Molecular signatures of extracellular vesicles in oral fluids of periodontitis patients

Objective: To characterize extracellular vesicles (EVs) in gingival crevicular fluid (GCF) and saliva samples from healthy/gingivitis and periodontitis patients and correlate them with clinical inflammatory periodontal parameters. Material and Method: An exploratory study, including 86 subjects, was conducted. Clinical and periodontal data were recorded, and oral fluid samples were obtained. EVs were precipitated by ExoQuick-TC™ and characterized by nanoparticle tracking (NanoSight™), Western blot (WB), transmission electron microscopy (TEM), and ELISA analysis. Results: TEM showed nanoparticles morphologically compatible with EVs, and WB analysis revealed bands of specific EV markers (CD9, TSG101, and Alix) in both oral fluids of periodontitis and healthy/gingivitis subjects. The total concentration of EVs in GCF was increased in periodontitis patients compared to healthy/gingivitis subjects (p =.017). However, we did not observe differences in the EV concentration of saliva samples (p =.190). The size of GCF-EVs was 144.2 nm in periodontitis and 160.35 nm in healthy/gingivitis patients (p =.038). The CD63 exosome marker was increased in GCF of periodontitis patients (p =.00001). The total concentration of EVs in GCF was correlated with bleeding on probing (rho = 0.63, p =.002), periodontal probing depth (rho = 0.56, p =.009), and clinical attachment level (rho = 0.48, p =.030). Conclusion: Periodontitis patients have an increased concentration of EVs in GCF, and their role in periodontitis should be clarified.