Abstract
Qualitative and quantitative studies on proteins in gingival fluid (GF) have greatly contributed to a better understanding of its nature and significance.
In this study, microscale multisample two- dimensional (2-D) electrophoresis in the absence of denaturing agent developed by Manabe et al. (1982) was applied for analyzing the whole proteins present in GF from various periodontal diseases.
The samples collected from 28 individuals, males and females aged 18 to 69 years, were classified into four groups according to their clinical findings. GF was collected from anterior tooth and also serum and whole saliva were sampled for comparison. Silver staining was used for both GF and whole saliva and Coomassie-Blue dye was applied for serum.
Some gels were transferred to nitrocellulose sheets for identification of GF protein according to western blotting and these sheets were immuno chemically stained by using anti-human serum, antihuman S-IgA and anti-human whole saliva, respectively. Both nitrocellulose sheets and polyacrylamide gels were imaged by TV-camera and analyzed by microcomputer system.
The results indicated that the majority of protein fraction in GF were mainly originated from serum, and that unidentified fractions whose counterparts are not in serum were particularly observed in GF from severe periodontitis. Furthermore, free hemoglobin and haptoglobin-hemoglobin complex were often detected in severe periodontitis.
Consequently, it was revealed that microscale multisample 2-D electrophoresis would provide valuable informations for diagnosis of periodontal disease.
