ObjectiveTo 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 MethodAn 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.ResultsTEM 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).ConclusionPeriodontitis patients have an increased concentration of EVs in GCF, and their role in periodontitis should be clarified.