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Journal articles on the topic 'Biofilm cariogenicity'
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1
Schwendicke, Falk, Franziska Korte, Christof E. Dörfer, Susanne Kneist, Karim Fawzy El-Sayed, and Sebastian Paris. "Inhibition of Streptococcus mutans Growth and Biofilm Formation by Probiotics in vitro." Caries Research 51, no. 2 (2017): 87–95. http://dx.doi.org/10.1159/000452960.
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To exert anticaries effects, probiotics are described to inhibit growth and biofilm formation of cariogenic bacteria such as Streptococcus mutans (SM). We screened 8 probiotics and assessed how SM growth or biofilm formation inhibition affects cariogenicity of probiotic-SM mixed-species biofilms in vitro. Growth inhibition was assessed by cocultivating probiotics and 2 SM strains (ATCC 20532/25175) on agar. Probiotics were either precultured before SM cultivation (exclusion), or SM precultured prior to probiotic cultivation (displacement). Inhibition of SM culture growth was assessed visually. Inhibition of SM biofilm formation on bovine enamel was assessed using a continuous-flow short-term biofilm model, again in exclusion or displacement mode. The cariogenicity of mixed-species biofilms of SM with the most promising growth and biofilm formation inhibiting probiotic strains was assessed using an artificial mouth model, and enamel mineral loss (ΔZ) was measured microradiographically. We found limited differences in SM growth inhibition in exclusion versus displacement mode, and in inhibition of SM 20532 versus 25175. Results were therefore pooled. Lactobacillus acidophilus LA-5 inhibited significantly more SM culture growth than most other probiotics. L. casei LC-11 inhibited SM biofilm formation similarly to other alternatives but showed the highest retention of probiotics in the biofilms (p < 0.05). Mineral loss from SM monospecies biofilms (ΔZ = 9,772, 25th/75th percentiles: 6,277/13,558 vol% × µm) was significantly lower than from mixed-species SM × LA-5 biofilms (ΔZ = 24,578, 25th/75th percentiles: 19,081/28,768 vol% × µm; p < 0.01) but significantly higher than from SM × LC-11 biofilms (ΔZ = 4,835, 25th/75th percentiles: 263/7,865 vol% × µm; p < 0.05). Probiotics inhibiting SM culture growth do not necessarily reduce the cariogenicity of SM-probiotic biofilms. Nevertheless, SM biofilm formation inhibition may be relevant in the reduction of cariogenicity.
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Fernández, Constanza E., Rodrigo A. Giacaman, Livia M. Tenuta, and Jaime A. Cury. "Effect of the Probiotic Lactobacillus rhamnosus LB21 on the Cariogenicity of Streptococcus mutans UA159 in a Dual-Species Biofilm Model." Caries Research 49, no. 6 (2015): 583–90. http://dx.doi.org/10.1159/000439315.
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Despite promising results using probiotics, evidence of the preventive effect on enamel demineralization is insufficient and the cariogenic potential of probiotics is still controversial. Probiotics could affect biofilm formation and interfere with adherence, growth or coaggregation with Streptococcus mutans in biofilms. However, most of the studies have been conducted using planktonic bacteria. Hence, the aim of the study was to assess the effect of probiotic bacteria on the cariogenicity of S. mutans using an in vitro biofilm caries model on enamel. Single-species biofilms (S. mutans UA159, SM or Lactobacillus rhamnosus LB21, LB) or dual-species biofilms simultaneously inoculated (SM + LB) or LB inoculated 8 h after SM (SM → LB) were grown for 96 h. Biofilms were formed on bovine enamel saliva-coated slabs of known surface hardness (SH) and immersed in culture media. Biofilms were exposed 8 times per day to 10% sucrose. Medium pH was monitored twice daily as a biofilm acidogenicity indicator. After 96 h, biofilms were collected to determine biomass and bacteria viability. Slab demineralization was calculated as percentage of SH loss (%SHL). Additionally, the model was tested with different concentrations of the initial inoculum (103, 106, 108 cells/ml) and different adhesion times (2 or 8 h). The dual-species biofilm revealed no LB effects on SM cariogenicity, without changes in acidogenicity or %SHL among groups (p > 0.05, n = 12). Lack of activity of LB on SM cariogenicity persisted even when 105 times higher concentration of the probiotic was tested. Coaggregation was not observed. In conclusion, findings suggest that LB does not reduce cariogenicity of SM in a validated experimental caries model.
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Giacaman, Rodrigo A., Vanesa Pailahual, and Natalia Díaz-Garrido. "Cariogenicity induced by commercial carbonated beverages in an experimental biofilm-caries model." European Journal of Dentistry 12, no. 01 (January 2018): 027–35. http://dx.doi.org/10.4103/ejd.ejd_188_17.
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ABSTRACT Objectives Frequent consumption of sugars-containing carbonated beverages has been associated with caries, but the consequences on the dental biofilm remain unclear. The aim was to evaluate the effect of commercial carbonated beverages and their sugar-free version on enamel and dentine demineralization and on the cariogenic properties of Streptococcus mutans biofilms. Materials and Methods Biofilms of S. mutans UA159 were grown on enamel and dentin slabs and exposed 3 times/ day for 5 min, to a commercial cola or orange-flavored carbonated beverage or to their sugar-free version. Biofilms/slabs were recovered to assess biomass, viable microorganisms, protein content and polysaccharides. Demineralization was estimated by the variation of Knoop surface microhardness. Results Exposures to the biofilm with sugars-containing carbonated beverages resulted in similar biomass, viable microorganisms, proteins, and polysaccharides than sucrose (P < 0.05). The sugar-free cola and orange-flavored drink showed lower effect on the biofilm, as compared with sucrose or their sugared version (P < 0.05). All of the products tested, included the sugar-free, showed higher demineralization than the negative control (P < 0.05). Conclusions: Sugars-containing carbonated beverages enhance cariogenic activity of S. mutans biofilms, comparable with sucrose. Sugar-free carbonated beverages also have a high demineralizing potential, without affecting biofilm properties.
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Sampaio, Aline A., Samilly E. Souza, Antônio P. Ricomini-Filho, Altair A. Del Bel Cury, Yuri W. Cavalcanti, and Jaime A. Cury. "Candida albicans Increases Dentine Demineralization Provoked by Streptococcus mutans Biofilm." Caries Research 53, no. 3 (November 16, 2018): 322–31. http://dx.doi.org/10.1159/000494033.
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Streptococcus mutans are considered the most cariogenic bacteria, but it has been suggested that Candida albicans could increase their cariogenicity. However, the effect of this dual-species microorganisms’ combination on dentine caries has not been experimentally evaluated. Biofilms of C. albicans, S. mutans and C. albicans + S. mutans (n = 12/biofilm) were grown in ultra-filtered tryptone yeast extract broth culture medium for 96 h on root dentine slabs of known surface hardness and exposed 8 times per day for 3 min to 10% sucrose. The medium was changed 2 times per day (after the 8 cariogenic challenges and after the overnight period of famine), and aliquots were analyzed to determinate the pH (indicator of biofilm acidogenicity). After 96 h, the biofilms were collected to determine the wet weight, colony-forming units, and the amounts of extracellular polysaccharides (soluble and insoluble). Dentine demineralization was assessed by surface hardness loss (% SHL). The architecture of the biofilms was analyzed by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Data were analyzed by ANOVA followed by Tukey’s test (α = 0.05). The dual-species C. albicans + S. mutans biofilm provoked higher % SHL on dentine (p < 0.05) than the S. mutans and C. albicans biofilm. This was supported by the results of biofilm acidogenicity and the amounts of soluble (6.4 ± 2.14 vs. 4.0 ± 0.94 and 1.9 ± 0.97, respectively) and insoluble extracellular polysaccharides (24.9 ± 9.22 vs. 18.9 ± 5.92 and 0.7 ± 0.48, respectively) (p < 0.05). The C. albicans biofilm alone presented low cariogenicity. The images by CLSM and TEM, respectively, suggest that the C. albicans + S. mutans biofilm is more voluminous than the S. mutans biofilm, and S. mutans cells interact with C. albicans throughout polysaccharides from the biofilm matrix. These findings show that C. albicans enhances the cariogenic potential of the S. mutans biofilm, increasing dentine demineralization.
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Spinola, Manuela S., Diego Figueiredo Nóbrega, Altair Antoninha Del Bel Cury, Antonio Pedro Ricomini Filho, Jaime Aparecido Cury, and Livia Maria Andaló Tenuta. "Fluoride Penetration and Clearance Are Higher in Exopolysaccharide-Containing Bacterial Pellets." Caries Research 53, no. 1 (June 6, 2018): 16–23. http://dx.doi.org/10.1159/000488596.
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Extracellular polysaccharides (EPS) could increase the penetration of fluoride through dental biofilm, reducing its cariogenicity. We measured the concentration of fluoride in EPS-containing (EPS+) or not-containing (EPS–) Streptococcus mutans bacterial pellets resembling test biofilms, before and up to 60 min after a 0.05% NaF rinse in situ. Fluoride penetration and clearance were higher in EPS+ bacterial pellets. The data suggest that EPS enhances fluoride penetration, but also accelerates fluoride clearance from dental biofilms.
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Díaz-Garrido, Natalia, Carla Lozano, and Rodrigo A. Giacaman. "Frequency of sucrose exposure on the cariogenicity of a biofilm-caries model." European Journal of Dentistry 10, no. 03 (July 2016): 345–50. http://dx.doi.org/10.4103/1305-7456.184163.
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ABSTRACT Objective: Although sucrose is considered the most cariogenic carbohydrate in the human diet, the question of how many exposures are needed to induce damage on the hard dental tissues remains unclear. To approach this question, different frequencies of daily sucrose exposure were tested on a relevant biological caries model. Materials and Methods: Biofilms of the Streptococcus mutans were formed on enamel slabs and exposed to cariogenic challenges with 10% sucrose for 5 min at 0, 1, 3, 5, 8, or 10 times per day. After 5 days, biofilms were retrieved to analyze biomass, protein content, viable bacteria, and polysaccharide formation. Enamel demineralization was evaluated by percentage of microhardness loss (percentage surface hardness loss [%SHL]). Results: Biomass, protein content, polysaccharide production, acidogenicity of the biofilm, and %SHL proportionally increased with the number of daily exposures to sucrose (P < 0.05). One daily sucrose exposure was enough to induce 20% more demineralization than the negative unexposed control. Higher frequencies induced greater demineralization and more virulent biofilms, but eight and ten exposures were not different between them in most of the analyzed variables (P > 0.05). Conclusions: Higher sucrose exposure seems to increase cariogenicity, in a frequency-dependent manner, by the modification of bacterial virulent properties.
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Lei, L., B. Zhang, M. Mao, H. Chen, S. Wu, Y. Deng, Y. Yang, H. Zhou, and T. Hu. "Carbohydrate Metabolism Regulated by Antisense vicR RNA in Cariogenicity." Journal of Dental Research 99, no. 2 (December 10, 2019): 204–13. http://dx.doi.org/10.1177/0022034519890570.
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Streptococcus mutans is a major cariogenic pathogen that resides in multispecies oral microbial biofilms. The VicRK 2-component system is crucial for bacterial adaptation, virulence, and biofilm organization and contains a global and vital response regulator, VicR. Notably, we identified an antisense vicR RNA (AS vicR) associated with an adjacent RNase III–encoding ( rnc) gene that was relevant to microRNA-size small RNAs (msRNAs). Here, we report that ASvicR overexpression significantly impeded bacterial growth, biofilm exopolysaccharide synthesis, and cariogenicity in vivo. Transcriptome analysis revealed that the AS vicR RNA mainly regulated carbohydrate metabolism. In particular, overproducing AS vicR demonstrated a reduction in galactose and glucose metabolism by monosaccharide composition analysis. The results of high-performance gel permeation chromatography revealed that the water-insoluble glucans isolated from AS vicR presented much lower molecular weights. Furthermore, direct evidence showed that total RNAs were disrupted by rnc-encoded RNase III. With the coexpression of T4 RNA ligase, putative msRNA1657, which is an rnc-related messenger RNA, was verified to bind to the 5′-UTR regions of the vicR gene. Furthermore, AS vicR regulation revealed a sponge regulatory-mediated network for msRNA associated with adjacent RNase III–encoding genes. There was an increase in AS vicR transcript levels in clinical S. mutans strains from caries-free children, while the expression of AS vicR was decreased in early childhood caries patients; this outcome may be explored as a potential strategy contributing to the management of dental caries. Taken together, our findings suggest an important role of AS vicR-mediated sponge regulation in S. mutans, indicating the characterization of lactose metabolism by a vital response regulator in cariogenicity. These findings have a number of implications and have reshaped our understanding of bacterial gene regulation from its transcriptional conception to the key roles of regulatory RNAs.
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Leme, A. F. Paes, H. Koo, C. M. Bellato, G. Bedi, and J. A. Cury. "The Role of Sucrose in Cariogenic Dental Biofilm Formation—New Insight." Journal of Dental Research 85, no. 10 (October 2006): 878–87. http://dx.doi.org/10.1177/154405910608501002.
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Dental caries is a biofilm-dependent oral disease, and fermentable dietary carbohydrates are the key environmental factors involved in its initiation and development. However, among the carbohydrates, sucrose is considered the most cariogenic, because, in addition to being fermented by oral bacteria, it is a substrate for the synthesis of extracellular (EPS) and intracellular (IPS) polysaccharides. Therefore, while the low pH environment triggers the shift of the resident plaque microflora to a more cariogenic one, EPS promote changes in the composition of the biofilms’ matrix. Furthermore, it has recently been shown that the biofilm formed in the presence of sucrose presents low concentrations of Ca, Pi, and F, which are critical ions involved in de- and remineralization of enamel and dentin in the oral environment. Thus, the aim of this review is to explore the broad role of sucrose in the cariogenicity of biofilms, and to present a new insight into its influence on the pathogenesis of dental caries.
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Mao, Meng-Ying, Meng Li, Lei Lei, Jia-Xin Yin, Ying-Ming Yang, and Tao Hu. "The Regulator Gene rnc Is Closely Involved in Biofilm Formation in Streptococcus mutans." Caries Research 52, no. 5 (2018): 347–58. http://dx.doi.org/10.1159/000486431.
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Streptococcus mutans is an important factor in the etiology and pathogenesis of dental caries, largely owing to its ability to form a stable biofilm. Previous animal studies have indicated that rnc could decrease the amount of sulcal caries, and that the downregulation of cariogenicity might be due to its capacity to disrupt biofilm formation. However, the biofunctions by which rnc is involved in biofilm formation remain to be elucidated. In this study, we further investigate the role of rnc based on the study of mature biofilm. Scanning electron microscopy and the crystal violet assay were used to detect the biofilm forming ability. The production and distribution of exopolysaccharides within biofilm was analyzed by exopolysaccharide staining. Gel permeation chromatography was used to perform molecular weight assessment. Its adhesion force was measured by atomic force microscopy. The expression of biofilm formation-associated genes was analyzed at the mRNA level by qPCR. Here, we found that rnc could occur and function in biofilm formation by assembling well-structured, exopolysaccharide-encased, stable biofilms in S. mutans. The weakened biofilm forming ability of rnc-deficient strains was associated with the reduction of exopolysaccharide production and bacterial adhesion. Over all, these data illustrate an interesting situation in which an unappreciated regulatory gene acquired for virulence, rnc, most likely has been coopted as a potential regulator of biofilm formation in S. mutans. Further characterization of rnc may lead to the identification of a possible pathogenic biofilm-specific treatment for dental caries.
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Pagotto, Leonardo Libardi, Manuela da Silva Spinola, Diego Figueiredo Nóbrega, Jaime Aparecido Cury, and Livia Maria Andaló Tenuta. "Fluoride Binding to Streptococcus mutans Pellets Rich in Extracellular Polysaccharides." Caries Research 55, no. 3 (2021): 234–37. http://dx.doi.org/10.1159/000515115.
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Extracellular polysaccharides (EPS), mainly the insoluble ones, increase the cariogenicity of dental biofilm, but whether they interfere with the binding and retention of fluoride is unknown. EPS-rich (EPS+) and EPS-poor (EPS–) pellets of <i>Streptococcus mutans</i> were formed and treated with increasing fluoride concentrations (0, 0.1, 1, or 10 mM). A concentration-dependent fluoride binding was observed in both EPS– and EPS+ pellets, but the presence of EPS did not affect the retention of fluoride in the pellets. In conclusion, the data suggest that a matrix of dental biofilm rich in EPS does not affect fluoride retention in the biofilm.
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Ricomini Filho, Antonio P., Ana Camila M. de Assis, Bárbara E. Costa Oliveira, and Jaime A. Cury. "Cariogenic Potential of Human and Bovine Milk on Enamel Demineralization." Caries Research 55, no. 4 (2021): 260–67. http://dx.doi.org/10.1159/000516090.
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The higher cariogenicity of human milk when compared with bovine milk is still a debatable subject. Therefore, we evaluated the effect of human or bovine milk exposure on biofilm composition and enamel demineralization using a validated cariogenic biofilm model. <i>Streptococcus mutans</i> UA159 biofilms (<i>n</i> = 8) were grown on human saliva-coated bovine enamel slabs of known surface hardness. The biofilms were exposed 8×/day to 0.9% NaCl (negative control), human milk, bovine milk, 7.0% lactose (active human milk control), 4.5% lactose (active bovine milk control), or 10% sucrose (positive control). The culture medium was changed twice daily, and the pH was analyzed as an indicator of biofilm acidogenicity. After 120 h of growth, biofilms were harvested to evaluate viable cells, and soluble and insoluble extracellular polysaccharides (EPS). Enamel demineralization was assessed by the percentage of surface hardness loss (%SHL). Data were analyzed by one-way ANOVA/Tukey’s test (α = 5%). In terms of %SHL, negative control (7.7 ± 3.1), human milk control (13.3 ± 7.5), bovine milk control (15.3 ± 8.2), human milk (7.5 ± 5.0), and bovine milk (8.7 ± 6.3) did not differ among them (<i>p</i> > 0.05) but differed (<i>p</i> < 0.05) from sucrose (55.1 ± 5.4). The findings of enamel demineralization (%SHL) were statistically supported by the data of biofilm acidogenicity, bacterial counts and EPS biofilm composition. This experimental study suggests that human and bovine milk have low cariogenic potential to provoke caries lesions in enamel.
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Signori, Cácia, Françoise H. van de Sande, Tamires T. Maske, Elenara F. de Oliveira, and Maximiliano S. Cenci. "Influence of the Inoculum Source on the Cariogenicity of in vitro Microcosm Biofilms." Caries Research 50, no. 2 (2016): 97–103. http://dx.doi.org/10.1159/000443537.
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This study investigated the cariogenic potential of biofilms originating from different types of inoculum (saliva and dental plaque) from caries-active and caries-free individuals. Ten volunteers were selected from each caries condition for the paired collection of saliva and dental plaque. Microcosm biofilms were grown in triplicate from each inoculum on enamel specimens in 24-well plates under cariogenic challenge. After 10 days, the biofilms were collected for analysis of outcome variables: percentage of surface hardness change (%SHC) and microbiological composition of biofilms. Statistical analysis was performed using the t test, the linear multivariate analysis model and Pearson's correlation coefficients (α = 0.05). A comparative analysis between microbiological baseline data showed higher counts of mutans streptococci in plaque samples within caries-active individuals; a comparative analysis of colony-forming unit (CFU) counts between individuals with different caries status showed higher counts of acid-tolerant microorganisms and mutans streptococci in dental plaque and of acid-tolerant microorganisms in saliva. After 10 days of biofilm growth, the CFU values for total microorganisms, lactobacilli, mutans streptococci and acid-tolerant bacteria, as well as for SHC, were not statistically significant, considering the type of inoculum and caries condition (p > 0.05). A positive correlation was found for %SHC and CFU counts of acid-tolerant bacteria (r = 0.406) and lactobacilli (r = 0.379). Under the limits of this study, the cariogenic potential of biofilms, formed under identical conditions in vitro, is similar, regardless of baseline differences between the source and type of inoculum.
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Ren, Zhi, Tao Cui, Jumei Zeng, Lulu Chen, Wenling Zhang, Xin Xu, Lei Cheng, et al. "Molecule Targeting Glucosyltransferase Inhibits Streptococcus mutans Biofilm Formation and Virulence." Antimicrobial Agents and Chemotherapy 60, no. 1 (October 19, 2015): 126–35. http://dx.doi.org/10.1128/aac.00919-15.
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ABSTRACTDental plaque biofilms are responsible for numerous chronic oral infections and cause a severe health burden. Many of these infections cannot be eliminated, as the bacteria in the biofilms are resistant to the host's immune defenses and antibiotics. There is a critical need to develop new strategies to control biofilm-based infections. Biofilm formation inStreptococcus mutansis promoted by major virulence factors known as glucosyltransferases (Gtfs), which synthesize adhesive extracellular polysaccharides (EPS). The current study was designed to identify novel molecules that target Gtfs, thereby inhibitingS. mutansbiofilm formation and having the potential to prevent dental caries. Structure-based virtual screening of approximately 150,000 commercially available compounds against the crystal structure of the glucosyltransferase domain of the GtfC protein fromS. mutansresulted in the identification of a quinoxaline derivative, 2-(4-methoxyphenyl)-N-(3-{[2-(4-methoxyphenyl)ethyl]imino}-1,4-dihydro-2-quinoxalinylidene)ethanamine, as a potential Gtf inhibitor.In vitroassays showed that the compound was capable of inhibiting EPS synthesis and biofilm formation inS. mutansby selectively antagonizing Gtfs instead of by killing the bacteria directly. Moreover, thein vivoanti-caries efficacy of the compound was evaluated in a rat model. We found that the compound significantly reduced the incidence and severity of smooth and sulcal-surface cariesin vivowith a concomitant reduction in the percentage ofS. mutansin the animals' dental plaque (P< 0.05). Taken together, these results represent the first description of a compound that targets Gtfs and that has the capacity to inhibit biofilm formation and the cariogenicity ofS. mutans.
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Kuang, Xinyi, Tao Yang, Chenzi Zhang, Xian Peng, Yuan Ju, Chungen Li, Xuedong Zhou, Youfu Luo, and Xin Xu. "Repurposing Napabucasin as an Antimicrobial Agent against Oral Streptococcal Biofilms." BioMed Research International 2020 (November 20, 2020): 1–9. http://dx.doi.org/10.1155/2020/8379526.
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Objectives. Disruption of microbial biofilms is an effective way to control dental caries. Drug resistance and side effects of the existing antimicrobials necessitate the development of novel antibacterial agents. The current study was aimed at investigating the antibacterial activities of the repurposed natural compound napabucasin against oral streptococci. Methods. The minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibition concentration, and minimum biofilm reduction concentration of Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis were examined by a microdilution method. Cytotoxicity of napabucasin against human oral keratinocytes, human gingival epithelia, and macrophage RAW264.7 was evaluated by CCK8 assays. The dead/live bacterium and exopolysaccharide in the napabucasin-treated multispecies biofilms were evaluated by confocal laser scanning microscopy. Microbial composition within the napabucasin-treated biofilms was further visualized by fluorescent in situ hybridization and qPCR. And the cariogenicity of napabucasin-treated biofilms was evaluated by transverse microradiography. Results. Napabucasin exhibited good antimicrobial activity against oral streptococcal planktonic cultures and biofilms but with lessened cytotoxicity as compared to chlorhexidine. Napabucasin reduced the cariogenic S. mutans and increased the proportion of the commensal S. gordonii in the multispecies biofilms. More importantly, napabucasin significantly reduced the demineralization capability of biofilms on tooth enamels. Conclusion. Napabucasin shows lessened cytotoxicity and comparable antimicrobial effects to chlorhexidine. Repurposing napabucasin may represent a promising adjuvant for the management of dental caries.
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Xu, Xin, Xue D. Zhou, and Christine D. Wu. "The Tea Catechin Epigallocatechin Gallate Suppresses Cariogenic Virulence Factors ofStreptococcus mutans." Antimicrobial Agents and Chemotherapy 55, no. 3 (December 13, 2010): 1229–36. http://dx.doi.org/10.1128/aac.01016-10.
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ABSTRACTStreptococcus mutans, the primary etiologic agent of dental caries, possesses a series of virulence factors associated with its cariogenicity. Alternatives to traditional antimicrobial treatment, agents selectively inhibiting the virulence factors without necessarily suppressing the resident oral species, are promising. The anticariogenic properties of tea have been suggested in experimental animals and humans. Tea polyphenols, especially epigallocatechin gallate (EGCg), have been shown to inhibit the growth and glucosyltransferases activity ofS. mutans. However, their effects on biofilm and cariogenic virulence factors of oral streptococci other than glucosyltransferases have not been well documented. In this study, we investigated the biological effect of EGCg on the virulence factors ofS. mutansassociated with its acidogenicity and acidurity. The antimicrobial effects of EGCg onS. mutansbiofilm grown in chemically defined medium were also examined. EGCg inhibited growth ofS. mutansplanktonic cells at an MIC of 31.25 μg/ml and a minimal bactericidal concentration (MBC) of 62.5 μg/ml. EGCg also inhibitedS. mutansbiofilm formation at 15.6 μg/ml (minimum concentration that showed at least 90% inhibition of biofilm formation) and reduced viability of the preformed biofilm at 625 μg/ml (sessile MIC80). EGCg at sub-MIC levels inhibited acidogenicity and acidurity ofS. mutanscells. Analysis of the data obtained from real-time PCR showed that EGCg significantly suppressed theldh,eno,atpD, andaguDgenes ofS. mutansUA159. Inhibition of the enzymatic activity of F1Fo-ATPase and lactate dehydrogenase was also noted (50% inhibitory concentration between 15.6 and 31.25 μg/ml). These findings suggest that EGCg is a natural anticariogenic agent in that it exhibits antimicrobial activity againstS. mutansand suppresses the specific virulence factors associated with its cariogenicity.
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Dotsey, Roger P., Elizabeth A. S. Moser, George J. Eckert, and Richard L. Gregory. "Effects of Cola-Flavored Beverages and Caffeine on Streptococcus mutans Biofilm Formation and Metabolic Activity." Journal of Clinical Pediatric Dentistry 41, no. 4 (January 1, 2017): 294–99. http://dx.doi.org/10.17796/1053-4628-41.4.294.
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Objective: To examine the effects of cola-flavored beverages and caffeine on growth and metabolism of Streptococcus mutans biofilm. This study was designed to determine if carbonated beverages or caffeine can increase S. mutans growth and biofilm formation and metabolic activity in vitro, potentially leading to increased S. mutans-associated cariogenicity in children that consume them. Study Design: Six different cola-flavored products, plus pure caffeine, and pure high fructose corn syrup (HFCS), at different concentrations similar to those in the beverages were tested. A 16-hour culture of S. mutans was treated with different dilutions in bacteriological media. To test for the effect on biofilm formation, the biofilm was stained with crystal violet. The absorbance was determined to evaluate biofilm growth. Biofilm metabolic activity was measured based on biofilm having the ability to reduce XTT to a water-soluble orange compound. Results: The inclusion of HFCS in the beverages, as well as pure HFCS, significantly enhanced bacterial biofilm formation and metabolic activity. Pure caffeine and the presence of caffeine in beverages did not significantly increase biofilm formation, but pure caffeine significantly increased metabolism, and Diet Coke had significantly greater metabolic activity than Caffeine-Free Diet Coke. Conclusions: HFCS increases both the biofilm formation and metabolism of S. mutans, and caffeine in some cases increases metabolism of S. mutans.
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Zhu, Lin, Jens Kreth, Sarah E. Cross, James K. Gimzewski, Wenyuan Shi, and Fengxia Qi. "Functional characterization of cell-wall-associated protein WapA in Streptococcus mutans." Microbiology 152, no. 8 (August 1, 2006): 2395–404. http://dx.doi.org/10.1099/mic.0.28883-0.
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Streptococcus mutans is known as a primary pathogen responsible for dental caries. One of the virulence factors of S. mutans in cariogenicity is its ability to attach to the tooth surface and form a biofilm. Several surface proteins have been shown to be involved in this process. A 29 kDa surface protein named wall-associated protein A (WapA, antigen A or antigen III), was previously used as a vaccine in animal studies for immunization against dental caries. However, the function of WapA in S. mutans is still not clear. This study characterized the function of WapA in cell surface structure and biofilm formation. Compared to the wild-type, the wapA mutant had much-reduced cell chain length, diminished cell–cell aggregation, altered cell surface ultrastructure, and unstructured biofilm architecture. Furthermore, in vivo force spectroscopy revealed that the cell surface of the wapA mutant was less sticky than that of the wild-type cells. More interestingly, these phenotypic differences diminished as sucrose concentration in the medium was increased to 0.5 %. Real-time RT-PCR analysis demonstrated that sucrose strongly repressed wapA gene expression in both planktonic and biofilm cells. These results suggest that the WapA protein plays an important structural role on the cell surface, which ultimately affects sucrose-independent cell–cell aggregation and biofilm architecture.
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Hwang, Minseon, Howon Park, Juhyun Lee, Hyunwoo Seo, and Siyoung Lee. "Comparison of Cariogenicity of Bovine Milk and Low-fat Milk on Streptococcus mutans Biofilm." JOURNAL OF THE KOREAN ACADEMY OF PEDTATRIC DENTISTRY 44, no. 2 (May 25, 2017): 170–79. http://dx.doi.org/10.5933/jkapd.2017.44.2.170.
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Giacaman, Rodrigo A., Pascale Jobet-Vila, and Cecilia Muñoz-Sandoval. "Fatty acid effect on sucrose-induced enamel demineralization and cariogenicity of an experimental biofilm–caries model." Odontology 103, no. 2 (April 11, 2014): 169–76. http://dx.doi.org/10.1007/s10266-014-0154-5.
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Castro, Ramiro J., Rodrigo A. Giacaman, Rodrigo A. Arthur, and Marisa Maltz. "Cariogenicity of a Milk-Based Drink Used as a Dietary Supplement for Older Adults Using a Root Caries Experimental Model." Caries Research 53, no. 1 (June 26, 2018): 76–83. http://dx.doi.org/10.1159/000489569.
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The aim of this study was to evaluate the cariogenicity of a milk-based drink intended for older adults that was used as part of a governmental initiative in Chile to improve their nutritional conditions. This drink contains a high concentration of sugars, which can contribute to root caries development. To test this hypothesis, an experimental biofilm/caries model was used. Dentin slabs were used to grow biofilms of Streptococcus mutans UA159. Slabs/biofilms were exposed 3× per day to bovine milk with different fat content, the milk-based drink, and the milk-based drink supplemented with 10 g of sucrose added per serving. Slabs exposed to 10% sucrose or 0.9% NaCl were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial counts and acidogenicity. Dentin demineralization was estimated by the loss of surface microhardness and integrated mineral loss. Results were compared by analysis of variance and Tukey’s test. The milk-based drink showed higher acidogenicity than milk with its entire (whole) or reduced total fat content (skim). The milk-based drink supplemented with sucrose had similar acidogenicity as the 10% sucrose positive control (p = 0.506). Whole milk exposure elicited lower bacterial counts than the positive control, the milk-based drink, and the milk-based drink supplemented with sucrose (p = 0.002; 0.006 and 0.014 respectively). Although skim milk induced higher demineralization than whole milk, both milk types produced lower demineralization than the milk-based drink. Regarding integrated mineral loss, demineralization induced by the milk-based drink and the milk-based drink supplemented with sucrose was similar to that induced by the positive control and skim milk (p > 0.05). Sugar-containing milk-based drinks used as dietary supplements for older adults may be highly cariogenic and could represent a potential risk for root caries.
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Tanzer, J. M., L. Grant, A. Thompson, L. Li, J. D. Rogers, E. M. Haase, and F. A. Scannapieco. "Amylase-binding proteins A (AbpA) and B (AbpB) differentially affect colonization of rats' teeth by Streptococcus gordonii." Microbiology 149, no. 9 (September 1, 2003): 2653–60. http://dx.doi.org/10.1099/mic.0.26022-0.
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Streptococcus gordonii produces two α-amylase-binding proteins, AbpA and AbpB, that have been extensively studied in vitro. Little is known, however, about their significance in oral colonization and cariogenicity (virulence). To clarify these issues, weanling specific pathogen-free Osborne-Mendel rats, TAN : SPFOM(OM)BR, were inoculated either with wild-type strains FAS4-S or Challis-S or with strains having isogenic mutations of abpA, abpB, or both, to compare their colonization abilities and persistence on the teeth. Experiments were done with rats fed a sucrose-rich diet containing low amounts of starch or containing only starch. The mutants and wild-types were quantified in vivo and carious lesions were scored. In 11 experiments, S. gordonii was a prolific colonizer of the teeth when rats were fed the sucrose (with low starch)-supplemented diet, often dominating the flora. Sucrose-fed rats had several-fold higher recoveries of inoculants than those eating the sucrose-free, starch-supplemented diet, regardless of inoculant type. The strain defective in AbpB could not colonize teeth of starch-only-eating rats, but could colonize rats if sucrose was added to the diet. Strains defective in AbpA surprisingly colonized better than their wild-types. A double mutant deficient in both AbpA and AbpB (abpA/abpB) colonized like its wild-type. Wild-types FAS4-S and Challis-S had no more than marginal cariogenicity. Notably, in the absence of AbpA, cariogenicity was slightly augmented. Both the rescue of colonization by the AbpB− mutant and the augmentation of colonization by AbpA− mutant in the presence of dietary sucrose suggested additional amylase-binding protein interactions relevant to colonization. Glucosyltransferase activity was greater in mutants defective in abpA and modestly increased in the abpB mutant. It was concluded that AbpB is required for colonization of teeth of starch-eating rats and its deletion is partially masked if rats eat a sucrose-starch diet. AbpA appears to inhibit colonization of the plaque biofilm in vivo. This unexpected effect in vivo may be associated with interaction of AbpA with glucosyltransferase or with other colonization factors of these cells. These data illustrate that the complex nature of the oral environment may not be adequately modelled by in vitro systems.
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Palmer, Sara R., Paula J. Crowley, Monika W. Oli, M. Adam Ruelf, Suzanne M. Michalek, and L. Jeannine Brady. "YidC1 and YidC2 are functionally distinct proteins involved in protein secretion, biofilm formation and cariogenicity of Streptococcus mutans." Microbiology 158, no. 7 (July 1, 2012): 1702–12. http://dx.doi.org/10.1099/mic.0.059139-0.
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Chen, Xiuqin, Eric Banan-Mwine Daliri, Ramachandran Chelliah, and Deog-Hwan Oh. "Isolation and Identification of Potentially Pathogenic Microorganisms Associated with Dental Caries in Human Teeth Biofilms." Microorganisms 8, no. 10 (October 16, 2020): 1596. http://dx.doi.org/10.3390/microorganisms8101596.
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Dental caries is attributed to the predominance of cariogenic microorganisms. Cariogenic microorganisms are pathological factors leading to acidification of the oral microenvironment, which is related to the initiation and progression of caries. The accepted cariogenic microorganism is Streptococcus mutans (S. mutans). However, studies have found that caries could occur in the absence of S. mutans. This study aimed to assess the presence of potentially cariogenic microorganisms in human teeth biofilm. The microorganisms were isolated from human mouth and freshly extracted human maxillary incisors extracted for reasons of caries. The isolates were sorted based on their acidogenic and aciduric properties, and the S. mutans was used as the reference strain. Four potentially cariogenic strains were selected. The selected strains were identified as Streptococcus salivarius (S. salivarius), Streptococcus anginosus (S. anginosus), Leuconostoc mesenteroides (L. mesenteroides), and Lactobacillus sakei (L. sakei) through morphological analysis followed by 16S rRNA gene sequence analysis. The cariogenicity of isolates was analyzed. We show, for the first time, an association between L. sakei (present in fermented food) and dental caries. The data provide useful information on the role of lactic acid bacteria from fermented foods and oral commensal streptococci in dental caries.
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Lee, Eun-Song, Si-Mook Kang, Hae-Youn Ko, Ho-Keun Kwon, and Baek-Il Kim. "Association between the cariogenicity of a dental microcosm biofilm and its red fluorescence detected by Quantitative Light-induced Fluorescence-Digital (QLF-D)." Journal of Dentistry 41, no. 12 (December 2013): 1264–70. http://dx.doi.org/10.1016/j.jdent.2013.08.021.
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Smith, Daniel J., William F. King, and Ronald Godiska. "Passive Transfer of Immunoglobulin Y Antibody toStreptococcus mutans Glucan Binding Protein B Can Confer Protection against Experimental Dental Caries." Infection and Immunity 69, no. 5 (May 1, 2001): 3135–42. http://dx.doi.org/10.1128/iai.69.5.3135-3142.2001.
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ABSTRACT Active immunization with Streptococcus mutans glucan binding protein B (GBP-B) has been shown to induce protection against experimental dental caries. This protection presumably results from continuous secretion of salivary antibody to GBP-B, which inhibits accumulation of S. mutans within the oral biofilm. The purpose of this study was to explore the influence of short-term (9- or 24-day) passive oral administration of antibody to S. mutans GBP-B on the longer-term accumulation and cariogenicity ofS. mutans in a rat model of dental caries. Preimmune chicken egg yolk immunoglobulin Y (IgY) or IgY antibody to S. mutans GBP-B was supplied in lower (experiment 1) and higher (experiment 2) concentrations in the diet and drinking water of rats for 9 (experiment 1) or 24 (experiment 2) days. During the first 3 days of IgY feeding, all animals were challenged with 5 × 106 streptomycin-resistant S. mutans strain SJ-r organisms. Rats remained infected with S. mutans for 78 days, during which rat molars were sampled for the accumulation ofS. mutans SJ-r bacteria and total streptococci. Geometric mean levels of S. mutans SJ-r accumulation on molar surfaces were significantly lower in antibody-treated rats on days 16 and 78 of experiment 2 and were lower on all but the initial (day 5) swabbing occasions in both experiments. Relative to controls, the extent of molar dental caries measured on day 78 was also significantly decreased. The decrease in molar caries correlated with the amount and duration of antibody administration. This is the first demonstration that passive antibody to S. mutans GBP-B can have a protective effect against cariogenic S. mutans infection and disease. Furthermore, this decrease in infection and disease did not require continuous antibody administration for the duration of the infection period. This study also indicates that antibody to components putatively involved only in cellular aggregation can have a significant effect on the incorporation of mutans streptococci in dental biofilm.
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Ahn, Sang-Joon, and Kelly C. Rice. "Understanding the Streptococcus mutans Cid/Lrg System through CidB Function." Applied and Environmental Microbiology 82, no. 20 (August 12, 2016): 6189–203. http://dx.doi.org/10.1128/aem.01499-16.
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ABSTRACTTheStreptococcus mutanslrgABandcidABoperons have been previously described as a potential model system to dissect the complexity of biofilm development and virulence ofS. mutans. Herein, we have attempted to further characterize the Cid/Lrg system by focusing on CidB, which has been shown to be critical for the ability ofS. mutansto survive and persist in a nonpreferred oxygen-enriched condition. We have found that the expression level ofcidBis critical to oxidative stress tolerance ofS. mutans, most likely by impactinglrgexpression. Intriguingly, the impaired aerobic growth phenotype of thecidBmutant could be restored by the additional loss of either CidA or LrgA. Growth-dependent expression ofcidandlrgwas demonstrated to be tightly under the control of both CcpA and the VicKR two-component system (TCS), regulators known to play an essential role in controlling major catabolic pathways and cell envelope homeostasis, respectively. RNA sequencing (RNA-Seq) analysis revealed that mutation ofcidBresulted in global gene expression changes, comprising major domains of central metabolism and virulence processes, particularly in those involved with oxidative stress resistance. Loss of CidB also significantly changed the expression of genes related to genomic islands (GI) TnSmu1 and TnSmu2, the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas system, and toxin-antitoxin (T/A) modules. Taken together, these data show that CidB impinges on the stress response, as well as the fundamental cellular physiology ofS. mutans, and further suggest a potential link between Cid/Lrg-mediated cellular processes,S. mutanspathogenicity, and possible programmed growth arrest and cell death mechanisms.IMPORTANCEThe ability ofStreptococcus mutansto survive a variety of harmful or stressful conditions and to emerge as a numerically significant member of stable oral biofilm communities are essential elements for its persistence and cariogenicity. In this study, the homologouscidABandlrgABoperons, previously identified as being highly balanced and coordinated duringS. mutansaerobic growth, were further characterized through the functional and transcriptomic analysis of CidB. Precise control of CidB levels is shown to impact the expression oflrg, oxidative stress tolerance, major metabolic domains, and the molecular modules linked to cell death and lysis. This study advances our understanding of the Cid/Lrg system as a key player in the integration of complex environmental signals (such as oxidative stress) into the regulatory networks that modulateS. mutansvirulence and cell homeostasis.
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Lei, L., Y. Yang, Y. Yang, S. Wu, X. Ma, M. Mao, and T. Hu. "Mechanisms by Which Small RNAs Affect Bacterial Activity." Journal of Dental Research 98, no. 12 (September 23, 2019): 1315–23. http://dx.doi.org/10.1177/0022034519876898.
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The oral cavity contains a distinct habitat that supports diverse bacterial flora. Recent observations have provided additional evidence that sRNAs are key regulators of bacterial physiology and pathogenesis. These sRNAs have been divided into 5 functional groups: cis-encoded RNAs, trans-encoded RNAs, RNA regulators of protein activity, bacterial CRISPR (clustered regularly interspaced short palindromic repeat) RNAs, and a novel category of miRNA-size small RNAs (msRNAs). In this review, we discuss a critical group of key commensal and opportunistic oral pathogens. In general, supragingival bacterial sRNAs function synergistically to fine-tune the regulation of cellular processes and stress responses in adaptation to environmental changes. Particularly in the cariogenic bacteria Streptococcus mutans, both the antisense vicR RNA and msRNA1657 can impede the metabolism of bacterial exopolysaccharides, prevent biofilm formation, and suppress its cariogenicity. In Enterococcus faecalis, selected sRNAs control the expression of proteins involved in diverse cellular processes and stress responses. In subgingival plaques, sRNAs from periodontal pathogens can function as novel bacterial signaling molecules that mediate bacterial-human interactions in periodontal homeostasis. In Porphyromonas gingivalis, the expression profiles of putative sRNA101 and sRNA42 were found to respond to hemin availability after hemin starvation. Regarding Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), a major periodontal pathogen associated with aggressive periodontitis, the predicted sRNAs interact with several virulence genes, including those encoding leukotoxin and cytolethal distending toxin. Furthermore, in clinical isolates, these associated RNAs could be explored not only as potential biomarkers for oral disease monitoring but also as alternative types of regulators for drug design. Thus, this emerging subspecialty of bacterial regulatory RNAs could reshape our understanding of bacterial gene regulation from their key roles of endogenous regulatory RNAs to their activities in pathologic processes.
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Azevedo, M. S., F. H. van de Sande, A. R. Romano, and M. S. Cenci. "Microcosm Biofilms Originating from Children with Different Caries Experience Have Similar Cariogenicity under Successive Sucrose Challenges." Caries Research 45, no. 6 (2011): 510–17. http://dx.doi.org/10.1159/000331210.
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Rezende, Gabriela, Rodrigo A. Arthur, Debora Grando, and Lina N. Hashizume. "Cariogenic Potential of Sucrose Associated with Maltodextrin on Dental Enamel." Caries Research 51, no. 2 (2017): 129–35. http://dx.doi.org/10.1159/000453623.
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Maltodextrin is a hydrolysate of cornstarch and has been widely used in the food industry associated with sucrose. The addition of starch can increase the cariogenic potential of sucrose; however, there are sparse data regarding the cariogenicity of sucrose associated with maltodextrin. Therefore, the aim of this study was to test in situ if maltodextrin could increase the cariogenic potential of sucrose. This was an in situ, randomized, crossover, split-mouth, and double-blind study. Volunteers wore palatal appliances containing bovine enamel blocks for 2 periods of 14 days. They dripped the following solutions on the enamel blocks 8 times per day: deionized distilled water (DDW), maltodextrin (M), sucrose + maltodextrin (S+M), or sucrose (S). At the end of each experimental period, biofilms were collected and analyzed for microbiological (mutans streptococci, lactobacilli, and total microorganisms counts) and biochemical (calcium, inorganic phosphate, fluoride, and insoluble extracellular polysaccharides concentrations) compositions. The enamel demineralization was assessed by microhardness. Treatments S and S+M resulted in a lower inorganic composition and higher concentration of insoluble extracellular polysaccharides in the biofilms, and higher enamel mineral loss compared to DDW and M. It can be concluded that the cariogenic potential of sucrose is not changed when this carbohydrate is associated with maltodextrin (dextrose equivalent 13-17).
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Miller, James H., Alejandro Avilés-Reyes, Kathy Scott-Anne, Stacy Gregoire, Gene E. Watson, Edith Sampson, Ann Progulske-Fox, et al. "The Collagen Binding Protein Cnm Contributes to Oral Colonization and Cariogenicity of Streptococcus mutans OMZ175." Infection and Immunity 83, no. 5 (March 2, 2015): 2001–10. http://dx.doi.org/10.1128/iai.03022-14.
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Streptococcus mutansis the etiological agent of dental caries and one of the many bacterial species implicated in infective endocarditis. The expression of the collagen-binding protein Cnm byS. mutanshas been associated with extraoral infections, but its relevance for dental caries has only been theorized to date. Due to the collagenous composition of dentinal and root tissues, we hypothesized that Cnm may facilitate the colonization of these surfaces, thereby enhancing the pathogenic potential ofS. mutansin advancing carious lesions. As shown for extraoral endothelial cell lines, Cnm mediates the invasion of oral keratinocytes and fibroblasts byS. mutans. In this study, we show that in the Cnm+native strain, OMZ175, Cnm mediates stringent adhesion to dentinal and root tissues as well as collagen-coated surfaces and promotes both cariogenicity and carriagein vivo. In vitro,ex vivo, andin vivoexperiments revealed that while Cnm is not universally required forS. mutanscariogenicity, it contributes to (i) the invasion of the oral epithelium, (ii) enhanced binding on collagenous surfaces, (iii) implantation of oral biofilms, and (IV) the severity of caries due to a native Cnm+isolate. Taken together, our findings reveal that Cnm is a colonization factor that contributes to the pathogenicity of certainS. mutansstrains in their native habitat, the oral cavity.
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Díaz-Garrido, Natalia, Carla P. Lozano, Jens Kreth, and Rodrigo A. Giacaman. "Competition and Caries on Enamel of a Dual-Species Biofilm Model with Streptococcus mutans and Streptococcus sanguinis." Applied and Environmental Microbiology 86, no. 21 (August 21, 2020). http://dx.doi.org/10.1128/aem.01262-20.
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ABSTRACT Imbalances within the dental biofilm trigger dental caries, currently considered a dysbiosis and the most prevalent noncommunicable disease. There is still a gap in knowledge about the dynamics of enamel colonization by bacteria from the dental biofilm in caries. The aim, therefore, was to test whether the sequence of enamel colonization by a typically commensal and a cariogenic species modifies biofilm’s cariogenicity. Dual-species biofilms of Streptococcus mutans and Streptococcus sanguinis on saliva-coated enamel slabs were inoculated in different sequences: S. mutans followed by S. sanguinis (Sm-Ss), S. sanguinis followed by S. mutans (Ss-Sm), S. mutans and S. sanguinis inoculated at the same time (Sm=Ss), and the single-species controls S. mutans followed by S. mutans (Sm-Sm) and S. sanguinis followed by S. sanguinis (Ss-Ss). Biofilms were exposed to 10% sucrose 3 times per day for 5 days, and the slabs/biofilms were retrieved to assess demineralization, viable cells, biomass, proteins, polysaccharides, and H2O2 production. Compared with Sm-Sm, primary inoculation with S. sanguinis reduced demineralization (P < 0.05). Both Ss-Sm and Sm=Ss sequences showed reduction in biomass, protein, and polysaccharide content (P < 0.05). The highest S. sanguinis viable count and H2O2 production level and the lowest acidogenicity were observed when S. sanguinis colonized enamel before S. mutans (P < 0.05). Initial enamel adherence with commensal biofilms seems to induce more intense competition against more typically cariogenic species, reducing cariogenicity. IMPORTANCE The concept of caries as an ecological disease implies the understanding of the intricate relationships among the populating microorganisms. Under frequent sugar exposure, some bacteria from the dental biofilm develop pathogenic traits that lead to imbalances (dysbiosis). Depending on which microorganism colonizes the dental surface first, different competition strategies may be developed. Studying the interactions in the entire dental biofilm is not an easy task. In this study, therefore, we modeled the interplay among these microorganisms using a caries-inducing species (S. mutans) and a health-associated species (S. sanguinis). Initial enamel adherence with S. sanguinis seems to induce more intense competition against typically caries-inducing species. Besides continuous exposure with sugars, early colonization of the enamel by highly cariogenic species like S. mutans appears to be needed to develop caries lesions as well. Promoting early colonization by health-associated bacteria such as S. sanguinis could help to maintain oral health, delaying dysbiosis.
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Niu, Yumei, Kun Wang, Sainan Zheng, Yufei Wang, Qian Ren, Haoran Li, Longjiang Ding, Wei Li, and Linglin Zhang. "Antibacterial Effect of Caffeic Acid Phenethyl Ester on Cariogenic Bacteria and Streptococcus mutans Biofilms." Antimicrobial Agents and Chemotherapy 64, no. 9 (June 15, 2020). http://dx.doi.org/10.1128/aac.00251-20.
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ABSTRACT Dental caries is the most common disease in the human mouth. Streptococcus mutans is the primary cariogenic bacterium. Propolis is a nontoxic natural product with a strong inhibitory effect on oral cariogenic bacteria. The polyphenol-rich extract from propolis inhibits S. mutans growth and biofilm formation, as well as the genes involved in virulence and adherence, through the inhibition of glucosyltransferases (GTF). However, because the chemical composition of propolis is highly variable and complex, the mechanism of its antimicrobial action and the active compound are controversial and not completely understood. Caffeic acid phenethyl ester (CAPE) is abundant in the polyphenolic compounds from propolis, and it has many pharmacological effects. In this study, we investigated the antibacterial effects of CAPE on common oral cariogenic bacteria (Streptococcus mutans, Streptococcus sobrinus, Actinomyces viscosus, and Lactobacillus acidophilus) and its effects on the biofilm-forming and cariogenic abilities of S. mutans. CAPE shows remarkable antimicrobial activity against cariogenic bacteria. Moreover, CAPE also inhibits the formation of S. mutans biofilms and their metabolic activity in mature biofilms. Furthermore, CAPE can inhibit the key virulence factors of S. mutans associated with cariogenicity, including acid production, acid tolerance, and the bacterium’s ability to produce extracellular polysaccharides (EPS), without affecting bacterial viability at subinhibitory levels. In conclusion, CAPE appears to be a new agent with anticariogenic potential, not only via inhibition of the growth of cariogenic bacteria.
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Guan, Chunru, Faai Che, Huoxiang Zhou, Yiwei Li, Yaru Li, and Jinpu Chu. "Effect of Rubusoside, a Natural Sucrose Substitute, on Streptococcus mutans Biofilm Cariogenic Potential and Virulence Gene Expression In Vitro." Applied and Environmental Microbiology 86, no. 16 (June 5, 2020). http://dx.doi.org/10.1128/aem.01012-20.
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ABSTRACT Dental caries is a biofilm-mediated disease in which Streptococcus mutans is the main pathogenic microorganism, and its incidence is closely related to sucrose. Rubusoside is a natural nonnutritive sweetener isolated from Rubus suavissimus S. Lee. This study was designed to determine the effect of this sucrose substitute on the cariogenic properties and virulence gene expression of S. mutans biofilms. S. mutans was exposed to brain heart infusion (BHI) medium (as a control), 1% sucrose-supplemented medium, 1% rubusoside-supplemented medium, and 1% xylitol-supplemented medium. The growth curve of the biofilm was monitored by crystal violet staining, and the pH was measured every 24 h. After 5 days, the biofilms formed on the glass coverslips were recovered to determine the biomass (dry weight and total amount of soluble proteins), numbers of CFU, and amounts of intra- and extracellular polysaccharides. Biofilm structural imaging was performed using a scanning electron microscope (SEM). Virulence gene expression (gtfB, gtfC, gtfD, ftf, spaP, gbpB, ldh, atpF, vicR, and comD) was determined by reverse transcription-quantitative PCR. Growth in rubusoside resulted in lower levels of acid production than observed during growth in sucrose, xylitol, and the control, while it also reduced the level of biofilm accumulation and bacterial viability and even reduced the level of production of extracellular polysaccharides. By SEM, the levels of biofilm formation and extracellular matrix during growth in rubusoside were lower than these levels during growth in sucrose and xylitol. From the perspective of virulence genes, growth in rubusoside and xylitol significantly inhibited the expression of virulence genes compared with their levels of expression after growth in sucrose. Among these genes, gtfB, gtfC, gbpB, ldh, and comD downregulation was found with growth in rubusoside compared with their expression with growth in xylitol. Therefore, rubusoside appears to be less potentially cariogenic than sucrose and xylitol and may become an effective sucrose substitute for caries prevention. Further studies are needed to deepen these findings. IMPORTANCE Dental caries is a major public health challenge and places heavy biological, social, and financial burdens on individuals and health care systems. To palliate the deleterious effect of sucrose on the virulence factors of S. mutans, massive commercial efforts have been oriented toward developing products that may act as sucrose substitutes. Rubusoside, a natural sucrose substitute, is a plant extract with a high level of sweetness. Although some studies have shown that rubusoside does not produce acids or inhibit the growth of S. mutans, little attention has been paid to its effect on dental biofilm and the underlying mechanisms. Our study focuses on the effect of rubusoside on the formation and structure of biofilms and the expression of virulence genes. The results confirm that rubusoside can inhibit accumulation, bacterial viability, polysaccharide production by the biofilm, and related gene expression. These results provide further insight into the cariogenicity of S. mutans biofilms and demonstrate a new perspective for studying the impact of sucrose substitutes on caries.
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Chen, J., A. Zhang, Z. Xiang, M. Lu, P. Huang, T. Gong, Y. Pan, Y. Lin, X. Zhou, and Y. Li. "EpsR Negatively Regulates Streptococcus mutans Exopolysaccharide Synthesis." Journal of Dental Research, March 20, 2021, 002203452110006. http://dx.doi.org/10.1177/00220345211000668.
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Streptococcus mutans is considered the primary etiological agent of human dental caries. Glucosyltransferases (Gtfs) from S. mutans play important roles in the formation of biofilm matrix and the development of cariogenic oral biofilm. Therefore, Gtfs are considered an important target to prevent the development of dental caries. However, the role of transcription factors in regulating gtf expression is not yet clear. Here, we identify a MarR (multiple antibiotic resistance regulator) family transcription factor named EpsR (exopolysaccharide synthesis regulator), which negatively regulates gtfB expression and exopolysaccharide (EPS) production in S. mutans. The epsR in-frame deletion strain grew slowly, aggregated more easily in the presence of dextran, and displayed different colony morphology and biofilm structure. Notably, epsR deletion resulted in altered 3-dimensional biofilm architecture, increased water-insoluble EPS production, and upregulated GtfB protein content and activity. In addition, global gene expression profiling revealed differences in the expression levels of 69 genes in which gtfB was markedly upregulated. The conserved DNA motif for EpsR binding was determined by electrophoretic mobility shift assay and DNase I footprinting assays. Moreover, analysis of β-galactosidase activity suggested that EpsR acted as a repressor and inhibited gtfB expression. Taken together, our findings indicate that EpsR is an important transcription factor that regulates gtfB expression and EPS production in S. mutans. These results add new aspects to the complexity of regulating the expression of genes involved in the cariogenicity of S. mutans, which might lead to novel strategies to prevent the formation of cariogenic biofilm that may favor diseases.
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Priya, Arumugam, Chandra Bose Manish Kumar, Alaguvel Valliammai, Anthonymuthu Selvaraj, and Shunmugiah Karutha Pandian. "Usnic acid deteriorates acidogenicity, acidurance and glucose metabolism of Streptococcus mutans through downregulation of two-component signal transduction systems." Scientific Reports 11, no. 1 (January 14, 2021). http://dx.doi.org/10.1038/s41598-020-80338-6.
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AbstractThe principal etiological agent of human dental caries, Streptococcus mutans is a multi-virulent pathogen that can transform commensal oral microbial community to plaque biofilms. Major virulence factors that are associated with the cariogenicity of S. mutans include adhesion, acidogenicity and acidurity. All these pathogenic traits coordinate and alter the dental plaque ecology which provide room for interaction with other similar acidogenic and aciduric bacteria. This cariogenic flora increases the possibility of enamel demineralization which headway to caries development. The present study was aimed at evaluating the antimicrobial and antiinfective potential of a lichen secondary metabolite usnic acid (UA) against S. mutans. Minimum inhibitory concentration (MIC), Minimum bactericidal concentration (MBC) and growth kinetics were evaluated to determine the antimicrobial potential of UA against S. mutans. UA at 5 µg mL−1 and 10 µg mL−1 concentration were considered as MIC and MBC respectively. Effect on biofilm formation was microscopically assessed and found to be reduced in a concentration dependent manner. Gene expression of gtfB, gtfC, gtfD, vicR, ComDE and smu0630 was found to be downregulated upon treatment with sub-MIC of UA. Acidogenicity, acidurity, eDNA synthesis and response to oxidative stress were found to be attenuated by the influence of UA. It was also demonstrated to act on preformed mature biofilm of S. mutans. Moreover, UA was shown to possess very low frequency to acquire spontaneous resistance development in S. mutans. Besides, no morphological aberrations or toxic effect was instigated by UA in the human buccal epithelial cells as well as to the oral commensals. Altogether, these results demonstrate the therapeutic potential of usnic acid in the treatment of S. mutans infection.
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Chen, Lulu, Brinta Chakraborty, Jing Zou, Robert A. Burne, and Lin Zeng. "Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci andStreptococcus mutans." Applied and Environmental Microbiology 85, no. 10 (March 15, 2019). http://dx.doi.org/10.1128/aem.00370-19.
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ABSTRACTN-Acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 ofStreptococcus gordoniiagainst the caries pathogenStreptococcus mutans. Here, we examine how amino sugars affect the interaction of five low-passage-number clinical isolates of abundant commensal streptococci withS. mutansby utilizing a dual-species biofilm model. Compared to that for glucose, growth on GlcN or GlcNAc significantly reduced the viability ofS. mutansin cocultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2was increased in most commensals when growing on amino sugars, and inhibition ofS. mutansbyStreptococcus cristatus,Streptococcus oralis, orS. gordoniiwas enhanced by amino sugars on agar plates. All commensals exceptS. oralishad higher arginine deiminase activities when grown on GlcN and, in some cases, GlcNAc. Inex vivobiofilms formed using pooled cell-containing saliva (CCS), the proportions ofS. mutanswere drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production byS. mutanson agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism byS. mutansin cocultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage-number commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.IMPORTANCEDental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars GlcNAc and GlcN in promoting the antagonistic properties of a health-associated oral bacterium,Streptococcus gordonii, in competition with the major caries pathogenStreptococcus mutans. Here, we investigated 5 low-passage-number clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiplein vitromodels, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.
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Rainey, Katherine, Landon Wilson, Stephen Barnes, and Hui Wu. "Quantitative Proteomics Uncovers the Interaction between a Virulence Factor and Mutanobactin Synthetases in Streptococcus mutans." mSphere 4, no. 5 (September 25, 2019). http://dx.doi.org/10.1128/msphere.00429-19.
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ABSTRACT Streptococcus mutans, the primary etiological agent of tooth decay, has developed multiple adhesion and virulence factors which enable it to colonize and compete with other bacteria. The putative glycosyltransferase SMU_833 is important for the virulence of S. mutans by altering the biofilm matrix composition and cariogenicity. In this study, we further characterized the smu_833 mutant by evaluating its effects on bacterial fitness. Loss of SMU_833 led to extracellular DNA-dependent bacterial aggregation. In addition, the mutant was more susceptible to oxidative stress and less competitive against H2O2 producing oral streptococci. Quantitative proteomics analysis revealed that SMU_833 deficiency resulted in the significant downregulation of 10 proteins encoded by a biosynthetic gene cluster responsible for the production of mutanobactin, a compound produced by S. mutans which helps it survive oxidative stress. Tandem affinity purification demonstrated that SMU_833 interacts with the synthetic enzymes responsible for the production of mutanobactin. Similar to the smu_833 mutant, the deletion of the mutanobactin gene cluster rendered the mutant less competitive against H2O2-producing streptococci. Our studies revealed a new link between SMU_833 virulence and mutanobactin, suggesting that SMU_833 represents a new virulent target that can be used to develop potential anticaries therapeutics. IMPORTANCE Streptococcus mutans is the major bacterium associated with dental caries. In order to thrive on the highly populated tooth surface and cause disease, S. mutans must be able to protect itself from hydrogen peroxide-producing commensal bacteria and compete effectively against the neighboring microbes. S. mutans produces mutacins, small antimicrobial peptides which help control the population of competing bacterial species. In addition, S. mutans produces a peptide called mutanobactin, which offers S. mutans protection against oxidative stress. Here, we uncover a new link between the putative glycosyltransferase SMU_833 and the mutanobactin-synthesizing protein complex through quantitative proteomic analysis and a tandem-affinity protein purification scheme. Furthermore, we show that SMU_833 mediates bacterial sensitivity to oxidative stress and bacterial ability to compete with commensal streptococci. This study has revealed a previously unknown association between SMU_833 and mutanobactin and demonstrated the importance of SMU_833 in the fitness of S. mutans.
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SIGNORI, Cácia, Andréia Drawanz HARTWIG, Ivam Freire da SILVA-JÚNIOR, Marcos Britto CORREA, Marina Sousa AZEVEDO, and Maximiliano Sérgio CENCI. "The role of human milk and sucrose on cariogenicity of microcosm biofilms." Brazilian Oral Research 32 (October 11, 2018). http://dx.doi.org/10.1590/1807-3107bor-2018.vol32.0109.
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