Sarah Batschkus1, Goekhan Cingoez2, Henning Urlaub3,4, Nicolai Miosge5, Christian Kirschneck6, Philipp Meyer-Marcotty2, Christof Lenz3,4. 1. Department for Orthodontics, Center for Dentistry, Oral Medicine and Craniomaxillofacial Surgery, University Medical Center, Georg August University, Robert-Koch-Str. 40, 37075, Göttingen, Germany. sarah.batschkus@med.uni-goettingen.de. 2. Department for Orthodontics, Center for Dentistry, Oral Medicine and Craniomaxillofacial Surgery, University Medical Center, Georg August University, Robert-Koch-Str. 40, 37075, Göttingen, Germany. 3. Max Planck Institute for Biophysical Chemistry, Bioanalytical Mass Spectrometry Group, Am Fassberg 11, 37077, Goettingen, Germany. 4. Institute of Clinical Chemistry, Bioanalytics Group, University Medical Center Goettingen, Robert-Koch-Straße 40, 37075, Goettingen, Germany. 5. Tissue Regeneration Group, Department of Prosthodontics, University Medical Center Goettingen, Robert-Koch-Straße 40, 37075, Goettingen, Germany. 6. Department of Orthodontics, University Medical Center Regensburg, Franz-Josef-Strauß-Allee 11, 93053, Regensburg, Germany.
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.
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 periodontitispatients 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.
Entities:
Keywords:
Gingival crevicular fluid; Mass spectrometry; Orthodontic tooth movement; Periodontitis; Serum albumin
Authors: Ignat V Shilov; Sean L Seymour; Alpesh A Patel; Alex Loboda; Wilfred H Tang; Sean P Keating; Christie L Hunter; Lydia M Nuwaysir; Daniel A Schaeffer Journal: Mol Cell Proteomics Date: 2007-05-27 Impact factor: 5.911