Comparative Analysis for Glycosaminoglycan (GAG) Level in Biological Samples by Three Different Assay Methods

• Published in: Nihon Shishubyo Gakkai Kaishi (Journal of the Japanese Society of Periodontology) Vol. 40; no. 3; pp. 340 - 349
• Main Authors: Kou, Yoshitada, Kawabata, Yoshikatsu, Shiraki, Masafumi, Iwayama, Yukio
• Format: Journal Article
• Language: Japanese
• Published: JAPANESE SOCIETY OF PERIODONTOLOGY 1998
• Subjects: Electrophoresis; Enzyme-linked immunosorbent assay; Glycosaminoglycan; High-performance liquid chromatography; Proteoglycan
• ISSN: 0385-0110; 1880-408X
• DOI: 10.2329/perio.40.3_340

Glycosaminoglycans (GAG) have been found in higher level in the gingival crevicular fluid associated with increased metabolic activity, such as periodontal inflammation or alveolar bone remodeling, and have also been proposed as a maker of periodontal disease activity. There appears, however, to be some confusion about the different levels of GAG depending or the assay method used. To compare three assay methods for GAG and glycosaminoglycan-linked proteoglycans (GAGPG) in biological samples, we analyzed three samples: from gingival homogenate supernatant, synovial fluid, and carrageenin-induced exdate. The isolated GAG were separated by electrophoresis on a cellulose acetate membrane and stained with Alcian blue. Substrate-dye complex was evaluated in a scanning densitometer. Unsaturated disaccharide isomers of chondroitin sulfate, ob taineol following chondroitinase digestion from isolated GAG, were analyzed by high-performance liquid chromatography. For comparison, a small amount of GAG-PG was assayed using the enzymelinked immunosorbent assay (ELISA) method, with a combination of monoclonal antibodies and specific enzyme digestion. Results were that the level of GAG obtained by the electrophoretic method was similar to that obtained by high-performance liquid chromatographic assay, while the GAG-PG level obtained using the ELISA method was 3-80 times higher than that by the other two methods. Dermatan sulfate was not detected by biochemical analysis in the samples from synovial fluid and carrageenin-induced exudate. This indicates that an assay of GAG-PG dose not necessarily substitute for GAG assay by biochemical methods. These findings may be useful in selecting the assay methods for GAG and help discuss ions of the results for biological samples by different assay methods.