title: Application of quantitative proteomic analysis using tandem mass tags for discovery and identification of novel biomarkers in periodontal disease.
abstract: Periodontal disease is a bacterial infection that destroys the gingiva and surrounding tissues of the oral cavity. Gingival crevicular fluid (GCF) is extracted from the gingival sulcus and pocket. Analysis of biochemical markers in GCF, which predict the progression of periodontal disease, may facilitate disease diagnosis. However, no useful GCF biochemical markers with high sensitivity for detecting periodontal disease have been identified. Thus, the search for biochemical markers of periodontal disease is of continued interest in experimental and clinical periodontal disease research. Using tandem mass tag (TMT) labeling, we analyzed GCF samples from healthy subjects and patients with periodontal disease, and identified a total of 619 GCF proteins based on proteomic analysis. Of these, we focused on two proteins, matrix metalloproteinase (MMP)-9 and neutrophil gelatinase-associated lipocalin (LCN2), which are involved in the progression of periodontal disease. Western blot analysis revealed that the levels of MMP-9 and LCN2 were significantly higher in patients with periodontal disease than in healthy subjects. In addition, ELISA also detected significantly higher levels of LCN2 in patients with periodontal disease than in healthy subjects. Thus, LC-MS/MS analyses of GCF using TMT labeling led to the identification of LCN2, which may be a promising GCF biomarker for the detection of periodontal disease.
authors.affiliation: Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan.
authors.forename: Sachio
authors.initials: S
authors.lastname: Tsuchida
authors.forename: Mamoru
authors.initials: M
authors.lastname: Satoh
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kawashima
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Sogawa
authors.forename: Sayaka
authors.initials: S
authors.lastname: Kado
authors.forename: Setsu
authors.initials: S
authors.lastname: Sawai
authors.forename: Motoi
authors.initials: M
authors.lastname: Nishimura
authors.forename: Mayumi
authors.initials: M
authors.lastname: Ogita
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kobyashi
authors.forename: Akira
authors.initials: A
authors.lastname: Aoki
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Kodera
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsushita
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Izumi
authors.forename: Fumio
authors.initials: F
authors.lastname: Nomura
mesh_terms: D000209:Acute-Phase Proteins / Q000032:analysis / Q000378:metabolism; D000328:Adult; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005883:Gingival Crevicular Fluid / Q000737:chemistry; D006801:Humans; D000071068:Lipocalin-2; D054834:Lipocalins / Q000032:analysis / Q000378:metabolism; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000378:metabolism; D008875:Middle Aged; D010510:Periodontal Diseases / Q000378:metabolism*; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000209:Acute-Phase Proteins; D015415:Biomarkers; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D011506:Proteins; D020543:Proteome; D011518:Proto-Oncogene Proteins; D020780:Matrix Metalloproteinase 9
keywords: Biomedicine; Gingival crevicular fluid; Periodontal disease; Tandem mass tag