The Effect of Liquid-Phase Exfoliated Graphene Film on Neurodifferentiation of Stem Cells from Apical Papilla

Abstract

Background: Dental stem cells, which originate from the neural crest, due to their easy accessibility might be good candidates in neuro-regenerative procedures, along with graphene-based nanomaterials shown to promote neurogenesis in vitro. We aimed to explore the potential of liquid-phase exfoliated graphene (LPEG) film to stimulate the neuro-differentiation of stem cells from apical papilla (SCAP). Methods: The experimental procedure was structured as follows: (1) fabrication of graphene film; (2) isolation, cultivation and SCAP stemness characterization by flowcytometry, multilineage differentiation (osteo, chondro and adipo) and quantitative PCR (qPCR); (3) SCAP neuro-induction by cultivation on polyethylene terephthalate (PET) coated with graphene film; (4) evaluation of neural differentiation by means of several microscopy techniques (light, confocal, atomic force and scanning electron microscopy), followed by neural marker gene expression analysis using qPCR. Results: SCAP demonstrated exceptional stemness, as judged by mesenchymal markers’ expression (CD73, CD90 and CD105), and by multilineage differentiation capacity (osteo, chondro and adipo-differentiation). Neuro-induction of SCAP grown on PET coated with graphene film resulted in neuron-like cellular phenotype observed under different microscopes. This was corroborated by the high gene expression of all examined key neuronal markers (Ngn2, NF-M, Nestin, MAP2, MASH1). Conclusions: The ability of SCAPs to differentiate toward neural lineages was markedly enhanced by graphene film.

Influence of Graphene-Based Nanocomposites in Neurogenesis and Neuritogenesis: A Brief Summary

“…Consequently, the PU/Gr (pluronic coated) hydrogel may eventually be used as a brain construct since it provides cells with a soft environment that promotes proliferation and differentiation . Liquid-phase exfoliated graphene (LPEG) sheets were shown to have the potential to promote the neural development of stem cells derived from the apical papilla (SCAP) . Over 95% of mesenchymal stem cells (MSCs) are found in the soft tissue called the apical papilla, which is located close to the crown of developing teeth. , The cells that were grown on LPEG exhibited a higher quantity of neural markers.…”

“…65 Liquid-phase exfoliated graphene (LPEG) sheets were shown to have the potential to promote the neural development of stem cells derived from the apical papilla (SCAP). 66 Over 95% of mesenchymal stem cells (MSCs) are found in the soft tissue called the apical papilla, which is located close to the crown of developing teeth. 67,68 The cells that were grown on LPEG exhibited a higher quantity of neural markers.…”

Surface Area of Graphene Governs Its Neurotoxicity

“…The electroactive and conformational properties of graphene stimulate and guide neural cells and increase neural proliferation and differentiation. , However, most conductive materials are toxic and prone to biodegradation. GO nanoparticles, ,− 2D graphene films, − and three-dimensional (3D) graphene scaffolds , have been found promising in the regeneration of nerve cells. However, graphene scaffolds are not biodegradable and the effects of graphene on neurons remain unclear.…”

“… 24 , 25 However, most conductive materials are toxic and prone to biodegradation. GO nanoparticles, 24 , 26 − 29 2D graphene films, 30 − 32 and three-dimensional (3D) graphene scaffolds 33 , 34 have been found promising in the regeneration of nerve cells. However, graphene scaffolds are not biodegradable and the effects of graphene on neurons remain unclear.…”

Editorial for Special Issue: “Supramolecular Nanomaterials for Biomedical Application”

“…Zhu et al synthesized water-dispersible glutathione-modified CdTe quantum dots (GSH-CdTe QDs) by covalently bonding Cd atoms on the surface of CdTe QDs and thiol groups of the GSH, which further reacted with the Ru (II) complex via electrostatic adsorption to synthesize the QDs-Ru complexes to achieve the rapid and sensitive identification of plasma cell-free DNA (cfDNA) biomarkers in cancer patients [ 11 ]. Considering that graphene-based nanomaterials can facilitate the process of neurogenesis in vitro, using different microscopy techniques and real-time gene-expression analysis, Simonovic et al explored the ability of liquid-phase-exfoliated graphene films to induce and stimulate the neural differentiation of stem cells from apical papilla (SCAP) [ 12 ]. Lee et al utilized simple and scalable top-down and bottom-up approaches to prepare reduced-graphene quantum dots (RGQDs) and hyaluronic acid–graphene quantum dots (HGQDs), which possessed substantial near-infrared (NIR) absorption and fluorescence throughout the visible and NIR ranges, showing significant photothermal performance as well as NIR and fluorescence-imaging capabilities in HeLa cells [ 13 ].…”