Engineering
cellular microenvironment on a functional platform
using various biophysical cues to modulate stem cell fate has been
the central theme in regenerative engineering. Among the various biophysical
cues to direct stem cell differentiation, the critical role of physiologically
relevant electric field (EF) stimulation was established in the recent
past. The present study is the first to report the strategy to switch
EF-mediated differentiation of human mesenchymal stem cells (hMSCs)
between neuronal and glial pathways, using tailored functional properties
of the biomaterial substrate. We have examined the combinatorial effect
of substrate functionalities (conductivity, electroactivity, and topography)
on the EF-mediated stem cell differentiation on polyvinylidene–difluoride
(PVDF) nanocomposites in vitro, without any biochemical
inducers. The functionalities of PVDF have been tailored using conducting
nanofiller (multiwall-carbon nanotube, MWNT) and piezoceramic (BaTiO3, BT) by an optimized processing approach (melt mixing-compression
molding-rolling). The DC conductivity of PVDF nanocomposites was tuned
from ∼10–11 to ∼10–4 S/cm and the dielectric constant from ∼10 to ∼300.
The phenotypical changes and genotypical expression of hMSCs revealed
the signatures of early differentiation toward neuronal pathway on
rolled-PVDF/MWNT and late differentiation toward glial lineage on
rolled-PVDF/BT/MWNT. Moreover, we were able to distinguish the physiological
properties of differentiated neuron-like and glial-like cells using
membrane depolarization and mechanical stimulation. The excitability
of the EF-stimulated hMSCs was also determined using whole-cell patch-clamp
recordings. Mechanistically, the roles of intracellular reactive oxygen
species (ROS), Ca2+ oscillations, and synaptic and gap
junction proteins in directing the cellular fate have been established.
Therefore, the present work critically unveils complex yet synergistic
interaction of substrate functional properties to direct EF-mediated
differentiation toward neuron-like and glial-like cells, with distinguishable
electrophysiological responses.