In Vitro Dental Plaque Culture Model for Biofilm Structural Analyses.
In Vitro Dental Plaque Culture Model for Biofilm Structural Analyses.
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用于生物膜结构分析的体外牙菌斑培养模型。
DOI：
10.1002/cpz1.902
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Valm,AlexM
中科院分区：
文献类型：
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作者：
Lemus,AlexA;Valm,AlexM
广泛的研究集中在从健康到疾病的过渡过程中牙菌斑微生物群落的组成变化，称为生态失调。然而，在从健康到疾病的整个过程中，这些社区的空间组成的变化仍然未被探索。我们描述了一种体外牙菌斑模型，用于培养接种有人类志愿者牙菌斑的口腔生物膜。我们的模型概括了体内环境的重要特征，包括唾液流过牙齿引起的剪切力和居住在牙齿的龈上和龈下方面之间的过渡区的微生物所经历的营养环境。重要的是，我们的模型适用于多重荧光标记和多光谱成像，用于测试有关系统级社区结构和功能的特定假设。该模型允许精确操纵各种环境条件，如流速和营养物质的可用性，以调查其对生物膜的发展和空间结构的影响。此外，该模型可用于测试各种治疗干预的效果，例如，该技术是一种新的生物膜分析方法，可在微米至毫米尺度上对生物膜的组成和结构进行分析，使其成为研究牙菌斑介导疾病的分子和细胞基础以及对新的治疗干预措施进行基准测试的有价值的工具。© 2023 Wiley Periodicals LLC.基本方案1：牙菌斑芯片体外模型培养系统支持方案：龈缘（GM）培养基澄清基本方案2：微观世界标记和多光谱图像采集
Extensive research has focused on the compositional changes in dental plaque microbiome communities during the transition from health to disease, known as dysbiosis. However, alterations in the spatial composition of these communities throughout the progression from health to disease remain under‐explored. We describe an in vitro dental plaque model for culturing oral biofilms seeded with dental plaque from human volunteers. Our model recapitulates important features of the in vivo environment including shear force induced by salivary flow over teeth and the nutritional milieu experienced by microbes that inhabit the transitional zone between supragingival and subgingival aspects of the teeth. Importantly, our model is amenable to multiplex fluorescent labeling and multispectral imaging for testing specific hypotheses regarding systems‐level community structure and function. The model allows for precise manipulation of various environmental conditions, such as flow rate and nutrient availability to investigate their effects on biofilm development and spatial structure. Furthermore, this model can be used to test the effects of various therapeutic interventions, e.g., antimicrobial agents, on the biofilm composition and structure at the micron to millimeter scale, making it a valuable tool for studying the molecular and cellular basis of dental plaque‐mediated diseases and for benchmarking new therapeutic interventions. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Dental plaque‐on‐a‐chip in vitro model culture systemSupport Protocol: Gingival margin (GM) medium preparationBasic Protocol 2: Microcosm labeling and multispectral image acquisition