Toward Spontaneous Neuronal Differentiation of SH-SY5Y Cells Using Novel Three-Dimensional Electropolymerized Conductive Scaffolds

Abstract

Neuroblastoma-derived SH-SY5Y cells have become an excellent model for nervous system regeneration to treat neurodegenerative disorders. Many approaches achieved a mature population of derived neurons in in vitro plates. However, the importance of the third dimension in tissue regeneration has become indispensable to achieve a potential implant to replace the damaged tissue. Therefore, we have prepared porous 3D structures composed uniquely of carbon nanotubes (CNT) and poly(3,4ethylenedioxythiophene) (PEDOT) that show great potential in the tridimensional differentiation of SH-SY5Y cells into mature neurons. The scaffolds have been manufactured through electropolymerization by applying 1.2 V in a three-electrode cell using a template of sucrose/CNT as a working electrode. By this method, PEDOT/CNT 3D scaffolds were obtained with homogeneous porosities and high conductivity. In vitro analyses showed that an excellent biocompatibility of the scaffold and the presence of high amount of β-tubulin class III and MAP-II target proteins that mainly expresses in neurons, suggesting the differentiation into neuronal cells already after a week of incubation.

Biomimetic Scaffolds for Spinal Cord Applications Exhibit Stiffness‐Dependent Immunomodulatory and Neurotrophic Characteristics

“…Substrate stiffness within the HyA‐Coll‐IV/Fn groups significantly affected neuronal viability while also altering the morphology of the neuronal network within the scaffold. Furthermore, in contrast to the poor neurite extension recently reported in SH‐SY5Y neurons, [ 58 ] ECM‐functionalization led to dramatic changes in neuronal behavior. Increased metabolic activity of SH‐SY5Y neurons in softer or ECM scaffolds may either be a result of increased proliferation through the ECM‐functionalized surfaces within the scaffold (a process associated with activation of MAP‐kinase pathway via integrin binding) or alternatively, a sign of increased neurite outgrowth.…”

Organic Bioelectronics for In Vitro Systems

“…Immunofluorescence staining confirmed the hypothesis that cells seeded onto PEDOT/CNT expressed more β-tubulin class III (Figure 46b(ii)) and MAP II compared to the PEDOT scaffolds after 7 days. 400 Electroactive scaffolds can be also fabricated by in situ polymerization of an already prepared scaffold by VPP. This technique allows the coating of the scaffolds in a controllable manner by vaporization of a conducting monomer.…”

“…Many of the studies utilizing electroactive substrates have examined neural populations on PEDOT-based, ,,,,,,− PPy-based, ,,,,,− and PANI-based substrates, − as well as other substrates including pentacene, melanin, P13, and P3HT. ,− In many of these cases, researchers will often examine neurite outgrowth as an indicator of neural cell compatibility with a given substrate. A neurite is a protrusion from a neuronal cell, sometimes synonymous with an axonal or dendritic projection but generally referring to any observed cellular projection.…”

“…The addition of biopolymers to CP scaffolds could result in compromised electrical properties compared to the pristine CP scaffold. Along these lines, incorporation of additives that enhance conductivity, such as CNTs, ,, has proven a good strategy to boost the electrochemical properties of the scaffold. In a recent study, different ratios of PEDOT:PSS/MWCNT scaffolds were prepared by the freeze-drying technique and characterized by EIS.…”

Biomolecular Electron Controller Composed of Nanobiohybrid with Electrically Released Complex for Spatiotemporal Control of Neuronal Differentiation

“…However, there is still room to be improved to demonstrate spatiotemporal differentiation control, such as the precise positioning at the cellular level using magnetic fields. [ 20–22 ]…”