Electrospun Biomaterial Scaffolds for Dopaminergic Differentiation of Human Neural Progenitor Cells

Project Description

Parkinson’s disease (PD) presents a significant challenge in neurodegenerative research due to its progressive nature and the degeneration of dopaminergic (DA) neurons in the central nervous system (CNS). Understanding the early stages and progression of PD is crucial for improving disease management and enhancing patients’ quality of life. A promising approach for studying the pathogenesis of PD is in vitro modeling. DA neurons obtained from neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (hiPSCs) can be cultured on electrospun (ES) nanofibrous polycaprolactone (PCL) scaffolds to promote their attachment and guide their specific transformation into DA neurons. However, since PCL is hydrophobic, in this study, we aimed to study the effectiveness of ES PCL scaffolds coated with hydrophilic extracellular matrix (ECM)-based biomaterials, including Cell Basement Membrane (CBM) proteins, Matrigel, and Fibrin (in varying concentrations), towards promoting the differentiation and maturation of NPCs into DA neurons. We hypothesized that fibrin-coated scaffolds would maximize the differentiation of NPCs into DA neurons, based on published studies that have demonstrated fibrin’s potential in neuronal tissue engineering, particularly in promoting cell adhesion and differentiation. The uncoated and coated ES PCL scaffolds underwent characterization using Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Fourier Transform Infrared Spectroscopy (FTIR-ATR), and Dynamic Mechanical Analysis (DMA) to assess their physical, chemical, and mechanical properties. For cell behavior analysis, NPCs were seeded onto the ES PCL scaffolds and then induced to differentiate into DA neurons. Confirmation of their differentiation into DA neurons was conducted through immunocytochemistry targeting Tyrosine Hydroxylase (TH) expression, along with SEM to observe morphological changes. Additionally, electrophysiology via a microelectrode array (MEA) technique was employed to measure neuronal firing from the differentiated and matured DA neurons. Our findings revealed neurite extension and increased TH expression in cells cultured on fibrin-coated scaffolds, along with active electrical activity in the differentiated cells. Moreover, scaffolds coated with diluted fibrin exhibited a higher degree of neuronal differentiation and maturation compared to other coatings. This study has contributed to the development of an innovative tissue-on-a-chip platform for studying neurodegenerative diseases using DA neurons.

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