Histopathological and immunohistochemical evaluation of the effect of tacrolimus on nerve regeneration following crushed sciatic nerve in mice

Subject Areas : Veterinary Clinical Pathology

fereshteh khomejani farahani ¹ (D.V.M, DVS.c. Graduate, Department of Clinical Science, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
Hamidreza Fattahian ² (Assistant Professor, Department of Clinical Science, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
Ahmad Asghari ³ (Associate Professor, Department of Clinical Science, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
pejman Mortazavi ⁴ (Associate Professor, Department of Pathobiology, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.)

Received: 2021-06-15 Accepted : 2021-10-30 Published : 2021-10-23

Keywords: Mice, Histopathology, Immunohistochemistry, Sciatic Nerve, Tacrolimus,

Abstract :

Studies have been conducted to find effective agents for nerve regeneration. The present study was conducted on sixteen adult male Syrian mice with the aim of assessment of the effects of tacrolimus on crushed sciatic nerve injury and its comparison with spontaneous repair. After exposure of the left sciatic nerve, crushing was performed using a 2 mm wide mosquito hemostatic forceps tip for 10 seconds. Animals were randomized into two groups of treatment group (Group I), and the control (Group II) with eight mice each. The treatment group received tacrolimus (5 mg/kg, q24h, SC) until the end of the study, and the control group received no therapeutic agents. Histopathological and immunohistochemical assessment was performed on second and fourth post-operative weeks, and suggested that perineurium formation did not show a significant difference between the study groups (p>0.05). At the end of the second week in the treatment group, the reduction of axon inflammation and swelling, and the increase in axonal count showed a significant difference with the control group (p<0.05).  At the end of the fourth week, the difference in inflammation severity between the groups was not significant (p>0.05) but the decrease in axonal swelling and increase in axonal count in the treatment group showed a significant difference with the control group (p<0.05).  Increase of glial fibrillary acidic protein (GFAP) expression showed significant difference between the treatment group and the control group at the end of the study (p<0.05). Based on the present results, the use of tacrolimus was responsible for reduction of inflammation in a two week period, and at the end of the fourth week, increase of axonal count and edema suppression caused regeneration of injured nerve.

References:

• Alvites, R., Caseiro, A.R., Pedrosa, S.S., Branquinho, M.V., Ronchi, G., Geuna, S., et al. (2018). Peripheral nerve injury and axonotmesis: State of the art and recent advances. Cogent Medicine, 5(1): 1-45.


• Blackshaw, J.K., Fenwick, D.C., Beattie, A.W. and Allan, D.J. (1988). The behavior of chicken, mice and rats during euthanasia with chloroform, carbon dioxide and ether. Laboratory Animals, 22(1): 67-75.


• Caner, B., Kafa, M.I., Bekar, A., Kurt, M.A., Karli, N., Cansev, M., et al. (2012). Intraperitoneal administration of CDP- choline or a combination of cytidine plus choline improves nerve regeneration and functional recovery in a rat model of sciatic nerve injury. Neuro Research, 34(3): 238-245.


• Danzi, M.C., Motti, D., Avison, D.L., Bixby, J.L. and Lemmon, V.P. (2016). Treatment with analgesics after mouse sciatic nerve injury does not alter expression of wound healing-associated genes. Neural Regeneration Research, 11(1): 144-149.


• Darzian Rostami, Z., Asghari, A., Jahandideh A., Mortazavi, P. and Akbarzadeh, A. (2020). Effect of oat (Avena Sativa L.) extraction on experimental sciatic nerve injury in rats. Archive of Razi Institute, 75(2): 249-256.


• Devesa, P., Gelabert, M., Gonzlez-Mosquera, T., Gallego, R., Relova, J.L., Devesa, J., et al. (2012). Growth hormone treatment enhances the functional recovery of sciatic nerves after transection and repair. Muscle & Nerve, 45(3): 385-392.


• Dewey, C.W. and Coates, J.R. (2003). Miscellaneous spinal condition and peripheral nerve injuries. In: Textbook of Small Animal Surgery. Sharp, N.J.H. editor. 3rd ed., USA, Philadelphia: Elsevier Science, pp: 1209-1226.


• Fargo, K.N., Foecking, EM., Jones, K.J. and Sengelaub, D.R. (2009). Neuroprotective action of androgen on motoneurons. Frontiers in Neuroendocrinology, 30(2): 130-141.


• Grand, A.G., Myckatyn, T.M., Mackinnon, S.E. and Hunter, D.A. (2002). Axonal regeneration after cold preservation of nerve allografts and immunosuppression with tacrolimus in mice. Journal of Neurosurgery, 96 (5): 944-932.


• Islamov, R.R., Hendricks, W.A., Jones, R.J., Lyall, G.J., Spanier, N.S. and Murashov, A.K. (2002). 17B- estradiol stimulation regeneration of sciatic nerve in female mice. Brain Research, 943(2): 283- 286.


• Islamov, R.R., Hendricks, W.A., Katwa, L.C., McMurray, R.J., Pak, E.S., Spanier, N.S., et al. (2003). Effect of 17B- estradiol in lumbar spinal cord following sciatic nerve crush injury in ovariectomized mice. Brain Research, 966(1): 65-75.


• Mahloujiyan, A., Jahandideh, A., Asghari, A. and Mortazavi, P. (2020). Experimental evaluation of the effect of folic acid on corneal burn ulcer healing in New Zealand white rabbit. Journal of Veterinary Clinical Pathology, 13(52): 371-383. [In Persian]


• Mekaj, A.Y., Morina, A.A., Bytyqi, C.I., Mekaj, Y.H. and Duci, S.B. (2014). Application of topical pharmacological agents at the site of peripheral nerve injury and methods used for evaluating the success of the regenerative process. Journal of Orthopaedic Surgery and Research, 9(94): 1-7.


• Mesquit Coutinho, P.R., Cristante, A.F., Barros Filho, T.E.P., Ferreira, R. and Santos, G.B. (2016). Effect of tacrolimus and erythropoietin in experimental spinal cord lesion in rats: functional and histological evaluation. Spinal Cord, 54(6): 439- 444.


• Platt, S.R. and Costa, R.C. (2012). Cervical Spine. In: Veterinary Surgery: Small Animal. Tobias, K.M. and Johnston, S.A. editors. First ed., USA, Maryland: Elsevier Saunders, pp: 410-448.


• Reyhani Rad, S., Mohajeri, D., Mousavi, Gh., Mahmoudi, J., Rezaie, A. and Yazdchi, S. (2009). Histometric and histopathological evaluation of the effect of Sertraline following cutaneous surgical trauma in the rat. Journal of Veterinary Clinical Pathology, 3(2): 465-477. [In Persian]


• Shabeed, D., Najafi, M., Keshavarz, M., Musa. A.E, Hassanzadeh, G., Hadian. M.R., et al. (2018). Recent finding in repair of peripheral nerve lesions using pharmacological agents: Common methods for evaluating the repair process. Central Nervous System Agents in Medicinal Chemistry, 18(3): 1-12.


• Suchyta, M.A., Sabbagh, M.D., Morsy, M., Mardini, S. and Moran, S.L. (2017). Advances in peripheral nerve regeneration as it relates to VCA. Vascularized Composite Allotransplantation, 3(1,2): 75-88.


• Sun, W., Sun, C., Lin, H., Zhao, H., Wang, J., Ma, H., et al. (2009). The effect of collagen-binding NGF-β on the promotion of sciatic nerve regeneration in a rat sciatic nerve crush injury model. Biomaterials, 30(27): 4649-4656.


• Wallemacq P.E. and Reding R. (1993). FK506 (Tacrolimus), a novel immunosuppressant in organ transplantation: clinical, biomedical, and analytical aspects. Clinical Chemistry, 39(11): 2219- 2228.


• Wang, M.S., Zeleny-Pooley, M. and Gold, B.G. (1997). Comparative dose- dependence study of Fk506 and cyclosporine A on the rate of axonal regeneration in the Rat Sciatic Nerve. The Journal of Pharmacology and Experimental Therapeutics, 282(2): 1084-­


• Yadegar, O., Asghari, A. and Hesaraki, S. (2013). Evaluation of wound healing activity of Commiphoramyrrha extract compared with silver sulfadiazine on experimental skin burn healing in rat. Journal of Veterinary Clinical Pathology, 7(3): 173-182. [In Persian]


• Yang, R.K., Lowe, J.B., Sobol, J.B., Sen, S.K., Hunter, D.A. and Mackinnon, S.E. (2001). Dose- dependent effects of Fk506 on neuroregeneration in a rat model. Plastic and Reconstructive Surgery, 112(7): 1832-1840.