Cutting-edge technologies in neural regeneration

Abstract

Neural regeneration stands at the forefront of neuroscience, aiming to repair and restore function to damaged neural tissues, particularly within the central nervous system (CNS), where regenerative capacity is inherently limited. However, recent breakthroughs in biotechnology, especially the revolutions in genetic engineering, materials science, multi-omics, and imaging, have promoted the development of neural regeneration. This review highlights the latest cutting-edge technologies driving progress in the field, including optogenetics, chemogenetics, three-dimensional (3D) culture models, gene editing, single-cell sequencing, and 3D imaging. Prospectively, the advancements in artificial intelligence (AI), high-throughput in vivo screening, and brain-computer interface (BCI) technologies promise to accelerate discoveries in neural regeneration further, paving the way for more precise, efficient, and personalized therapeutic strategies. The convergence of these multidisciplinary approaches holds immense potential for developing transformative treatments for neural injuries and neurological disorders, ultimately improving functional recovery.

Keywords: 3D cell culture; 3D imaging; Chemogenetics; Gene editing; Neural regeneration; Optogenetics; Organoid; Single-cell sequencing.

Fig. 1

Principles of optogenetics (left) and chemogenetics (right). The application of optogenetic and chemogenetic tools allows for precise manipulation of neuronal functions, facilitating the study of molecular mechanisms and strategies for axonal regeneration. Created with BioRender

Fig. 2

3D culture of neurons. The use of microphysiological systems and biomaterials facilitates the 3D culture of neuronal cells, organoids, or tissues, and carries out modeling of approximate physiological states, mechanism research, and drug screening. For example, the development of hydrogel materials, microfluidic chips, and organoid technology provides solutions for constructing a similar in vivo microenvironmental neuronal culture system. Created with BioRender

Fig. 3

Principles of gene editing technology. Gene editing technology utilizes sequence-specific nucleases to recognize specific DNA target sequences to create double-strand breaks (DSBs) and then repair the broken DNA by homologous recombination (HR) and nonhomologous end joining (NHEJ) to enable specific genetic modification. The initial generation of gene editing instruments, zinc finger nucleases (ZFNs), fuses the non-specific cleavage domain of FokI endonuclease with a zinc finger domain, offering a general way to deliver a site-specific DSB to the genome. The second-generation tool, transcription activator-like effector nucleases (TALENs), consists of a nonspecific DNA-cleaving FokI nuclease domain fused to a customizable DNA-binding domain (TALE) to introduce targeted DSBs. The CRISPR-Cas9 system, known as the third-generation gene editing technology, involves an endonuclease guided by a single guide RNA (sgRNA), which binds to the target DNA site of the protospacer adjacent motif (PAM) and produces DSBs for targeted genome modification. Recently, a variety of novel editing tools have been developed based on the CRISPR strategy, including CRISPRi and CRISPRa for gene suppression and activation, CRISPRoff and CRISPRon for epigenetic editing, base editors, and prime editors for long fragment editing. Created with BioRender

Fig. 4

scRNA-seq workflow. The scRNA-seq procedure includes the following steps: single-cell suspension preparation, cell sorting, adding barcodes, library construction, high-throughput sequencing, and data analysis. This workflow enables comprehensive transcriptomic profiling at single-cell resolution, facilitating the investigation of cellular heterogeneity and gene expression dynamics within neural tissues. Created with BioRender

Fig. 5

3D imaging of neural tissues using advanced tissue clearing and Light-sheet microscopy. The combination of tissue clearing methods with high-resolution Light-sheet microscopy enables multiscale 3D reconstruction of regenerated axons. This cutting-edge imaging platform offers a powerful tool for investigating structural and functional relationships within complex neural networks at cellular and subcellular resolution. Created with BioRender