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

来自人类牙菌斑的多物种聚集体在唾液涂层表面上形成高度多样化的空间结构口腔生物膜

• 批准号: 10679723
• 负责人: Alex Lemus
• 金额: $ 3.57万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679723
• 关键词: 16S ribosomal RNA sequencing; Address; Affect; Age; Algorithms; Anaerobic Bacteria; Antibiotic Therapy; Bacteria; Biochemical; Biological Assay; Biological Models; Cell Communication; Cell Survival; Cells; Chemicals; Clinical; Communities; Complex; DNA sequencing; Data; Dental Hygiene; Dental Plaque; Dental caries; Development; Disease; Event; Excision; Fluorescent in Situ Hybridization; Foundations; Genes; Genomics; Growth; Health; Human; Image; In Vitro; Individual; Knowledge; Lead; Left; Mediating; Methods; Microbial Biofilms; Modeling; Modernization; Neighborhoods; Oral; Oral cavity; Periodontal Diseases; Periodontitis; Persons; Protocols documentation; Recurrence; Resolution; Role; Saliva; Sampling; Structure; Structure-Activity Relationship; Surface; System; Taxon; Taxonomy; Testing; Work; aged; beta diversity; confocal imaging; dental biofilm; differential expression; dysbiosis; experimental study; human disease; in vitro Model; member; microbial; microbial composition; microorganism; microscopic imaging; multiplexed imaging; oral biofilm; oral microbiome; preservation; response; spectrograph; surface coating; tooth surface; virtual

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.

摘要 根据CDC，牙周炎影响47.2%的30岁及以上的人。到了65岁， 患牙周炎的人数上升到70.1%。生态失调，生物膜组成的变化导致 微生物组成的不平衡是大范围人类疾病的前兆， （龋齿）和牙周炎。生物膜空间结构的变化如何影响从健康到 疾病未知。我们开发了一种体外牙菌斑培养系统， 通过牙线清洁从健康的捐赠者获得，用于播种复杂的社区。定量分析 荧光原位杂交（FISH）、共聚焦显微镜和光谱成像证实了该模型 生物膜是高度多样的、空间结构化的，并且在空间结构上出乎意料地不均匀。我们 观察到来自健康供体的牙菌斑接种物由单细胞和大细胞组成， 多物种共聚集体。我们假设这些多物种介导的早期随机事件 共聚集体导致高度空间结构化和异质的生物膜，这取决于这些生物膜的位置。 相互作用的群落落在新生的表面上。为了验证我们的假设，我们制定了一个协议， 破坏斑块共聚集体。我们观察到生物膜来源于牙菌斑接种物，其中 在接种体外培养物之前，聚集体被破坏，导致生物膜的多样性降低， 增加空间的均匀性。生物膜衍生的解聚菌斑接种物缺乏许多革兰氏阴性， 专性厌氧菌群落成员。我们的研究结果证明了一个以前未知的作用多物种 在构建口腔生物膜中聚集，并且可能在口腔中具有临床重要性。