Exploring the efficacy of in-vitro low-temperature plasma treatment on single and multispecies dental cariogenic biofilms.

The primary aim of this study was to investigate the impact of treatment with low-temperature plasma (LTP) for varying exposure durations on a multispecies cariogenic biofilm comprising C. albicans, L. casei, and S. mutans, as well as on single-species biofilms of L. casei and C. albicans, cultured on hydroxyapatite discs. Biofilms were treated with LTP-argon at a 10 mm distance for 30 s, 60 s, and 120 s. Chlorhexidine solution (0.12%) and NaCl (0.89%) were used as positive (PC) and negative controls (NC), respectively. Argon flow only was also used as gas flow control (F). Colony-forming units (CFU) recovery and confocal laser scanning microscopy (CLSM) were used to analyze biofilm viability. LTP starting at 30 s of application significantly reduced the viability of multispecies biofilms by more than 2 log10 in all treated samples (p < 0.0001). For single-species biofilms, L. casei showed a significant reduction compared to PC and NC of over 1 log10 at all exposure times (p < 0.0001). In the case of C. albicans biofilms, LTP treatment compared to PC and NC resulted in a significant decrease in bacterial counts when applied for 60 and 120 s (1.55 and 1.90 log10 CFU/mL, respectively) (p < 0.0001). A significant effect (p ≤ 0.05) of LTP in single-species biofilms was observed to start at 60 s of LTP application compared to F, suggesting a time-dependent effect of LTP for the single-species biofilms of C. albicans and L. casei. LTP is a potential mechanism in treating dental caries by being an effective anti-biofilm therapy of both single and multispecies cariogenic biofilms.

Riesgo microbiológico de caries y factores predisponentes en mujeres gestantes / Microbiological caries risk and predisposing factors in pregnant women

La caries es una enfermedad multifactorial que afecta los tejidos duros del diente, con la evaluación del riesgo de caries es posible determinar la probabilidad de su incidencia durante un determinado periodo de tiempo. El objetivo consistió en determinar el estado de salud bucal en mujeres gestantes y su relación con factores de riesgo de caries. Se realizó un estudio descriptivo observacional clínico. La población estuvo constituida por mujeres gestantes que concurrieron al Hospital Ángela Llano de la cuidad de Corrientes para su atención. Se confeccionó una historia clínica y se determinó el estado de salud bucal y de riesgo de caries según protocolo CAMBRA. Para la carac- terización y descripción de los datos se utilizaron estadísticas univariadas. Los resultados alcanzados fueron: Índice de Caries de 4.5 a 6.5, en el índice de Placa la media ± DS 56,8% ± 39,9% Min: 28,5%/ Max: 90,9%. Respecto al valor de pH de 7,11 ±0,43, Min.: 7,00, Max.: 8,67. Flujo salival: media ± DS: 0,58ml/m ± 0,28 ml/m Min.:0,2 ml/m Max.: 0,62 ml/m; Edad promedio: 22años. Min.:18 Max.: 33 años. Recuento de Strepto- coccus mutans (SM) con periodo gestacional 1° y 2° trimestre, el conteo de bacterias: 2.3x105 +/- 3.2x 105 UFC/ml de saliva y 3° Trimestre el conteo de 1.2x 106 +/-2.02 x 106 UFC/ml de saliva. La presencia de SM puede aumentar considerable- mente el riesgo de caries, si los mecanismos de defen- sa del huésped no anulan la bacteria (AU)

Caries is a multifactorial disease that affects the hard tissues of the tooth. By evaluating the risk of caries, it is possible to determine the probability of its incidence during a certain period of time. The objective was to determine the oral health status of pregnant women and its relationship with caries risk factors. A clinical observational descriptive study was carried out. The population was made up of pregnant women who attended the Ángela Llano Hospital in the city of Corrientes for care. A clinical history was prepared, and the oral health status and caries risk were determined according to the CAMBRA protocol. Univariate statistics were used to characterize and describe the data. The results achieved were Caries Index from 4.5 to 6.5, in the Plaque index the mean ± SD 56.8% ± 39.9% Min: 28.5%/ Max: 90.9%. Regarding the pH value of 7.11 ±0.43, Min.: 7.00, Max.: 8.67. Salivary flow: mean ± SD: 0.58 ml/m ± 0.28 ml/m Min.: 0.2 ml/m Max.: 0.62 ml/m; Average age: 22 years. Min.:18 Max.: 33 years. Streptococcus mutans (SM) count with gestational period 1st and 2nd trimester, bacteria count: 2.3x105 +/- 3.2x 105 CFU/ml of saliva and 3rd Trimester count 1.2x 106 +/- 2.02 x 106 CFU/ml of saliva. The presence of SM can considerably increase the risk of caries, if the host defense mechanisms do not override the bacteria (AU)

Dual-species biofilm of Streptococcus mutans and Candida albicans produces subsurface caries lesions on bovine enamel.

OBJECTIVES: To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro. DESIGN: Biofilms were grown on bovine enamel specimens in artificial saliva (AS) for seven days. After 24 h of formation, the AS was supplemented or not with fluoride (F) using sodium fluoride (0.005 or 0.008 ppm F), and the biofilms were exposed or not to a 20 % sucrose solution (reproducing a cariogenic challenge) once/day. On the seventh day, the biofilms were harvested and had their extracellular polysaccharides (EPS) and inorganic components analyzed. The specimens were subjected to computed X-ray microtomography analysis to determine their mineral concentration. Data were compared using two-way analyses of variance, followed by Fisher's LSD or Student-Newman-Keuls tests (p < 0.05). RESULTS: Biofilms exposed to the cariogenic challenge had significantly higher EPS concentrations than those not exposed, regardless of the presence of F. For biofilms grown with 0.008 ppm F, those exposed to the cariogenic challenge had lower F levels than those not exposed. For biofilms exposed to the cariogenic challenge, those grown with 0.008 ppm F had lower lesion depths and integrated mineral loss, and higher outer layers than those grown without F. CONCLUSIONS: The dual biofilm model assessed was able to create subsurface caries lesions in bovine enamel in vitro, which was influenced by the presence of F in the culture medium and exposure to sucrose.

Comparison of biofilm models for producing artificial active white spot lesions.

OBJECTIVE: This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models. METHODOLOGY: In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey's post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001). RESULTS: In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols. CONCLUSION: The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth.

In vitro effect of TiF4/NaF solution on the development of radiation-induced dentin caries.

OBJECTIVE: To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. METHODOLOGY: bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. RESULTS: All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Reduction of microorganisms in carious dentin by photodynamic therapy mediated by potassium iodide added to methylene blue and red laser.

OBJECTIVE: To evaluate the effect of the association of potassium iodide to antimicrobial photodynamic therapy on human carious dentin produced with a microcosm biofilm model. METHODS: A microcosm biofilm model was used to generate a caries lesion on human dentin. Pooled human saliva diluted with glycerol was used as an inoculum on specimens immersed on McBain artificial saliva enriched with 1 % sucrose (24 h at 37 °C in 5 % CO2). After refreshing culture media for 7 days, the dentin specimens were divided in 5 groups (3 specimens per group, in triplicate; n = 9): C (NaCl 0.9 %), CX (2 % chlorhexidine), PKI (0.01 % methylene blue photosensitizer+50 mM KI), L (laser at 15 J, 180 s, 22.7 J/cm2), and PKIL (methylene blue + KI + Laser). After the treatments, dentin was collected, and a 10-fold serial dilution was performed. The number of total microorganisms, total lactobacilli, total streptococci, and Streptococcus mutans was analyzed by microbial counts (CFU/mL). After normality and homoscedasticity analysis, the Welch's ANOVA and Dunnett's tests were used for CFU. All tests used a 5 % significance level. RESULTS: CX and PKIL groups showed significant bacterial decontamination of dentin, compared to group C (p < 0.05) reaching reductions up to 3.8 log10 for CX for all microorganisms' groups and PKIL showed 0.93, 1.30, 1.45, and 1.22 log10 for total microorganisms, total lactobacilli, total streptococci, and S. mutans, respectively. CONCLUSION: aPDT mediated by the association of KI and methylene blue with red laser reduced the viability of microorganisms from carious dentin and could be a promising option for cavity decontamination.

Enhanced anti-biofilm and anti-caries potential of arginine combined with calcium glycerophosphate and fluoride.

OBJECTIVE: The aim of this work was to evaluate the antibiofilm and anticaries properties of the association of arginine (Arg) with calcium glycerophosphate (CaGP) and fluoride (F). METHODS: An active attachment, polymicrobial biofilm model obtained from saliva and bovine teeth discs were used. After the initial biofilm growth period, the enamel discs were transferred to culture medium. The treatment solutions were added to the culture media to achieve the desired final concentration. The following groups were used: negative control (Control); F (110 ppm F); CaGP (0.05 %); Arg (0.8 %) and their associations (F + CaGP; Arg + F; Arg + CaGP; Arg +F + CaGP). The following analyses were carried out: bacterial viability (total bacteria, aciduric bacteria and mutans streptococci), pH assessment of the spent culture medium, dry weight quantification, evaluation of surface hardness loss (%SH) and subsurface mineral content. Normality and homoscedasticity were tested (Shapiro-Wilk and Levene's test) and the following tests were applied: two-way ANOVA (acidogenicity), Kruskall-Wallis (microbial viability) and one way ANOVA (dry weight, %SH, mineral content). RESULTS: The association Arg + F + CaGP resulted in the lowest surface hardness loss in tooth enamel (-10.9 ± 2.3 %; p < 0.05). Arg +F + CaGP exhibited highest values of subsurface mineral content (10.1 ± 2.9 gHAP/cm3) in comparison to Control and F (p < 0.05). In comparison to Control and F, Arg +F + CaGP promoted the highest reduction in aciduric bacteria and mutans streptococci (5.7 ± 0.4; 4.4 ± 0.5 logCFU/mL, p < 0.05). CONCLUSIONS: The Arg-F-Ca association demonstrated to be the most effective combination in protecting the loss of surface hardness and subsurface mineral content, in addition to controlling important virulence factors of the cariogenic biofilm. CLINICAL SIGNIFICANCE: Our findings provide evidence that the Arg-F-Ca association showed an additive effect, particularly concerning protection against enamel demineralization. The combination of these compounds may be a strategy for patients at high risk of caries.

Assessment of dental and periodontal indices and Streptococcus mutans virulence in fragile X syndrome patients.

INTRODUCTION: Fragile X syndrome (FXS) is the most common cause of hereditary genetic disorder in a single gene characterised by intellectual disability. Behavioural features such as autism, hyperactivity and anxiety disorder may be present. Biofilm development and pathogenicity of Streptococcus mutans may be altered because FXS renders the dental approach and oral hygiene more complex. OBJECTIVES: The purpose of this study was to compare the levels of transcripts for VicRK and CovR of S. mutans isolated from FXS patients with the levels of transcripts for VicRK and CovR of standard strain ATCC, using a quantitative polymerase chain reaction (qPCR). METHODS: The caries experience index was assessed by the International Caries Detection and Assessment System (ICDAS), Periodontal Condition Index (PCI) and Invasive Dental Treatment Need Index (INI). RESULTS: The clinical index findings revealed a high rate of caries cavities and bleeding on probing of FXS patients. When VicRK and CovR transcript levels were compared with the reference strain, Fragile X patients were found to have significantly higher values. CONCLUSION: The present study demonstrated that FXS patients have more adverse clinical conditions, with increased biofilm accumulation and virulence. When combined with behavioural abnormalities, these patients become even more vulnerable to dental caries.

Association of Root Biofilm Bacteriome with Root Caries Lesion Severity and Activity.

INTRODUCTION: This research aimed to assess the association of root biofilm bacteriome with root caries lesion severity and activity in institutionalised Colombian elderlies and was conducted to gather data on the root caries bacteriome in this population. METHODS: A bacteriome evaluation of biofilm samples from sound and carious root surfaces was performed. Root caries was categorised (ICDAS Root criteria) based on severity (sound surfaces, initial: non-cavitated, moderate/extensive combined: cavitated) and activity status (active and inactive). DNA was extracted and the V4 region of the 16S rRNA gene was sequenced; afterwards the classification of features was conducted employing amplicon sequence variants and taxonomic assignment via the Human Oral Microbiome Database (HOMD). Bacterial richness, diversity (Simpson's and Shannon's indices), and relative abundance estimation were assessed and compared based on root caries severity and activity status (including Sound surfaces). RESULTS: A total of 130 biofilm samples were examined: sound (n = 45) and with root caries lesions (n = 85; by severity: initial: n = 41; moderate/extensive: n = 44; by activity: active: n = 60; inactive: n = 25). Species richness was significantly lower in biofilms from moderate/extensive and active groups compared to sound sites. There was a higher relative abundance of species like Lechtotricia wadei, Capnocytophaga granulosa, Cardiobacterium valvarum, Porphyromonas pasteri - in sound sites; Dialister invisus, Streptococcus mutans, Pseudoramibacter alactolyticus and Bacteroidetes (G-5) bacterium 511 - in moderate/extensive lesions, and Fusobacterium nucleatum subsp. animalis, Prevotella denticola, Lactobacillus fermentum, Saccharibacteria (TM7) (G-5)bacterium HMT 356 - in active lesions. CONCLUSION: Root caries bacteriome exhibited differences in species proportions between the compared groups. Specifically, cavitated caries lesions and active caries lesions showed higher relative abundance of acidogenic bacteria.

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.

Serotipos prevalentes de Streptococcus mutans en América Latina / Prevalent serotypes of Streptococcus mutans in Latin America

La microbiota oral está conformada por diversas especies bacterianas que en condiciones normales desempeñan una función protectora del huésped; sin embargo, cuando existe un desequilibrio en el ecosistema, estos microorganismos son capaces de producir diversas manifestaciones como lo es el caso de la caries dental, enfermedad infecciosa producida principalmente por Streptococcus mutans, patógeno capaz de desmineralizar los tejidos duros del diente mediante la fermentación de hidratos de carbono obtenidos de la dieta. Se ha identificado en la pared celular de este microorganismo ocho serotipos que intervienen en la adhesión, agregación y coagregación bacteriana. En los seres humanos S. mutans presenta los serotipos c, e y f, siendo el serotipo c el más prevalente a nivel mundial, el cual se conoce que está asociado a pacientes sanos, a diferencia del e y f que son capaces de invadir las células endoteliales de las arterias coronarias. No obstante, en los últimos años se ha logrado identificar el serotipo k que de igual manera presenta alta capacidad de invadir el endotelio humano, actuando en la patogénesis de las enfermedades cardiovasculares. El objetivo de la presente revisión bibliográfica es lograr cuantificar los serotipos prevalentes de S. mutans en América Latina (AU)

The oral microbiota is made up of various bacterial species that under normal conditions perform a protective function of the host, however, when there is an imbalance in the ecosystem, these microorganisms are capable of producing various manifestations such as caries, an infectious disease. produced mainly by Streptococcus mutans, a pathogen capable of demineralizing the hard tissues of the tooth through the fermentation of carbohydrates obtained from the diet. Eight serotypes involved in bacterial adhesion, aggregation and coaggregation have been identified in the cell wall of this microorganism. In humans, S. mutans presents serotypes c, e, and f, serotype c being the most prevalent worldwide, which is known to be associated with healthy patients, unlike e and f, which are capable of invading the endothelial cells of the coronary arteries. However, in recent years it has been possible to identify serotype k, which also has a high capacity to invade the human endothelium, acting in the pathogenesis of cardiovascular diseases. The objective of this literature review is to quantify the prevalent serotypes of S. mutans in Latin America (AU)

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.

Chapter 9.3: Current Concepts of Caries Removal in Daily Practice.

Although the discussion about the amount of carious dentin to be removed during cavity preparation is quite old, concepts for caries removal have evolved and changed considerably over the last decades. The antiquate understanding that it was necessary to eliminate the microbial contamination of a cavity before placing the restoration was replaced by the current knowledge that maintaining contaminated dentin beneath restorations is inevitable and is not associated with treatment failure. This chapter brings together the body of evidence behind carious dentin removal to indicate a conservative treatment, aiming to preserve both tooth vitality and structure. Studies that evaluated the effects of sealing contaminated dentin are described, which are focused on different outcomes, such as microbiological counts, clinical characteristics, laboratory analysis, and radiographic findings. Long-term studies and randomized clinical trials also support the current recommendations. After addressing the available literature on this topic, this chapter concludes that (1) the amount of carious dentin to be removed should be defined by lesion depth; (2) sealing and/or selective caries removal to firm dentin is recommended for the management of shallow and moderate lesions; (3) the selective caries removal to soft dentin in a single session is indicated for deep caries lesions aiming to preserve tooth vitality; and (4) the use of a cavity liner after selective caries removal seems to be an unnecessary clinical step.

Bypass gastroplasty impacts oral health, salivary inflammatory biomarkers, and microbiota: a controlled study.

OBJECTIVES: Knowledge about the impact of gastroplasty on oral health and salivary biomarkers is limited. The aim was to prospectively evaluate oral health status, salivary inflammatory markers, and microbiota in patients undergoing gastroplasty compared with a control group undergoing a dietary program. MATERIALS AND METHODS: Forty participants with obesity class II/III were included (20 individuals in each sex-matched group; 23-44 years). Dental status, salivary flow, buffering capacity, inflammatory cytokines, and uric acid were assessed. Salivary microbiological analysis (16S-rRNA sequencing) assessed the abundance of genus, species, and alpha diversity. Cluster analysis and mixed-model ANOVA were applied. RESULTS: Oral health status, waist-to-hip ratio, and salivary alpha diversity were associated at baseline. A subtle improvement in food consumption markers was observed, although caries activity increased in both groups, and the gastroplasty group showed worse periodontal status after three months. IFNγ and IL10 levels decreased in the gastroplasty group at 3 months, while a decrease was observed in the control group at 6 months; IL6 decreased in both groups (p < 0.001). Salivary flow and buffering capacity did not change. Significant changes in Prevotella nigrescens and Porphyromonas endodontalis abundance were observed in both groups, while alpha diversity (Sobs, Chao1, Ace, Shannon, and Simpson) increased in the gastroplasty group. CONCLUSIONS: Both interventions changed in different degrees the salivary inflammatory biomarkers and microbiota, but did not improve the periodontal status after 6 months. CLINICAL RELEVANCE: Although the observed discrete improvement in dietary habits, caries activity increased with no clinical improvement in the periodontal status, emphasizing the need of oral health monitoring during obesity treatment.

Nicotine is a potent extracellular polysaccharide inducer in Fusobacterium nucleatum biofilms

The purpose of this in vitro study was to analyze the influence of nicotine on the extracellular polysaccharides in Fusobacterium nucleatum biofilm. Methods: F. nucleatum (ATCC 10953) biofilms supplemented with different concentrations of nicotine (0, 0.5, 1, 2, 4, and 8 mg/mL) were grown in two different BHI broth conditions [no sucrose and 1% sucrose]. Extracellular polysaccharides assay, pH measurements, and a spectrophotometric assay were performed. Data were submitted for ANOVA and Tukey honestly significant difference analyses (HSD) tests (α =.05). Results: Extracellular polysaccharides synthesis was influenced by an interaction between nicotine concentrations and growth medium solution containing sucrose (P<.05). The pH values declined in the sucrose-exposed biofilm were greater than in the group exposed only to nicotine (P<.05). The biofilm exposed to sucrose and nicotine had a higher total biofilm growth (P<.05) than the nicotine-treated biofilm without sucrose. Conclusions: Regardless of sucrose exposure, biofilms exposed to different nicotine concentrations influenced the amount of extracellular polysaccharides

Overexpression of proinflammatory cytokines in dental pulp tissue and distinct bacterial microbiota in carious teeth of Mexican Individuals.

The prevalence of dental caries in the Mexican adult population aged 20 to 85 years is around 93.3%, and 50% in Mexican children and adolescents. Worldwide, it is the most common non-communicable disease. One of the main etiological factors for dental caries is the oral microbiome and changes in its structure and function, with an expansion of pathogenic bacteria like Streptococcus mutans. The exposed dental pulp tissue triggers an innate immune response to counteract this bacterial invasion. The relation between oral dysbiosis and innate immune responses remains unclear. We aimed to understand the relationship between innate immune response and the oral microbiota by quantifying the expression of Toll-like receptors (TLRs) and proinflammatory markers (cytokines and a chemokine) in dental pulp tissue, either exposed or not to carious dentin, and to correlate this information with the oral microbiome found in healthy teeth and those with moderate caries. RNA was purified from pulp tissue, subjected to RT-qPCR and analysed with the ΔΔCt method. Supragingival dental plaque of non-carious teeth and dentin of carious teeth were subjected to 16S targeted sequencing. Principal coordinate analysis, permutational multivariate ANOVA, and linear discriminant analysis were used to assess differences between non-carious and carious teeth. Correlations were assessed with Spearman´s test and corrected for multiple comparisons using the FDR method. The relative abundance (RA) of Lactobacillus, Actinomyces, Prevotella, and Mitsuokella was increased in carious teeth; while the RA of Haemophilus and Porphyromonas decreased. Olsenella and Parascardovia were only detected in carious teeth. Significant overexpression of interleukin 1 beta (IL1 ß), IL6, and CXCL8 was detected in pulp tissue exposed to carious dentin. IL1ß correlated positively with TLR2 and Actinomyces; yet negatively with Porphyromonas. These findings suggest that immune response of pulp tissue chronically exposed to cariogenic microbiome is triggered by proinflammatory cytokines IL1ß and IL6 and the chemokine CXCL8.

Does high sugar intake really alter the oral microbiota?: A systematic review.

OBJECTIVES: Diet is one of the main factors influencing the diversity and interactions of the oral microbiota. The purpose of this study is to determine the impact of sugar intake on the microbial diversity and bacteria that predominate under these conditions. MATERIAL AND METHODS: A systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guide, using the PubMed, Scopus, and Science Direct databases and combinations of the words "microbiota," "microbiology," "bacteria," "sugars," "dysbiosis," "caries," "microbiome," "oral microbial," and "oral microbiota profile pattern." The selection criteria included year, language, type of publication, comparison of microbiota during low and high sugar intake, and bacterial identification by molecular sequencing of the 16S subunit of ribosomal RNA. RESULTS: Out of a total of 374 papers that came up after the initial search, 8 met the criteria for this review. The papers included research on populations comprising children, young adults, and adults, with most of the studies reporting selection criteria for the participants and using validated instruments to determine sugar intake. Apart from one study, all others reported for high sugar intake conditions a significant decrease in microbial diversity of the oral microbiome and the predominance of several bacterial genera or species, including Streptococcus, Scardovia, Veillonella, Rothia, Actinomyces, and Lactobacillus. CONCLUSIONS: Sugar-rich diets have a significantly unfavorable effect on the diversity and balance of oral microbiota; however, further studies are required to determine the exact role of sugar in microbial interactions.

Ultrasound device as a minimally invasive approach for caries dentin removal.

The aim of this study was to evaluate the efficacy of an ultrasound device and the dentin surface morphology after removal of the caries dentin lesions by removal rate and scanning electron microscopy (SEM). The Knoop hardness test on the bovine dentin blocks (n = 20, 4x4x2mm) was performed to standardize the samples and only those with 38 ± 2 KHN were included. The dentin blocks were submitted to induction of artificial caries lesions, using the bacterial model. Strains of Streptococcus mutans and Lactobacillus acidophilus were used for 7 days. The caries dentin lesion was removed for 1 min, according to two methods: G1 - carbide bur under low-speed rotation (control group) and G2 - ultrasound device under refrigeration. For the removal rate, the samples were weighed 3 times: T0 (before induction), T1 (after induction) and T2 (after removal). Morphology evaluation of the residual dentin surface was performed by SEM. Data normality was verified by Shapiro-Wilk test (p ≥ 0.240). T-test for independent samples was applied to evaluate the removal rate. A significance level of 5% was adopted. G2 provided lower removal rate than G1 (G1: 3.68 mg and G2 = 2.26 mg). SEM images showed different morphological characteristics between the groups. G2 showed absent of smear layer, while G1 showed a visible smear layer over the surface. We concluded that ultrasound device provides minimally invasive removal with residual dentin exhibiting open dentin tubules and no smear layer formation and no bacteria, which infer the removal of the infected tissue.

Antimicrobial potential of plant extracts from the Brazilian Cerrado.

Bacteria are related do different oral diseases, such as dental caries and periodontal disease. Therefore, the control or/and eradication of microorganisms and their by-products is primordial for the success of their treatment. An alternative for decrease bacterial load is the use of plant extracts used in popular medicine. The cytotoxicity and antimicrobial action of extracts of Cariniana rubra Gardiner ex Miers, Senna martiniana, Anadenanthera colubrina (Vell.) Brenan and Spiranthera odoratissima St. Hil. against strains of Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Aggregatibacter actinomyces- tencomitans and Candida albicans were investigated. Cytotoxicity was assessed at concentrations of 1, 10, 40, 80, 100 and 1000 µg/mL by means of the MTT test and compared to a control group with untreated cells. Those with acceptable cytotoxicity had the antimicrobial action measured by the XTT test. As a positive control, sodium hypochlorite was used. Cariniana rubra Gardiner ex Miers had the highest citototoxicity results while Spiranthera odoratissima St. Hil. had the best results, but all extracts showed acceptable cytotoxicity at different concentrations. The plant extracts showed higher activity against A. actinomycetencomitans: Anadenanthera columbrina (Vell.) Brenan (80.52%) at 40 µg/mL, Spiranthera odoratissima St. Hil (78.48%) in 1 µg/mL, Senna martiniana (73.28%) in the concentration of 40 µg/mL and Cariniana rubra Gardiner ex Miers (70.50%) in 10 µg/mL. All extracts analyzed showed acceptable cytotoxicity at different concentrations and were promising for inhibition of the pathogenic microorganisms studied.