Analysis of hyaluronic acid in human gingival crevicular fluid using high-performance liquid chromatography

We determined the hyaluronic acid disaccharides, delta Di-HA, in the gingival crevicular fluid (GCF) and whole saliva of patients with periodontal disease, and in the peri-implant sulcus fluid (PISF) from sites around titanium osseointegrated implants, and compared these values with those in the GCF and whole saliva of controls. We also determined values for chondroitin sulfate disaccharide isomers at the same time. Glycosaminoglycans were extracted by digestion with Pronase E, followed by digestion of GAGs with hyaluronidase SD and chondroitinase ACII. Unsaturated disaccharide isomers produced from hyaluronic acid and chondroitin sulfate were analyzed by high-performance liquid chromatography (HPLC). The hyaluronic acid disaccharide delta Di-HA was found in all samples of GCF, PISF and whole saliva. The concentration of delta Di-HA in both GCF and whole saliva of the periodontitis group was greater than that in the controls. There was no difference in the concentration of delta Di-HA between the PISF and GCF of the controls. The ratios of hyaluronic acid to chondroitin sulfate in the GCF and in the whole saliva of the periodontitis group were significantly lower than that of the controls. There was no difference between the ratios in PISF and those in GCF of the controls. These results indicate that checking hyaluronic acid in GCF and whole saliva using HPLC is a useful means of assessing the condition of periodontal tissues, and that assaying hyaluronic acid in PISF may also be effective for monitoring the condition of tissues around dental implants.