Effect of cells in the epimysium conduit on the regeneration of peripheral nerve_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

FENG Weifeng , LU Haibin , XU Zhuqiu , CHEN Lulu ,  YANG Xiaonan ,  QI Zuoliang

Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100144, P.R.China;

Corresponding author：

YANG Xiaonan, Email: yxnan@aliyun.com; QI Zuoliang, Email: qizuoliang1h@163.com

Keywords：

Epimysium conduit; nerve regeneration; cell migration inhibition; decellularization

DOI：

10.7507/1002-1892.201712092

Video：

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ObjectiveTo investigate the effect of cells in the epimysium conduit (EMC) on the regeneration of sciatic nerve of mice.MethodsThe epimysium of the 8-week-old male C57BL/6J enhanced green fluorescent protein (EGFP) mouse was trimmed to a size of 5 mm×3 mm, and prepared in a tubular shape (ie, EMC). Some epimysia were treated with different irradiation doses (0, 15, 20, 25, 30, 35 Gy) to inhibit cells migration. Then the number of migrating cells were counted, and the epimysia with the least migrating cells were selected to prepare EMC. Some epimysia were subjected to decellularization treatment and prepared EMC. HE and Masson staining were used to identify the decellularization effect. Twenty-four C57BL/6J wild-type mice were used to prepare a 3-mm-long sciatic nerve defect of right hind limb model and randomly divided into 3 groups (n=8). EMC (group A), EMC after cell migration inhibition treatment (group B), and decellularized EMC (group C) were used to repair defects. At 16 weeks after operation, the midline of the regenerating nerve was taken for gross, toluidine blue staining, immunofluorescence staining, and transmission electron microscopy.ResultsAt 15 days, the number of migrating cells gradually decreased with the increase of irradiation dose. There was no significant difference between 30 Gy group and 35 Gy group (P>0.05); there were significant differences between the other groups (P<0.05). The epimysium after treatment with 35 Gy irradiation dose was selected for thein vivo experiment. After the decellularization of the epimysium, no nucleus was found in the epimysium and the epimysium could be sutured to prepare EMC. At 16 weeks after operation, the nerves in all groups were recanalized. The sciatic nerve was the thickest in group A, followed by group B, and the finest in group C. Immunofluorescence staining showed that the EGFP cells in group A were surrounded by regenerated axons. Toluidine blue staining and transmission electron microscopy observation showed that the number of regenerated axons and the thickness of regenerated myelin sheath in group A were significantly better than those in groups B and C (P<0.05). There was no significant difference between groups B and C (P>0.05).ConclusionThe cellular components of the epimysium participate in and promote the regeneration of the sciatic nerve in mice.

Citation： FENG Weifeng, LU Haibin, XU Zhuqiu, CHEN Lulu, YANG Xiaonan, QI Zuoliang. Effect of cells in the epimysium conduit on the regeneration of peripheral nerve. Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(5): 617-624. doi: 10.7507/1002-1892.201712092 Copy

1.	Hu Y, Wu Y, Gou Z, et al. 3D-engineering of cellularized conduits for peripheral nerve regeneration. Sci Rep, 2016, 6: 32184.
2.	Niu Y, Li L, Chen KC, et al. Scaffolds from alternating block polyurethanes of poly (ε-caprolactone) and poly(ethylene glycol) with stimulation and guidance of nerve growth and better nerve repair than autograft. J Biomed Mater Res A, 2015, 103(7): 2355-2364.
3.	Prabhakaran MP, Ghasemi-Mobarakeh L, Jin G, et al. Electrospun conducting polymer nanofibers and electrical stimulation of nerve stem cells. J Biosci Bioeng, 2011, 112(5): 501-507.
4.	Binan L, Tendey C, De Crescenzo G, et al. Differentiation of neuronal stem cells into motor neurons using electrospun poly-L-lactic acid/gelatin scaffold. Biomaterials, 2014, 35(2): 664-674.
5.	Junka R, Valmikinathan CM, Kalyon DM, et al. Laminin functionalized biomimetic nanofibers for nerve tissue engineering. J Biomater Tissue Eng, 2013, 3(4): 494-502.
6.	林强, 蔡杨庭, 李皓莘. 负载浓度梯度 NGF 的周围神经导管修复大鼠周围神经缺损的实验研究. 中国修复重建外科杂志, 2014, 28(2): 167-172.
7.	Pereira T, Gärtner A, Amorim I, et al. Promoting nerve regeneration in a neurotmesis rat model using poly (DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton’s jelly: in vitro and in vivo analysis. Biomed Res Int, 2014, 2014: 302659.
8.	Saito K, Tamaki T, Hirata M, et al. Reconstruction of multiple facial nerve branches using skeletal muscle-derived multipotent stem cell sheet-pellet transplantation. PLoS One, 2015, 10(9): e0138371.
9.	Li G, Xiao Q, Zhang L, et al. Nerve growth factor loaded heparin/chitosan scaffolds for accelerating peripheral nerve regeneration. Carbohydr Polym, 2017, 171: 39-49.
10.	Cao J, Xiao Z, Jin W, et al. Induction of rat facial nerve regeneration by functional collagen scaffolds. Biomaterials, 2013, 34(4): 1302-1310.
11.	Hu N, Wu H, Xue C, et al. Long-term outcome of the repair of 50 mm long median nerve defects in rhesus monkeys with marrow mesenchymal stem cells-containing, chitosan-based tissue engineered nerve grafts. Biomaterials, 2013, 34(1): 100-111.
12.	Gu X, Ding F, Williams DF. Neural tissue engineering options for peripheral nerve regeneration. Biomaterials, 2014, 35(24): 6143-6156.
13.	Yang XN, Jin YQ, Wei W, et al. Peripheral nerve repair with epimysium conduit. Biomaterials, 2013, 34(22): 5606-5616.
14.	Iliakis G, Wang Y, Guan J, et al. DNA damage checkpoint control in cells exposed to ionizing radiation. Oncogene, 2003, 22(37): 5834-5847.
15.	Rofstad EK, Mathiesen B, Galappathi K. Increased metastatic dissemination in human melanoma xenografts after subcurative radiation treatment: radiation-induced increase in fraction of hypoxic cells and hypoxia-induced up-regulation of urokinase-type plasminogen activator receptor. Cancer Res, 2004, 64(1): 13-18.
16.	Nehls P, van Beuningen D, Karwowski M. After X-irradiation a transient arrest of L929 cells in G2-phase coincides with a rapid elevation of the level of O6-alkylguanine-DNA alkyltransferase. Radiat Environ Biophys, 1991, 30(1): 21-31.
17.	Storme G, Mareel M, De Bruyne G. Influence of cells number on directional migration of MO4 cells in vitro. Arch Geschwulstforsch, 1981, 51(1): 45-50.
18.	Weber B, Emmert MY, Schoenauer R, et al. Tissue engineering on matrix: future of autologous tissue replacement. Semin Immunopathol, 2011, 33(3): 307-315.
19.	Piccoli M, Urbani L, Alvarez-Fallas ME, et al. Improvement of diaphragmatic performance through orthotopic application of decellularized extracellular matrix patch. Biomaterials, 2016, 74: 245-255.
20.	Gilbert TW. Strategies for tissue and organ decellularization. J Cell Biochem, 2012, 113(7): 2217-2222.
21.	Jessen KR, Mirsky R. The origin and development of glial cells in peripheral nerves. Nat Rev Neurosci, 2005, 6(9): 671-682.
22.	Tamaki T, Okada Y, Uchiyama Y, et al. Clonal multipotency of skeletal muscle-derived stem cells between mesodermal and ectodermal lineage. Stem Cells, 2007, 25(9): 2283-2290.
23.	Bollyky PL, Falk BA, Wu RP, et al. Intact extracellular matrix and the maintenance of immune tolerance: high molecular weight hyaluronan promotes persistence of induced CD4+CD25+ regulatory T cells. J Leukoc Biol, 2009, 86(3): 567-572.
24.	Bollyky PL, Wu RP, Faik BA, et al. ECM components guide IL-10 producing regulatory T-cell (TR1) induction from effector memory T-cell precursors. Proc Natl Acad Sci U S A, 2011, 108(19): 7938-7943.
25.	Morwood SR, Nicholson LB. Modulation of the immune response by extracellular matrix proteins. Arch Immunol Ther Exp (Warsz), 2006, 54(6): 367-374.
26.	Brown BN, Valentin JE, Stewart-Akers AM, et al. Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. Biomaterials, 2009, 30(8): 1482-1491.
27.	Beattie AJ, Gilbert TW, Guyot JP, et al. Chemoattraction of progenitor cells by remodeling extracellular matrix scaffolds. Tissue Eng Part A, 2009, 15(5): 1119-1125.
28.	Reing JE, Zhang L, Myers-Irvin J, et al. Degradation products of extracellular matrix affect cell migration and proliferation. Tissue Eng Part A, 2009, 15(3): 605-614.

1. Hu Y, Wu Y, Gou Z, et al. 3D-engineering of cellularized conduits for peripheral nerve regeneration. Sci Rep, 2016, 6: 32184.
2. Niu Y, Li L, Chen KC, et al. Scaffolds from alternating block polyurethanes of poly (ε-caprolactone) and poly(ethylene glycol) with stimulation and guidance of nerve growth and better nerve repair than autograft. J Biomed Mater Res A, 2015, 103(7): 2355-2364.
3. Prabhakaran MP, Ghasemi-Mobarakeh L, Jin G, et al. Electrospun conducting polymer nanofibers and electrical stimulation of nerve stem cells. J Biosci Bioeng, 2011, 112(5): 501-507.
4. Binan L, Tendey C, De Crescenzo G, et al. Differentiation of neuronal stem cells into motor neurons using electrospun poly-L-lactic acid/gelatin scaffold. Biomaterials, 2014, 35(2): 664-674.
5. Junka R, Valmikinathan CM, Kalyon DM, et al. Laminin functionalized biomimetic nanofibers for nerve tissue engineering. J Biomater Tissue Eng, 2013, 3(4): 494-502.
6. 林强, 蔡杨庭, 李皓莘. 负载浓度梯度 NGF 的周围神经导管修复大鼠周围神经缺损的实验研究. 中国修复重建外科杂志, 2014, 28(2): 167-172.
7. Pereira T, Gärtner A, Amorim I, et al. Promoting nerve regeneration in a neurotmesis rat model using poly (DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton’s jelly: in vitro and in vivo analysis. Biomed Res Int, 2014, 2014: 302659.
8. Saito K, Tamaki T, Hirata M, et al. Reconstruction of multiple facial nerve branches using skeletal muscle-derived multipotent stem cell sheet-pellet transplantation. PLoS One, 2015, 10(9): e0138371.
9. Li G, Xiao Q, Zhang L, et al. Nerve growth factor loaded heparin/chitosan scaffolds for accelerating peripheral nerve regeneration. Carbohydr Polym, 2017, 171: 39-49.
10. Cao J, Xiao Z, Jin W, et al. Induction of rat facial nerve regeneration by functional collagen scaffolds. Biomaterials, 2013, 34(4): 1302-1310.
11. Hu N, Wu H, Xue C, et al. Long-term outcome of the repair of 50 mm long median nerve defects in rhesus monkeys with marrow mesenchymal stem cells-containing, chitosan-based tissue engineered nerve grafts. Biomaterials, 2013, 34(1): 100-111.
12. Gu X, Ding F, Williams DF. Neural tissue engineering options for peripheral nerve regeneration. Biomaterials, 2014, 35(24): 6143-6156.
13. Yang XN, Jin YQ, Wei W, et al. Peripheral nerve repair with epimysium conduit. Biomaterials, 2013, 34(22): 5606-5616.
14. Iliakis G, Wang Y, Guan J, et al. DNA damage checkpoint control in cells exposed to ionizing radiation. Oncogene, 2003, 22(37): 5834-5847.
15. Rofstad EK, Mathiesen B, Galappathi K. Increased metastatic dissemination in human melanoma xenografts after subcurative radiation treatment: radiation-induced increase in fraction of hypoxic cells and hypoxia-induced up-regulation of urokinase-type plasminogen activator receptor. Cancer Res, 2004, 64(1): 13-18.
16. Nehls P, van Beuningen D, Karwowski M. After X-irradiation a transient arrest of L929 cells in G2-phase coincides with a rapid elevation of the level of O6-alkylguanine-DNA alkyltransferase. Radiat Environ Biophys, 1991, 30(1): 21-31.
17. Storme G, Mareel M, De Bruyne G. Influence of cells number on directional migration of MO4 cells in vitro. Arch Geschwulstforsch, 1981, 51(1): 45-50.
18. Weber B, Emmert MY, Schoenauer R, et al. Tissue engineering on matrix: future of autologous tissue replacement. Semin Immunopathol, 2011, 33(3): 307-315.
19. Piccoli M, Urbani L, Alvarez-Fallas ME, et al. Improvement of diaphragmatic performance through orthotopic application of decellularized extracellular matrix patch. Biomaterials, 2016, 74: 245-255.
20. Gilbert TW. Strategies for tissue and organ decellularization. J Cell Biochem, 2012, 113(7): 2217-2222.
21. Jessen KR, Mirsky R. The origin and development of glial cells in peripheral nerves. Nat Rev Neurosci, 2005, 6(9): 671-682.
22. Tamaki T, Okada Y, Uchiyama Y, et al. Clonal multipotency of skeletal muscle-derived stem cells between mesodermal and ectodermal lineage. Stem Cells, 2007, 25(9): 2283-2290.
23. Bollyky PL, Falk BA, Wu RP, et al. Intact extracellular matrix and the maintenance of immune tolerance: high molecular weight hyaluronan promotes persistence of induced CD4+CD25+ regulatory T cells. J Leukoc Biol, 2009, 86(3): 567-572.
24. Bollyky PL, Wu RP, Faik BA, et al. ECM components guide IL-10 producing regulatory T-cell (TR1) induction from effector memory T-cell precursors. Proc Natl Acad Sci U S A, 2011, 108(19): 7938-7943.
25. Morwood SR, Nicholson LB. Modulation of the immune response by extracellular matrix proteins. Arch Immunol Ther Exp (Warsz), 2006, 54(6): 367-374.
26. Brown BN, Valentin JE, Stewart-Akers AM, et al. Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. Biomaterials, 2009, 30(8): 1482-1491.
27. Beattie AJ, Gilbert TW, Guyot JP, et al. Chemoattraction of progenitor cells by remodeling extracellular matrix scaffolds. Tissue Eng Part A, 2009, 15(5): 1119-1125.
28. Reing JE, Zhang L, Myers-Irvin J, et al. Degradation products of extracellular matrix affect cell migration and proliferation. Tissue Eng Part A, 2009, 15(3): 605-614.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of cells in the epimysium conduit on the regeneration of peripheral nerve

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