Italian Institute of Technology, Center for Bio-Molecular Nanotechnology, Via Barsanti-73010 Arnesano, Lecce, Italy.
ACS Nano. 2011 Mar 22;5(3):1865-76. doi: 10.1021/nn102692m. Epub 2011 Feb 23.
Bacterial adhesion onto inorganic/nanoengineered surfaces is a key issue in biotechnology and medicine, because it is one of the first necessary steps to determine a general pathogenic event. Understanding the molecular mechanisms of bacteria-surface interaction represents a milestone for planning a new generation of devices with unanimously certified antibacterial characteristics. Here, we show how highly controlled nanostructured substrates impact the bacterial behavior in terms of morphological, genomic, and proteomic response. We observed by atomic force microscopy (AFM) and scanning electron microscopy (SEM) that type-1 fimbriae typically disappear in Escherichia coli adherent onto nanostructured substrates, as opposed to bacteria onto reference glass or flat gold surfaces. A genetic variation of the fimbrial operon regulation was consistently identified by real time qPCR in bacteria interacting with the nanorough substrates. To gain a deeper insight into the molecular basis of the interaction mechanisms, we explored the entire proteomic profile of E. coli by 2D-DIGE, finding significant changes in the bacteria adherent onto the nanorough substrates, such as regulations of proteins involved in stress processes and defense mechanisms. We thus demonstrated that a pure physical stimulus, that is, a nanoscale variation of surface topography, may play per se a significant role in determining the morphological, genetic, and proteomic profile of bacteria. These data suggest that in depth investigations of the molecular processes of microorganisms adhering to surfaces are of great importance for the design of innovative biomaterials with active biological functionalities.
细菌在无机/纳米工程表面的黏附是生物技术和医学中的一个关键问题,因为它是决定一般致病事件的第一步。理解细菌-表面相互作用的分子机制是规划具有一致认证抗菌特性的新一代设备的一个里程碑。在这里,我们展示了高度可控的纳米结构基底如何影响细菌在形态、基因组和蛋白质组响应方面的行为。我们通过原子力显微镜(AFM)和扫描电子显微镜(SEM)观察到,在纳米结构基底上黏附的大肠杆菌中,通常会消失类型-1菌毛,而在参考玻璃或平整金表面上黏附的细菌则不会。通过实时 qPCR 一致鉴定了菌毛操纵子调节的遗传变异,这些细菌与纳米粗糙基底相互作用。为了更深入地了解相互作用机制的分子基础,我们通过 2D-DIGE 探索了大肠杆菌的整个蛋白质组谱,发现黏附在纳米粗糙基底上的细菌有明显变化,例如涉及应激过程和防御机制的蛋白质的调节。因此,我们证明了纯物理刺激,即表面形貌的纳米级变化,本身可能在决定细菌的形态、遗传和蛋白质组谱方面发挥重要作用。这些数据表明,深入研究微生物黏附在表面上的分子过程对于设计具有主动生物学功能的创新生物材料非常重要。