Laminin-conjugated aligned nanofiber yarns for topographical and biochemical guidance of neurite outgrowth and branching regulation

BACKGROUND: Aligned nanofiber scaffolds have shown great promise in promoting neurite orientation and elongation, making them a key design strategy in peripheral nerve tissue engineering. However, most current designs lack biochemical specificity and long-term stability, limiting their regenerative efficacy. RESULTS: Herein, we developed structurally aligned nanofiber yarns (NFYs) scaffold conjugated with laminin via covalently incorporated N-hydroxysuccinimide (NHS) ester groups. The NHS-functionalized NFYs were fabricated using a dry-wet electrospinning approach based on a copolymer of N-succinimidyl acrylate (AA-NHS) and N-isopropyl acrylamide (NIPAM), blended with polycaprolactone (PCL) and multi-walled carbon nanotubes (MW-CNTs). This scaffold integrates both topographical and biochemical guidance, forming a stable biomimetic extracellular matrix (ECM) interface that supports long-range neurite extension. Laminin-conjugated NFY-NHS scaffolds significantly enhanced neurite alignment and elongation in PC12 cells and dissociated dorsal root ganglion (DRG) neurons, while simultaneously reducing excessive neurite branching. In DRG explant models, the scaffolds supported fasciculated neurite bundles that extended unidirectionally along the fiber orientation, mimicking native nerve architecture. These findings demonstrate that laminin-conjugated aligned NFYs provide a dual-function platform for guiding neurite outgrowth and regulating branching morphology. CONCLUSIONS: This strategy offers a promising approach for engineering physiologically relevant neural scaffolds with applications in peripheral nerve regeneration and in vitro nerve model development.