Three‐dimensional‐printed polycaprolactone/polypyrrole conducting scaffolds for differentiation of human olfactory ecto‐mesenchymal stem cells into Schwann cell‐like phenotypes and promotion of neurite outgrowth

Abstract

Implantation of a suitable nerve guide conduit (NGC) seeded with sufficient Schwann cells (SCs) is required to improve peripheral nerve regeneration efficiently. Given the limitations of isolating and culturing SCs, using various sources of stem cells, including mesenchymal stem cells (MSCs) obtained from nasal olfactory mucosa, can be desirable. Olfactory ecto-MSCs (OE-MSCs) are a new population of neural crestderived stem cells that can proliferate and differentiate into SCs and can be considered a promising autologous alternative to produce SCs. Regardless, a biomimetic physicochemical microenvironment in NGC such as electroconductive substrate can affect the fate of transplanted stem cells, including differentiation toward SCs and nerve regeneration. Therefore, in this study, the effect of 3D printed polycaprolactone (PCL)/polypyrrole (PPy) conductive scaffolds on differentiation of human OE-MSCS into functional SC-like phenotypes was investigated. Biological evaluation of 3D printed scaffolds was examined by in vitro culturing the OE-MSCs on samples surfaces, and conductivity showed no effect on increased cell attachment, proliferation rate, viability, and distribution. In contrast, immunocytochemical staining and real-time polymerase chain reaction analysis indicated that 3D structures coated with PPy could provide a favorable microenvironment for OE-MSCs differentiation. In addition, it was found that differentiated OE-MSCs within PCL/PPy could secrete the highest amounts of nerve growth factor and brain-derived neurotrophic factor neurotrophic factors compared to pure PCL and 2D culture. After co-culturing with PC12 cells, a significant increase in neurite outgrowth on PCL/PPy conductive scaffold seeded with differentiated OE-MSCs. These findings indicated that 3D printed PCL/PPy conductive scaffold could support differentiation of OE-MSCs into SC-like phenotypes to promote neurite outgrowth, suggesting their potential for neural tissue engineering applications.

Implantation of a nerve protector embedded with human GMSC-derived Schwann-like cells accelerates regeneration of crush-injured rat sciatic nerves

“…[ 33 , 49 ]. In recent years, various approaches have been developed to promote induction of MSCs into Schwann-like cells, particularly the use of certain small molecules or growth factors [ 48 , 50 ] and engineered substrates or scaffolds such as graphene oxide substrate [ 51 ], 3D-printed polycaprolactone/polypyrrole (PCL/PPy) conductive scaffold [ 52 ], 3D-printed bionic polycaprolactone (PCL) polymer scaffold [ 53 ], and aligned nanofibers [ 54 ]. Consistent with our recent studies [ 37 ], we, herein, have also demonstrated that GMSCs could directly and rapidly convert into Schwann-like cells when they were encapsulated in methacrylated 3D-collagen hydrogel and cultured under the regular MSC culture medium without the additional introduction of special nutritional and growth factor supplements (Fig.…”

Conducting polymer-based scaffolds for neuronal tissue engineering

“…PC12 cells co-cultured with stem cells exhibit extended neurite outgrowth, suggesting their potential for neuronal regeneration. 130 Compared to regular PPy homopolymers, the amine-functionalized polypyrrole (APPy) exhibits an average hydrophilic surface and electrical activity. In addition, APPy showed superior attachment to Schwann cells and human dermal fibroblasts under serum-containing and serum-free conditions.…”

Development and In Vitro Differentiation of Schwann Cells

“…Similar effects were reported from 3D-bioprinted cultures with aligned fibrin fibers in print strands [ 325 , 342 ]. Furthermore, polymer fibers manufactured from conductive materials and enabling electrostimulation of cells enhanced cell proliferation, migration, and differentiation [ 355 , 371 , 372 , 373 , 374 ]. Besides the composition and micropatterning, the stiffness is another important parameter of 3D culture materials.…”