Rj. Waddington et al., IMMUNOCHEMICAL DETECTION OF THE PROTEOGLYCANS DECORIN AND BIGLYCAN INHUMAN GINGIVAL CREVICULAR FLUID FROM SITES OF ADVANCED PERIODONTITIS, Archives of oral biology, 43(4), 1998, pp. 287-295
This study characterized proteoglycan metabolites present in gingival
crevicular fluid (GCF) collected from sites with clinical evidence of
advanced periodontal disease. The metabolites were purified by anion-e
xchange chromatography from which a chondroitin sulphate rich fraction
was identified by cellulose acetate electrophoresis. Sodium dodecylsu
lphate-polyacrylamide gel electrophoresis of this fraction revealed a
broad silver-staining band with mel. wt 55-65 k and Western blotting s
uggested that this band was immunoreactive with CS-56, a monoclonal an
tibody for chondroitin sulphate. Digestion of the metabolite with chon
droitinase ABC (protease-free) led to the loss of the silver-staining
band. Dot-blot analysis identified components in this fraction that we
re immunoreactive for the monoclonal/polyclonal antibodies against the
C-termino of decorin and biglycan. Amino acid analysis revealed the c
omposition of the proteoglycan metabolite to be rich in glycine, serin
e and glutamic acid. Immunochemical and biochemical analyses were comp
ared with those of proteoglycan purified from human alveolar bone. Cha
nges in the amino acid composition were noted, suggesting the proteogl
ycan metabolite has undergone extensive modification and fragmentation
to the protein core. The results suggest that the proteoglycan metabo
lite from GCF represented a degradation product originating from the a
ctive destruction of the alveolar bone. They provide further support f
or the proposal that the appearance of proteoglycan metabolites in GCF
is a biomarker for active destruction of alveolar bone, the biochemic
al analysis of which provides important information on mechanisms invo
lved in the pathology of periodontal diseases. (C) 1998 Elsevier Scien
ce Ltd. All rights reserved.