Measuring Sulfated Glycosaminoglycans in Gingival Crevicular Fluid Based on Dimethylmethylene Blue

概要

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The information about periodontal conditions is necessary for regeneration of periodontal tissues. The measurement of sulfated glycosaminoglycans (S-GAGs) in gingival crevicular fluid (GCF) is important in the search for disease biomarkers. To analyze the amount of S-GAGs in GCF quickly and accurately, we have modified a microplate method previously used for urinary S-GAGs to estimate the content of S-GAGs in GCF without interference of other GCF components. GCF samples were collected from six maxillary anterior teeth and diluted GCF samples were divided into two groups (20 μl each), with or without chondroitinase ABC treatment, mixed with a dimethylmethylene blue (DMB) dye solution (180 μl) to detect S-GAG. Absorbance at 540 nm was measured using a microplate reader and the amount of S-GAG was calculated from calibration curves for standard S-GAGs where a linear relationship existed in the concentration range (0-3 μg/100μl). Other GCF components such as glycoprotein and serum constituents did not influence the measurement of S-GAGs. Although trace amounts of S-GAGs were found in controls with a healthy periodontium (0.05μg/100μl ± 0.04, mean±S.E., n=5), considerable amounts of S-GAGs were found in patients with advanced periodontal disease (0.42 ± 0.11, n=6, p<0.05). The accuracy of this simple and rapid method of measuring S-GAGs in GCF makes it useful for screening periodontal conditions.
日本再生歯科医学会の論文
2006-09-30