Quantitative gingival crevicular fluid proteome in health and periodontal disease using stable isotope chemistries and mass spectrometry

Abstract

Aim Application of quantitative stable-isotope-labeling chemistries and mass spectrometry (MS) to determine alterations in gingival crevicular fluid (GCF) proteome in periodontal disease. Materials and Methods Quantitative proteome of GCF from 40 healthy individuals versus 40 patients with periodontal disease was established using 320 GCF samples and stable-isotope-labeling reagents, ICAT and mTRAQ, with MS technology and validated by enzyme-linked immunosorbent methods. Results We have identified 238 distinct proteins of which 180 were quantified in GCF of both healthy and periodontal patients with additional 26 and 32 distinct proteins that were found only in GCF of healthy or periodontal patients. In addition, 42 pathogenic bacterial proteins and 11 yeast proteins were quantified. The data highlighted a series of proteins not quantified previously by large-scale MS approaches in GCF with relevance to periodontal disease, such as host derived Ig alpha-2 chain C, Kallikrein-4, S100-A9, transmembrane proteinase 13, peptidase S1 domain, several collagen types and pathogenic bacterial proteins e.g., formamidase, leucine amidopeptidase and virulence factor OMP85. Conclusions The innovative analytical approaches provided detailed novel changes in both host and microbial derived GCF proteomes of periodontal patients. The study defined 50 host and 16 pathogenic bacterial proteins significantly elevated in periodontal disease most of which were novel with significant potential for application in the clinical arena of periodontal disease.

Influence of storage conditions on MALDI‐TOF MS profiling of gingival crevicular fluid: Implications on the role of S100A8 and S100A9 for clinical and proteomic based diagnostic investigations

“…This was the first study which explored and preliminarily demonstrated the potential use of MALDI‐TOF MS technology in the field of periodontal disease diagnostics. The recent evidence for GCF diagnostic capabilities are also highlighted and corroborated by the numerous proteomics studies in the last decade . The attractiveness of GCF as a diagnostic tool, however, must pose important critical issues, among which the requirement of the proteome stability for this oral fluid in such comparative proteomic studies.…”

Fetuin‐A, serum amyloid A and tumor necrosis factor alpha levels in periodontal health and disease

“…GCF is a serum exudate found in the gingival sulcus that can be readily collected in multiple sites within the oral cavity simultaneously (McCulloch, ; Kinney et al , ). In GCF, the inflammation‐related molecules may give dependable information pertaining to the condition of PD activity and systemic diseases (Offenbacher et al , ; Buduneli and Kinane, ; Carneiro et al , ). Further, serum supplies information about periodontal pathogen‐induced inflammation and response (Buduneli et al , ).…”

Novel isotopic N,N-Dimethyl Leucine (iDiLeu) Reagents Enable Absolute Quantification of Peptides and Proteins Using a Standard Curve Approach

“…This feature avoids the quantitative suppression that sometimes occurs in reporter ion quantification due to co-isolated precursor ions. Some groups have also used mTRAQ for candidate discovery in relative quantification studies [20–22], but one convincing report showed that while mTRAQ quantification is more accurate than AB SCIEX's isobaric label, iTRAQ, it also has a greater standard deviation [23]. Furthermore, multiplexing samples with the triplex mTRAQ kit creates samples with three times as many precursor ions to select for MS 2 events.…”