Department of Periodontology, College of Dentistry, University of Florida, Gainesville, FL, 32610, USA.
Research Laboratory for Archaeology and the History of Art, University of Oxford, Oxford, OX1 3QY, UK.
BMC Microbiol. 2018 Jul 11;18(1):70. doi: 10.1186/s12866-018-1212-x.
Several in vitro oral biofilm growth systems can reliably construct oral microbiome communities in culture, yet their stability and reproducibility through time has not been well characterized. Long-term in vitro growth of natural biofilms would enable use of these biofilms in both in vitro and in vivo studies that require complex microbial communities with minimal variation over a period of time. Understanding biofilm community dynamics in continuous culture, and whether they maintain distinct signatures of health and disease, is necessary to determine the reliability and applicability of such models to broader studies. To this end, we performed next-generation sequencing on biofilms grown from healthy and disease-site subgingival plaque for 80 days to assess stability and reliability of continuous oral biofilm growth.
Biofilms were grown from subgingival plaque collected from periodontitis-affected sites and healthy individuals for ten eight-day long generations, using hydroxyapatite disks. The bacterial community in each generation was determined using Human Oral Microbe Identification by Next-Generation Sequencing (HOMINGS) technology, and analyzed in QIIME. Profiles were steady through the ten generations, as determined by species abundance and prevalence, Spearman's correlation coefficient, and Faith's phylogenetic distance, with slight variation predominantly in low abundance species. Community profiles were distinct between healthy and disease site-derived biofilms as demonstrated by weighted UniFrac distance throughout the ten generations. Differentially abundant species between healthy and disease site-derived biofilms were consistent throughout the generations.
Healthy and disease site-derived biofilms can reliably maintain consistent communities through ten generations of in vitro growth. These communities maintain signatures of health and disease and of individual donors despite culture in identical environments. This subgingival oral biofilm growth and perpetuation model may prove useful to studies involving oral infection or cell stimulation, or those measuring microbial interactions, which require the same biofilms over a period of time.
有几种体外口腔生物膜生长系统可以可靠地在培养物中构建口腔微生物组群落，但它们的稳定性和随时间的重现性尚未得到很好的描述。长期的体外生物膜生长将使这些生物膜能够用于需要在一段时间内具有最小变化的复杂微生物群落的体外和体内研究。了解连续培养中的生物膜群落动态，以及它们是否保持健康和疾病的独特特征，对于确定这些模型在更广泛的研究中的可靠性和适用性是必要的。为此，我们对从健康和疾病部位龈下菌斑中生长的生物膜进行了下一代测序，以评估连续口腔生物膜生长的稳定性和可靠性。
使用羟基磷灰石盘从牙周炎部位和健康个体采集的龈下菌斑培养了十个为期八天的生物膜世代。使用下一代测序的人类口腔微生物鉴定（HOMINGS）技术确定每个世代的细菌群落，并在 QIIME 中进行分析。通过物种丰度和流行率、Spearman 相关系数和 Faith 系统发育距离来确定十个世代的生物膜特征是稳定的，只有少量的变化主要发生在低丰度的物种中。通过整个十个世代的加权 UniFrac 距离来证明健康和疾病部位来源的生物膜的群落特征是不同的。健康和疾病部位来源的生物膜之间差异丰富的物种在整个世代中是一致的。
健康和疾病部位来源的生物膜可以通过十代体外生长可靠地维持一致的群落。这些群落保持了健康和疾病以及个体供体的特征，尽管在相同的环境中培养。这种龈下口腔生物膜生长和维持模型可能对涉及口腔感染或细胞刺激的研究有用，或者对需要在一段时间内使用相同生物膜的微生物相互作用研究有用。
