Proteomic Analysis of Gingival Crevicular Fluid During Tooth Eruption

Abstract

Objective: Gingival crevicular fluid (GCF) is a biological fluid that has the unique capacity to reflect changes in periodontium to its protein composition, making it ideal for potential biomarkers. There is limited information about the mechanism of tooth eruption, for which GCF might provide valuable knowledge. This study aimed to provide a proteomic approach to investigate the composition of GCF obtained from two different supraosseous tooth eruption stages of permanent molars changes. Methods: GCF samples were taken from a total of 26 healthy children, whose permanent molar just emerged from the gingiva (5-8 years old) and the occlusal equilibrium stage (9-13 years old). Proteins were extracted with Bio-Rad Rehydration Buffer followed by ZebaTM Spin Desalting Column. GCF samples were separated with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) followed by mass spectrometry- based protein identification. Results: A new optimized protocol with enriched protein extraction from GCF samples was developed. Several proteins related to tooth eruption were detected. It was determined that keratin type II cytoskeletal 4 and keratin type I cytoskeletal 9 decreased and albumin increased in GCF protein content in erupting teeth compared to those in the occlusal equilibrium phase. Conclusion: This methodology, which we have applied for 2D-PAGE of GCF, can also be a source for other studies. There is huge diagnostic potential in mass spectrometry technologies, this study can be carried forward by using other approaches.