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.

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

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

Evaluation of Dental Enamel Mass in Prolonged Supervised Dental Bleaching Protocol with Calcium-Containing Gel

ABSTRACT: The objective of this study was to determine the enamel mass variation after a prolonged bleaching treatment using a calcium-containing 4% hydrogen peroxide gel Twenty sound bovine incisors were randomly assigned to two groups (n=10) stored in G1) distilled water and G2) artificial saliva. An electronic analytic scale (.000 grams measurements) was used to determine the enamel mass variation before and after the bleaching procedures at the following evaluation times: T0) before the bleaching procedures; T1) 14 days of treatment, as instructed by the manufacturer; T2) 21 days of treatment, 50 % beyond what is instructed by the manufacturer; and T3) 28 days of treatment, 100 % beyond what is instructed by the manufacturer. The highest mean was observed at T2/G2 (0.3259 g) and the lowest at T2/G1 (0.3265 g). The specimens stored in distilled water (G1) showed 6 % mass reduction when T0 (0.3277 g) was compared to T3 (0.3277 g). On the other hand, the specimens stored in artificial saliva exhibited a significant mass increase of 19 % when T0 (0.3521 g) was compared to T3 (0.3528 g). Prolonged bleaching therapy using 4 % hydrogen peroxide with calcium resulted in a massive reduction when water was used as a storage medium. When the specimens were stored in artificial saliva, an increase in mass was observed, probably due to the mineralizing properties of the artificial saliva.

RESUMEN: El objetivo de este estudio fue determinar la variación de la masa del esmalte después de un tratamiento de blanqueamiento prolongado utilizando calcio conteniendo un gel de peróxido de hidrógeno al 4 %. Veinte incisivos bovinos intactos fueron asignados aleatoriamente a dos grupos (n-10) almacenados en G1 - agua destilada y G2- saliva artificial. Se utilizó una escala analítica electrónica (.000 gramos) para determinar la variación de la masa de esmalte antes y después de los procedimientos de blanqueamiento en los siguientes tiempos de evaluación: T0) antes de los procedimientos blanqueadores; T1) 14 días de tratamiento, según las instrucciones del fabricante; T2) 21 días de tratamiento, 50 % más allá del tiempo indicado por el fabricante; y T3) 28 días de tratamiento, 100 % más allá del tiempo indicado por el fabricante. La media más alta se observó en T2/G2 (0,3259 g) y la más baja en T2/G1 (0,3265 g). Los especímenes almacenados en agua destilada (G1) mostraron una reducción de masa del 6 % cuando se comparó T0 (0,3277 g) con T3 (0,3277 g). Por otro lado, los dientes almacenados en saliva artificial mostraron un aumento significativo de masa del 19 % cuando se comparó T0 (0,3521 g) con T3 (0,3528 g). La terapia de blanqueamiemto prolongado con calcio conteniendo un gel de peróxido de hidrógeno al 4 % condujo a una reducción masiva cuando se utilizó agua como medio de almacenamiento, mientras que los dientes almacenados en saliva artificial mostraron un aumento en la masa, probablemente debido a las propiedades remineralizadoras de la saliva artificial.

Influence of Prolonged Tooth Bleaching on Enamel Mass Variation / Influencia del Blanqueo Dental Prolongado en la Variación de Masa del Esmalte Dental

ABSTRACT: The objective evaluate was the influence of prolonged tooth bleaching with 10 % carbamide peroxide (10CP) on tooth enamel mass variation. Ten healthy bovine incisor teeth were divided (n = 5) into G1 - storage in distilled water and G2 - storage in artificial saliva. The samples were weighed in an electronic analytical balance at the following times: T0 - before application of the bleaching gel, T1 - after 14 days of bleaching (the time recommended by the manufacturer), T2 - after 21 days of bleaching (50 % increase in the time recommended by the manufacturer), and T3 - after 28 days of bleaching (100 % increase in the time recommended by the manufacturer). The data were subjected to ANOVA for related samples (p = 0.05). The highest mean was observed in G2 (0.5982 g) and the lowest mean was observed in G1 (0.3074 g) at T2 and T3, respectively. Significant differences were observed between the groups at all times. Overall, 10CP caused variation in the enamel mass after a 100 % increase in the use time recommended by the manufacturer, with a decrease in mass when distilled water was used as the storage medium and an increase when artificial saliva was used.

RESUMEN: El objetivo fue investigar la influencia del blanqueamiento dental prolongado con peróxido de carbamida al 10 % (10CP) sobre la variación de masa del esmalte dental. Las muestras se dividieron en dos grupos: G1, diez dientes sanos de los incisivos bovinos (n = 5) en agua destilada, y G2, almacenamiento en saliva artificial. Las muestras se midieron en una escala analítica electrónica de precisión en los siguientes tiempos: T0-antes de la aplicación del gel blanqueador, T1-después de 14 días de blanqueo (el tiempo recomendado por el fabricante), T2-después de 21 días de blanqueo (aumento de 50 % en el tiempo recomendado por el fabricante), y T3-después de 28 días de blanqueo (aumento de 100 % en el tiempo recomendado por el fabricante). Los datos se presentaron al ANOVA para las muestras relacionadas (P = 0,5). La media más alta se observó en G2 (0,5982 g) y la media más baja se observó en G1 (0,3074 g) en T2 y T3, respectivamente. Se observaron diferencias significativas entre los grupos en todo momento. En general, la 10 CP causó variación en la masa del esmalte después de un aumento de 100 % en el tiempo de uso recomendado por el fabricante, con una disminución en la masa cuando el agua destilada se utilizó como medio de almacenamiento y aumentó cuando se usó saliva artificial.