ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Antimicrobials, Resistance and Chemotherapy
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1675322
This article is part of the Research TopicInnovative Antibiofilm Strategies: Advancing the Management of Microbial Biofilm InfectionsView all articles
Inhibition of Streptococcus mutans biofilm formation and virulence by natural extract stevioside
Provisionally accepted
- The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China
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Objective: This study aimed to investigate the effect of the natural extract of Stevioside on biofilm formation and cariogenic virulence factors of Streptococcus mutans (S. mutans), and to explore its mechanism of action preliminarily, with the goal of identifying a safer and more effective non-cariogenic sweetener. Methods: The inhibitory effect of Stevioside on the growth of S. mutans biofilm was detected by crystal violet staining. The acid production capacity of S. mutans biofilms was evaluated by measuring the pH values and lactic acid contents. The bacterial viability within the biofilms was determined using the plate counting method.Scanning electron microscopy (SEM) and laser confocal microscopy (CLSM) were used to observe the biofilm structure, and the fluorescence intensity of live and dead bacteria and biofilm thickness were further analyzed. The anthrone sulfuric acid method quantified the production of Soluble Extracellular Polysaccharide (SEPS)and Insoluble Extracellular Polysaccharide (IEPS) in biofilm. Furthermore, real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of genes related to biofilm growth, acid production, acid tolerance, exopolysaccharide synthesis, two-component signal and quorum sensing. Results: Stevioside significantly inhibited the formation of S. mutans biofilm, and reduced acid production, bacterial activity and EPS production. SEM and CLSM confirmed the reduction of the dense three-dimensional structure of biofilm. In addition, compared with sucrose, the expression of related genes was down-regulated in Stevioside. Conclusion: Stevioside inhibits S. mutans virulence factors and biofilms and is a promising natural sucrose substitute for preventing dental caries.
Keywords: Stevioside, S. mutans, Biofilm, virulence factor, Gene Expression
Received: 29 Jul 2025; Accepted: 18 Sep 2025.
Copyright: © 2025 Ma, Wang, Zhang, Wang and Chu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Jinpu Chu, cjp605@163.com
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