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Electroaddressing Functionalized Polysaccharides as Model Biofilms for Interrogating Cell Signaling

机译：电寻址功能化多糖作为模型生物膜的询问细胞信号。

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摘要

Bacteria often reside at surfaces as complex biofilms in which an exopolysac-charide matrix entraps the population while allowing access to its chemical environment. There is a growing awareness that the biofilm structure and activity are integral to a wide array of properties important to health (the microbiome), disease (drug resistance) and technology (fouling). Despite the importance of bacterial biofilms, few experimental platforms and systems are available to assemble complex populations and monitor their activities. Here, a functionalized alginate composite material for creating in vitro model biofilms suitable for cell-cell signaling studies by entrapping bacterial cells in situ is reported. Biofilm assembly is achieved using device-imposed electrical signals to electrodeposit the stimuli-responsive polysaccharide alginate. This electrodeposition mechanism is versatile in that it allows control of the bacterial population density and distribution. For instance, it is demonstrated that a mixed population can be homogeneously distributed throughout the biofilm or can be assembled as spatially segregated populations within a stratified biofilm. The "electroaddressable" biofilms are visualized using both a planar 2D chip with patterned electrodes and a microfluidic bioMEMS device with sidewall electrodes. Specifically, it is observed that bacteria entrapped within the model biofilm recognize and respond to chemical stimuli imposed from the fluidic environment. Finally, reporter cells are used to demonstrate that bacteria entrapped within this model biofilm engage in intercellular quorum sensing. This work demonstrates the functionality of the stimuli-responsive polysaccharide by biofabricating pseudo-3D cell-gel biocomposites, mimicking the formation of biofilms, for interrogating phenotypes of E. coli bacterial populations. In addition to controlling assembly, the microfluidic device allows the biofilm to be monitored through the fluorescence methods commonly used in biological research. This platform technology should be able to be exploited for monitoring biofilm development, as well as for extending the understanding of the interactions between various bacterial species arranged in controlled patterns.
机译：细菌通常以复杂的生物膜形式存在于表面，在其中生物膜多糖基质捕获了种群，同时允许进入其化学环境。人们日益认识到，生物膜的结构和活性是对健康（微生物组），疾病（抗药性）和技术（结垢）重要的各种特性必不可少的。尽管细菌生物膜很重要，但很少有实验平台和系统可用于组装复杂的种群并监测其活动。在此，报道了一种功能化的藻酸盐复合材料，该材料用于通过原位捕获细菌细胞来创建适用于细胞信号研究的体外模型生物膜。使用设备施加的电信号电沉积刺激响应性多糖藻酸盐可实现生物膜组装。这种电沉积机理是通用的，因为它可以控制细菌的种群密度和分布。例如，证明了混合种群可以均匀地分布在整个生物膜中，或者可以组装成分层生物膜中的空间分隔种群。使用具有图案化电极的平面2D芯片和具有侧壁电极的微流体bioMEMS器件，均可看到“可电寻址”生物膜。具体地，观察到截留在模型生物膜内的细菌识别并响应由流体环境施加的化学刺激。最后，记者细胞被用来证明这种模型生物膜中截留的细菌参与细胞间群体感应。这项工作通过生物伪3D细胞凝胶生物复合材料，模仿生物膜的形成，用于询问大肠杆菌细菌种群的表型，证明了刺激反应性多糖的功能。除了控制组装，微流控设备还允许通过生物研究中常用的荧光方法监控生物膜。该平台技术应该能够用于监测生物膜的发育，以及扩展对以受控模式排列的各种细菌之间相互作用的理解。

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  • 来源
    《Advanced Functional Materials》 |2012年第3期|p.519-528|共10页
  • 作者

    Yi Cheng; Chen-Yu Tsao; Hsuan-Chen Wu; Xiaolong Luo; Jessica L. Terrell; Jordan Betz; Gregory F. Payne; William E. Bentley; Gary W. Rubloff;

  • 作者单位

    Institute for Systems Research University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Systems Research University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Bioscience and Biotechnology Research (IBBR) University of Maryland College Park, MD 20742, USA,Fischell Department of Bioengineering University of Maryland College Park, MD 20742, USA;

    Institute for Systems Research University of Maryland College Park, MD 20742, USA,Department of Materials Science and Engineering University of Maryland College Park, MD 20742, USA;

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