Abstract
Background: Anesthetic-induced preconditioning (AIP) with volatile anesthetics is a well-known experimental technique to protect tissues from ischemic injury or oxidative stress. Additionally, plasmatic extracellular vesicle (EV) populations and their cargo are known to be affected by AIP in vitro, and to provide organ protective properties via their cargo. We investigated whether AIP would affect the generation of EVs in an in vivo rat model. Methods: Twenty male Sprague Dawley rats received a repetitive treatment with either isoflurane or with sevoflurane for a duration of 4 or 8 weeks. EVs from blood plasma were characterized by nanoparticle tracking analysis, transmission electron microscopy (TEM) and Western blot. A scratch assay (H9C2 cardiomyoblast cell line) was performed to investigate the protective capabilities of the isolated EVs. Results: TEM images as well as Western blot analysis indicated that EVs were successfully isolated. The AIP changed the flotillin and CD63 expression on the EV surface, but not the EV concentration. The scratch assay did not show increased cell migration and/or proliferation after EV treatment. Conclusion: AIP in rats changed the cargo of EVs but had no effect on EV concentration or cell migration/proliferation. Future studies are needed to investigate the cargo on a miRNA level and to investigate the properties of these EVs in additional functional experiments.
Highlights
Preconditioning is a commonly used experimental method to protect organs and tissues from injury, related to hypoxic or ischemic insults [1]
Volatile anesthetics have an effect on the composition of extracellular vesicle (EV) derived from numerous cell types such as cardiomyocytes and fibroblasts [3]
No differences in the concentration or the appearance of EVs between the experimental groups were shown (Figure 1). Since this method is not well suited for characterizing the size and concertation of EVs on a larger scale, these characteristics were investigated by Nanoparticle Tracking Analysis (NTA)
Summary
Preconditioning is a commonly used experimental method to protect organs and tissues from injury, related to hypoxic or ischemic insults [1]. A variety of experimental in vitro and in vivo models target the heart and the brain as the organs of interest in different animal- or cell-culture AIP or IPC models. In vitro AIP or IPC models are characterized as repetitive incubation cycles of cardiomyocyte cell cultures; for instance, with volatile anesthetics [3] or noble gases [4]. Anesthetic-induced preconditioning (AIP) with volatile anesthetics is a well-known experimental technique to protect tissues from ischemic injury or oxidative stress. Conclusion: AIP in rats changed the cargo of EVs but had no effect on EV concentration or cell migration/proliferation. Future studies are needed to investigate the cargo on a miRNA level and to investigate the properties of these EVs in additional functional experiments
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