DISACCHARIDE ANALYSIS OF CHONDROITIN SULFATE IN HUMAN GINGIVAL CREVICULAR FLUID USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

Gingival crevicular fluid (GCF) was collected into capillary tubes fro m healthy gingiva and sites of advanced periodontitis. Following diges tion with Pronase E, the glycosaminoglycans were isolated by successiv e precipitation into 5% cetylpyridinium chloride and 95% ethanol. Unsa turated disaccharide isomers of chondroitin sulphate, obtained followi ng chondroitinase ACII digestion, were analysed by high-performance li quid chromatography. Chondroitin sulphate was found in all GCF samples , with greater amounts in patients with periodontal disease than at co ntrol sites with a relatively healthy periodontium. The predominant is omer in the periodontal diseased group was Delta Di-4S, while that in the control group and serum samples was Delta Di-0S. Comparison of the relative proportions of the unsaturated disaccharides in GCF with pre viously reported values for alveolar bone, cementum, gingiva and perio dontal ligament, as well as for serum, indicates that the chondroitin sulphate present in GCF of patients with periodontal disease originate d from the mineralized connective tissues of the periodontium, notably alveolar bone, possibly with some contributions from soft connective tissues of gingiva and periodontal ligament and from serum.