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

项目总结 口腔包含不同的微环境，即牙齿的不脱落面和 上皮粘膜，口腔屏障和微生物群落共存的地方。互动与平衡 在这两个群落之间负责口腔组织的动态平衡或生物失调，最终 主宰健康或疾病。打破这种平衡是迈向慢性疾病的必要的第一步 慢性牙周炎的炎症和永久性组织损伤。当前的实验 动物和体外模型并不完全类似于人类的情况。以改善临床结果和 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最初的 寄主-病原体失衡。以前开发的体外系统已被用于测试刺激性 新型牙科材料、牙膏和口腔护理消费品的反应，但无法维持 口腔病原体群落组织的复杂性，由于缺乏天然氧气和 新陈代谢状况。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、为不同社区提供营养并提供 抗菌功能。因此，重述包括氧气在内的口腔生理状况 梯度、生理剪切力和唾液缓冲将增强人性化的 体外持续牙龈组织模型研究初始宿主与病原体的相互作用。我们正在提议 设计一种基于人工唾液的生理培养系统，以支持以后的长期培养 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 并评估宿主唾液对缓冲和向口腔社区提供营养的贡献，以及 有助于健康的上皮成熟和维持的生理剪切力。这个 3D牙龈组织模型已被证明可以调节一系列细胞因子的产生 以及趋化因子与来自健康患者的接种斑块样本的相互作用。至 评估这种反应的临床相关性，我们将研究选定的细胞因子在 不同的情况；对添加来自健康和疾病(牙周炎)的菌斑的反应 将患者与从同一患者池中提取的脑脊液进行比较。这些努力将澄清 负责口腔组织的这两个群落之间的初始相互作用和平衡 动态平衡或生物失调，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 工程师、微生物学家和牙周病医生。他们是宿主-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。

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.