Role of extracellular vesicles on inflammatory process related with atherosclerosis
Our research is focused on the evaluation of circulating extracellular vesicles in proinflammatory processes related to metabolic diseases such as obesity and diabetes, and their implication with atherosclerosis development. We characterize and analyze the impact of circulating extracellular vesicles isolated from the pathological condition on the function of different cell types and tissues. One part of our aim is the characterization of these extracellular vesicles to induce the activation and polarization of macrophages to proinflammatory subtypes. Also, we are interested in the evaluation of the impact on vascular function of extracellular vesicles subtypes isolated from monocytes and macrophages after exposure to a different proinflammatory stimulus. We also use different knock-out murine models to evaluate their impact on atherosclerosis development and the inflammatory process related to it.
Animal and cellular models
KO models for atherosclerosis (ApoE KO) and sterile inflammation (NLRP3 KO).
Human and murine vascular cells (endothelial and smooth muscle cells), monocyte and macrophage cell lines, and others.
Technics and methods:
Isolation of different subtypes of extracellular vesicles (both, large- and small-EVs) from human and murine plasma, cell culture supernatant, and cardiovascular tissues by high-speed centrifugations.
EV characterization and numeration using flow-cytometry and Nano Tracking Analysis.
Cell origin characterization of large-EVs by flow-cytometer.
EV-specific markers expression analysis by western-blot.
Isolation and culture of CD14-positive isolated cells from peripheral blood mononuclear cells.
Macrophage polarization and phenotype characterization by flow cytometry.