Uptake and permeability studies of BBB-targeting immunoliposomes using the hCMEC/D3 cell line.
The targeting potential of OX-26-decorated immunoliposomes was investigated, using the human brain endothelial cell line hCMEC/D3 as a model of the blood-brain barrier (BBB). Immuno-nanoliposomes were prepared by the biotin/streptavidin ligation strategy, and their uptake by hCMEC/D3 cells and permeability through cell monolayers was studied. In order to elucidate the mechanisms of uptake, pH-sensitive fluorescence signal of HPTS was used, while transport was measured using double labeled immunoliposomes (with aqueous and lipid membrane fluorescent tags). PEGylated and non-specific-IgG-decorated liposomes were studied under identical conditions, as controls. CHO-K1 cells (which do not overexpress the transferrin receptor) were studied in some cases for comparative purposes. Experimental results reveal that hCMEC/D3 cells are good models for in vitro screening of BBB-targeting nanoparticulate drug delivery systems. Uptake and transcytosis of immunoliposome-associated dyes by cell monolayers was substantially higher compared to those of control liposomes. HPTS-entrapping OX-26-immunoliposome uptake indicated lysosomal localization and receptor-mediated mechanism. The ratio of aqueous/lipid label transport is affected by pre-incubation with antibody, or use of high lipid doses, suggesting that vesicles are transported intact after lysosome saturation. Co-decoration with a second ligand slightly decreases OX-26-decorated vesicle uptake, but not transcytosis, proving that the biotin-streptavidin technique can be applied for the generation of dual-targeting nanoliposomes.
Laboratory of Pharmaceutical Technology, Department of Pharmacy, University of Patras, Rio, Greece.
Eleni
E
Markoutsa
Georgios
G
Pampalakis
Anna
A
Niarakis
Ignacio A
IA
Romero
Babette
B
Weksler
Pierre-Olivier
PO
Couraud
Sophia G
SG
Antimisiaris
D000818:Animals; D000911:Antibodies, Monoclonal; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D055598:Chemical Phenomena; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G; D008081:Liposomes / Q000737:chemistry / Q000276:immunology* / Q000493:pharmacokinetics*; D051379:Mice; D053758:Nanoparticles; D010316:Particle Size; D010539:Permeability; D011990:Receptors, Transferrin / Q000378:metabolism; D057900:Transcytosis; D014168:Transferrin / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008081:Liposomes; D011990:Receptors, Transferrin; D014168:Transferrin