Volume 30, Issue 1 (March 2026)                   Physiol Pharmacol 2026, 30(1): 81-97 | Back to browse issues page

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Ahmadnia H, Pashazadeh M P, Moradi A, Mahdavi-Shahri N, Behnam-Rassouli M, Riahi E. An optimized sondell decellularization method for sciatic nerve graft. Physiol Pharmacol 2026; 30 (1) :81-97
URL: http://ppj.phypha.ir/article-1-2588-en.html
An optimized sondell decellularization method for sciatic nerve graft
Hananeh Ahmadnia , Muhammad Parsa Pashazadeh , Ali Moradi , Naser Mahdavi-Shahri , Morteza Behnam-Rassouli , Esmail Riahi
Abstract:   (1444 Views)

Introduction: 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.
Methods: 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.
Results: 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’s modulus of the OSD grafts (p < 0.05). Histological studies, including hematoxylin and eosin, Masson’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. 
Conclusion: 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.

Keywords: Peripheral Nerve Injuries, Decellularized Extracellular Matrix, Scaffold, Sciatic Nerve
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Type of Manuscript: Experimental research article | Subject: Neurophysiology/Pharmacology
References
1. Ahearne M, Coyle A. Application of UVA-riboflavin crosslinking to enhance the mechanical properties of extracellular matrix derived hydrogels. Journal of the Mechanical Behavior of Biomedical Materials 2016; 54: 259-267. [DOI:10.1016/j.jmbbm.2015.09.035]
2. Alizadeh M, Rezakhani L, Soleimannejad M, Sharifi E, Anjomshoa M, Alizadeh A. Evaluation of vacuum washing in the removal of SDS from decellularized bovine pericardium: method and device description. Heliyon 2019; 5. [DOI:10.1016/j.heliyon.2019.e02253]
3. Allodi I, Udina E, Navarro X. Specificity of peripheral nerve regeneration: interactions at the axon level. Progress in Neurobiology 2012; 98: 16-37. [DOI:10.1016/j.pneurobio.2012.05.005]
4. Alvites R, Rita Caseiro A, Santos Pedrosa S, Vieira Branquinho M, Ronchi G, Geuna S, et al. Peripheral nerve injury and axonotmesis: State of the art and recent advances. Cogent Medicine 2018; 5: 1466404. [DOI:10.1080/2331205X.2018.1466404]
5. Arthur-Farraj P J, Latouche M, Wilton D K, Quintes S, Chabrol E, Banerjee A, et al. c-Jun reprograms Schwann cells of injured nerves to generate a repair cell essential for regeneration. Neuron 2012; 75: 633-647. [DOI:10.1016/j.neuron.2012.06.021]
6. Bae J-Y, Park S Y, Shin Y H, Choi S W, Kim J K. Preparation of human decellularized peripheral nerve allograft using amphoteric detergent and nuclease. Neural Regeneration Research 2021; 16: 1890-1896. [DOI:10.4103/1673-5374.306091]
7. Chen L, Wei L, Su X, Qin L, Xu Z, Huang X, et al. Preparation and characterization of biomimetic functional scaffold with gradient structure for osteochondral defect repair. Bioengineering 2023; 10: 213. [DOI:10.3390/bioengineering10020213]
8. Crapo P M, Gilbert T W, Badylak S F. An overview of tissue and whole organ decellularization processes. Biomaterials 2011; 32: 3233-3243. [DOI:10.1016/j.biomaterials.2011.01.057]
9. El Soury M, García-García Ó D, Moretti M, Perroteau I, Raimondo S, Lovati A B, et al. Comparison of decellularization protocols to generate peripheral nerve grafts: a study on rat sciatic nerves. International Journal of Molecular Sciences 2021; 22: 2389. [DOI:10.3390/ijms22052389]
10. Faroni A, Mobasseri S A, Kingham P J, Reid A J. Peripheral nerve regeneration: experimental strategies and future perspectives. Advanced Drug Delivery Reviews 2015; 82: 160-167. [DOI:10.1016/j.addr.2014.11.010]
11. Faroni A, Smith R J, Reid A J. Adipose derived stem cells and nerve regeneration. Neural Regeneration Research 2014; 9: 1341-1346. [DOI:10.4103/1673-5374.137585]
12. García-Gareta E, Abduldaiem Y, Sawadkar P, Kyriakidis C, Lali F, Greco K V. Decellularised scaffolds: just a framework? Current knowledge and future directions. Journal of Tissue Engineering 2020; 11: 2041731420942903. [DOI:10.1177/2041731420942903]
13. Gilbert T W, Sellaro T L, Badylak S F. Decellularization of tissues and organs. Biomaterials 2006; 27: 3675-3683. [DOI:10.1016/j.biomaterials.2006.02.014]
14. Gilpin A, Yang Y. Decellularization strategies for regenerative medicine: from processing techniques to applications. Biomed Research International 2017; 2017: 9831534. [DOI:10.1155/2017/9831534]
15. Golebiowska A A, Intravaia J T, Sathe V M, Kumbar S G, Nukavarapu S P. Decellularized extracellular matrix biomaterials for regenerative therapies: advances, challenges and clinical prospects. Bioactive Materials 2024; 32: 98-123. [DOI:10.1016/j.bioactmat.2023.09.017]
16. He Z, Jin Y. Intrinsic control of axon regeneration. Neuron 2016; 90: 437-451. [DOI:10.1016/j.neuron.2016.04.022]
17. Hillebrandt K H, Everwien H, Haep N, Keshi E, Pratschke J, Sauer I M. Strategies based on organ decellularization and recellularization. Transplant International 2019; 32: 571-585. [DOI:10.1111/tri.13462]
18. Hudson T W, Zawko S, Deister C, Lundy S, Hu C Y, Lee K, et al. Optimized acellular nerve graft is immunologically tolerated and supports regeneration. Tissue Engineering 2004; 10: 1641-1651. [DOI:10.1089/ten.2004.10.1641]
19. Hussain G, Wang J, Rasul A, Anwar H, Qasim M, Zafar S, et al. Current status of therapeutic approaches against peripheral nerve injuries: a detailed story from injury to recovery. International Journal of Biological Sciences 2020; 16: 116. [DOI:10.7150/ijbs.35653]
20. Kasravi M, Ahmadi A, Babajani A, Mazloomnejad R, Hatamnejad M R, Shariatzadeh S, et al. Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine. Biomaterials Research 2023; 27: 10. [DOI:10.1186/s40824-023-00348-z]
21. Keane T J, Swinehart I T, Badylak S F. Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance. Methods 2015; 84: 25-34. [DOI:10.1016/j.ymeth.2015.03.005]
22. Kuffler D P, Foy C. Restoration of neurological function following peripheral nerve trauma. International Journal of Molecular Sciences 2020; 21: 1808. [DOI:10.3390/ijms21051808]
23. Liu Y-Y, Blazquez J P F, Yin G-Z, Wang D-Y, Llorca J, Echeverry-Rendón M. A strategy to tailor the mechanical and degradation properties of PCL-PEG-PCL based copolymers for biomedical application. European Polymer Journal 2023; 198: 112388. [DOI:10.1016/j.eurpolymj.2023.112388]
24. Lovati A B, D’Arrigo D, Odella S, Tos P, Geuna S, Raimondo S. Nerve repair using decellularized nerve grafts in rat models. A review of the literature. Frontiers in Cellular Neuroscience 2018; 12: 427. [DOI:10.3389/fncel.2018.00427]
25. McInnes A D, Moser M A, Chen X. Preparation and use of decellularized extracellular matrix for tissue engineering. Journal of Functional Biomaterials 2022; 13: 240. [DOI:10.3390/jfb13040240]
26. Moradi A, Ataollahi F, Sayar K, Pramanik S, Chong P P, Khalil A A, et al. Chondrogenic potential of physically treated bovine cartilage matrix derived porous scaffolds on human dermal fibroblast cells. Journal of Biomedical Materials Research Part A 2016; 104: 245-256. [DOI:10.1002/jbm.a.35561]
27. Moreno-Manzano V, Mellado-López M, Morera-Esteve M J, Alastrue-Agudo A, Bisbal-Velasco V, Forteza-Vila J, et al. Human adipose-derived mesenchymal stem cells accelerate decellularized neobladder regeneration. Regenerative Biomaterials 2020; 7: 161-169. [DOI:10.1093/rb/rbz049]
28. Philips C, Campos F, Roosens A, Sánchez-Quevedo M d C, Declercq H, Carriel V. Qualitative and quantitative evaluation of a novel detergent-based method for decellularization of peripheral nerves. Annals of Biomedical Engineering 2018; 46: 1921-1937. [DOI:10.1007/s10439-018-2082-y]
29. Rosadas M, Silva I V, Costa J B, Ribeiro V P, Oliveira A L. Decellularized dermal matrices: unleashing the potential in tissue engineering and regenerative medicine. Frontiers in Materials 2024; 10: 1285948. [DOI:10.3389/fmats.2023.1285948]
30. Sondell M, Lundborg G, Kanje M. Regeneration of the rat sciatic nerve into allografts made acellular through chemical extraction. Brain Research 1998; 795: 44-54. [DOI:10.1016/S0006-8993(98)00251-0]
31. Sridharan R, Reilly R B, Buckley C T. Decellularized grafts with axially aligned channels for peripheral nerve regeneration. Journal of the Mechanical Behavior of Biomedical Materials 2015; 41: 124-135. [DOI:10.1016/j.jmbbm.2014.10.002]
32. Sun Y, Zhang R, Mao X, Zhang M. Research of acellular xenogeneic nerve combined with adipose-derived stem cells and platelet rich plasma in repair of rabbit facial nerve injury. Zhongguo xiu fu Chong Jian wai ke za zhi= Zhongguo Xiufu Chongjian Waike Zazhi= Chinese Journal of Reparative and Reconstructive Surgery 2018; 32: 736-744.
33. Terenzio M, Koley S, Samra N, Rishal I, Zhao Q, Sahoo P K, et al. Locally translated mTOR controls axonal local translation in nerve injury. Science 2018; 359: 1416-1421. [DOI:10.1126/science.aan1053]
34. Vijayavenkataraman S. Nerve guide conduits for peripheral nerve injury repair: A review on design, materials and fabrication methods. Acta Biomaterialia 2020; 106: 54-69. [DOI:10.1016/j.actbio.2020.02.003]
35. Wang S, Liu X, Wang Y. Evaluation of platelet-rich plasma therapy for peripheral nerve regeneration: a critical review of literature. Frontiers in Bioengineering and Biotechnology 2022; 10: 808248. [DOI:10.3389/fbioe.2022.808248]
36. Wong M L, Griffiths L G. Immunogenicity in xenogeneic scaffold generation: antigen removal vs. decellularization. Acta Biomaterialia 2014; 10: 1806-1816. [DOI:10.1016/j.actbio.2014.01.028]
37. Yang X, Xue P, Chen H, Yuan M, Kang Y, Duscher D, et al. Denervation drives skeletal muscle atrophy and induces mitochondrial dysfunction, mitophagy and apoptosis via miR-142a-5p/MFN1 axis. Theranostics 2020; 10: 1415. [DOI:10.7150/thno.40857]
38. Zaminy A, Sayad-Fathi S, Kasmaie F M, Jahromi Z, Zendedel A. Decellularized peripheral nerve grafts by a modified protocol for repair of rat sciatic nerve injury. Neural Regeneration Research 2021; 16: 1086-1092. [DOI:10.4103/1673-5374.300449]
39. Zhang F, Zhang N, Xu Q, Zhang L, Zhang C, Liu H, et al. Decellularized nerve extracellular matrix/chitosan crosslinked by genipin to prepare a moldable nerve repair material. Cell and Tissue Banking 2021; 22: 419-430. [DOI:10.1007/s10561-020-09889-2]
40. Zilic L, Wilshaw S P, Haycock J W. Decellularisation and histological characterisation of porcine peripheral nerves. Biotechnology and Bioengineering 2016; 113: 2041-2053. [DOI:10.1002/bit.25964]
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