In vitro 3D human gingival tissue model to study oral microbiome

用于研究口腔微生物组的体外 3D 人类牙龈组织模型

• 批准号: 10303383
• 负责人: Chiara Ghezzi
• 金额: $ 17.42万
• 依托单位: UNIVERSITY OF MASSACHUSETTS LOWELL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10303383
• 关键词: 16S ribosomal RNA sequencing; 3-Dimensional; Air; Anatomy; Animals; Artificial Saliva; Bacteria; Behavior; Biological; Biological Markers; Biopolymers; Bone Resorption; Buffers; CCL2 gene; Calibration; Cells; Chronic; Clinical; Collagen; Communities; Complex; Conditioned Reflex; Cytokeratin; DNA; Dental Materials; Dental Plaque; Dentifrices; Direct Costs; Disease; Disease model; Ecosystem; Environment; Epithelial; Epithelial Cells; Equilibrium; Exposure to; Exudate; Facilities and Administrative Costs; Gingiva; Gingival Pocket; Gingivitis; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Health; Histologic; Homeostasis; Host Defense; Human; IL8 gene; Immune response; Immunohistochemistry; Immunology; In Vitro; Infection; Inflammation; Inflammatory; Interferon Type II; Interleukin-1 alpha; Interleukin-10; Interleukin-17; Interleukin-2; Interleukin-4; Interleukin-6; Irritants; Lead; Liquid substance; Longitudinal Studies; Maintenance; Medicine; Membrane; Metabolic; Modeling; Monitor; Mucous Membrane; Nutrient; Oral; Oral cavity; Organism; Outcome; Oxygen; Pathogenesis; Pathogenicity; Patients; Periodontal Diseases; Periodontitis; Physiological; Play; Production; Resistance; Role; Saliva; Sampling; Scientist; Silk; Stratified Epithelium; Stromal Cells; Structure; Symptoms; System; TNF gene; Testing; Therapeutic; Time; Tissue Engineering; Tissue Model; Tissue Viability; Tissues; Tooth Loss; Ursidae Family; Visualization; anakinra; antimicrobial; base; cell immortalization; chemokine; clinically relevant; community organizations; consumer product; cost estimate; cytokine; design; dysbiosis; effective therapy; improved; in vitro Model; in vivo; member; metabolic profile; microbial; microbial community; microbiome; multiplex assay; nutrition; oral bacteria; oral care; oral commensal; oral condition; oral microbial community; oral microbiome; oral pathogen; oral plaque; oral tissue; pathogen; polycarbonate; reconstitution; reconstruction; response; scaffold; shear stress; stress management; tissue culture; tool; tooth surface; transcriptome sequencing; translational impact; two-dimensional

PROJECT SUMMARY The oral cavity contains different microenvironments, i.e. the non-shedding surface of the teeth and the epithelial mucosa, where oral barriers and microbial communities coexist. The interactions and balances between these two communities are responsible for oral tissue homeostasis or dysbiosis, that ultimately dictate health or disease. Disruption of this equilibrium is the first necessary step towards chronic inflammation and permanent tissue damage in the case of chronic periodontitis. Current experimental animal and in vitro models do not fully resemble the human condition. To improve clinical outcomes and design effective treatments, new humanized experimental tools are needed to further elucidate these initial host-pathogens unbalances. Previously developed in vitro systems have been used to test irritant responses of new dental materials, dentifrices, and oral care consumer products, but are unable to maintain the complexity of the oral pathogen community organization, due to the lack of the native oxygen and metabolic conditions. In addition, host saliva contributes to the maintenance of the overall oral system stability by buffering the oral environment, providing nutrition to the different communities and delivering antimicrobial features. Therefore, the recapitulation of physiological oral conditions, including oxygen gradients, physiological shear stress, and buffering from saliva will enhance the functions of a humanized sustained gingival tissue model to study initial host-pathogen interactions in vitro. We are proposing to design a physiological culture system based on artificial saliva in order to support long-term culture after inoculation with oral microbiota derived from healthy patients. We will replicate the stability of the ecosystem and evaluate the contribution of host saliva to buffer and provide nutrition to the oral community, as well as physiological shear stress that contributes to the maturation and maintenance of a healthy epithelium. The 3D anatomical gingival tissue model has been shown to modulate the production of a range of cytokines and chemokines in response to interactions with inoculated plaque samples from healthy patients. To assess the clinical relevance of this response, we will study the production of selected cytokines under different conditions; response to the addition of plaque derived from healthy and diseased (gingivitis) patients in comparison to crevicular fluid extracted from the same pool of patients. The efforts will elucidate the initial interactions and balances between these two communities that are responsible for the oral tissue homeostasis or dysbiosis, that ultimately dictates healthy or diseased tissue states. The interdisciplinary team involved in the project has established collaborative activities and includes material scientists, tissue engineers, microbiologists, and periodontists. They are experts in host-material interactions, advanced in vitro tissue and culture models, microbial pathogenesis, host defense, periodontal immunology, and periodontal inflammation.

项目概要 口腔包含不同的微环境，即牙齿的非脱落表面和牙齿的表面。 上皮粘膜，口腔屏障和微生物群落共存。相互作用和平衡 这两个群落之间的相互作用负责口腔组织稳态或生态失调，最终 决定健康或疾病。打破这种平衡是迈向慢性病的第一步。 慢性牙周炎时会出现炎症和永久性组织损伤。当前实验 动物和体外模型并不完全类似于人类的状况。为了改善临床结果和 设计有效的治疗方法，需要新的人性化实验工具来进一步阐明这些初步的结果 宿主与病原体不平衡。先前开发的体外系统已用于测试刺激物 新牙科材料、洁牙剂和口腔护理消费品的反应，但无法维持 由于缺乏原生氧气和 代谢条件。此外，宿主唾液有助于维持整个口腔系统 通过缓冲口腔环境、为不同群体提供营养并传递稳定性 抗菌功能。因此，重现口腔生理状况，包括氧气 梯度、生理剪切应力和唾液缓冲将增强人源化的功能 持续的牙龈组织模型来研究体外初始宿主-病原体相互作用。我们提议 设计基于人工唾液的生理培养系统，以支持术后长期培养 接种来自健康患者的口腔微生物群。我们将复制生态系统的稳定性 并评估宿主唾液对口腔群落缓冲和提供营养的贡献，以及 生理剪切应力有助于健康上皮的成熟和维持。这 3D 牙龈组织解剖模型已被证明可以调节一系列细胞因子的产生 和趋化因子响应与来自健康患者的接种斑块样本的相互作用。到 评估这种反应的临床相关性，我们将研究所选细胞因子的产生 不同的条件；对添加源自健康和患病（牙龈炎）的牙菌斑的反应 患者与从同一组患者中提取的裂隙液进行比较。这些努力将阐明 这两个负责口腔组织的群落之间最初的相互作用和平衡 体内平衡或生态失调，最终决定健康或患病的组织状态。跨学科 参与该项目的团队已经建立了协作活动，包括材料科学家、组织科学家 工程师、微生物学家和牙周病学家。他们是主体材料相互作用方面的专家，在 体外组织和培养模型、微生物发病机制、宿主防御、牙周免疫学和 牙周炎症。