Overcoming the Blood-Brain-Tumour Barrier Using Superparamagnetic Iron Oxide Nanoparticles in Vitro
Author
Odgaard, Frederik Hegner
Term
4. term
Publication year
2024
Abstract
Glioblastoma multiforme (GBM) is an aggressive brain tumor in which the blood–brain barrier (BBB) and the tumor-altered blood–brain–tumor barrier (BBTB) hinder drug delivery to the brain. This thesis evaluates whether superparamagnetic iron oxide nanoparticles can traverse the BBTB in vitro and whether they affect barrier integrity. Transwell models were built using porcine or human brain endothelial cells (PBECs/HBMECs) co-cultured with T10 GBM cells; U87 cells and a monoculture BBB served as controls. Treatments included nanoparticles with or without an external magnetic field and untreated controls. Nanoparticle passage and cellular uptake were assessed by fluorescence microscopy and Perl’s Prussian blue staining, and barrier integrity by transendothelial electrical resistance (TEER); cytotoxicity was also evaluated. FluidMAG-ARA nanoparticles were non-cytotoxic to PBECs and HBMECs (p>0.05) but reduced viability of U87 cells (p<0.05). TEER was unchanged (p>0.05), immunocytochemistry showed no uptake by endothelial cells, and no nanoparticle passage across BBB or BBTB models was detected, with or without magnetic exposure. In contrast, nanoparticles crossed cell-free insert membranes, with a clear difference between inserts with cells and without cells (p<0.0001). In summary, an in vitro BBTB was established, and contrary to expectations the nanoparticles did not cross the barrier under the tested conditions, but they were well tolerated by endothelial cells, indicating that the potential for targeted drug delivery remains.
Glioblastoma multiforme (GBM) er en aggressiv hjernetumor, hvor den blod-hjerne-barriere (BBB) og den tumormodificerede blod-hjerne-tumor-barriere (BBTB) begrænser lægemiddeltransport til hjernen. Denne afhandling undersøger, om superparamagnetiske jernoxid-nanopartikler kan krydse BBTB in vitro, og om de påvirker barriereintegriteten. Der blev opbygget transwell-modeller med enten porcint eller humant hjerneendotel (PBEC/HBMEC) i samkultur med T10 GBM-celler; U87-celler og en monokultur BBB-model fungerede som kontroller. Behandlinger omfattede nanopartikler med og uden ekstern magnetpåvirkning samt ubehandlede kontroller. Passage og celleoptag blev vurderet ved fluorescensmikroskopi og Perl’s Prussian blue-farvning, og barriereintegritet blev målt med transendotelial elektrisk modstand (TEER); cytotoksicitet blev også evalueret. FluidMAG-ARA-nanopartikler var ikke cytotoksiske for PBEC- eller HBMEC-celler (p>0,05), men reducerede levedygtigheden af U87-celler (p<0,05). TEER blev ikke påvirket (p>0,05), immunocytokemi viste ingen optag i endotelceller, og der blev ikke påvist passage af nanopartikler gennem BBB- eller BBTB-modellerne, hverken med eller uden magnetfelt. Derimod passerede nanopartiklerne indsatsmembraner uden celler, og der var en klar forskel mellem indsats med og uden celler (p<0,0001). Samlet blev et in vitro BBTB etableret, og i modsætning til forventningen krydsede nanopartiklerne ikke barrieren under de testede betingelser, men de var vel tolereret af endotelcellerne, hvilket indikerer, at potentialet for målrettet lægemiddellevering fortsat består.
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