Engineering 2D approaches fibrous platform incorporating turmeric and polyaniline nanoparticles to predict the expression of βIII-Tubulin and TREK-1 through qRT-PCR to detect neuronal differentiation of PC12 cells

Highlights: • Development of fibrous substrate from PCL with turmeric and PANI • The substrate as promising platform for mimicking neuronal extracellular matrix • Low hydrophilicity and anti-oxidizing behavior of substrate enhancing cells-cell interactions. • Secretion of neurotrophic proteins like βIII-Tubulin, MAP2 and TREK-1 are expressed. The bioengineering electroactive construct of a nerve-guided conduit for repairing and restoring injured nerves is an exciting biomedical endeavor that has implications for the treatment of peripheral nerve injury. In this study, we report the development the polycaprolactone (PCL) nanofibrous substrate consisting of turmeric (TUR) and polyaniline nanoparticles (PANINPs) exhibits topological and biological features that mimics the natural extracellular matrix (ECM) for nerve cells. We evaluated the morphology of 2-dimensional (2D) fibrous substrates, and their ability of stem cell adhesion, growth and proliferation rate were influenced by use of various concentrations of turmeric in PCL–TUR substrates. The results showed that 0.62 wt% of TUR and 0.28 wt% of PANINPs in PCL nanofibers substrate exhibited the optimal cellular microenvironment to accelerate PC12 cellular activities. The in vitro experiments revealed that PCL–PCL–TUR@PANI substrates significantly stimulated the proliferation, differentiation, and spontaneous outgrowth and extension of neurites from the cells. The substrate has the capacity to respond directly to neuronal markers with significant upregulation of βIII-Tubulin and TREK-1 through myelination, and also trigger neurotrophic protein expression, which was confirmed via immunocytochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. This study provides a new technique to design substrate of nerve tissue-specific microenvironment for peripheral nerve cell regeneration and could offer promising biomaterials for in vivo peripheral nerve repair.