Multivariate Spectroscopic Analysis of Protein Secondary Structures in Gingival Crevicular Fluid: Insights from FTIR Amide III Band Across Oral Disease Stages.

This study applies multivariate data analysis to deconvolute the spectral profiles of the Amide III region in the infrared spectra of gingival crevicular fluid (GCF). This reveals the impact of major oral diseases, such as dental caries and periodontal diseases, on the transformation of the secondary structure of GCF proteins. A two-stage analytical approach was employed: first, principal component analysis (PCA) was performed to establish the main factors of variation in the data, followed by pairwise comparisons of the samples based on the results of the Amide III profile deconvolution. The analysis also accounted for comorbidities, such as oncological and gastrointestinal diseases. This approach allowed for the identification of subtle differences in the composition and conformation of the secondary structure of GCF proteins while accounting for the superposition of multiple influencing factors. This methodology was effective in identifying biomarkers of oral diseases in GCF. For the first time, it has been demonstrated that the relative content of the β-sheet-associated component in the spectral profile of the secondary structure element of the protein fraction of GCF serves as a statistically significant marker for dental caries, regardless of the presence or absence of other diseases. Additionally, a significant decrease in the relative content of α-helix structures was observed in GCF from patients with oncological diseases. The changes in the spectral profile of the Amide III band of GCF identified in this study have not been previously detected using molecular spectroscopy, correlated with the secondary structure of proteins, or analyzed using multivariate analysis methods.