en Neurophysiology/Pharmacology Neurophysiology/Pharmacology Experimental research article Experimental research article <p><span style="color:#000000;"><span style="font-size:12px;"><span style="font-family:Arial;"><strong>Introduction:</strong> In the realm of nerve injury repair, acellularized nerve grafts (ANGs) are gaining recognition as a promising tissue engineering approach, offering better regeneration outcomes compared to traditional nerve conduits. This study, conducted on rat and chicken sciatic nerves (R-ANGs and C-ANGs, respectively), seeks to explore an optimized nerve decellularization technique that achieves effective decellularization while preserving the extracellular matrix (ECM) integrity.<br /> <strong>Methods:</strong> The Sondell decellularization (SD) protocol was evaluated against an optimized Sondell decellularization (OSD) method improved by DNase treatment and extra-washing with ultrapure water, and then MTT, DNA quantification, mechanical tensile, and histological evaluations were performed.<br /> <strong>Results:</strong> Results showed that both R-ANGs and C-ANGs prepared by the OSD method had better biocompatibility, measured by MTT, than those prepared by the SD (p < 0.001). DNA quantification of samples obtained from the OSD method showed an improved elimination of residual DNA fragments (p < 0.0001). Mechanical tensile test also revealed a higher Young&rsquo;s modulus of the OSD grafts (p < 0.05). Histological studies, including hematoxylin and eosin, Masson&rsquo;s trichrome, Luxol fast blue, DAPI, and scanning electron microscopy (FESEM), confirmed that the OSD was superior to the SD in preserving ECM while equally removing cell nuclei and myelinated fibers.<br /> <strong>Conclusion: </strong>The OSD protocol yielded promising results for more reliable in vivo trials, as in vitro tests revealed better preservation of ECM components compared to the SD method. Notably, the OSD provided higher biocompatibility and improved viscoelastic integrity of the R-ANGs and C-ANGs.</span></span></span></p> Peripheral Nerve Injuries,Decellularized Extracellular Matrix,Scaffold,Sciatic Nerve 81 97 http://ppj.phypha.ir/browse.php?a_code=A-10-2627-2&slc_lang=en&sid=1 Hananeh Ahmadnia h.ahmadnia1990@mail.um.ac.ir 3200319475328460039086 3200319475328460039086 No Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran Muhammad Parsa Pashazadeh pashazadehmuhammadparsa@gmail.com 3200319475328460039087 3200319475328460039087 No Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran.Department of Physiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Ali Moradi ralimoradi@gmail.com 3200319475328460039088 3200319475328460039088 No Orthopedic Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.The Clinical Research Development Unit, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran. Naser Mahdavi-Shahri Mahdavin@um.ac.ir 3200319475328460039089 3200319475328460039089 No Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran. Morteza Behnam-Rassouli behnam@um.ac.ir 3200319475328460039090 0000000347349886 No Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran Esmail Riahi riahi@tums.ac.ir 3200319475328460039091 0000000209941195 Yes Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran.Department of Physiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.