Using Eggshell Membrane as Nerve Guide Channels in Peripheral Nerve Regeneration


1 Neurophysiology Research Center, Department of Anatomy, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran

2 Neurophysiology Research Center, Department of Physiology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran

3 Department of Anatomy, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran

4 Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran


Objective(s):  The aim of this study was to evaluate the final outcome of nerve regeneration across the eggsell membrane (ESM) tube conduit in comparison with autograft.
Materials and Methods: Thirty adult male rats (250-300 g) were randomized into (1) ESM conduit, (2) autograft, and (3) sham surgery groups. The eggs submerged in 5% acetic acid. The decalcifying membranes were cut into four pieces, rotated over the teflon mandrel and dried at   37°C. The left sciatic nerve was surgically cut. A 10-mm nerve segment was cut and removed. In the ESM group, the proximal and distal cut ends of the sciatic nerve were telescoped into the nerve guides. In the autograft group, the 10 mm nerve segment was reversed and used as an autologous nerve graft. All animals were evaluated by sciatic functional index (SFI) and electrophysiology testing. 
Results:The improvement in SFI from the first to the last evalution in ESM and autograft groups were evaluated. On days 49 and 60 post-operation, the mean SFI of ESM group was significantly greater than the autograft group (P< 0.05). On day 90, the mean nerve conduction velocity (NCV) of ESM group was greater than autograft group, although the difference was not statistically significant (P> 0.05).
Conclusion:These findings demonstrate that ESM effectively enhances nerve regeneration and promotes functional recovery in injured sciatic nerve of rat.


1. Sunderland S. Nerves and nerve injuries. New York: Churchill-Livingstone; 1978. Chap. 5, p. 68.

2. Valentini RF. Nerve guide channels. In: Bronzino JD, editor.The biomedical engineering handbook. Boca Raton. FL: CRC Press; 1995. p. 1985-1996.

3. Hudson TW, Evans GRD, Schmidt CE. Engineering strategies for peripheral nerve repair. Clin Plast Surg 1999; 26:617-628.

4. Johnson EO, Zoubos AB, Soucacos PN. Regeneration and repair of peripheral nerves. Injury 2005; 365:S24-S29.

5. Pannunzio ME, Jou I, Long A, Wind TC, Beck G, Balian G. A new method of selecting Schwann cells from adult mouse sciatic nerve. J Neurosci Methods 2005; 149:74-81.

6. Eppley BL, Snyders RV, Winkelmann TM, Roufa DG. Efficacy of nerve growth factor in regeneration of the mandibular nerve: A Preliminary Report. J Oral Maxillofac Surg 1991; 49:61-68.

7. Dahlin LB. The biology of nerve injury and repair. JHS 2004; 4:143-155.

8. Cordeiro CM, Hincke MT. Recent patents on eggshell: shell and membrane applications. Recent Pat Food Nutr Agric 2011; 3:1-8.

9. Nakano T, Ikawa NI, Ozimek L. Chemical composition of chicken eggshell and shell membranes. Poult Sci  2003; 82:510-514. 

10. Ruff KJ, Winkler A, Jackson RW, DeVore DP, Ritz BW. Eggshell membrane in the treatment of pain and stiffness from osteoarthritis of the knee: a randomized, multicenter, double-blind, placebo-controlled clinical study. Clin Rheumatol 2009; 28:907-914. 

11. Tavassoli M. Effect of the substratum on the growth of CFU-c in continuous marrow culture. Experientia 1983; 39:411-412.  

12. Maeda K, Sasaki Y. An experience of hen-egg membrane as a biological dressing burns. Incl Therm Inj 1982; 8:313-316.  

13. Yi F, Guo Z, Zhang L, Yu J, Li Q. Soluble eggshell membrane protein: preparation, characterization and biocompatibility. Biomaterials  2004; 25:4591-4599.

14. Jia J, Duan YY, Yu J, Lu JW. Preparation and immobilization of soluble eggshell membrane protein on the electrospun nanofibers to enhance cell adhesion and growth. J Biomed Mater Res A  2008; 86:346-373.

15. Arias JI, Gonzalez A, Fernandez Ms, Gonzalez C, Sez D, Arias IL. Eggshell membrane as a biodegradable bone regeneration inhibitor. J Tissue Regen Med 2008; 2:228-235.

16. Leighton J, Mansukhani S, Estes L. Decalcifiied eggshell membrane, a supporting substrate for electron microscopic cross sections of monolayers of epithelial cell line MDCK. InVitro 1971; 6:251-252.  

17. Bain JR, Mackinnon SE, Hunter DA. Functional evaluation of complete sciatic, peroneal, and posterior tibial nerve lesions in the rat. Plast Reconstr Surg 1989; 83: 129-138.   

18. Menovsky T, Beek JF. Laser, fibrin glue, or suture repair of peripheral nerves: a comparative functional, hstological, and morphometric study in the rat sciatic nerve. J Neurosurg 2001; 95:694-699.

19. Seniuk NA. Neurotrophic factors: role in peripheral neuron survival and axonal repair. J Reconstr Microsurg 1992; 8:399-404. 

20. Upman PJ, Anderson J, Tasse L. An analysis of ISO intracutaneous reactivity test results to justify a reduction in animal requirements. Lab Anim (NY) 2003; 32: 26-27.   

21. Nakano T, Ikawa N, Ozimek L. Extraction of glycosaminoglycans from chicken eggshell poultry. Science  2001; 80:681-684.

22. Wang KK, Nemeth IR, Seckel BR, Chakalis-Haley DP, Swann DA, Kuo JW, et al. Hyaluronic acid enhances peripheral nerve regeneration in vivo. Microsurgery 1998; 18:270-275.

23. Miksik L, Eckhadt A, Sedlakova P, Mikulikova K. Proteins of insoluble matrix of avian (gallus gallus) eggshell. Connect Tissue Res 2007; 48:1-8. 

24. Arias JL, Carrino DA, Fernandez MS, Rodriguez JP, Dennis JE, Caplan AL. Partial biochemical and immunochemical characterization of avian eggshell extracellular matrices. Arch Biochem Biophys 1992; 298:293-302.

25. Kitahara A K, Suzuki Y, Nishimura Y, Suzuki K, Kiyotani T, Takimoto Y, et al. Evalution of collagen nerve guide in facial nerve regeneration. J Artif Organs 1998; 1:22-27.

26. Mohammad J, Shenaq J, Rabinovsky E, Shenag S. Modulation of peripheral nerve regeneration: a tissue-engineering approach. The role of amnion tube nerve conduit across a 1- centimeter nerve gap. Plast Reconstr Surg 2000; 105:660-666.

27. Hare GMT, Evans PJ, Mackinnon SE, Best TJ, Bain JR, Szalai JP, et al. Walking track analysis: a long-term assessment of peripheral nerve recovery. Plast Reconstr surg 1992; 89:251-258.

28. Reynolds JL, Urbanchek MS, Asato H, Kuzon WM JR. Deletion of individual muscles alters rat walking-track parameters. J Reconstr Microsurg 1996; 12:461-466.   

29. Ansselin AD, Fink T, Davey DF. Peripheral nerve regeneration through nerve guides seeded with adult Schwann cells. Neuropathol Appl Neurobiol 1997; 23:    387-398.

30. Castaneda F, Kinne RK. Omental graft improves functional recovery of transected peripheral nerve. Muscle Nerve 2002; 26:527-532. 

31. Munro CA, Szalai JP, Mackinnon SE, Midha R. Lack of association between outcome measures of nerve regeneration. Muscle Nerve 1998; 21:1095-1097.  

32. Kanaya F, Firrell JC, Breidenbach WC. Sciatic functional index, nerve conduction tests, muscle contraction, and axon morphometry as indication of regeneration. Plast Reconstr Surg 1996; 98:1264-1271.

33. Mohammadi R, Azizi S, Delirezh N, Hobbenaghi R, Amini K. Functional recovery of sciatic nerve through inside-out vein graft in rats. Chin J Traumatol 2011; 14:46-52.

34. Labrador RO, Buti M, Navarro X. Influence of collagen and laminin gels concentration on nerve regeneration after resection and tube repair.Exp Neurol 1998; 149:243-252.