Preparation of Decellularized Human Peripheral Nerve Grafts Using Sodium Hydroxide and Sodium Deoxycholate

Peripheral nerve injuries often require graft materials that provide structural guidance for axonal regeneration. Although autologous nerve grafting remains the clinical gold standard, its use is limited by donor-site morbidity and restricted tissue availability. These limitations have driven increasing interest in decellularized human peripheral nerve grafts as an alternative biomaterial for nerve regeneration. However, commonly used detergent-based decellularization protocols may disrupt extracellular matrix (ECM) architecture and leave cytotoxic residues that can compromise graft quality. In this article, a standardized protocol is described for preparing decellularized human peripheral nerve grafts using an alkaline-assisted decellularization approach. The procedure begins with sodium hydroxide (NaOH) treatment to promote efficient cellular disruption, followed by a mild sodium deoxycholate (SDC) step that removes remaining cellular and immunogenic components while preserving the native ECM organization. This strategy limits detergent exposure while supporting preservation of the native ECM architecture. The workflow includes donor nerve preparation, controlled NaOH treatment, subsequent mild SDC treatment, and extensive washing under continuous solution exchange using a peristaltic pump-driven system to facilitate removal of residual processing agents. Further post-processing steps are implemented to maintain structural integrity and prepare the grafts for final packaging and sterilization. Decellularization efficiency is assessed through histological analysis and quantification of residual DNA, and the preservation of ECM structure is examined using scanning electron microscopy. The absence of residual SDC and the biocompatibility of processed grafts were confirmed through cytotoxicity testing. This protocol provides a practical approach for producing decellularized human peripheral nerve grafts for peripheral nerve regeneration research and translational applications.