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Cellular Vesicles: New Insights in Engineering Methods, Interaction with Cells and Potential for Brain Targeting
A Marazioti1, K Papadia1, M Kannavou1
1Foundation for Research and Technology Hellas, Institute of Chemical Engineering Sciences, FORTH/ICE-HT, Rio, Greece (A.M., M.K., A.B., S.G.A.); Laboratory of Pharmaceutical Technology, Department of Pharmacy (K.P., M.K., A.B., S.G.A.) and Laboratory for Molecular Respiratory Carcinogenesis, Department of Physiology, Faculty of Medicine (M.Sp., G.T.S.), University of Patras, Rio, Greece; Laboratory of Immunohematology, Division of Hematology, Department of Internal Medicine, Medical School, University of Patras, Patras, Greece (A.-L.d.L., M.R., A.M.); B.S.R.C. Alexander Fleming, Vari, Attica, Greece (M.Sa., G.P.); and Comprehensive Pneumology Center and Institute for Lung Biology and Disease, University Hospital, Ludwig-Maximilians University and Helmholtz Center Munich, Member of the German Center for Lung Research, Munich, Bavaria, Germany (G.T.S.).
Cellular vesicles (CVs) show promise for drug delivery, with hCMEC/D3-derived CVs demonstrating superior blood-brain barrier penetration and brain localization in vivo. Proteomics identified components influencing organotropism.
Area of Science:
- Biotechnology
- Nanomedicine
- Drug Delivery Systems
Background:
- Cellular vesicles (CVs) are emerging as potential alternatives to exosomes for targeted drug delivery.
- CVs can be engineered from various cell sources, including HEK-293, B16F10 melanoma, and hCMEC/D3 cells.
Purpose of the Study:
- To characterize CVs for morphology, cytotoxicity, and cell uptake.
- To evaluate the brain-targeting potential of CVs in vitro and in vivo.
- To investigate the impact of cell origin and culture conditions on CV properties and brain localization.
Main Methods:
- CVs were prepared using liposome technology methods and characterized by size, ζ-potential, and calcein loading/latency.
- Dehydration-rehydration method was optimized for calcein loading.
- In vitro blood-brain barrier (BBB) model (hCMEC/D3) and in vivo/ex vivo studies were used to assess brain targeting.
- Proteomics was employed to analyze CV composition.
Main Results:
- CVs ranged from 135-285 nm with negative ζ-potential.
- The dehydration-rehydration method yielded optimal calcein loading and latency.
- Pegylation improved CV integrity and reduced liver uptake, correlating with biodistribution predictions.
- hCMEC/D3-derived CVs exhibited the highest in vitro BBB permeability and in vivo brain localization.
- Different culture media for hCMEC/D3 cells affected CV interaction with brain cells and localization, linked to proteomic differences.
Conclusions:
- CVs derived from hCMEC/D3 cells show significant potential for brain-targeted drug delivery.
- CV integrity and biodistribution can be predicted using calcein leakage assays.
- Proteomic analysis offers a method to identify CV components responsible for organotropism, enabling targeted delivery optimization.
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