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. 2023 Jun 20;12(6):848.
doi: 10.3390/pathogens12060848.

Dose-Dependent Inhibitory Effect of Probiotic Lactobacillus plantarum on Streptococcus mutans- Candida albicans Cross-Kingdom Microorganisms

Jianhang Bao  1   2 , Xinyan Huang  1   2 , Yan Zeng  1 , Tong Tong Wu  3 , Xingyi Lu  3 , Gina Meng  4 , Yanfang Ren  1 , Jin Xiao  1
Affiliations

Dose-Dependent Inhibitory Effect of Probiotic Lactobacillus plantarum on Streptococcus mutans- Candida albicans Cross-Kingdom Microorganisms

Jianhang Bao et al. Pathogens. .

Abstract

Dental caries is one of the most common chronic diseases worldwide. Streptococcus mutans and Candida albicans are two major pathogens associated with dental caries. Several recent studies revealed that Lactobacillus plantarum inhibits S. mutans and C. albicans in biofilms and in a rodent model of dental caries. The aim of this study was to investigate the dose-dependent effect of L. plantarum against S. mutans and C. albicans in a planktonic model that simulated a high-caries-risk clinical condition. Mono-, dual-, and multi-species models were utilized, with five doses of L. plantarum (ranging from 1.0 × 104 to 1.0 × 108 CFU/mL). Real-time PCR was used to assess the expression of the virulence genes of C. albicans and S. mutans and the genes of L. plantarum. Student's t-tests and one-way ANOVA, followed by post hoc tests, were employed to compare the cell viability and gene expression among groups. A dose-dependent inhibition on C. albicans and S. mutans was observed with increased dosages of L. plantarum. L. plantarum at 108 CFU/mL demonstrated the highest antibacterial and antifungal inhibitory effect in the dual- and multi-species models. Specifically, at 20 h, the growth of C. albicans and S. mutans was suppressed by 1.5 and 5 logs, respectively (p < 0.05). The antifungal and antibacterial effects were attenuated in lower doses of L. plantarum (104-107 CFU/mL). The expression of C. albicans HWP1 and ECE 1 genes and S. mutans lacC and lacG genes were significantly downregulated with an added 108 CFU/mL of L. plantarum (p < 0.05). The addition of 108 CFU/mL L. plantarum further inhibited the hyphae or pseudohyphae formation of C. albicans. In summary, L. plantarum demonstrated dose-dependent antifungal and antibacterial effects against C. albicans and S. mutans. L. plantarum emerged as a promising candidate for the creation of novel antimicrobial probiotic products targeting dental caries prevention. Further research is warranted to identify the functional metabolites produced by L. plantarum at different dosages when interacting with C. albicans and S. mutans.

Keywords: Candida albicans; Lactobacillus plantarum; Streptococcus mutans; cross-kingdom interaction; fungi; probiotic.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 3
Figure 3
Effect of L. plantarum on the expression of C. albicans and S. mutans genes in multi-species model at 20 h. qRT-PCR was performed for S. mutans and C. albicans genes of interest for mixed-species culture at 20 h. S. mutans (A) and C. albicans (B) gene expression ratios are shown, and the comparison is relative to S. mutans and C. albicans dual-species. p values were determined by one-way ANOVA with post hoc test. * p < 0.05, ** p < 0.01, **** p < 0.0001.
Figure 4
Figure 4
Dose-related expression of L. plantarum gene in mono-species model. qRT-PCR was performed for L. plantarum genes of interest for mono-species culture at 20 h. L. plantarum gene expression ratio is shown, and the comparison is relative to 104 CFU/mL L. plantarum mono-species group. p values were determined by one-way ANOVA with post hoc test. * p < 0.05, *** p < 0.001, **** p < 0.0001.
Figure 5
Figure 5
Expression of L. plantarum genes when interacting with S. mutans and C. albicans in multi-species model at 20 h. qRT-PCR was performed for L. plantarum genes of interest for mixed-species culture at 20 h. L. plantarum gene expression ratio is shown (A–O), and the comparison is relative to L. plantarum mono-species group. p values were determined by t-test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Figure 1
Figure 1
Dynamic changes in the viable cells of C. albicans, S. mutans, and L. plantarum in mono-, dual-, and multi-species models. (A,B) C. albicans viable cells in dual- and multi-species conditions. (C,D) S. mutans viable cells in dual- and multi-species conditions. (E) L. plantarum viable cells in mono-species condition. (F) L. plantarum viable cells in C. albicans presence dual-species condition. (G) L. plantarum viable cells in S. mutans presence dual-species condition. (H) L. plantarum viable cells in multi-species condition. Overall, dose-dependent antimicrobial and antifungal effects were seen for L. plantarum; 108 CFU/mL of L. plantarum showed inhibition on the growth of C. albicans and S. mutans. The dotted line represents the control groups in each model.
Figure 2
Figure 2
Changes in species composition in multi-species. The composition of each microorganism in multi-species condition is shown. (A–E) The composition of C. albicans, S. mutans, and L. plantarum in multi-species condition.
Figure 6
Figure 6
Dose-dependent inhibition of C. albicans hyphae formation by L. plantarum gene in multi-species model at × 100 magnification. (A) S. mutans and C. albicans grown in dual-species model at 20 h. (B–F) S. mutans and C. albicans grown with addition of various doses of L. plantarum at 20 h. The addition of 108 CFU/mL L. plantarum inhibited the growth of C. albicans and the transition from yeast to hyphae or pseudohyphae form. These are representative images of multiple fields of view.
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