<jats:title>Abstract</jats:title><jats:sec><jats:title>Objective</jats:title><jats:p>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.</jats:p></jats:sec><jats:sec><jats:title>Material and Method</jats:title><jats:p>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.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>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 (<jats:italic>p = .017</jats:italic>). However, we did not observe differences in the EV concentration of saliva samples (<jats:italic>p = .190</jats:italic>). The size of GCF‐EVs was 144.2 nm in periodontitis and 160.35 nm in healthy/gingivitis patients (<jats:italic>p = .038</jats:italic>). The CD63 exosome marker was increased in GCF of periodontitis patients (<jats:italic>p</jats:italic> = <jats:italic>.00001</jats:italic>). The total concentration of EVs in GCF was correlated with bleeding on probing (rho = 0.63, <jats:italic>p = .002</jats:italic>), periodontal probing depth (rho = 0.56, <jats:italic>p = .009</jats:italic>), and clinical attachment level (rho = 0.48, <jats:italic>p = .030</jats:italic>).</jats:p></jats:sec><jats:sec><jats:title>Conclusion</jats:title><jats:p>Periodontitis patients have an increased concentration of EVs in GCF, and their role in periodontitis should be clarified.</jats:p></jats:sec>