Proteomic analysis of endodontic infections by liquid chromatography-tandem mass spectrometry.

R Nandakumar, N Madayiputhiya, A F Fouad
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Abstract

Introduction: Endodontic infections are very prevalent and have a polymicrobial etiology characterized by complex interrelationships between endodontic microorganisms and the host defenses. Proteomic analysis of endodontic infections can provide global insights into the invasion, pathogenicity mechanisms, and multifactorial interactions existing between root canal bacteria and the host in the initiation and progression of apical periodontitis. The purpose of this study was to apply proteomic techniques such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the identification of proteins of bacterial origin present in endodontic infections.

Methods: Endodontic specimens were aseptically obtained from seven patients with root canal infections. Protein mixtures were subjected to tryptic in-solution digestion and analysed by reverse-phase nano-LC-MS/MS followed by a database search.

Results: Proteins, mainly of cell wall or membrane origin, from endodontic bacteria especially Enterococcus faecalis, Enterococcus faecium, Porphyromonas gingivalis, Fusobacterium nucleatum, and Treponema denticola were identified from all the samples tested. Identified proteins included adhesins, autolysins, proteases, virulence factors, and antibiotic-resistance proteins.

Conclusions: LC-MS/MS offers a sensitive analytical platform to study the disease processes in the root canal environment. The array of proteins expressed in endodontic infections reflects the complex microbial presence and highlights the bacterial species involved in the inflammatory process.

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用液相色谱-串联质谱法分析牙髓感染的蛋白质组学。
牙髓感染非常普遍,具有多微生物病因,其特点是牙髓微生物与宿主防御之间存在复杂的相互关系。根管感染的蛋白质组学分析可以全面了解根管细菌与宿主在根尖牙周炎发生和发展过程中的侵袭、致病机制以及多因素相互作用。本研究的目的是应用蛋白质组学技术,如液相色谱-串联质谱(LC-MS/MS)来鉴定牙髓感染中细菌来源的蛋白质。方法:对7例根管感染患者无菌采集根管标本。蛋白质混合物经胰酶溶酶切,通过反相纳米lc -MS/MS进行分析,并进行数据库检索。结果:检出的蛋白主要来自于牙髓细菌,特别是粪肠球菌、粪肠球菌、牙龈卟啉单胞菌、核梭杆菌和牙密螺旋体。鉴定的蛋白质包括粘附素、自溶素、蛋白酶、毒力因子和抗生素抗性蛋白。结论:LC-MS/MS为研究根管环境下的病变过程提供了一个灵敏的分析平台。在牙髓感染中表达的一系列蛋白质反映了复杂的微生物存在,并突出了参与炎症过程的细菌种类。
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Distribution and hydrolytic enzyme characteristics of Candida albicans strains isolated from diabetic patients and their non-diabetic consorts. Automutanolysin disrupts clinical isolates of cariogenic streptococci in biofilms and planktonic cells. Scavenger receptor A is expressed by macrophages in response to Porphyromonas gingivalis, and participates in TNF-alpha expression. Helicobacter pylori in the oral cavity is associated with gastroesophageal disease. Progression of chronic periodontitis can be predicted by the levels of Porphyromonas gingivalis and Treponema denticola in subgingival plaque.
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