PuSH - Publication Server of Helmholtz Zentrum München: Engineered versus hybrid cellular vesicles as efficient drug delivery systems: A comparative study with brain targeted vesicles.

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Kannavou, M.* ; Marazioti, A.* ; Stathopoulos, G.T. ; Antimisiaris, S.G.*

Engineered versus hybrid cellular vesicles as efficient drug delivery systems: A comparative study with brain targeted vesicles.

Drug Deliv. Transl. Res. 11, 547-565 (2021)

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Herein we elaborated on methods to load cellular vesicles (CVs) and to incorporate cholesterol (Chol) and PEG lipids in their membrane, for enhancing the potential of such engineered CVs (e-CVs) as drug carriers. Hybrids formed by fusion between PEGylated liposomes (PEG-LIP) and CVs were evaluated as alternatives to e-CV, for the first time. Freeze-thawing cycles (FT) and incubation protocols were tested, and vesicle fusion was monitored by FRET dilution. B16F10, hCMEC/D3, and LLC cells were used for e-CV or hybrid development, and FITC-dextran as a model hydrophilic drug. Results show that dehydration rehydration vesicle (DRV) method is optimal for highest CV loading and integrity, while optimal protocols for Chol/PEG enrichment were identified. FT was found to be more efficient than incubation for hybrid formation. Interestingly, despite their high Chol content, CVs had very low integrity that was not increased by enrichment with Chol, but only after PEG coating; e-CVs demonstrated higher integrity than hybrids. Vesicle uptake by hCMEC cells is in the order: LIP < e-CVs < Hybrids ≤ CVs (verified by confocal microscopy); the higher PEG content of e-CVs is possibly the reason for their reduced cell uptake. While CV and hybrid uptake are highly caveolin-dependent, e-CVs mostly follow clathrin-dependent pathways. In vivo and ex vivo results show that brain accumulation of hybrids is only slightly higher that of CVs, indicating that the surface PEG content of hybrids is not sufficient to prevent uptake by macrophages of the reticuloendothelial system. Taking together with the fact that subjection of CVs to FT cycles reduced their cellular uptake, it is concluded that PEGylated e-CVs are better than hybrids as brain-targeted drug carriers.

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