing as biofilms on biotic and abiotic surfaces. Understanding the interaction among oral microbiota counterparts is pivotal for clarifying the pathogenesis of oral diseases. Candida spp. is one of the most abundant fungi in the oral mycobiome with the ability to cause severe soft tissue lesions under certain conditions. Paracoccidioides spp., the causative agent of paracoccidioidomycosis, may also colonize the oral cavity leading to soft tissue damage. It was hypothesized that both fungi can interact with each other, increasing the growth of the biofilm and its virulence, which in turn can lead to a more aggressive infectivity. Therefore, this study aimed to evaluate the dynamics of mono- and dual-species biofilm growth of Paracoccidioides brasiliensis and Candida albicans and their infectivity using the Galleria mellonella model. Biomass and fungi metabolic activity were determined by the crystal violet and the tetrazolium salt reduction tests (XTT), respectively, and the colony-forming unit (CFU) was obtained by plating. Biofilm structure was characterized by both scanning electronic- and confocal laser scanning- microscopy techniques. Survival analysis of G. mellonella was evaluated to assess infectivity. Our results showed that dual-species biofilm with P. brasiliensis plus C. albicans presented a higher biomass, higher metabolic activity and CFU than their mono-species biofilms. Furthermore, G. mellonella larvae infected with P. brasiliensis plus C. albicans presented a decrease in the survival rate compared to those infected with P. brasiliensis or C. albicans, mainly in the form of biofilms. Our data indicate that P. brasiliensis and C. albicans co-existence is likely to occur on oral mucosal biofilms, as per in vitro and in vivo analysis. These data further widen the knowledge associated with the dynamics of fungal biofilm growth that can potentially lead to the discovery of new therapeutic strategies for these infections.

Candida and dental caries in children, adolescents and adults: A systematic review and meta-analysis.

OBJECTIVE: This systematic review and meta-analysis aimed to investigate whether the presence of fungi of the genus Candida in oral cavity is associated with dental caries in children and adolescents (from 6 to 18 years) or in adults (older than 18 years). DESIGN: Electronic search was carried out in MEDLINE, EMBASE and LILACS databases. Studies that evaluated the presence of Candida spp. and dental caries on those individuals were included. Data extraction and evidence quality assessment were performed by two independent investigators. Prevalence ratio (PR) was calculated considering 95 % confidence interval (CI). Meta-analysis was performed using random effect Mantel-Haenszel model. RESULTS: Thirty out of 123 publications selected for full-reading were included in the systematic review. Prevalence of Candida spp. in the oral cavity ranged from 7.7 % to 78 %. Prevalence of dental caries in individuals harbouring Candida spp. ranged from 27.2%-100% and was higher than in individuals not harbouring Candida spp. (PR = 1.72 [1.46-2.02]). Prevalence of dental caries was 2.3 times higher in individuals harbouring Candida spp. in mucosa. Moreover, dental caries prevalence was 80 % and 48 % higher in children/adolescents and adults harbouring Candida spp., respectively (95 % CI [1.44-2.25] and [1.20-1.83]). The quality of evidence of most studies (n = 21) was graded as fair. CONCLUSIONS: These results suggest that, regardless of age or sample site, individuals with the presence of Candida spp. have a higher prevalence of dental caries when compared to individuals without these microorganisms in the oral cavity.