Multispecies aggregates from human dental plaque nucleate highly diverse spatially structured oral biofilms on saliva coated surfaces

摘要 根据疾控中心的数据，在30岁及以上的人群中，牙周炎的发病率为47.2%。到65岁时， 受牙周炎影响的人数跃升至70.1%。生物失调，生物膜组成的变化导致 微生物组成的失衡是广泛传播的人类疾病的前兆，如龋齿 (龋齿)和牙周炎。生物膜空间结构的变化如何影响从健康到 疾病是未知的。我们开发了一种牙菌斑体外培养系统，在该系统中，菌斑样本 通过牙线从健康捐赠者那里获得的牙线被用来播种复杂的社区。定量分析方法： 荧光原位杂交(FISH)、共聚焦显微镜和光谱成像证实了该模型 生物膜具有高度的多样性、空间结构和出人意料的空间结构异质性。我们 观察到来自健康捐赠者的牙菌斑接种物由单细胞和大细胞组成 多物种共聚体。我们假设早期的随机事件是由这些多物种 共聚体导致高度空间结构和异质生物膜，这取决于这些 相互作用的社区落在新生的表面上。为了验证我们的假设，我们开发了一个协议，用于 破坏斑块聚集物。我们观察到，来自牙菌斑接种物的生物膜 在体外培养中，聚集体在播种前被破坏，导致生物膜的多样性降低和 提高了空间同质性。生物膜来源的解聚菌斑接种物缺乏许多革兰氏阴性菌， 有义务成为厌氧菌社区成员。我们的结果显示了一个以前未知的多物种角色。 聚集体在组织口腔生物膜中的作用，并可能在口腔中具有临床重要性。

Abstract According to the CDC, Periodontitis affects 47.2% of individuals aged 30 and over. By the age of 65, the number of people impacted by periodontitis jumps to 70.1%. Dysbiosis, shifts in biofilm composition leading to imbalances in microbial composition are precursors of widespread human diseases such as tooth decay (dental caries) and periodontitis. How shifts in biofilm spatial structure impact the progression from health to disease is unknown. We have developed an in vitro dental plaque culture system in which plaque samples obtained from healthy donors via flossing are used to seed complex communities. Quantitative analysis by Fluorescence in situ Hybridization (FISH), confocal microscopy, and spectral imaging demonstrated that model biofilms are highly diverse, spatially structured, and unexpectedly heterogeneous in spatial structure. We observed that the dental plaque inocula from healthy donors were composed of single cells and large multispecies coaggregates. We hypothesize that early stochastic events mediated by these multispecies coaggregates lead to highly spatially structured and heterogeneous biofilms depending on where these interacting communities land on nascent surfaces. To test our hypothesis, we developed a protocol for disrupting plaque coaggregates. We observed that biofilms derived from dental plaque inocula in which aggregates were disrupted before seeding in vitro cultures resulted in biofilms with decreased diversity and increased spatial homogeneity. Biofilms derived de-aggregated plaque inocula lacked many gram-negative, obligate anaerobe community members. Our results demonstrate a previously unknown role multispecies aggregates in structuring oral biofilms and may have clinical importance in the mouth.