RESUMO
Objective@# To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines.@*Methods@# pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. @*Results @#1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. @*Conclusion@#P.m played an important role in the transcriptome of oral keratinocytes.
RESUMO
El presente estudio tuvo como propósito conocer la asociación entre la presencia de pigmentaciones negras extrínsecas en superficies dentarias y la frecuencia de caries dental en niños con dentición mixta del Servicio de Odontopediatría del Hospital Central Fuerza Aérea del Perú (n=185), de los cuales el 6,49% (12 niños) presentaban dichas pigmentaciones. Para el análisis de los datos se emplearon frecuencias, porcentajes, medidas de tendencia central, medidas de dispersión, Chi-cuadrado y para conocer la asociación se empleó la prueba U de Mann Whitney. Al analizar la asociación entre las pigmentaciones negras extrínsecas y la frecuencia de caries dental, los resultados mostraron en los niños que presentaban dichas pigmentaciones un CEO+CPO de 1,33±1,07; mientras que los niños que no presentaban dichas pigmentaciones negras extrínseca sun CEO+CPO de 3,39±2,39. Sobre la base del análisis estadístico se concluye que el CEO+CPO en los niños con pigmentaciones negras extrínsecas fue menor que el CEO+CPO en los niños que no tenían dichas pigmentaciones siendo esta diferencia estadísticamente significativa con un p=0,001. Cabe resaltar que se encontró asociación entre la presencia de estas pigmentaciones y el consumo de suplementos dietéticos a base de fierro siendo estadísticamente significativa con un p=0,027.
The aim of the present study was to explore the association between the presence of extrinsic black stains on tooth surfaces and ithe frequency of dental caries in children from the Pediatric Clinic of the Peruvian Air Force Central Hospital (n=185). Among the children studied, 6,49% (12 children) presented those pigmentations. For the analysis of the data, frequencies, percentages, measures of central tendency, measures of dispersion and Chi-square were used and to explore the association Mann-Whitney U test was used, processed and analyzed using SPSS version 12 program. After the analysis of the relationship between the extrinsic black stains and the frequency of dental caries, the results showed that in children presenting such pigmentations the DMF+def index was of 1.33±1.07, while in children who did not present such black extrinsic pigmentations a DMF+def of 3.39±2.39 was found. Based on the statistical analysis it is concluded that the DMF+def in children with extrinsic black stains was lower than the DMF+def on children who had not such pigmentations and this difference was statistically significant with p=0.001. It is worth mentioning that an association found between the presence of these pigmentations and consumption of dietary supplements based on iron was statistically significant with p=0.027.