Abstract
Cellular vesicles (CVs) have been proposed as alternatives to exosomes for targeted drug delivery. CVs, prepared from HEK-293, B16F10 and hCMEC/D3 cells, by liposome technology methods were characterized for properties, morphology, cytotoxicity and cell uptake. CV brain-targeting potential was evaluated in vitro on the hCMEC/D3 BBB-model, and in vivo/ ex vivo. CV sizes were between 135-285 nm; zeta-potential was negative. The Dehydration Rehydration method conferred highest calcein loading and latency to CVs, compared to other methods. Increased calcein-leakage from CVs compared to liposomes, indicated their poor integrity, which was increased by pegylation. In vivo results confirmed lower liver uptake by PEG-CVs (compared to non-pegylated) proving that calcein integrity test is useful for prediction of CV-biodistribution, as used for liposomes. Cell-uptake of homologous-origin CVs was not always higher compared to that of non-homologous. Nevertheless, CVs from hCMEC/D3 demonstrated highest BBB permeability (in vitro), compared to OX-26 targeted liposomes, and brain localization (in vivo). CVs from hCMEC/D3 cells grown in different media demonstrated decreased interaction with brain cells and brain localization; significant differences in proteome of the two latter CV types were identified by proteomics, suggesting a potential methodology for identification of organotropism-determining CV-components
SIGNIFICANCE STATEMENT N/A
- blood-brain barrier
- cholesterol
- drug delivery
- drug targeting
- endothelial cells
- liposomal drug delivery
- nanoparticles
- permeability
- pharmacokinetics
- proteomics
- The American Society for Pharmacology and Experimental Therapeutics