NanoBioAnalytical (NBA) Platform to Decipher Extracellular Vesicles Secreted by Microvascular Endothelial Cells Under...

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Toxicology

Background:

• Extracellular vesicles (EVs) show promise as biomarkers for disease and therapeutics.
• Characterizing EV surface and cargo proteins for phenotyping is challenging.
• Benzo[a]pyrene (B[a]P), a toxic polycyclic aromatic hydrocarbon, can affect cellular function.

Purpose of the Study:

• To characterize EVs from human microvascular endothelial cells (HMEC-1) exposed to B[a]P.
• To develop and apply a multiplex biosensor for EV profiling.
• To investigate B[a]P-induced phenotypic and morphological changes in EVs.

Main Methods:

• A multiplex biosensor in µarray format was developed.
• The biosensor combined surface plasmon resonance imaging (SPRi) and in situ atomic force microscopy (AFM).
• EVs from B[a]P-treated and control HMEC-1 cells were analyzed using specific ligands.

Main Results:

• SPRi detected significant differences in EV capture with ligands like Anti-CD36, Anti-CD81, and Anti-ApoA between treated and control groups.
• AFM revealed size differences in EVs from B[a]P-exposed cells.
• Ligands such as Annexin-V, Anti-CD36, and Anti-VEGFR1 identified potential cytotoxicity biomarkers.

Conclusions:

• B[a]P exposure may increase EV size and alter marker expression, indicating cytotoxic stress.
• The combined SPRi-AFM approach provides insights into EV subset heterogeneity under stress.
• EVs show potential as specific toxicological markers for cytotoxic insults.