BMC oral health · 2025
In vitro evaluation of tolerance ability of cross-kingdom biofilm towards oral dynamic fluctuations.
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Li Yijun, Huang Shan, Du Jingyun, Huang Jing, Huang Xiaojing
Аннотация
OBJECTIVES: The aim of this study was to investigate the resilience of Streptococcus mutans (S. mutans) and Candida albicans (C. albicans) cross-kingdom biofilms in response to environmental stresses.
MATERIALS AND METHODS: The growth kinetics of S. mutans, C. albicans, and their co-culture in planktonic form were assessed using a BioScreen system. Biofilms were established on 96-well plates for a duration of 48 h, after which microbial counts were determined. The biofilms were then subjected to different stress conditions, including oxidative, acid, osmotic, and heat stress, for 2 h. The survival and structural integrity of the biofilms were evaluated through colony-forming unit (CFU) counting and fluorescence microscopy, respectively. Additionally, the transcriptional levels of genes concerning matrix formation, acid tolerance, oxidative tolerance were determined by quantitative real-time PCR (RT-PCR). The data was analyzed by one-way ANOVA, and post hoc Tukey's test (α = 0.05).
RESULTS: Co-culturing S. mutans and C. albicans resulted in an extended logarithmic growth phase compared to monocultures. Dual-species biofilm had higher microorganism counts after biofilm formation, displayed higher surviving cells, and a more complex structure after exposure to various stresses when compared to monospecies biofilm (p<0.05). The transcriptional levels of genes concerning matrix formation (gtfB, gtfC, ftf, bcr1, hwp1), acid tolerance (atpD, fabM, phr1), oxidative tolerance (nox, sodA, sod1, and trx1) were upregulated in dual-species biofilm (p<0.05).
CONCLUSIONS: Dual-species biofilm present higher tolerance to various stresses in the oral cavity. The upregulation of genes involved in matrix formation and stress tolerance may partially account for this increased resilience.
MATERIALS AND METHODS: The growth kinetics of S. mutans, C. albicans, and their co-culture in planktonic form were assessed using a BioScreen system. Biofilms were established on 96-well plates for a duration of 48 h, after which microbial counts were determined. The biofilms were then subjected to different stress conditions, including oxidative, acid, osmotic, and heat stress, for 2 h. The survival and structural integrity of the biofilms were evaluated through colony-forming unit (CFU) counting and fluorescence microscopy, respectively. Additionally, the transcriptional levels of genes concerning matrix formation, acid tolerance, oxidative tolerance were determined by quantitative real-time PCR (RT-PCR). The data was analyzed by one-way ANOVA, and post hoc Tukey's test (α = 0.05).
RESULTS: Co-culturing S. mutans and C. albicans resulted in an extended logarithmic growth phase compared to monocultures. Dual-species biofilm had higher microorganism counts after biofilm formation, displayed higher surviving cells, and a more complex structure after exposure to various stresses when compared to monospecies biofilm (p<0.05). The transcriptional levels of genes concerning matrix formation (gtfB, gtfC, ftf, bcr1, hwp1), acid tolerance (atpD, fabM, phr1), oxidative tolerance (nox, sodA, sod1, and trx1) were upregulated in dual-species biofilm (p<0.05).
CONCLUSIONS: Dual-species biofilm present higher tolerance to various stresses in the oral cavity. The upregulation of genes involved in matrix formation and stress tolerance may partially account for this increased resilience.