Angelika Silbereisen1, Saeed Alassiri2, Kai Bao1, Jonas Grossmann3, Paolo Nanni3, Claudia Fernandez3, Taina Tervahartiala2, Gustavo G Nascimento4, Georgios N Belibasakis1, Anna-Maria Heikkinen2, Rodrigo Lopez4, Timo Sorsa1,2, Nagihan Bostanci1. 1. Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden. 2. Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki University Central Hospital, P.O. Box 41 (Mannerheimintie 172), 00014, Helsinki, Finland. 3. Functional Genomics Center Zurich, ETH Zurich and University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland. 4. Section of Periodontology, Department of Dentistry and Oral Health, Aarhus University, Vennelyst Boulevard 9, 8000, Aarhus C, Denmark.
Abstract
PURPOSE: This study aims to validate label-free quantitative proteomics (LFQ) against antibody-based methods for quantifying established periodontal disease biomarkers in saliva. EXPERIMENTAL DESIGN: In an experimental gingivitis model, healthy volunteers (n = 10) provide saliva at baseline (d0), during the induction (d7, d14, d21) and resolution (d35) of gingival inflammation (total n = 50). Biomarker levels are analyzed by LFQ and time-resolved immunofluorometric assay (IFMA) or enzyme-linked immunosorbent assay (ELISA). Molecular matrix metalloproteinase (MMP)-8 forms are assessed by Western blot (WB) analysis. RESULTS: LFQ detects significantly (p < 0.05) elevated MMP-8 (d21vsd7, d35vsd7) and tissue inhibitor of matrix metalloproteinases (TIMP)-1 (d35vsd7). Latent MMP-8 (70-80 kDa) is present (d0-d35), but not active MMP-8 (50-60 kDa). LFQ and immunoassay data significantly correlate for MMP-8 (r = 0.36), myeloperoxidase (r = 0.39), polymorphonuclear leukocyte elastase (r = 0.33), and TIMP-1 (r = -0.24). CONCLUSION AND CLINICAL RELEVANCE: LFQ can quantify enzyme levels in saliva, however lacks the ability to measure enzymatic activity. WB analysis reveals that MMP-8 may not be activated during induction of gingival inflammation. Significant but weak correlations between IFMA or ELISA and LFQ suggest a limited capacity of available antibodies to reliably quantify salivary biomarkers for periodontal diseases. Novel "anti-peptide" antibodies designed by newer targeted mass spectrometry-based approaches can help to overcome these drawbacks.
PURPOSE: This study aims to validate label-free quantitative proteomics (LFQ) against antibody-based methods for quantifying established periodontal disease biomarkers in saliva. EXPERIMENTAL DESIGN: In an experimental gingivitis model, healthy volunteers (n = 10) provide saliva at baseline (d0), during the induction (d7, d14, d21) and resolution (d35) of gingival inflammation (total n = 50). Biomarker levels are analyzed by LFQ and time-resolved immunofluorometric assay (IFMA) or enzyme-linked immunosorbent assay (ELISA). Molecular matrix metalloproteinase (MMP)-8 forms are assessed by Western blot (WB) analysis. RESULTS: LFQ detects significantly (p < 0.05) elevated MMP-8 (d21vsd7, d35vsd7) and tissue inhibitor of matrix metalloproteinases (TIMP)-1 (d35vsd7). Latent MMP-8 (70-80 kDa) is present (d0-d35), but not active MMP-8 (50-60 kDa). LFQ and immunoassay data significantly correlate for MMP-8 (r = 0.36), myeloperoxidase (r = 0.39), polymorphonuclear leukocyte elastase (r = 0.33), and TIMP-1 (r = -0.24). CONCLUSION AND CLINICAL RELEVANCE: LFQ can quantify enzyme levels in saliva, however lacks the ability to measure enzymatic activity. WB analysis reveals that MMP-8 may not be activated during induction of gingival inflammation. Significant but weak correlations between IFMA or ELISA and LFQ suggest a limited capacity of available antibodies to reliably quantify salivary biomarkers for periodontal diseases. Novel "anti-peptide" antibodies designed by newer targeted mass spectrometry-based approaches can help to overcome these drawbacks.
Authors: Kehinde Adesola Umeizudike; Hanna Lähteenmäki; Ismo T Räisänen; John J Taylor; Philip M Preshaw; Susan M Bissett; Taina Tervahartiala; Solomon O Nwhator; Pirjo Pärnänen; Timo Sorsa Journal: J Periodontal Res Date: 2022-03-19 Impact factor: 3.946
Authors: Anna M Heikkinen; Teija Raivisto; Ismo Räisänen; Taina Tervahartiala; Nagihan Bostanci; Timo Sorsa Journal: Clin Exp Dent Res Date: 2022-06-08
Authors: Pune N Paqué; Christopher Herz; Daniel B Wiedemeier; Konstantinos Mitsakakis; Thomas Attin; Kai Bao; Georgios N Belibasakis; John P Hays; Joël S Jenzer; Wendy E Kaman; Michal Karpíšek; Philipp Körner; Johannes R Peham; Patrick R Schmidlin; Thomas Thurnheer; Florian J Wegehaupt; Nagihan Bostanci Journal: J Pers Med Date: 2021-03-23