Genotypic Diversity and Virulence Traits of Streptococcus mutans

The aim of this study was to compare the genotypic diversity and virulence traits of Streptococcus mutans isolated from carious dentin before and after partial dentin caries removal (PDR) and sealing. Carious dentin samples were obtained three months before and after the PDR and cavity sealing. Up to seven isolates of each morphological type of S. mutans were selected and strain identity was confirmed using gtfB primer. Genotyping was performed by arbitrary primer-PCR (AP-PCR). Acidogenesis and acidurance of the genotypes were evaluated as virulence traits. A paired t-test and a Wilcoxon test were used to compare the virulence of genotypes. A total of 48 representative S. mutans isolates were genotyped (31 before and 17 after the sealing). At least one of the genotypes found before the sealing was also found on dentin after the sealing. The number of genotypes found before the sealing ranged from 2 to 3 and after the sealing from 1 to 2 genotypes. No difference was observed in the acidogenesis and acidurance between genotypes isolated before and after the sealing. In conclusion, genotypic diversity of S. mutans decreased after the PDR and sealing, but the virulence traits of S. mutans remained unchangeable

Role of Candida albicans on enamel demineralization and on acidogenic potential of Streptococcus mutans in vitro biofilms

There is growing evidence that C. albicans is associated with dental caries, but its role on caries development needs to be better clarified. Objective:To evaluate at the hard tissue level the effect of C. albicans on the cariogenic potential of S. mutans biofilms focusing on the mineral profile of induced carious lesions. This study also aimed to evaluate the effect of C. albicans on the acidogenic potential of S. mutans biofilms. Methodology: Dual-species (CA+SM) and single-species biofilms (CA or SM) were grown on the surface of enamel slabs in the presence of glucose/sucrose supplemented culture medium for 24, 48 and 72 hours. Demineralization was evaluated through percentage of surface microhardness change (%SMC) and transversal microradiography analysis (ILM and LD) and pH of the spent medium was recorded daily. Data were analyzed by two-way ANOVA followed by Bonferroni correction. Results: %SMC was statistically different among the biofilms at each time point being the highest for SM biofilms and the lowest for CA biofilms which also differed from CA+SM biofilms [SM (24 h: 47.0±7.3; 48 h: 66.3±8.3; 72 h: 75.4±3.9); CA (24 h: 7.3±3.3; 48 h: 7.1±6.4; 72 h: 6.6±3.6); CA+SM (24 h: 35.9±7.39.1; 48 h: 47.2±9.5; 72 h: 47.6±9.5)]. pH of spent medium was statistically lower for SM biofilms compared to the other biofilms at each time point and remained constant over time while pH values increased from 24 to 72 h for both CA and CA+SM biofilms [SM (24 h: 4.4±0.1; 48 h: 4.4±0.1; 72 h: 4.5±0.1); CA (24 h: 6.9±0.3; 48 h: 7.2±0.2; 72 h: 7.5±0.2); CA+MS (24 h: 4.7±0.2; 48 h: 5.1±0.1; 72 h: 6.1±0.6)]. IML and LD for SM biofilms increased over time while no difference was observed from 24 to 72 h for the other biofilms. Conclusions: The present data suggest that C. albicans has low enamel demineralization potential and the presence of C. albicans can reduce both the cariogenic and acidogenic potentials of S. mutans biofilms

Effect of Stevia rebaudianaBertoni infusion on enamel demineralization and dental biofilm formation : an in-situ study

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 BertoniObjetivo: 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 Berto

Role of Candida albicans on enamel demineralization and on acidogenic potential of Streptococcus mutans in vitro biofilms

There is growing evidence that C. albicans is associated with dental caries, but its role on caries development needs to be better clarified. Objective: To evaluate at the hard tissue level the effect of C. albicans on the cariogenic potential of S. mutans biofilms focusing on the mineral profile of induced carious lesions. This study also aimed to evaluate the effect of C. albicans on the acidogenic potential of S. mutans biofilms. Methodology: Dual-species (CA+SM) and single-species biofilms (CA or SM) were grown on the surface of enamel slabs in the presence of glucose/sucrose supplemented culture medium for 24, 48 and 72 hours. Demineralization was evaluated through percentage of surface microhardness change (%SMC) and transversal microradiography analysis (ILM and LD) and pH of the spent medium was recorded daily. Data were analyzed by two-way ANOVA followed by Bonferroni correction. Results: %SMC was statistically different among the biofilms at each time point being the highest for SM biofilms and the lowest for CA biofilms which also differed from CA+SM biofilms [SM (24 h: 47.0±7.3; 48 h: 66.3±8.3; 72 h: 75.4±3.9); CA (24 h: 7.3±3.3; 48 h: 7.1±6.4; 72 h: 6.6±3.6); CA+SM (24 h: 35.9±7.39.1; 48 h: 47.2±9.5; 72 h: 47.6±9.5)]. pH of spent medium was statistically lower for SM biofilms compared to the other biofilms at each time point and remained constant over time while pH values increased from 24 to 72 h for both CA and CA+SM biofilms [SM (24 h: 4.4±0.1; 48 h: 4.4±0.1; 72 h: 4.5±0.1); CA (24 h: 6.9±0.3; 48 h: 7.2±0.2; 72 h: 7.5±0.2); CA+MS (24 h: 4.7±0.2; 48 h: 5.1±0.1; 72 h: 6.1±0.6)]. IML and LD for SM biofilms increased over time while no difference was observed from 24 to 72 h for the other biofilms. Conclusions: The present data suggest that C. albicans has low enamel demineralization potential and the presence of C. albicans can reduce both the cariogenic and acidogenic potentials of S. mutans biofilms

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. SARSCoV-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

Enamel Carious Lesion Development in Response to Sucrose and Fluoride Concentrations and to Time of Biofilm Formation: An Artificial-Mouth Study

The aim of this study was to evaluate both sucrose and fluoride concentrations and time of biofilm formation on enamel carious lesions induced by an in vitro artificial-mouth caries model. For Study 1, biofilms formed by streptococci and lactobacilli were grown on the surface of human enamel slabs and exposed to artificial saliva containing 0.50 or 0.75 ppmF (22.5 h/d) and broth containing 3 or 5% sucrose (30 min; 3x/d) over 5 d. In Study 2, biofilms were grown in the presence of 0.75 ppmF and 3% sucrose over 3 and 9 days. Counts of viable cells on biofilms, lesion depth (LD), and the integrated mineral loss (IML) on enamel specimens were assessed at the end of the tested conditions. Counts of total viable cells and L. casei were affected by sucrose and fluoride concentrations as well as by time of biofilm formation. Enamel carious lesions were shallower and IML was lower in the presence of 0.75 ppmF than in the presence of 0.50 ppmF (P 0.25). Additionally, deeper lesions and higher IML were found after 9 d of biofilm formation (P < 0.005). Distinct sucrose concentrations did not affect enamel carious lesion development. The severity of enamel demineralization was reduced by the presence of the higher fluoride concentration. Additionally, an increase in the time of biofilm formation produced greater demineralization. Our results also suggest that the present model is suitable for studying aspects related to caries lesion development

Effect of silver nanoparticles on the physicochemical and antimicrobial properties of an orthodontic adhesive

Orthodontic treatment with fixed brackets plays a major role on the formation of white spot lesions. Objective This study aimed to incorporate silver nanoparticle solutions (AgNP) in an orthodontic adhesive and evaluate its physicochemical and antimicrobial properties. Material and Methods Silver nanoparticle solutions were added to a commercial adhesive in different concentrations (w/w): 0%, 0.11%, 0.18%, and 0.33%. Shear bond strength (SBS) test was performed after bonding metal brackets to enamel. Raman spectroscopy was used to analyze in situ the degree of conversion (DC) of the adhesive layer. The surface free energy (SFE) was evaluated after the measurement of contact angles. Growth inhibition of Streptococcus mutans in liquid and solid media was determined by colony-forming unit count and inhibition halo, respectively. One-way ANOVA was performed for SBS, DC, SFE, and growth inhibition. Results The incorporation of AgNP solution decreased the SBS (

Effect of silver nanoparticles on the physicochemical and antimicrobial properties of an orthodontic adhesive

Ortodontiv treatment with fixed brackets plays a mayor role on the formation of white spot lesions. Objective:Tadhesive and evaluate its physicochemical and antimicrobial properties. Material and Methods: Silver nanoparticle solutions were added to a commercial adhesive in different concentrations (w/w): 0%, 0.11%, 0.18%, and 0.33%. Shear bond strength (SBS) test was performed after bonding metal brackets to enamel. Raman spectroscopy was used to analyze in situ the degree of conversion (DC) of the adhesive layer. The surface free energy (SFE) was evaluated after the measurement of contact angles. Growth inhibition of Streptococcus mutans in liquid and solid media was determined by colony-forming unit count and inhibition halo, respectively. One-way ANOVA was performed for SBS, DC, SFE, and growth inhibition. Results: The incorporation of AgNP solution decreased the SBS (p<0.001) and DC in situ (p<0.001) values. SFE decreased after addition of 0.18% and 0.33% AgNP. Growth inhibition of S. mutans in liquid media was obtained after silver addition (p<0.05). Conclusions: The addition of AgNP solutions to Transbond™ XT adhesive primer inhibited S. mutans growth. SBS, DC, and SFE values decreased after incorporation up to 0.33% AgNP solution without compromising the chemical and physical properties of the adhesive

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