Laboratoire de Chimie Physique et Microbiologie pour les Matériaux et l'Environnement, LCPME, UMR 7564, Université de Lorraine-CNRS, 405, rue de Vandoeuvre, 54600 Villers-lès-Nancy, France.
Institut Jean Lamour, IJL, UMR 7198, Université de Lorraine-CNRS, Campus Artem, 2 allée André Guinier, 54000 Nancy, France.
ACS Appl Mater Interfaces. 2020 Apr 1;12(13):14933-14945. doi: 10.1021/acsami.0c01335. Epub 2020 Mar 19.
Bacteria grow on surfaces and form communities called biofilms. Bacterial adhesion and properties of the derived biofilms depend on, among others, the nature of the supporting substrate. Here, we report how the surface properties of the substrate affect the biofilm growth of probiotic GG (LGG). Hydrophilic (OH), hydrophobic (CH), and positively charged (NH) surfaces were obtained by the functionalization of a ZnSe crystal with alkanethiol self-assembled monolayers (SAM). The self-assembly of alkanethiols onto ZnSe was studied using infrared spectroscopy in attenuated total reflection mode (ATR-FTIR). The organization of grafted SAMs was analyzed based on the results of ATR-FTIR, high-energy elastic backscattering spectrometry, and contact angle measurements. The kinetics and adhesion strength of LGG initial attachment as well as its physiological state on surfaces terminated by the different functional groups were assessed by the combination of ATR-FTIR, force measurements based on atomic force microscopy, and fluorescent staining of bacteria. The strength of interactions between LGG and the surface was strongly affected by the terminal group of the alkanethiol chain. The -NH groups displayed the highest affinity with LGG at the first stage of interaction. The surface properties also played an important role when LGG biofilms were further grown in a nutritive medium for 24 h under flow conditions. Notably, the analysis of the infrared spectra recorded during the biofilm cultivation revealed differences in the kinetics of growth and in the polysaccharide features of the biofilm depending on the substrate functionality. LGG biofilm was stable only on the positively charged surface upon rinsing. Findings of this work clearly show that the adhesion features and the growth of LGG biofilms are substrate-dependent.
细菌在表面生长并形成称为生物膜的群落。细菌的粘附和衍生生物膜的特性取决于支撑基底的性质等。在这里,我们报告了基底表面性质如何影响益生菌 GG(LGG)的生物膜生长。通过用烷硫醇自组装单层(SAM)对 ZnSe 晶体进行功能化,获得了亲水性(OH)、疏水性(CH)和正电性(NH)表面。使用衰减全反射模式(ATR-FTIR)的红外光谱研究了烷硫醇在 ZnSe 上的自组装。基于 ATR-FTIR、高能弹性背散射光谱和接触角测量的结果分析了接枝 SAM 的组织。通过 ATR-FTIR、基于原子力显微镜的力测量和细菌荧光染色的组合评估了 LGG 初始附着的动力学和粘附强度及其在不同官能团终止表面上的生理状态。LGG 与表面之间相互作用的强度受烷硫醇链末端基团强烈影响。-NH 基团在相互作用的第一阶段与 LGG 显示出最高的亲和力。在营养培养基中进一步生长 24 小时并在流动条件下形成生物膜时,表面性质也起着重要作用。值得注意的是,在生物膜培养过程中记录的红外光谱分析表明,根据基底功能,生物膜的生长动力学和多糖特征存在差异。仅在正电荷表面上,LGG 生物膜在冲洗后是稳定的。这项工作的结果清楚地表明,LGG 生物膜的粘附特性和生长是依赖于基底的。