Laboratory of Biomechanics and Biomedical Engineering, Department of Mechanical Engineering and Aeronautics, University of Patras, Rion, Patras, Greece.
Eur Cell Mater. 2012 Nov 17;24:386-402. doi: 10.22203/ecm.v024a28.
The assessment of biomaterial susceptibility to infection relies mainly on the analysis of macroscopic bacterial responses to material interactions, usually under static conditions. However, new technologies permit a more profound understanding of the molecular basis of bacteria-biomaterial interactions. In this study, we combine both conventional phenotypic analysis - using confocal microscopy - and genotypic analysis - using the relative reverse transcription polymerase chain reaction (RT-PCR) - to examine the interaction of bacteria with OH- and CH3-terminated glass surfaces, under dynamic flow conditions. Bacterial adhesion, as well as slime production and biofilm formation, was much higher on the CH3-terminated than on the OH-terminated glass - for four Staphylococcus epidermidis strains. This was in agreement with the icaA and icaD gene expression results that showed increased expression for the bacteria adhering to the CH3-terminated substrate, especially under the higher shear rate. Therefore, the combined effect of the surface chemistry and shear significantly influence the adhesion and phenotype of interacting bacterial cells, while there are putative links between phenotypic responses to bacteria-material interactions and gene-expression profile alterations. This indicates that analysis of gene expression not only can greatly refine our knowledge of bacteria-material interactions, but also yield novel biomarkers for potential use in biocompatibility assessment.
生物材料感染易感性的评估主要依赖于对材料相互作用下宏观细菌反应的分析,通常是在静态条件下进行。然而,新技术可以更深入地了解细菌-生物材料相互作用的分子基础。在这项研究中,我们将常规表型分析(使用共聚焦显微镜)和基因型分析(使用相对逆转录聚合酶链反应(RT-PCR))相结合,研究了在动态流动条件下,细菌与 OH 和 CH3 端玻璃表面的相互作用。与 OH 端玻璃相比,四种表皮葡萄球菌菌株在 CH3 端玻璃上的细菌黏附、胞外多糖产生和生物膜形成都要高得多。这与 icaA 和 icaD 基因表达结果一致,表明在较高的剪切速率下,黏附在 CH3 端底物上的细菌表达增加。因此,表面化学和剪切的综合作用显著影响相互作用的细菌细胞的黏附和表型,而细菌-材料相互作用的表型反应与基因表达谱改变之间存在潜在联系。这表明,基因表达分析不仅可以极大地完善我们对细菌-材料相互作用的认识,还可以为潜在的生物相容性评估提供新的生物标志物。