Human Gingival Crevicular Fluid Contains MRP8 (S100A8) and MRP14 (S100A9), Two Calcium-binding Proteins of the S100 Family

Abstract

Human gingival crevicular fluid contains unidentified proteins which might play a role as markers in periodontal diseases. Therefore, low-molecular-weight proteins found in human gingival crevicular fluid (GCF), but absent from serum, were identified in the present study by means of two-dimensional electrophoresis (2-D PAGE) analysis. GCF, serum, and whole saliva were collected from periodontitis and healthy subjects, as well as from edentulous and newborn subjects. Protein samples were separated by two-dimensional polyacrylamide gel electrophoresis, stained with silver, and compared with reference protein maps in the SWISS-2D PAGE database. In GCF and saliva from periodontitis patients and healthy subjects, four dominant low-molecular-mass (from 8 to 14 kDa) acidic spots were observed. They were not found in serum and were less visible in saliva from edentulous and newborn subjects. From N-terminal amino acid sequencing, the two 2-D protein spots of 8 kDa and isoelectric points between 6.5 and 7.0 were both identified as protein MRP8 (SI00A8), a member of the S100 family of calcium-binding proteins. Using peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), we identified the other two protein spots, with mass of 14 kDa and isoelectric points between 5.5 and 6.0, as protein MRP14 (S100A9), also belonging to the S100 family. The presence of MRP8 and MRP14 in GCF was confirmed by Western blot, with monoclonal antibodies. The two polypeptides, MRP8 and MRP14, identified in GCF represent the major difference between the 2-D PAGE patterns of serum and GCF, and we hypothesize that they may play an important role in the gingival sulcus and could represent possible markers for periodontal diseases.

Gingival Crevicular Fluid Calprotectin, Osteocalcin and Cross-Linked N-Terminal Telopeptid Levels in Health and Different Periodontal Diseases

“…The present data indicated that GCF calprotectin levels increased as the periodontal disease progresses from health to disease, but similar in G-AgP and CP. Our findings are consistent with the previous data showing the role of calprotectin as an inflammatory marker of periodontal disease [11,12,42,43]. Kaner et al [44] have demonstrated that GCF calprotectin levels are significantly correlated with the levels of periodontal inflammation and decreased after periodontal therapy in G-AgP patients.…”

“…It has been previously shown that calprotectin was present in human dental calculus and suggested that it might be derived from gingival crevicular fluid (GCF), saliva and dental plaque [10]. Studies have shown that calprotectin level in GCF from periodontitis patients is higher than that from healthy subjects and positively correlated with clinical and biochemical markers of periodontal inflammation [11][12][13][14].…”

Molar Incisor Hypomineralization: Morphological, Aetiological, Epidemiological and Clinical Considerations

“…(Farah et al, 2010a) In severe cases, the defective enamel is lost shortly after molar eruption, exposing underlying dentine favoring the tooth sensitivity and the dental carious lesion. (Kilpatrick, 2009) Lately, MIH is understood as a hypocalcified subtype of enamel defect with reduced mineral content, low residual content of amelogenins and the presence of more than 16 types of proteins in affected teeth, thirteen of which are found in saliva and crevicular fluid (Kojima et al, 2000, Denny et al, 2008 and the three others (hemoglobin, albumin, complement C3) are major components of blood. Moreover, protein composition of MIH enamel varies with severity of enamel defect.…”

Blockade of S100A8 and S100A9 Suppresses Neutrophil Migration in Response to Lipopolysaccharide

“…They are also expressed at very high levels in the synovial fluid and plasma of patients suffering from rheumatoid arthritis and gout (C. Ryckman and P. A. Tessier, unpublished observations). Local secretion of the proteins has also been detected in periodontal infections and in experimental murine abscesses (14,15). Recent results obtained in our laboratory demonstrate that S100A8 and S100A9 are also important for neutrophil migration in a murine model of acute gouty arthritis (C. Ryckman and P. A. Tessier, unpublished observations).…”