Department of Stomatology, Shanghai Hudong Hospital, Shanghai, China.
Department of Stomatology, Eye and Ent Hospital of Fudan University, Shanghai, China.
Technol Health Care. 2024;32(S1):371-383. doi: 10.3233/THC-248033.
Enterococcus faecalis biofilm was frequently found on the failed treated root canal wall, which survived by resisting disinfectant during endodontic treatment.Many researches have been conducted to explore the mechanisms of persistence of this pathogen in unfavorable conditions. However, no comprehensive proteomics studies have been conducted to investigate stress response in Enterococcus faecalis caused by alkali and NaOCl.
Enterococcus faecalis (E.f) has been recognized as a main pathogen of refractory apical periodontitis, its ability to withstand environmental pressure is the key to grow in the environment of high alkaline and anti-bacterial drug that causes chronic infection in the root canal. This study aims to focus on the protein expression patterns of E.f biofilm under extreme pressure environment".
Enterococcus faecalis biofilm model was established in vitro. Liquid Chromatograph-Mass Spectrometer (LC-MS/MS)-based label free quantitative proteomics approach was applied to compare differential protein expression under different environmental pressures (pH 10 and 5% sodium hypochlorite (NaOCl)). And then qPCR and Parallel Reaction Monitoring Verification (PRM) were utilized to verify the consequence of proteomics.
The number of taxa in this study was higher than those in previous studies, demonstrating the presence of a remarkable number of proteins in the groups of high alkaline and NaOCl. Proteins involved in ATP-binding cassette (ABC) transporter were significantly enriched in experimental samples. We identified a total of 15 highly expressed ABC transporters in the high alkaline environment pressure group, with 7 proteins greater than 1.5 times.
This study revealed considerable changes in expression of proteins in E.f biofilm during resistance to environmental pressures. The findings enriched our understanding of association between the differential expression proteins and environmental pressures.
粪肠球菌生物膜经常在根管治疗失败的根管壁上发现，它在根管治疗过程中通过抵抗消毒剂而存活。许多研究已经探索了这种病原体在不利条件下持续存在的机制。然而，目前还没有进行全面的蛋白质组学研究来研究碱性和次氯酸钠对粪肠球菌的应激反应。
粪肠球菌（E.f）已被认为是难治性根尖周炎的主要病原体，其耐受环境压力的能力是其在根管内高碱性和抗菌药物环境中生长的关键。本研究旨在关注粪肠球菌生物膜在极端压力环境下的蛋白质表达模式。
体外建立粪肠球菌生物膜模型。采用基于液相色谱-质谱联用（LC-MS/MS）的无标记定量蛋白质组学方法，比较不同环境压力（pH 10 和 5%次氯酸钠（NaOCl））下的差异蛋白质表达。然后利用 qPCR 和平行反应监测验证（PRM）来验证蛋白质组学的结果。
本研究中的分类群数量高于以往研究，表明在高碱性和 NaOCl 组中存在大量蛋白质。涉及三磷酸腺苷结合盒（ABC）转运体的蛋白质在实验样本中显著富集。我们在高碱性环境压力组中总共鉴定出 15 种高度表达的 ABC 转运蛋白，其中 7 种蛋白的表达量超过 1.5 倍。
本研究揭示了粪肠球菌生物膜在抵抗环境压力时蛋白质表达的显著变化。研究结果丰富了我们对差异表达蛋白与环境压力之间关系的理解。
