Background- Endometriosis is a debilitating gynaecological disease affecting at least 10% of menstruating people born with a uterus. Patients typically suffer for many years before diagnosis due to poor public and clinical awareness and unfortunately current treatments are often ineffective. Endometriosis is vastly under-researched and there is a great need to better understand its pathophysiology before more effective diagnostic and treatment methods can be developed. There have been limited studies on the role of extracellular vesicles (EVs) in endometriosis that meet the international guidelines for EV research.
Aims and methods- To optimise the isolation of EVs from the peritoneal fluid (PF) of patients being investigated for endometriosis by comparing three methods: a commercial size exclusion column (SEC), ultracentrifugation (UC), and a filtration device with a 20-nanometre pore (S20).
Results- From five PF samples, the SEC method yielded the highest concentration of nanoparticles (5.04 x 108 – 2.15 x 1010 particles/mL) and lowest concentration of co-isolated proteins (45.3 – 172.5 µg/mL) compared to the UC (9.15 x 107 – 1.74 x 1010 particles/mL; 44.0 – 599.6 µg/mL) or S20 methods (4.85 x 108 – 1.33 x 1010 particles/mL; 432.9 – 1723.1 µg/mL).
Future directions- EVs are being isolated from the PF of a further 50 endometriosis patients using the SEC method. These will be used to treat cell lines to determine their effects on proliferation, migration, invasion, and macrophage polarisation – typical endometriotic behaviours. Insight into the role of endometriotic EVs on cellular behaviour will provide unique understanding of endometriosis pathophysiology.