NSF/FDA SIR: A Microdialysis-Based Co-Culture System for Molecular Treatment and Assessment of Biofilms.

NSF/FDA SIR：基于微透析的共培养系统，用于生物膜的分子治疗和评估。

• 批准号: 1641166
• 负责人: Julie Stenken
• 金额: $ 10万
• 依托单位: University of Arkansas
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2018-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1641166&HistoricalAwards=false
• 关键词: NSF; FDA; SIR; Microdialysis; Based

Non-technical: This NSF/FDA Scholar-in-Residence award by the Biomaterials program in the Division of Materials Research to the University of Arkansas is to develop a microdialysis sampling system to assess the localized chemistry associated with biofilm/polymer interfaces. Biofilms are complex chemical matrices produced by communities of bacteria. In a biofilm, free-floating bacteria begin to form a community of bacteria and start synthesizing specialized chemical matrices in support of this bacterial community. The bacteria in the biofilm are far more antibiotic-resistant than the free-floating ones, and the biofilm formation adds enormous costs to healthcare and various industries. There is considerable interest in the improvement of scientific methods for early biofilm detection and its eradication. Bacteria communicate with each other through the biofilm via a process called quorum sensing. In quorum sensing, chemicals unique to the bacterial species are released, and 'talk' to each others present in the surrounding medium. This project will use the microdialysis sampling approach to collect the quorum sensing chemicals from the bacterial/material interface. The broader impact of the project is in the early detection of the biofilm formation on medical devices, and possible development of treatments to control the biofilm formation. The proposed approaches also could allow the implementation of various treatments to see how these affect the bacterial community, chemical communication and the biofilm structure.Technical: The primary goal for this NSF/FDA Scholar-in-Residence award is to create a dual culture system with microdialysis sampling that incorporates medically-relevant bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis) and macrophages (RAW 267.4). Microdialysis sampling provides a unique platform to quantify localized chemistry in situ at the bacterial colonization/material interface. Aim 1 of the project seeks to develop the microdialysis sampling/biofilm interface. In Aim 2, localized pH changes and quorum sensing signals will be quantified in real-time during the biofilm colonization process onto the dialysis polymeric membrane. Aim 3 seeks to develop a biofilm/macrophage co-culture system to address needed challenges in understanding initial host response to bacterial biofilms. Building on the strength of the investigator with macrophage biology and associated cytokine signaling measurements using microdialysis, Aim 3 will develop a system to determine system parameters and signaling output that affect the combination of macrophage cells and bacteria. The successful outcomes of this proposed work include: 1) A rapid and interchangeable method to assess how different prophylactic treatments affect biofilm formation and quorum sensing. 2) A method to assess the temporal chemical dynamics associated with biofilm formation thus allowing important basic science studies to affect various checkpoints in the biofilm process. 3) An easy-to-use, robust and laboratory transferrable basic science method for monitoring various aqueous chemical aspects of interfacial biofilm chemistry. 4) A novel co-culture system to begin to assess the dynamic role of the localized immunological response, particularly macrophages, toward fighting and/or eliminating the initial formation of a bacterial colony and thus associated biofilm and infection. 5) Potential for in vitro to in vivo translation.

非技术：由阿肯色大学材料研究部生物材料项目颁发的NSF/FDA驻场学者奖旨在开发一种微透析取样系统，以评估与生物膜/聚合物界面相关的局部化学。生物膜是由细菌群落产生的复杂的化学基质。在生物膜中，自由漂浮的细菌开始形成一个细菌群落，并开始合成专门的化学基质来支持这个细菌群落。生物膜中的细菌比自由漂浮的细菌更耐抗生素，生物膜的形成给医疗保健和各种行业增加了巨大的成本。人们对改善早期生物膜检测和根除的科学方法非常感兴趣。细菌通过一种称为群体感应的过程通过生物膜相互交流。在群体感应中，细菌种类特有的化学物质被释放出来，并在周围介质中相互“交谈”。该项目将使用微透析取样方法从细菌/材料界面收集群体感应化学物质。该项目更广泛的影响在于早期发现医疗器械上的生物膜形成，并可能开发控制生物膜形成的治疗方法。提出的方法还可以允许实施各种处理，以了解这些处理如何影响细菌群落，化学通讯和生物膜结构。技术：该NSF/FDA驻场学者奖的主要目标是创建一种双培养系统，微透析采样包含医学相关细菌（铜绿假单胞菌、金黄色葡萄球菌和表皮葡萄球菌）和巨噬细胞（RAW 267.4）。微透析取样提供了一个独特的平台，可以在细菌定植/材料界面上就地量化局部化学。该项目的目标1旨在开发微透析采样/生物膜界面。在Aim 2中，在生物膜定殖到透析聚合物膜上的过程中，将实时量化局部pH变化和群体感应信号。目的3旨在开发一种生物膜/巨噬细胞共培养系统，以解决理解宿主对细菌生物膜的初始反应所需要的挑战。基于巨噬细胞生物学和相关细胞因子信号测量的研究人员的力量，Aim 3将开发一个系统，以确定影响巨噬细胞和细菌结合的系统参数和信号输出。这项工作的成功成果包括：1)一种快速和可互换的方法来评估不同的预防性治疗如何影响生物膜的形成和群体感应。2)一种评估与生物膜形成相关的时间化学动力学的方法，从而使重要的基础科学研究能够影响生物膜过程中的各个检查点。3)一种易于使用，稳健和实验室可转移的基础科学方法，用于监测界面生物膜化学的各种水化学方面。4)一种新的共培养系统，开始评估局部免疫反应的动态作用，特别是巨噬细胞，对抗和/或消除细菌菌落的初始形成，从而相关的生物膜和感染。5)体外到体内翻译的潜力。