Engineering an Aβ-Induced 3D Alzheimer’s Disease Model via Magnetic Levitation for Neuroprotective Drug Screening

Abstract

Background: This study aims to develop an amyloid-beta-induced three-dimensional (3D) Alzheimer’s disease model using the Magnetic Levitation (MagLev) technique and rat pheochromocytoma (PC-12) (rat pheochromocytoma) cells. Methods: Initially, optimization studies were performed to determine the appropriate paramagnetic agent concentration and cell number required for the formation of 3D cellular structures. The formation of these structures was monitored by light microscopy, and cell viability was evaluated using a live/dead assay. Filamentous actin (F-actin) expression was assessed by immunostaining. Subsequently, amyloid beta 1–42 (Aβ1–42) aggregates were introduced into the 3D cellular structures to establish a 3D Alzheimer’s disease model. The 3D disease model was characterized by immunostaining of Choline Acetyltransferase (ChAT). Finally, the applicability of the developed model for therapeutic evaluation was demonstrated using curcumin as a representative neuroprotective compound. Results: The optimization studies successfully enabled the formation of 3D cellular structures using the magnetic levitation technique. Immunostaining analysis showed an increase in F-actin expression in 3D structures. Following the introduction of Aβ1–42 aggregates, a significant decline in cell viability was observed by day 21. ChAT immunostaining revealed a decrease in fluorescence intensity (F.I.) to 28% of the control level, corresponding to an approximately 72% reduction in ChAT expression. Curcumin treatment increased cell viability by 22%, 23%, and 13% on days 7, 15, and 21, respectively. Conclusion: Overall, the findings showed that the 3D Aβ 1–42 induced 3D Alzheimer’s disease model can provide a valuable experimental platform for investigating neurodegenerative diseases and evaluating potential therapeutic interventions.