Iranian Journal of Basic Medical Sciences

Iranian Journal of Basic Medical Sciences

Optimized peripheral nerve decellularization protocol with enhanced extracellular matrix integrity, mechanical strength, and biocompatibility vs Hudson’s standard method

Articles in Press

Document Type : Original Article

Authors
Hussein Abbood Shanan Al-Shafeay 1
Saber Zahri 1
Asadollah Asadi 1
Arash Abdolmaleki 2
1 Department of Biology, Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran
2 Department of Biophysics, Faculty of Advanced Technologies, University of Mohaghegh Ardabili, Namin, Iran
10.22038/ijbms.2026.95387.20565
Abstract
Objective(s): Decellularized nerve scaffolds are promising biomaterials for peripheral nerve regeneration. This study compared an optimized decellularization protocol with Hudson’s standard method to assess structural, mechanical, biochemical, and biological performance.
Materials and Methods: Rat nerve tissues underwent native and optimized decellularization protocols. Morphological evaluation was performed using H&E and DAPI staining, while biomechanical properties and degradation kinetics were assessed over 28 days. DNA content, S100B, NEFL, collagen, laminin, and glycosaminoglycans (GAGs) were quantified to evaluate extracellular matrix (ECM) preservation and cellular removal. Neurotrophic factors, including NGF, BDNF, and GDNF, were measured to assess regenerative potential. Biocompatibility was evaluated after transplantation of rat bone marrow-derived mesenchymal stem cells (BM-MSCs) using MTT and Caspase-3 assays. All experiments used n=5 specimens per group, with triplicate measurements.
Results: The optimized protocol achieved near-complete decellularization while preserving ECM architecture and significantly outperformed Hudson’s method (P<0.001). Optimized scaffolds exhibited intermediate stiffness (13.76 MPa), higher ultimate strength (2.3 MPa vs 1.62 MPa; P<0.01), and slower degradation (48.3% reduction; P<0.001). DNA content decreased by 96.9% in the optimized group compared with 90.5% in Hudson’s group. Neurotrophic factors were better preserved, with NGF reaching 56.2 pg/mg (P<0.001), and ECM components such as collagen and laminin improved significantly (P<0.05). Optimized scaffolds enhanced cell viability (OD 0.58 vs 0.43; P<0.001) and reduced Caspase-3 activity (P<0.01).
Conclusion: The optimized decellularization protocol surpasses Hudson’s method by achieving effective decellularization while preserving ECM structure, mechanical integrity, and BM-MSC viability, thereby supporting its potential application in peripheral nerve tissue engineering.
Keywords
Biocompatibility
Caspase-3
Decellularization
Extracellular matrix
Hudson’s method
Optimized protocol
Sciatic nerve
Subjects
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Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 17 October 2026