Probiotics alter biofilm formation and the transcription of Porphyromonas gingivalis virulence-associated genes

Abstract

Background and Objective: The potential of probiotics on the prevention and control of periodontitis and other chronic inflammatory conditions has been suggested. Lactobacillus and Bifidobacterium species influence P. gingivalis interaction with gingival epithelial cells (GECs) but may not act in a unique way. In order to select the most appropriate probiotic against P. gingivalis, we aimed to evaluate the effect of several strains on Porphyromonas gingivalis biofilm formation and transcription virulence-associated factors (PgVAFs). Methods: Cell-free pH neutralized supernatants (CFS) and living Lactobacillus spp. and Bifidobacterium spp. were tested against P. gingivalis ATCC 33277 and W83, in mono-and multi-species (with Streptococcus oralis and S. gordonii) biofilms. Relative transcription of P. gingivalis genes (fimA, mfa1, kgp, rgp, ftsH and luxS) was determined in biofilms and under GECs co-infection. Results: Probiotics CFS reduced P. gingivalis ATCC 33277 levels in mono-species biofilms and living probiotics reduced P. gingivalis abundance in multi-species biofilms. L. acidophilus LA5 down-regulated transcription of most PgVAFs in biofilms and GECs. Conclusions: Probiotics affect P. gingivalis biofilm formation by down-regulating overall PgVAFs with the most pronounced effect observed for L. acidophilus LA5.

Lactobacilli postbiotics reduce biofilm formation and alter transcription of virulence genes of Aggregatibacter actinomycetemcomitans

“…Lactobacilli cell-free supernatants (CFS) were obtained as previously described (Ishikawa et al, 2020). Briefly, tested lactobacilli were inoculated in BHI broth at 1 × 10 7 CFU/ml and incubated under anaerobiosis at 37°C, with gentle agitation for 24 hr.…”

“…The assays to evaluate the effects of CFS on biofilms were adapted from other studies (Ishikawa et al, 2020;Matsubara, Wang, et al, 2016). In order to access the effect of CFS on biofilm formation, A. actinomycetemcomitans was suspended into CFS diluted in fresh BHI at 1:2.5, 1:5, and 1:10 and inoculated in 96-wells flat-bottom polystyrene microtiter plates (Corning Inc.) at 1 × 10 7 CFU/well.…”

“…Living microorganisms which promote health benefits, known as probiotics (Hill et al, 2014), may be used in the control of periodontitis based on their ability to decrease colonization of pathogens and alter host immune response . Probiotic strains interfere with the ability of the periodontopathogen P. gingivalis to interact with host cells (Albuquerque-Souza et al, 2019), to express virulence factors, and to form biofilm (Ishikawa et al, 2020).…”

“…Thus, postbiotics comprise secreted bioactive metabolites such as bacteriocins, hydrogen peroxide, and short-chain fatty acids or microbial compounds, like teichoic acid (Collado et al, 2019;O'Grady et al, 2019;Sánchez et al, 2017;Wegh et al, 2019). Postbiotics derived from LAB were shown to inhibit biofilms of several organisms including pathogens such as Candida albicans (Matsubara, Wang et al, 2016), S. aureus (Koohestani et al, 2018), and P. gingivalis (Ishikawa et al, 2020). Their clinical application may be safer than the use of viable bacteria (Tsilingiri & Rescigno, 2013), and these compounds are usually stable, i.e., present a prolonged shelf life (Koohestani et al, 2018), suggesting their potential use in oral hygiene products.…”

Quantifying the antimicrobial activity of CRISPR-Cas9-accA modified ΔB. subtilismutants againstV. harveyiandE. Coli

“…However, dysbiosis or an imbalance in microflora can cause increased colonization and infection by pathogenic opportunistic microbes [16]. The rate of dysbiosis depends on the ratio between the number of disease-associated pathogenic species and health promoting species [17].…”