Proteomic analysis of gingival crevicular fluid for discovery of novel periodontal disease markers

Sachio Tsuchida, Mamoru Satoh, Hiroshi Umemura, Kazuyuki Sogawa, Yusuke Kawashima, Sayaka Kado, Setsu Sawai, Motoi Nishimura, Yoshio Kodera, Kazuyuki Matsushita, Fumio Nomura

Research output: Contribution to journalArticlepeer-review

42 Citations (Scopus)

Abstract

The protein composition of gingival crevicular fluid (GCF) may reflect the pathophysiology of periodontal diseases. A standard GCF proteomic pattern of healthy individuals would serve as a reference to identify biomarkers of periodontal diseases by proteome analyses. However, protein profiles of GCF obtained from apparently healthy individuals have not been well explored. As a step toward detection of proteomic biomarkers for periodontal diseases, we applied both gel-based and gel-free methods to analyze GCF obtained from healthy subjects as compared with supragingival saliva. To ensure optimized protein extraction from GCF, a novel protocol was developed. The proteins in GCF were extracted with high yield by urea buffer combined with ultrafiltration and the intensity of spots with supragingival saliva and GCF was compared using agarose two-dimensional electrophoresis. Eight protein spots were found to be significantly more intense in GCF. They included superoxide dismutase 1 (SOD1), apolipoprotein A-I (ApoA-I), and dermcidin (DCD). Moreover, GCF proteins from healthy subjects were broken down into small peptide fragments and then analyzed directly by LC-MS/MS analysis. A total of 327 proteins including ApoA-I, SOD1, and DCD were identified in GCF. These results may serve as reference for future proteomic studies searching for GCF biomarkers of periodontal diseases.

Original languageEnglish
Pages (from-to)2190-2202
Number of pages13
JournalProteomics
Volume12
Issue number13
DOIs
Publication statusPublished - Jul 2012

Keywords

  • Biomedicine
  • Gingival crevicular fluid
  • Periodontal disease

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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