Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however, this medium does not maintain the diversity of gram‐negative bacteria more associated with subgingival plaque. Here, we systematically modified SHI medium components to investigate the impacts of varying nutrients and develop a medium capable of supporting a specific disease‐state subgingival community. A diseased subgingival plaque sample was inoculated in SHI medium with increasing concentrations of sucrose (0%, 0.1%, 0.5%), fetal bovine serum (FBS) (0%, 10%, 20%, 30%, 50%), and mucin (0.1, 2.5, 8.0 g/L) and grown for 48 hrs, then the 16S rRNA profiles of the resulting biofilms were examined. In total, these conditions were able to capture 89 of the 119 species and 43 of the 51 genera found in the subgingival inoculum. Interestingly, biofilms grown in high sucrose media, although dominated by acidogenic Firmicutes with a low final pH, contained several uncultured taxa from the genus Treponema, information that may aid culturing these periodontitis‐associated fastidious organisms. Biofilms grown in a modified medium (here named subSHI‐v1 medium) with 0.1% sucrose and 10% FBS had a high diversity closest to the inoculum and maintained greater proportions of many gram‐negative species of interest from the subgingival periodontal pocket (including members of the genera Prevotella and Treponema, and the Candidate Phyla Radiation phylum Saccharibacteria), and therefore best represented the disease community.

中文翻译：

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改良的SHI培养基可在体外支持疾病状态的龈下多菌群落的生长

建立从口腔健康菌斑到患病菌斑的体外研究至关重要的口腔微生物群落实验室模型。SHI培养基是一种丰富的生长培养基，能够支持体外生物膜，其多样性与健康的龈上接种物相似；但是，这种培养基不能保持与牙龈下菌斑更相关的革兰氏阴性细菌的多样性。在这里，我们系统地修改了SHI培养基成分，以研究各种营养素的影响，并开发出能够支持特定疾病状态龈下群落的培养基。将病态的龈下菌斑样品接种在SHI培养基中，并添加浓度不断增加的蔗糖（0％，0.1％，0.5％），胎牛血清（FBS）（0％，10％，20％，30％，50％）和粘蛋白（0.1，2.5，8.0 g / L）并生长48小时，然后检查所得生物膜的16S rRNA图谱。总体而言，这些条件能够捕获龈下接种物中发现的119个物种中的89个物种和51个属中的43个物种。有趣的是，在高蔗糖培养基中生长的生物膜，虽然以最终酸碱度较低的产酸硬毛菌为主导，但仍含有一些未培养的类群。梅毒螺旋体，可能有助于培养这些与牙周炎相关的顽固性微生物的信息。在含有0.1％蔗糖和10％FBS的改良培养基（此处称为subSHI-v1培养基）中生长的生物膜具有最接近接种物的高度多样性，并保留了来自龈下牙周袋的许多革兰氏阴性菌（包括成员）的更大比例Prevotella和Treponema属，以及候选Phyla辐射门菌（Saccharibacteria）），因此最能代表疾病社区。