Pressure Cycling Technology‐Assisted Mass Spectrometric Quantification of Gingival Tissue Reveals Proteome Dynamics During the Initiation and Progression of Inflammatory Periodontal Disease
Understanding the progression of periodontal tissue destruction is at the forefront of periodontal research. We aimed to capture the dynamics of gingival tissue proteome during the initiation and progression of experimental (ligature‐induced) periodontitis in mice. Pressure cycling technology (PCT), a recently developed platform that uses ultra‐high pressure to disrupt tissues, was utilized to achieve efficient and reproducible protein extraction from ultra‐small amounts of gingival tissues in combination with liquid chromatography‐tandem mass spectrometry (MS). The MS data were processed using Progenesis QI and the regulated proteins were subjected to METACORE, STRING, and WebGestalt for functional enrichment analysis. A total of 1614 proteins with ≥2 peptides were quantified with an estimated protein false discovery rate of 0.06%. Unsupervised clustering analysis showed that the gingival tissue protein abundance is mainly dependent on the periodontitis progression stage. Gene ontology enrichment analysis revealed an over‐representation in innate immune regulation (e.g., neutrophil‐mediated immunity and antimicrobial peptides), signal transduction (e.g., integrin signaling), and homeostasis processes (e.g., platelet activation and aggregation). In conclusion, we applied a PCT‐assisted label‐free quantitative proteomics workflow that allowed cataloging the deepest gingival tissue proteome on a rapid timescale and provided novel mechanistic insights into host perturbation during periodontitis progression.