Institute of Dentistry, University of Turku, Lemminkäisenkatu 2, 20520, Turku, Finland.
Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, Netherlands.
BMC Microbiol. 2020 Jun 29;20(1):184. doi: 10.1186/s12866-020-01867-8.
Regular consumption of xylitol decreases the number of cariogenic streptococci in dental plaque. In vitro biofilm models to study the mechanism of xylitol action have been set-up, but the obtained results are contradictory. Biofilm growth is a dynamic process with time-specific characteristics that may remain undetected in conventional end-point biofilm tests. In this study we used an impedance spectroscopy instrument, xCELLigence Real Time Cell Analyzer (RTCA), that allows label-free, non-invasive real-time monitoring of biofilm formation, to explore effects of xylitol on biofilm formation by Streptococcus mutans. Based on the obtained information of biofilm dynamics, we assessed the number of viable bacteria, the polysaccharide content, and the expression levels of selected genes involved in glucan-mediated biofilm formation in different biofilm stages. Xylitol inhibition was compared with that of erythritol; another polyol suggested to have a positive impact on oral health.
Our results showed that real-time monitoring provided new information of polyol-induced changes in S. mutans biofilm formation dynamics. The inhibitory effect of polyols was more pronounced in the early stages of biofilm formation but affected also the measured total amount of formed biofilm. Effects seen in the real-time biofilm assay were only partially explained by changes in CFU values and polysaccharide amounts in the biofilms. Both xylitol and erythritol inhibited real-time biofilm formation by all the nine tested S. mutans strains. Sensitivity of the strains to inhibition varied: some were more sensitive to xylitol and some to erythritol. Xylitol also modified the expression levels of gbpB, gtfB, gtfC and gtfD genes that are important in polysaccharide-mediated adherence of S. mutans.
The erythritol- and xylitol- induced inhibition of biofilm formation was only partly explained by decrease in the number of viable S. mutans cells or the amount of polysaccharides in the biofilm matrix, suggesting that in addition to reduced proliferation also the matrix composition and thereby the surface attachment quality of biofilm matrix may be altered by the polyols.
经常食用木糖醇可以减少牙菌斑中致龋链球菌的数量。已经建立了体外生物膜模型来研究木糖醇作用的机制,但得到的结果却相互矛盾。生物膜的生长是一个具有时间特异性特征的动态过程,这在传统的终点生物膜试验中可能无法检测到。在这项研究中,我们使用阻抗谱仪,即 xCELLigence 实时细胞分析(RTCA),对变形链球菌生物膜形成进行无标记、非侵入性的实时监测,以探索木糖醇对生物膜形成的影响。基于获得的生物膜动力学信息,我们评估了不同生物膜阶段活细菌数量、多糖含量和参与葡聚糖介导生物膜形成的选定基因的表达水平。将木糖醇的抑制作用与赤藓糖醇的抑制作用进行了比较,赤藓糖醇被认为对口腔健康有积极影响。
我们的研究结果表明,实时监测提供了有关多醇诱导变形链球菌生物膜形成动力学变化的新信息。多醇在生物膜形成的早期阶段的抑制作用更为明显,但也影响了所测量的形成生物膜的总量。实时生物膜测定中观察到的作用仅部分解释了生物膜中 CFU 值和多糖含量的变化。木糖醇和赤藓糖醇都抑制了所有 9 株测试的变形链球菌的实时生物膜形成。菌株对抑制的敏感性不同:有些对木糖醇更敏感,有些对赤藓糖醇更敏感。木糖醇还改变了 gbpB、gtfB、gtfC 和 gtfD 基因的表达水平,这些基因在变形链球菌多糖介导的黏附中很重要。
赤藓糖醇和木糖醇诱导的生物膜形成抑制作用部分是由于变形链球菌活细胞数量或生物膜基质中多糖含量减少引起的,这表明除了增殖减少外,多醇还可能改变生物膜基质的组成,从而改变生物膜基质的表面附着质量。