Abstract

Background: Colostrum extracellular vesicles (C-EVs) are nanoscale, bioactive vesicles with therapeutic potential. The mechanisms of action include the control of cellular and tissue homeostasis. These make C-EVs a novel means to control inflammatory and cellular dysfunctions. However, a limitation for their broad use is the ease of C-EV isolation and in ensuring their stability. Methods: Standard ultracentrifugation and gradient techniques used for EV recovery were employed, which included ultracentrifugation. Exodus dual-frequency ultrasonic nanofiltration (UNF) was a comparator used to overcome standard limitations by recovering pure vesicles at high concentrations. Both systems were evaluated for their abilities to recover clinical-grade C-EVs with optimal vesicle structural integrity and intact biological functions. Results: This study affirms UNF C-EV recovery by demonstrating intact Alix, CD63, Tsg101, and Flotillin antigens. The EVs maintained an intact bilayer structure with sizes ranging from to 50–200 nm. Functional tests showed preservation of their anti-inflammatory activities by suppression of pro-inflammatory cytokines and the NLRP3 inflammasome, caspase 1, interleukin-1, and 18 and maintaining cellular homeostasis. Processing time, high yield, and functional responses controlled cellular function. Conclusions: These data support the notion that UNF C-EVs can be recovered safely, at high yields, and reproducibly for future clinical applications.

IPC Classification

Keywords