Rapid amyloid-β clearance and cognitive recovery by modulating BBB transport

Summary

A40-PO polymersome preparation and characterizationP[(OEG)10MA]20-PDPA120 and angiopep2-P[(OEG)10MA]20-PDPA120 copolymers were synthesized via atom transfer radical polymerization (ATRP) as previously reported.33 A40-POs were formulated adhering to the film-rehydration method, whereby P[(OEG)10MA]20-PDPA120 and a 1.88% molar ratio of angiopep2-P[(OEG)10MA]20-PDPA120 were dissolved in a mixture of methanol and chloroform (v/v, 3:1). The mixture was left to evaporate, allowing the organic solvent to completely dissipate and form a homogeneous polymer film at the bottom of the vial. PBS solution was added to rehydrate the polymer film and sonicated for 30 min. The solution was stirred continuously for 7 days at 4 °C via a magnetic stirrer at 1000 rpm. The morphology of the A40-POs was characterized via transmission electron microscopy (JEM-2100Plus, Japan). The diameter distribution was assessed via dynamic light scattering (Malvern Zetasizer Pro, UK).AnimalsAll the animal studies were conducted in accordance with the guidelines set forth by the West China Hospital Animal Care Committee (IACUC-approved project number: 20211475A). Considerable efforts were made to minimize the number of animals utilized in these studies and to alleviate any pain or discomfort experienced by the animals. In all the experiments, the animals were housed in a room where the temperature was regulated, with consistent alternating cycles of light and darkness.Immunohistochemistry (IHC)Paraffin-embedded sections (5 μm) of mouse brain tissue were deparaffinized with xylene and subjected to gradient alcohol hydration (100%, 80%, 50%, 30%). Endogenous peroxidase activity was quenched via the addition of 3% H2O2 (room temperature, 10 min) following antigen retrieval (Biosharp, 22315828). The tissues were blocked with 1% BSA (Aladdin, A104912) at room temperature for 30 min. Aβ-targeting primary antibodies were incubated overnight at 4 °C (Servicebio, GB13414-1, 1:200), followed by washing with PBS....