Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, Guangdong, China.
Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong, China.
Appl Microbiol Biotechnol. 2024 May 10;108(1):330. doi: 10.1007/s00253-024-13149-8.
A more optimized culture medium used in vitro to mimic the bacterial composition of original oral flora as similar as possible remains difficult at present, and the goal of this study is to develop a novel oral biofilm medium to restore the original oral microbiome. Firstly, we conducted a systematic literature review by searching PubMed and summarized the current reported culture media in vitro. Seven culture media were found. We used mixed saliva as the origin of oral species to compare the effects of the above media in culturing oral multispecies biofilms. Results indicated that among the seven media brain heart infusion containing 1% sucrose (BHIs) medium, PG medium, artificial saliva (AS) medium, and SHI medium could obviously gain large oral biofilm in vitro. The nutrients contained in different culture media may be suitable for the growth of different oral bacteria; therefore, we optimized several novel media accordingly. Notably, results of crystal violet staining showed that the biofilm cultured in our modified artificial saliva (MAS) medium had the highest amount of biofilm biomass. 16S rRNA gene sequencing showed that the operational taxonomic units (OTUs) and Shannon index of biofilm cultured in MAS medium were also the highest among all the tested media. More importantly, the 16S rRNA gene sequencing analysis indicated that the biofilm cultured in MAS medium was closer to the original saliva species. Besides, biofilm cultured by MAS was denser and produced more exopolysaccharides. MAS supported stable biofilm formation on different substrata. In conclusion, this study demonstrated a novel MAS medium that could culture oral biofilm in vitro closer to the original oral microbiome, showing a good application prospect. KEY POINTS: • We compare the effects of different media in culturing oral biofilms • A novel modified artificial saliva (MAS) medium was obtained in our study • The MAS medium could culture biofilm that was closer to oral microbiome.
一种更优化的体外培养基，用于尽可能模拟原始口腔菌群的细菌组成，目前仍然难以实现，本研究的目的是开发一种新型口腔生物膜培养基，以恢复原始口腔微生物组。首先，我们通过搜索 PubMed 进行了系统的文献回顾，并总结了目前体外报道的培养介质。发现了七种培养介质。我们使用混合唾液作为口腔物种的来源，比较了上述介质在培养口腔多物种生物膜中的效果。结果表明，在七种培养基中，含 1%蔗糖的脑心浸液（BHIs）培养基、PG 培养基、人工唾液（AS）培养基和 SHI 培养基可以明显在体外获得大量口腔生物膜。不同培养基中所含的营养物质可能适合不同口腔细菌的生长；因此，我们相应地对几种新型培养基进行了优化。值得注意的是，结晶紫染色结果表明，在我们改良的人工唾液（MAS）培养基中培养的生物膜具有最高的生物膜生物量。16S rRNA 基因测序结果表明，MAS 培养基中培养的生物膜的操作分类单元（OTUs）和 Shannon 指数也是所有测试培养基中最高的。更重要的是，16S rRNA 基因测序分析表明，MAS 培养基中培养的生物膜与原始唾液物种更为接近。此外，MAS 培养的生物膜更密集，产生更多的胞外多糖。MAS 支持在不同基质上形成稳定的生物膜。综上所述，本研究证明了一种新型 MAS 培养基，可以在体外培养更接近原始口腔微生物组的口腔生物膜，具有良好的应用前景。 关键点：• 我们比较了不同培养基在培养口腔生物膜中的效果• 本研究获得了一种新型改良人工唾液（MAS）培养基• MAS 培养基可以培养更接近口腔微生物组的生物膜。
