Novel flow cell model to study oral mucosa-polymicrobial biofilm interactions

描述(由申请人提供)：微生物不是单独存在的，而是动态的多微生物群落的一部分。人类拥有多个微生物群落，这些群落在不同的上皮表面附近形成生物膜。在口腔中，生物膜主要形成在牙齿或修复体上，并与口腔粘膜相邻。在易感宿主中，含有白色念珠菌或牙龈卟啉单胞菌等物种的口腔生物膜构成了感染和炎症性疾病(如念珠菌病和牙周病)的主要病原学。因此，确定根除口腔生物膜或干扰其发展的战略将有助于治疗或预防此类疾病。为了制定这样的策略，我们需要了解控制口腔多菌生物膜群落形成的因素，包括群落成员之间发生的相互作用和宿主在形成生物膜特征中的作用。在这项试点拨款中，我们将描述一种新型的体外模型系统，在该系统中，这些问题可以开始得到解答。所提出的模型由流动细胞装置组成，该流动细胞装置旨在容纳在口腔粘膜组织类似物存在下生长的多微生物生物膜。在目标A中，我们将通过确定包含口腔病原体(牙龈假单胞菌或白色念珠菌)的微生物组合组装多微生物生物膜的最佳实验条件来表征我们模型的生物膜成分。在目标B中，我们将通过评估在流动条件下存在和不存在生物膜的情况下细胞因子和抗菌肽的产生来表征我们的模型(口腔粘膜3D模拟物)的组织成分。最后，在目标C中，我们将研究相邻上皮在多菌生物膜组装中的作用，并通过微阵列鉴定在白念珠菌和牙龈假单胞菌中形成多菌群落的部分时受宿主差异调控的基因。我们设想，我们的新模型将导致发现影响口腔环境中生物膜发育的宿主因素。这些宿主因素可以进一步研究，以了解它们在口腔中形成多微生物生物膜生长和人类口腔生物膜相关疾病易感性特征中的作用。此外，更好地了解多菌生物膜与宿主的相互作用将导致开发新的治疗策略来干预生物膜的形成。

DESCRIPTION (provided by applicant): Microorganisms do not exist individually but are part of dynamic polymicrobial communities. Humans harbor polymicrobial communities that form biofilms in close proximity to different epithelial surfaces. In the oral cavity, biofilms mostly form on teeth or prosthetic devices and adjacent to the oral mucosa. In the susceptible host, oral biofilms containing species such as Candida albicans or Porphyromonas gingivalis constitute the primary etiology of infectious and inflammatory conditions such as candidiasis and periodontal disease. Therefore, the identification of strategies to eradicate oral biofilms or interfere with their development will aid in the treatment or prevention of such conditions. In order to develop such strategies we need to understand the factors that govern the formation of oral polymicrobial biofilm communities, including both the interactions occurring among community members and the role of the host in shaping biofilm characteristics. In this pilot grant we will characterize a novel in vitro model system where such questions can begin to be answered. The proposed model consists of a flow cell device designed to harbor a polymicrobial biofilm growing in the presence of an oral mucosa tissue analogue. In Aim A we will characterize the biofilm component of our model by determining the optimal experimental conditions for polymicrobial biofilm assembly by microbial consortia containing an oral pathogen (either P. gingivalis or C. albicans). In Aim B we will characterize the tissue component of our model (an oral mucosa 3-D analogue) by evaluating the production of cytokines and anti-microbial peptides in the presence and absence of biofilms under flow conditions. Finally, in Aim C we will examine the role of the adjacent epithelium on polymicrobial biofilm assembly and identify, via microarrays, genes differentially regulated by the host in C. albicans and P. gingivalis when forming part of a polymicrobial community. We envision that our new model will lead to the discovery of host factors that influence biofilm development in the oral environment. Such host factors can be examined further to understand their role in shaping polymicrobial biofilm growth in the oral cavity and oral biofilm-related disease susceptibility traits in humans. Furthermore, a better understanding of polymicrobial biofilm-host interactions will lead to the development of novel therapeutic strategies to interfere with biofilm formation.