AUTHOR=Ma Nan , Yang Wei , Chen Bairu , Bao Meihua , Li Yimin , Wang Meng , Yang Xiaopeng , Liu Junyi , Wang Chengyue , Qiu Lihong TITLE=Exploration of the primary antibiofilm substance and mechanism employed by Lactobacillus salivarius ATCC 11741 to inhibit biofilm of Streptococcus mutans JOURNAL=Frontiers in Cellular and Infection Microbiology VOLUME=Volume 15 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1535539 DOI=10.3389/fcimb.2025.1535539 ISSN=2235-2988 ABSTRACT=IntroductionLactobacillus salivarius serves as a probiotic potentially capable of preventing dental caries both in vitro and in vivo. This study focused on understanding the key antibiofilm agents and the mechanisms of action of the Lactobacilli supernatant against Streptococcus mutans.MethodsStreptococcus mutans biofilm was constructed and the cell-free supernatant of Lactobacillus salivarius was added. After the biofilm was collected, RNA-seq and qRT-PCR were then performed to get gene information. The influence of temperature, pH and other factors on the supernatant were measured and non-targeted metabolome analysis was performed to analyze the effective components.ResultsThe findings indicated that the supernatant derived from Lactobacillus salivarius could inhibit the biofilm formation of Streptococcus mutans at different times. Through transcriptome analysis, we discovered that the cell-free supernatant reduced biofilm formation, by suppressing phosphoenolpyruvate-dependent phosphotransferase systems along with two ATP-binding cassette transporters, rather than directly affecting the genes that code for glucosyltransferases; additionally, the supernatant was observed to diminish the expression of genes linked to two-component systems, polyketides/non-ribosomal peptides, acid stress response, quorum sensing, and exopolysaccharide formation. Non-targeted LC-MS/MS analysis was employed to discover a variety of potential active compounds present in the cellular filtrate of Lactobacillus salivarius that hinder the growth of S. mutans, including phenyllactic acid, sorbitol, and honokiol.DiscussionIn summary, our findings support the evaluation of Lactobacillus salivarius as a promising oral probiotic aimed at hindering the formation of biofilms by cariogenic pathogens and the development of dental caries.