Sciatic nerve regeneration by using collagen type I hydrogel containing naringin

  • Hadi Samadian
  • Ahmad Vaez
  • Arian Ehterami
  • Majid SalehiEmail author
  • Saeed Farzamfar
  • Hamed Sahrapeyma
  • Pirasteh Norouzi
Tissue Engineering Constructs and Cell Substrates Original Research
Part of the following topical collections:
  1. Tissue Engineering Constructs and Cell Substrates


In the present study, collagen hydrogel containing naringin was fabricated, characterized and used as the scaffold for peripheral nerve damage treatment. The collagen was dissolved in acetic acid, naringin added to the collagen solution, and cross-linked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide powder (EDC; 0.10 mM) to form the hydrogel. The microstructure, swelling behavior, biodegradation, and cyto/hemocompatibility of the fabricated hydrogels were assessed. Finally, the healing efficacy of the prepared collagen hydrogel loaded with naringin on the sciatic nerve crush injury was assessed in the animal model. The characterization results showed that the fabricated hydrogels have a porous structure containing interconnected pores with the average pore size of 90 µm. The degradation results demonstrated that about 70% of the primary weight of the naringin loaded hydrogel had been lost after 4 weeks of storage in PBS. The in vitro study showed that the proliferation of Schwann cells on the collagen/naringin hydrogel was higher than the control group (tissue culture plate) at both 48 and 72 h after cell seeding and even significantly higher than pure collagen 72 h after cell seeding (*p < 0.005, **p < 0.001). The animal study implied that the sciatic functional index reached to −22.13 ± 3.00 at the end of 60th days post-implantation which was statistically significant (p < 0.05) compared with the negative control (injury without the treatment) (−82.60 ± 1.06), and the pure collagen hydrogel (−59.80 ± 3.20) groups. The hot plate latency test, the compound muscle action potential, and wet weight-loss of the gastrocnemius muscle evaluation confirmed the positive effect of the prepared hydrogels on the healing process of the induced nerve injury. In the final, the histopathologic examinations depicted that the collagen/naringin hydrogel group reduced all the histological changes induced from the nerve injury and showed more resemblance to the normal sciatic nerve, with well-arranged fibers and intact myelin sheath. The overall results implied that the prepared collagen/naringin hydrogel can be utilized as a sophisticated alternative to healing peripheral nerve damages.



The authors gratefully acknowledge the research council of Kermanshah University of Medical Sciences (grant no. 980264) for financial support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Wang EW, Zhang J, Huang JH. Repairing peripheral nerve injury using tissue engineering techniques. Neural Regener Res. 2015;10:1393.CrossRefGoogle Scholar
  2. 2.
    Özkan HS, Karatas Silistreli O, Ergur B, İrkoren S. Repairing peripheral nerve defects by vein grafts filled with adipose tissue derived stromal vascular fraction: an experimental study in rats. Ulus Travma Acids Cerrahi Derg. 2016;22:7–11.Google Scholar
  3. 3.
    Wang H, Wu J, Zhang X, Ding L, Zeng Q. Study of synergistic role of allogenic skin-derived precursor differentiated Schwann cells and heregulin-1β in nerve regeneration with an acellular nerve allograft. Neurochem Int. 2016;97:146–53.CrossRefGoogle Scholar
  4. 4.
    Oprych KM, Whitby RL, Mikhalovsky SV, Tomlins P, Adu J. Repairing peripheral nerves: is there a role for carbon nanotubes? Adv Healthc Mater. 2016;5:1253–71.CrossRefGoogle Scholar
  5. 5.
    Massoumi B, Hatamzadeh M, Firouzi N, Jaymand M. Electrically conductive nanofibrous scaffold composed of poly (ethylene glycol)-modified polypyrrole and poly (ε-caprolactone) for tissue engineering applications. Mater Sci Eng: C. 2019;98:300–10.CrossRefGoogle Scholar
  6. 6.
    Mozaffari Z, Hatamzadeh M, Massoumi B, Jaymand M. Synthesis and characterization of a novel stimuli‐responsive magnetite nanohydrogel based on poly (ethylene glycol) and poly (N‐isopropylacrylamide) as drug carrier. J Appl Polym Sci. 2018;135:46657.CrossRefGoogle Scholar
  7. 7.
    Massoumi B, Mozaffari Z, Jaymand M. A starch-based stimuli-responsive magnetite nanohydrogel as de novo drug delivery system. Int J Biol Macromolecules. 2018;117:418–26.CrossRefGoogle Scholar
  8. 8.
    Poorgholy N, Massoumi B, Jaymand M. A novel starch-based stimuli-responsive nanosystem for theranostic applications. Int J Biol Macromolecules. 2017;97:654–61.CrossRefGoogle Scholar
  9. 9.
    Abbasian M, Massoumi B, Mohammad-Rezaei R, Samadian H, Jaymand M. Scaffolding polymeric biomaterials: are naturally occurring biological macromolecules more appropriate for tissue engineering? Int J Biol Macromolecules. 2019;134:673–94.CrossRefGoogle Scholar
  10. 10.
    Samadian H, Mobasheri H, Hasanpour S, Faridi-Majid R. Needleless electrospinning system, an efficient platform to fabricate carbon nanofibers. J Nano Res. 2017;50:78–89.CrossRefGoogle Scholar
  11. 11.
    Casolaro M, Casolaro I. Polyelectrolyte hydrogel platforms for the delivery of antidepressant drugs. Gels. 2016;2:24.CrossRefGoogle Scholar
  12. 12.
    Sgambato A, Cipolla L, Russo L. Bioresponsive hydrogels: chemical strategies and perspectives in tissue engineering. Gels. 2016;2:28.CrossRefGoogle Scholar
  13. 13.
    Adibi-Motlagh B, Lotfi AS, Rezaei A, Hashemi E. Cell attachment evaluation of the immobilized bioactive peptide on a nanographene oxide composite. Mater Sci Eng: C.2018;82:323–9.CrossRefGoogle Scholar
  14. 14.
    Ehterami A, Salehi M, Farzamfar S, Vaez A, Samadian H, Sahrapeyma H. In vitro and in vivo study of PCL/COLL wound dressing loaded with insulin-chitosan nanoparticles on cutaneous wound healing in rats model. Int J Biol Macromolecules. 2018;117:601–9.CrossRefGoogle Scholar
  15. 15.
    Ai A, Behforouz A, Ehterami A, Sadeghvaziri N, Jalali S, Farzamfar S. Sciatic nerve regeneration with collagen type I hydrogel containing chitosan nanoparticle loaded by insulin. Int J Polym Mater Polym Biomater. 2019;68:1–10.CrossRefGoogle Scholar
  16. 16.
    Yoshii S, Oka M. Collagen filaments as a scaffold for nerve regeneration. J Biomed Mater Res. 2001;56:400–5.CrossRefGoogle Scholar
  17. 17.
    Phillips JB, Bunting SC, Hall SM, Brown RA. Neural tissue engineering: a self-organizing collagen guidance conduit. Tissue Eng. 2005;11:1611–7.CrossRefGoogle Scholar
  18. 18.
    Kemp SW, Syed S, Walsh SK, Zochodne DW, Midha R. Collagen nerve conduits promote enhanced axonal regeneration, schwann cell association, and neovascularization compared to silicone conduits. Tissue Eng Part A. 2009;15:1975–88.CrossRefGoogle Scholar
  19. 19.
    Chen R, Qi Q-L, Wang M-T, Li Q-Y. Therapeutic potential of naringin: an overview. Pharm Biol. 2016;54:3203–10.CrossRefGoogle Scholar
  20. 20.
    Rong W, Wang J, Liu X, Jiang L, Wei F, Hu X. Naringin treatment improves functional recovery by increasing BDNF and VEGF expression, inhibiting neuronal apoptosis after spinal cord injury. Neurochem Res. 2012;37:1615–23.CrossRefGoogle Scholar
  21. 21.
    Rong W, Pan Y-W, Cai X, Song F, Zhao Z, Xiao S-H. The mechanism of Naringin-enhanced remyelination after spinal cord injury. Neural Regen Res. 2017;12:470.CrossRefGoogle Scholar
  22. 22.
    Wang D, Yan J, Chen J, Wu W, Zhu X, Wang Y. Naringin improves neuronal insulin signaling, brain mitochondrial function, and cognitive function in high-fat diet-induced obese mice. Cell Mol Neurobiol. 2015;35:1061–71.CrossRefGoogle Scholar
  23. 23.
    Salehi M, Naseri-Nosar M, Ebrahimi-Barough S, Nourani M, Vaez A, Farzamfar S. Regeneration of sciatic nerve crush injury by a hydroxyapatite nanoparticle-containing collagen type I hydrogel. J Physiological Sci. 2018;68:579–87.CrossRefGoogle Scholar
  24. 24.
    Ehterami A, Salehi M, Farzamfar S, Samadian H, Vaeez A, Ghorbani S. Chitosan/alginate hydrogels containing Alpha-tocopherol for wound healing in rat model. J Drug Deliv Sci Technol. 2019;51:204–213.CrossRefGoogle Scholar
  25. 25.
    Salehi M, Naseri-Nosar M, Azami M, Nodooshan SJ, Arish J. Comparative study of poly (L-lactic acid) scaffolds coated with chitosan nanoparticles prepared via ultrasonication and ionic gelation techniques. Tissue Eng Regener Med. 2016;13:498–506.CrossRefGoogle Scholar
  26. 26.
    Salehi M, Naseri‐Nosar M, Ebrahimi‐Barough S, Nourani M, Khojasteh A, Hamidieh AA. Sciatic nerve regeneration by transplantation of Schwann cells via erythropoietin controlled‐releasing polylactic acid/multiwalled carbon nanotubes/gelatin nanofibrils neural guidance conduit. J Biomed Mater Res Part B: Appl Biomater. 2018;106:1463–76.CrossRefGoogle Scholar
  27. 27.
    alehi M, Naseri-Nosar M, Ebrahimi-Barough S, Nourani M, Khojasteh A, Farzamfar S. Polyurethane/gelatin nanofibrils neural guidance conduit containing platelet-rich plasma and melatonin for transplantation of Schwann cells. Cell Mol Neurobiol. 2018;38:703–13.Google Scholar
  28. 28.
    Aurand ER, Lampe KJ, Bjugstad KB. Defining and designing polymers and hydrogels for neural tissue engineering. Neurosci Res. 2012;72:199–213.CrossRefGoogle Scholar
  29. 29.
    Woerly S. Porous hydrogels for neural tissue engineering. In: Materials Science Forum. Trans Tech Publ; 1997;250:53–68.CrossRefGoogle Scholar
  30. 30.
    Feng G, Nguyen TD, Yi X, Lyu Y, Lan Z, Xia J. Evaluation of long-term inflammatory responses after implantation of a novel fully bioabsorbable scaffold composed of poly-l-lactic acid and amorphous calcium phosphate nanoparticles. J Nanomater. 2018;2018:1–9.Google Scholar
  31. 31.
    Yin L, Cheng W, Qin Z, Yu H, Yu Z, Zhong M. Effects of naringin on proliferation and osteogenic differentiation of human periodontal ligament stem cells in vitro and in vivo. Stem Cells Int. 2015;2015:47–56.CrossRefGoogle Scholar
  32. 32.
    Dai K-R, Yan S-G, Yan W-Q, Chen D-Q, Xu Z-W. Effects of naringin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cell. Eur J Pharmacol. 2009;607:1–5.Google Scholar
  33. 33.
    Avia‐Saiz M, Busto MD, Pilar‐Izquierdo MC, Ortega N, Perez‐Mateos M, Muñiz P. Antioxidant properties, radical scavenging activity and biomolecule protection capacity of flavonoid naringenin and its glycoside naringin: a comparative study. J Sci Food Agriculture. 2010;90:1238–44.Google Scholar
  34. 34.
    Anuja G, Latha P, Suja S, Shyamal S, Shine V, Sini S, et al. Anti-inflammatory and analgesic properties of Drynaria quercifolia (L.) J. Smith. J Ethnopharmacol. 2010;132:456–60.CrossRefGoogle Scholar
  35. 35.
    Choe S-C, Kim H-S, Jeong T-S, Bok S-H, Park Y-B. Naringin has an antiatherogenic effect with the inhibition of intercellular adhesion molecule-1 in hypercholesterolemic rabbits. J Cardiovasc Pharmacol. 2001;38:947–55.CrossRefGoogle Scholar
  36. 36.
    Pereira JE, Costa LM, Cabrita AM, Couto PA, Filipe VM, Magalhães LG. Methylprednisolone fails to improve functional and histological outcome following spinal cord injury in rats. Exp Neurol. 2009;220:71–81.CrossRefGoogle Scholar
  37. 37.
    Ma X, Lv J, Sun X, Ma J, Xing G, Wang Y. Naringin ameliorates bone loss induced by sciatic neurectomy and increases Semaphorin 3A expression in denervated bone. Sci Rep. 2016;6:24562.Google Scholar
  38. 38.
    Kim HJ, Song JY, Park HJ, Park HK, Yun DH, Chung JH. Naringin protects against rotenone-induced apoptosis in human neuroblastoma SH-SY5Ycells. Korean J Physiol Pharmacol. 2009;13:281–5.CrossRefGoogle Scholar
  39. 39.
    Satou T, Nishida S, Hiruma S, Tanji K, Takahashi M, Fujita S. A morphological study on the effects of collagen gel matrix on regeneration of severed rat sciatic nerve in silicone tubes. Pathol Int. 1986;36:199–208.CrossRefGoogle Scholar
  40. 40.
    Chamberlain L, Yannas I, Hsu H, Strichartz G, Spector M. Collagen-GAG substrate enhances the quality of nerve regeneration through collagen tubes up to level of autograft. Exp Neurol. 1998;154:315–29.CrossRefGoogle Scholar
  41. 41.
    Goto E, Mukozawa M, Mori H, Hara M. A rolled sheet of collagen gel with cultured Schwann cells: model of nerve conduit to enhance neurite growth. J Biosci Bioeng. 2010;109:512–8.CrossRefGoogle Scholar
  42. 42.
    Kim HD, Jeong KH, Jung UJ, Kim SR. Naringin treatment induces neuroprotective effects in a mouse model of Parkinson’s disease in vivo, but not enough to restore the lesioned dopaminergic system. J Nutr Biochem. 2016;28:140–6.CrossRefGoogle Scholar
  43. 43.
    Rong W, Cai X, Pan Y, Song F, Zhang C, Xiao S. Combination therapy of chitosan conduit and naringin facilitate regeneration of injured sciatic nerve in rats. In: BIBE 2018; International Conference on Biological Information and Biomedical Engineering. VDE 2018. pp 1–4.Google Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Nano Drug Delivery Research Center, Health Technology InstituteKermanshah University of Medical SciencesKermanshahIran
  2. 2.Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran
  3. 3.Department of Mechanical and Aerospace Engineering, Science and Research BranchIslamic Azad UniversityTehranIran
  4. 4.Department of Tissue Engineering, School of MedicineShahroud University of Medical SciencesShahroudIran
  5. 5.Tissue Engineering and Stem Cells Research CenterShahroud University of Medical SciencesShahroudIran
  6. 6.Department of Biomedical Engineering, Science and Research BranchIslamic Azad UniversityTehranIran
  7. 7.Department of Physiology, School of MedicineShahroud University of Medical SciencesShahroudIran

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