Comparative gene expression analysis of Porphyromonas gingivalis ATCC 33277 in planktonic and biofilms states

Romero-Lastra, Patricia; Sánchez Beltrán, María Del Carmen; Ribeiro-Vidal, Honorato; Llama Palacios, María Arantxazu; Figuero Ruiz, Elena; Herrera González, David; Sanz Alonso, Mariano

Comparative gene expression analysis of Porphyromonas gingivalis ATCC 33277 in planktonic and biofilms states

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Official URL

https://doi.org/10.1371/journal.pone.0174669

Publication date

2017

Authors

Romero-Lastra, Patricia

Sánchez Beltrán, María Del Carmen

Ribeiro-Vidal, Honorato

Llama Palacios, María Arantxazu

Figuero Ruiz, Elena

Herrera González, David

Sanz Alonso, Mariano

Publisher

Public Library of Science

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URI

https://hdl.handle.net/20.500.14352/94547

Citation

Romero-Lastra P, Sánchez MC, Ribeiro-Vidal H, Llama-Palacios A, Figuero E, Herrera D, Sanz M. Comparative gene expression analysis of Porphyromonas gingivalis ATCC 33277 in planktonic and biofilms states. PLoS One. 2017 Apr 3;12(4):e0174669.

Abstract

Background and objective Porphyromonas gingivalis is a keystone pathogen in the onset and progression of periodontitis. Its pathogenicity has been related to its presence and survival within the subgingival biofilm. The aim of the present study was to compare the genome-wide transcription activities of P. gingivalis in biofilm and in planktonic growth, using microarray technology. Material and methods P. gingivalis ATCC 33277 was incubated in multi-well culture plates at 37˚C for 96 hours under anaerobic conditions using an in vitro static model to develop both the planktonic and biofilm states (the latter over sterile ceramic calcium hydroxyapatite discs). The biofilm development was monitored by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy (SEM). After incubation, the bacterial cells were harvested and total RNA was extracted and purified. Three biological replicates for each cell state were independently hybridized for transcriptomic comparisons. A linear model was used for determining differentially expressed genes and reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to confirm differential expression. The filtering criteria of ±2 change in gene expression and significance p-values of <0.05 were selected. Results A total of 92 out of 1,909 genes (4.8%) were differentially expressed by P. gingivalis growing in biofilm compared to planktonic. The 54 up-regulated genes in biofilm growth were mainly related to cell envelope, transport, and binding or outer membranes proteins. Thirty-eight showed decreased expression, mainly genes related to transposases or oxidative stress. Conclusion The adaptive response of P. gingivalis in biofilm growth demonstrated a differential gene expression.