Extracellular vesicles (EVs), detectable in all bodily fluids, mediate intercellular communication by transporting molecules between cells. The capacity of EVs to transport molecules between distant organs has drawn interest for clinical applications in diagnostics and therapeutics. Although EVs hold potential for nucleic-acid-based and other molecular therapeutics, the lack of standardized technologies, including isolation, characterization, and storage, leaves many challenges for clinical applications, potentially resulting in misinterpretation of crucial findings. Previously, several groups demonstrated the problems of commonly used storage methods that distort EV integrity. This work aims to evaluate the process to optimize the storage conditions of EVs and then characterize them according to the experimental conditions and the models used previously. Our study reports a highly efficient EV storage condition, focusing on EV capacity to protect their molecular cargo from biological, chemical, and mechanical damage. Compared with commonly used EV storage conditions, our EV storage buffer leads to less size and particle number variation at both 4 °C and −80 °C, enhancing the ability to protect EVs while maintaining targeting functionality.

Extracellular vesicles (EVs), detectable in all bodily fluids, mediate intercellular communication by transporting molecules between cells. The capacity of EVs to transport molecules between distant organs has drawn interest for clinical applications in diagnostics and therapeutics. Although EVs hold potential for nucleic acid-based and other molecular therapeutics, the lack of standardized technologies, including isolation, characterization, and storage, leaves many challenges for clinical applications, potentially resulting in misinterpretation of crucial findings. Previously, several groups demonstrated the problems of commonly used storage methods that distort EV integrity. This work aims to evaluate the process to optimize the storage conditions of EVs and then characterize them according to the experimental conditions and the models used previously. Our study reports a highly efficient EV storage condition, focusing on EVs’ capacity to protect their molecular cargo from biological, chemical, and mechanical damage. Compared with commonly used EV storage conditions, our EV storage buffer leads to less size and particle number variation at both 4°C and -80 °C, enhancing the ability to protect EVs while maintaining targeting functionality.

Impact de l'inflammation parodontale sur l'axe splénique cardiaque : étude des Neutrophils Extracellular Traps (NETs) et des microvésicules spléniques comme médiateurs des réponses procoagulantes et inflammatoires: P.gingivalis (PG) induit la parodontite et est associée au risque cardiovasculaire. Les microvésicules (MV) sont des effecteurs pro-inflammatoires et coagulants libérés des membranes plasmiques. L’effet à distance de PG sur le cœur et la rate de souris est détecté par immuno-marquage, mesure de ROS, et émission de MVs splénocytaires (SMVs). Les lésions gingivales par fils (LIG) imbibés ou non de PG (LIG-PG) sont comparées au gavage (gvg) et à l'injection intrapéritonéale (IP). LIG et LIG-PG réduisent les granulocytes, doublent les monocytes et les neutrophiles spléniques, sans réponse adaptative. IP triple les monocytes, multiplie par 10 les neutrophiles, réduit par 2 lymphocytes T ou B. L'inhibiteur de PAD-4 réduit les ROS, suggérant une Nétose. In vitro, les SMVs de LIG-PG, IP et gvg induisent la sénescence endothéliale. Dans le cœur, VCAM, Gasdermin-D, IL-6 et TNF- inflammatoires sont surexprimés pour IP et LIG-PG.En conclusion, PG induit l’activation splénique et cardiaque à distance, l'émission de SMVs pro-coagulantes et -sénescentes favorisant le risque cardiovasculaire.