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

非技术性：由阿肯色州大学材料研究部生物材料项目授予的NSF/FDA驻校学者奖旨在开发一种微透析采样系统，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化学基质。在生物膜中，自由漂浮的细菌开始形成细菌群落，并开始合成支持该细菌群落的专门化学基质。生物膜中的细菌比自由漂浮的细菌更具耐药性，生物膜的形成给医疗保健和各种行业增加了巨大的成本。有相当大的兴趣，在改善科学方法的早期生物膜检测和根除。细菌通过生物膜通过一种称为群体感应的过程相互交流。在群体感应中，细菌物种特有的化学物质被释放出来，并在周围介质中相互“交谈”。本计画将利用微透析取样法，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是早期检测医疗器械上的生物膜形成，以及可能开发控制生物膜形成的治疗方法。所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：这个NSF/FDA奖学金的主要目标是建立一个双培养系统，微透析采样，结合医学相关的细菌（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）。 微透析采样提供了一个独特的平台，以量化局部化学原位在细菌定植/材料界面。该项目的目标1旨在开发微透析采样/生物膜接口。在目标2中，在生物膜定植到透析聚合物膜上的过程中，将实时定量局部pH变化和群体感应信号。目的3寻求开发生物膜/巨噬细胞共培养系统，以解决理解细菌生物膜的初始宿主反应所需的挑战。基于研究者使用微透析进行巨噬细胞生物学和相关细胞因子信号传导测量的优势，Aim 3将开发一种系统，以确定影响巨噬细胞和细菌组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，从而允许重要的基础科学研究影响生物膜过程中的各种检查点。3)一种易于使用、强大且可在实验室中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，其开始评估局部免疫应答，特别是巨噬细胞对抗和/或消除细菌菌落的初始形成以及由此相关的生物膜和感染的动态作用。5)体外转体内的潜力。

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.