Liposomes functionalized to overcome the blood–brain barrier and to target amyloid-β peptide: the chemical design affects the permeability across an in vitro model
Elisa Salvati, Francesca Re, Silvia Sesana, Ilaria Cambianica, Giulio Sancini, Massimo Masserini, Maria GregoriDepartment of Health Sciences, University of Milano-Bicocca, Monza, ItalyPurpose: We investigated the ability of amyloid-β-targeting liposomes, decorated with an anti-transferrin r...
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| Autores principales: | , , , , , , |
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| Formato: | article |
| Lenguaje: | EN |
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Dove Medical Press
2013
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| Acceso en línea: | https://doaj.org/article/b5fa9f054e694b19b47660cb4d2cee87 |
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| Sumario: | Elisa Salvati, Francesca Re, Silvia Sesana, Ilaria Cambianica, Giulio Sancini, Massimo Masserini, Maria GregoriDepartment of Health Sciences, University of Milano-Bicocca, Monza, ItalyPurpose: We investigated the ability of amyloid-β-targeting liposomes, decorated with an anti-transferrin receptor antibody, to cross the blood–brain barrier (BBB), comparing two antibody ligation techniques.Methods: Fluorescent or radiolabeled liposomes composed of sphingomyelin/cholesterol and containing phosphatidic acid, known to bind amyloid-β, were further functionalized with the anti-transferrin receptor antibody RI7217. Two different techniques were used to attach RI7217 to the liposomes surface: biotin/streptavidin linkage or thiol–maleimide covalent ligation. Surface plasmon resonance (SPR) and immunoblotting were employed to assess the nanoparticles’ binding performances. Confocal microscopy and radiochemical techniques were used for uptake and permeability studies on an in vitro BBB model made of human brain capillary endothelial cells hCMEC/D3.Results: Immunoblotting experiments showed that RI7217-functionalized liposomes bind to transferrin receptor independently of the procedure employed to ligate their surface with the antibody, while SPR experiments showed a slightly higher affinity for covalently functionalized nanoliposomes. The functionalization with RI7217 did not affect the liposomes’ affinity for amyloid-β. The functionalization of liposomes with RI7217, independently of the ligation procedure, gave higher values of uptake and permeability across the barrier model in comparison to the nondecorated ones, without cell monolayer alterations. Of note, the best performing particles were those covalently coupled with the antibody. The ratios of the two radiolabeled lipids (3H-sphingomyelin and 14C-phosphatidic acid) present in the liposome bilayer were found to be similar in the apical and in the basolateral compartments of the barrier model, suggesting that liposomes were transported intact across the cell monolayer. Confocal experiments showed no co-localization of RI7217-liposomes with early/late endosomes or early lysosomes.Conclusion: Our results suggest that RI7217 promotes the in vitro barrier crossing of liposomes containing phosphatidic acid, targeting the Alzheimer’s disease amyloid-β peptide. Moreover, for the first time, we prove herein the superior efficiency of covalent coupling of RI7217 versus biotin/streptavidin ligation to facilitate liposomes in overcoming the BBB in vitro.Keywords: liposomes, blood–brain barrier, amyloid-β peptide, RI7217, antibody conjugation, surface plasmon resonance |
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