Direct detection of nano-scale extracellular vesicles derived from inflammation-triggered endothelial cells using surface plasmon resonance

- Extracellular vesicles (EV) having an approximate size range between 30 and 300 nm were successfully isolated from inflammation-induced vascular endothelial cells to mimic cardiovascular disease.
- The count and biomarker content of EV derived from endothelial cells are governed by cellular stress.
- The membrane bounded-biomarker profile of EV was verified using ultrastructural immunogold labeling, ELISA and SPR techniques.
- An unique procedure was established to isolate and to culture ex vivo vascular endothelial cells (VEC) from patients suffering from coronary heart disease (CHD) and from control persons having catheterization or vascular assessments for further EV studies.
- An SPR based method was optimized to discriminate EV derived from healthy and inflamed cells and for molecular profiling of EV-associated inflammation.
- This study is the first study completed with respect to develop an SPR platform to discover EV biomarkers in primary cells derived from patients with acute coronary heart disease (CHD).
- ICAM-1 and CD63 biomarkers have potential to discriminate between EV derived from healthy (hEV) and EV derived from inflammatory-triggered cell (tEV).

A major conceptual breakthrough in cell signaling has been the finding of EV as new biomarker shuttles in body fluids. Now, one of the major challenges in using these nanometer-sized biological entities as diagnostic marker is the development of translational methodologies to profile them. SPR offers a promising label-free and real time platform with a high potential for biomarker detection. Therefore, we aimed to develop a uniform SPR methodology to detect specific surface markers on EV derived from patient with CHD. EVs having an approximate size range between 30 and 100 nm (~48.5%) and 100-300 nm (~51.5%) were successfully isolated. The biomarker profile of EV was verified using immunogold labeling, ELISA and SPR. Using SPR, we demonstrated an increased binding of EV derived from patients with CHD to anti-ICAM-1 antibodies as compared to EV from healthy donors. Our current findings open up novel opportunities for in-depth and label-free investigation of EV.

Labeled and label free-based platforms to profile the nano-scale extracellular vesicles (EV) derived from inflammation-triggered endothelial cells and EV derived from patients with acute coronary heart disease (CHD).157