RESUMO

Transport of therapeutics across the blood-brain barrier (BBB) is a fundamental requirement for effective treatment of numerous brain diseases. However, most therapeutics (>500 Da) are unable to permeate through the BBB and do not achieve therapeutic doses. Nanoparticles (NPs) are being investigated to facilitate drug delivery to the brain. Here, we investigate the effect of nanoparticle stiffness on NP transport across an in vitro BBB model. To this end, fluorescently labeled poly(N-isopropylmethacrylamide) (p(NIPMAM)) nanogels' stiffness was varied by the inclusion of 1.5 mol% (NG1.5), 5 mol% (NG5), and 14 mol% (NG14) N,N'-methylenebis(acrylamide) (BIS) cross-linker and nanogel uptake and transcytosis was quantified. The more densely cross-linked p(NIPMAM) nanogels showed the highest level of uptake by polarized brain endothelial cells, whereas the less densely cross-linked nanogels demonstrated the highest transcytotic potential. These findings suggest that nanogel stiffness has opposing effects on nanogel uptake and transcytosis at the BBB.

Assuntos

Barreira Hematoencefálica , Nanogéis/química , Acrilamidas/química , Linhagem Celular , Endotélio Vascular/citologia , Corantes Fluorescentes/química , Humanos , Técnicas In Vitro , Polímeros/química

RESUMO

Evidence suggests that extracellular vesicles (EVs) act as mediators and biomarkers of neurodegenerative diseases. Two distinct forms of Alzheimer disease (AD) are known: a late-onset sporadic form (SAD) and an early-onset familial form (FAD). Recently, neurovascular dysfunction and altered systemic immunological components have been linked to AD neurodegeneration. Therefore, we characterized systemic-EVs from postmortem SAD and FAD patients and evaluated their effects on neuroglial and endothelial cells. We found increase CLN-5 spots with vesicular morphology in the abluminal portion of vessels from SAD patients. Both forms of AD were associated with larger and more numerous systemic EVs. Specifically, SAD patients showed an increase in endothelial- and leukocyte-derived EVs containing mitochondria; in contrast, FAD patients showed an increase in platelet-derived EVs. We detected a differential protein composition for SAD- and FAD-EVs associated with the coagulation cascade, inflammation, and lipid-carbohydrate metabolism. Using mono- and cocultures (endothelium-astrocytes-neurons) and human cortical organoids, we showed that AD-EVs induced cytotoxicity. Both forms of AD featured decreased neuronal branches area and astrocytic hyperreactivity, but SAD-EVs led to greater endothelial detrimental effects than FAD-EVs. In addition, FAD- and SAD-EVs affected calcium dynamics in a cortical organoid model. Our findings indicate that the phenotype of systemic AD-EVs is differentially defined by the etiopathology of the disease (SAD or FAD), which results in a differential alteration of the NVU cells implied in neurodegeneration.