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Clinical Oral Investigations

, Volume 22, Issue 3, pp 1375–1384 | Cite as

A new albumin-depletion strategy improves proteomic research of gingival crevicular fluid from periodontitis patients

  • Sarah Batschkus
  • Goekhan Cingoez
  • Henning Urlaub
  • Nicolai Miosge
  • Christian Kirschneck
  • Philipp Meyer-Marcotty
  • Christof Lenz
Original Article
  • 213 Downloads

Abstract

Objectives

Gingival crevicular fluid (GCF), the inflammatory infiltrate within the crevicular sulcus, is of great importance for diverse processes in the oral cavity and has a high impact in oral sciences. It is assumed to serve as a source of biomarkers for systemic or periodontal diseases and mediators of orthodontic tooth movement. In order to characterize the protein content of the GCF in an unbiased and complete approach, we employed mass spectrometry (MS), which allows not only the identification, but also the quantification of these proteins. In samples obtained from patients suffering from periodontitis, this method is often limited due to the presence of highly abundant serum albumin deriving from serum. The aim of this investigation was to employ a protein precipitation procedure for the efficient depletion of serum albumin from GCF samples.

Materials and methods

GFC samples collected from five adult periodontitis patients were fractionated by trichloroacetic acid/acetone precipitation and the resulting soluble and pelleted fractions were analyzed by SDS-PAGE and high-resolution mass spectrometry.

Results

Trichloroacetic acid/acetone precipitation was successfully employed as a protein precipitation procedure for the efficient depletion of serum albumin from GCF samples. Careful analysis revealed that the precipitation step reduced the serum albumin content efficiently, and increased subsequent protein identifications by 32%. Three hundred seventeen proteins could only be identified with this new approach.

Conclusion

The increased coverage of the GCF proteome will help improve our understanding of molecular mechanisms in the periodontium during pathogenesis of periodontitis.

Clinical relevance

Our new albumin depletion strategy combined with high-resolution mass spectrometry can be used to effectively monitor the molecular signals of the periodontium.

Keywords

Gingival crevicular fluid Mass spectrometry Serum albumin Periodontitis Orthodontic tooth movement 

Notes

Funding

There was no grant support or funding for this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was by the responsible ethics committee (University of Goettingen) with protocol number 23/7/15.

Informed consent

Informed consent was obtained from all individual participants included in this study.

Supplementary material

784_2017_2213_MOESM1_ESM.pdf (1.3 mb)
Supplementary Table 1 (PDF 1323 kb)
784_2017_2213_MOESM2_ESM.pdf (1.2 mb)
Supplementary Table 2 (PDF 1246 kb)

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Department for Orthodontics, Center for Dentistry, Oral Medicine and Craniomaxillofacial Surgery, University Medical CenterGeorg August UniversityGöttingenGermany
  2. 2.Max Planck Institute for Biophysical Chemistry, Bioanalytical Mass Spectrometry GroupGoettingenGermany
  3. 3.Institute of Clinical Chemistry, Bioanalytics GroupUniversity Medical Center GoettingenGoettingenGermany
  4. 4.Tissue Regeneration Group, Department of ProsthodonticsUniversity Medical Center GoettingenGoettingenGermany
  5. 5.Department of OrthodonticsUniversity Medical Center RegensburgRegensburgGermany

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