OBJECTIVE: Cell secretomes represent a promising strategy for periodontal and bone regeneration. The objective of this study was to characterize the secretome of human gingival fibroblasts (GF) from periodontally diseased tissues (GF-perio) using proteomics. MATERIALS AND METHODS: Conditioned media of GF-perio from periodontitis patients (n = 6, 48-h serum-free culture) were subjected to liquid chromatography with tandem mass spectrometry. Global profiles, differentially expressed proteins (DEPs), and functional/gene-set enrichment (FEA) were analyzed using bioinformatics. Selected bone regeneration-related proteins were additionally measured using a multiplex immunoassay. Conditioned media of GF from periodontally healthy subjects were used as a reference. RESULTS: Overall, 1833 proteins were detected in GF-perio secretomes, including several growth factors, cytokines, chemokines, and extracellular matrix proteins important for wound healing and regeneration. Key bone-related cytokines (FGF2, MCP1, GPNMB, MMP2, IL6, IL8) were confirmed by an immunoassay. Compared to the reference group, 127 exclusive proteins and 73 DEPs (p <
0.05) were identified in the GF-perio group. FEA revealed significant enrichment of "exosome" and "cytoplasm" related cellular components in GF-perio secretomes. CONCLUSION: The secretome of GF from periodontally diseased tissues may hold therapeutic potential, with several proteins important for wound healing and regeneration, especially those related to exosome functions.