Plant‐Derived Viral Nanoparticles Enable Simultaneous Guidance of Neuronal Cell Outgrowth and Targeting of Neurodifferentiation Pathways

Differentiating neuronal cells in vitro is a complex process that can be significantly enhanced by using a combination of functional peptides and nanostructured scaffolds in combination. However, applying these elements simultaneously remains challenging. Here, a novel neural tissue engineering approach that uses plant-derived viral nanoparticles (VNPs) to simultaneously promote neuronal differentiation and growth guidance is presented. It is hypothesized that the simultaneous alignment and promotion of neurodifferentiation could be achieved by using genetically engineered potato virus X and tobacco mosaic virus, which display high local concentrations of functional peptides derived from laminin (RGD and IKVAV) and brain-derived neurotrophic factor. Immunostaining, gene analysis, immunoprecipitation, and western blotting are employed to evaluate the effect of VNPs on neurodifferentiation and their mechanism of action via cell membrane receptors. 3D printing with sacrificial materials is used to align the VNPs, as confirmed by scanning electron microscopy. This approach creates an orientated microarchitecture that simultaneously combines growth guidance and pathway targeting. The incorporation of growth-factor-like peptides onto the VNP surface through genetic engineering represents a significant advancement in this area of research. This provides unparalleled control over neural cell differentiation and neurite outgrowth by utilizing plant-derived, bioactive, and biomimetic nanoparticles as a multifunctional scaffold base.