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 fluids samples were obtained. EVs were precipitated by ExoQuick-TC and characterized by nanoparticle tracking analysis (Nanosight), western-blot (WB), transmission electron microscopy (TEM) and ELISA. RESULTS: TEM showed nanoparticles morphologically compatible with EVs and WB analysis revealed bands of specific EVs markers (CD9, TSG101, 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=0.017). However, we did not observe differences in the EVs concentration of saliva samples (p=0.190). The size of GCF-EVs was 144.2nm in periodontitis and 160.35nm in healthy/gingivitis patients (p=0.038). The CD63 exosome marker was increased in GCF of periodontitis patients (p=0.00001). The total concentration of EVs in GCF was correlated with bleeding on probing (rho=0.63, p=0.002), periodontal probing depth (rho=0.56, p=0.009) and clinical attachment level (rho=0.48, p=0.030). CONCLUSION: periodontitis patients have increased concentration of EVs in GCF and their role in periodontitis should be clarified.