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Application of quantitative proteomic analysis using tandem mass tags for discovery and identification of novel biomarkers in periodontal disease

Authors

  • Sachio Tsuchida,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
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  • Mamoru Satoh,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
    3. Chemical Analysis Center, Chiba University, Chiba, Japan
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  • Yusuke Kawashima,

    1. Laboratory of Biomolecular Dynamics, Department of Physics, School of Science, Kitasato University, Kanagawa, Japan
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  • Kazuyuki Sogawa,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
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  • Sayaka Kado,

    1. Chemical Analysis Center, Chiba University, Chiba, Japan
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  • Setsu Sawai,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
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  • Motoi Nishimura,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
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  • Mayumi Ogita,

    1. Department of Periodontology, Graduate school, Tokyo Medical and Dental University, Bunko, Tokyo, Japan
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  • Yasuo Takeuchi,

    1. Department of Periodontology, Graduate school, Tokyo Medical and Dental University, Bunko, Tokyo, Japan
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  • Hiroaki Kobyashi,

    1. Department of Periodontology, Graduate school, Tokyo Medical and Dental University, Bunko, Tokyo, Japan
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  • Akira Aoki,

    1. Department of Periodontology, Graduate school, Tokyo Medical and Dental University, Bunko, Tokyo, Japan
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  • Yoshio Kodera,

    1. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
    2. Laboratory of Biomolecular Dynamics, Department of Physics, School of Science, Kitasato University, Kanagawa, Japan
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  • Kazuyuki Matsushita,

    1. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
    2. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
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  • Yuichi Izumi,

    1. Department of Periodontology, Graduate school, Tokyo Medical and Dental University, Bunko, Tokyo, Japan
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  • Fumio Nomura

    Corresponding author
    1. Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
    • Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan
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Correspondence: Dr. Fumio Nomura, Department of Molecular Diagnosis (F8), Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan

E-mail: fnomura@faculty.chiba-u.jp

Fax: +81-43-226-2169

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.

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