Fibrin-Polycaprolactone Scaffolds for the Differentiation\nof Human Neural Progenitor Cells into Dopaminergic Neurons

Parkinson’s\ndisease (PD), a progressive central nervous\nsystem disorder marked by involuntary movements, poses a significant\nchallenge in neurodegenerative research due to the gradual degeneration\nof dopaminergic (DA) neurons. Early diagnosis and understanding of\nPD’s pathogenesis could slow disease progression and improve\npatient management. In vitro modeling with DA neurons derived from\nhuman-induced pluripotent stem cell-derived neural progenitor cells\n(NPCs) offers a promising approach. These neurons can be cultured\non electrospun (ES) nanofibrous polycaprolactone (PCL) scaffolds,\nbut PCL’s hydrophobic nature limits cell adhesion. We investigated\nthe ability of ES PCL scaffolds coated with hydrophilic extracellular\nmatrix-based biomaterials, including cell basement membrane proteins,\nMatrigel, and Fibrin, to enhance NPC differentiation into DA neurons.\nWe hypothesized that fibrin-coated scaffolds would maximize differentiation\nbased on fibrin’s known benefits in neuronal tissue engineering.\nThe scaffolds both coated and uncoated were characterized using scanning\nelectron microscopy (SEM), transmission electron microscopy, Fourier\ntransform infrared spectroscopy-attenuated total reflectance, and\ndynamic mechanical analysis to assess their properties. NPCs were\nseeded on the coated scaffolds, differentiated, and matured into DA\nneurons. Immunocytochemistry targeting tyrosine hydroxylase (TH) and\nSEM confirmed DA neuronal differentiation and morphological changes.\nElectrophysiology via microelectrode array recorded their neuronal\nfiring. Results showed enhanced neurite extension, increased TH expression,\nand active electrical activity in cells on fibrin-coated scaffolds.\nDiluted fibrin coatings particularly promoted more pronounced neuronal\ndifferentiation and maturation. This study introduces a novel tissue-on-a-chip\nplatform for neurodegenerative disease research using DA neurons.