Journal of Oral Tissue Engineering

ORIGINAL ARTICLE
Measuring Sulfated Glycosaminoglycans in Gingival Crevicular Fluid Based on Dimethylmethylene Blue

Joji OKAZAKI1, Yutaka KOMASA1, Aiko KAMADA2, Rachel J. WADDINGTON3, Akiyo KAWAMOTO1, Ryoko KUSUNOKI4, Daigo SAKAI4, Norihiko KIMURA1, Takashi IKEO2, Ding-Ming HUANG5, Xue-Dong ZHOU5, Qin SU5 and Bing GUO6

1Department of Geriatric Dentistry, Osaka Dental University, Japan
2Department of Biochemistry, Osaka Dental University, Japan
3Dental Health and Biological Sciences, School of Dentistry, Cardiff University, UK
4Graduate School of Dentistry (Geriatric Dentistry), Osaka Dental University, Japan
5Department of Endodontics, West China College of Stomatology, Sichuan University, China
6Department of Geriatric Dentistry, West China College of Stomatology, Sichuan University, China


J Oral Tissue Engin 2006;4(1): 51-56

Full Text. DOI https://doi.org/10.11223/jarde.4.51

SYNOPSIS
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 (0-3µg/100 µl). 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 (0.05 µg/100 µl ± 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.

Key words: chondroitin sulfate, glycosaminoglycans, gingival crevicular fluid, dimethylmethylene blue