IN VIVO STUDY ON TISSUE ENGINEERED SKELETAL MUSCLE WITH HYPOGLOSSAL NERVE IMPLANTATION_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

TANG Xiufa ¹ , ZHANG Fugui ² , FENG Yang ² , ZHOU Wei ² , LIU Jiyuan ² , HUA Chengge ¹

1Department of Head &;
Neck Oncology, 2State Key Laboratory of Oral Diseases,West China School of Stomatology, Sichuan University, Chengdu Sichuan, 610041, P.R.China.;

Corresponding author：

Keywords：

Tissue engineered skeletal muscle; Glial cell derived neurotrophic factor; Acellular collagen sponge; Hypoglossal nerve; Myoblast Rat

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

【Abstract】 Objective To construct tissue engineered skeletal muscle in vivo using glial cell derived neurotrophic factor (GDNF) genetically modified myoblast (Mb) on acellular collagen sponge with hypoglossal nerve implantation, and to observe whether structural or functional connection could be established between engineered tissue and motor nerve or not. Methods Mbs were isolated from 7 male Lewis rats at age of 2 days, cultured and genetically modified by recombinant adenovirus carrying GDNF cDNA (MbGDNF). Calf skin-derived acellular collagen sponge was used as scaffold; cell adhesion was detected by scanning electron microscope after 24 hours. Hypoglossal nerve was implanted into Mb-scaffold complex (Mb group, n=27) or MbGDNF-scaffold complex (MbGDNF group, n=27) in 54 female Lewis rats at age of 8 weeks. HE staining was performed at 1, 6, and 12 weeks postoperatively, and immunohistochemistry staining and fluorescence in situ hybridization were used. Results MbGDNF could highly expressed GDNF gene. Mb and MbGDNF could adhere to the scaffold and grew well. HE staining showed tight junctions between implant and peripheral tissue with new muscle fiber and no distinguished line at 12 weeks in 2 groups. Immunohistochemistry staining showed that positive cells of myogenin and slow skeletal myosin were detected, as well as positive cells of actylcholine receptor α1 at 1, 6, and 12 weeks. The positive cells of Y chromosome decreased with time. At 1, 6, and 12 weeks, the positive neurons were 261.0 ± 6.6, 227.3 ± 8.5, and 173.3 ± 9.1, respectively in MbGDNF group, and were 234.7 ± 5.5, 196.0 ± 13.5, and 166.7 ± 11.7, respectively in Mb group; significant differences were found between 2 groups at 1 and 6 weeks (P lt; 0.05), no significant difference at 12 weeks (P gt; 0.05). Conclusion Connection can be established between engineered tissue and implanted hypoglossal nerve. Recombinant GDNF produced by MbGDNF might play a critical role in protecting central motor neurons from apoptosis by means of retrograde transportation.

Citation： TANG Xiufa,ZHANG Fugui,FENG Yang,ZHOU Wei,LIU Jiyuan,HUA Chengge. IN VIVO STUDY ON TISSUE ENGINEERED SKELETAL MUSCLE WITH HYPOGLOSSAL NERVE IMPLANTATION. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(3): 359-364. doi: Copy

1.	Kimata Y, Sakuraba M, Namba Y, et al. Functional reconstruction with free flaps following ablation of oropharyngeal cancer. Int J Clin Oncal, 2005, 10(4): 229-233.
2.	Slaughter K, Li H, Sokoloff AJ. Neuromuscular organization of the superior longitudinalis muscle in the human tongue. Cells Tissues Organs, 2005, 181(1): 51-64.
3.	Pfrieger FW. Roles of glial cells in synapse development. Cell Mol Life Sci, 2009, 66(13): 2037-2047.
4.	Witzemann V. Development of the neuromuscular junction. Cell Tissue Res, 2006, 326(2): 263-271.
5.	唐休发, 温玉明, 王大章. 舌下神经颏舌肌蒂植入失神经胸大肌瓣的实验研究. 实用口腔医学杂志, 2001, 17(5): 377-380.
6.	Ohishi M, Harada F, Rahman F, et al. GDNF expression in terminal schwann cells associated with the periodontal ruffini endings of the rat incisors during nerve regeneration. Anat Rec (Hoboken), 2009, 292(8): 1185-1191.
7.	Zhang L, Ma Z, Smith GM, et al. GDNF-enhanced axonal regeneration and myelination following spinal cord injury is mediated by primary effects on neurons. Glia, 2009, 57(11): 1178-1191.
8.	Schmidt CE, Leach JB. Neural tissue engineering: Strategies for repair and regeneration. Annu Rev Biomed Eng, 2003, 5: 293-347.
9.	Wang H, Hughes I, Planer W, et al. The timing and location of glial cell line-derived neurotrophic factor expression determine enteric nervous system structure and function. J Neurosci, 2010, 30(4): 1523-1538.
10.	蔡炜嵩, 赵红宇, 袁正伟. 胶质细胞源性神经营养因子对运动神经元发育及运动神经元性疾病的作用. 中国修复重建外科杂志, 2005, 19(6): 490-493.
11.	Li W, Brakefield D, Pan Y, et al. Muscle-derived but not centrally derived transgene GDNF is neuroprotective in G93A-SOD1 mouse model of ALS. Exp Neurol, 2007, 203(2): 457-471.
12.	Henderson CE, Phillips HS, Pollock RA, et al. GDNF: a potent survival factor for motoneurons present in periphery nerve and muscle. Science, 1994, 266(5187): 1062-1064.
13.	李军, 张洹, 刘革修. 小鼠胎盘组织细胞移植的抗衰老作用研究. 生物医学工程学杂志, 2010, 27(6): 1312-1316.
14.	Paxinos G, Waston C. 大鼠脑立体定位图谱. 诸葛启钏, 译. 3版. 北京: 人民卫生出版社, 2005: 1-26.
15.	赵硕, 张彩顺. 辣根过氧化物酶逆行示踪评价无细胞神经移植物复合骨髓间充质干细胞对坐骨神经缺损大鼠运动神经元的作用. 中国组织工程研究与临床康复, 2010, 14(19): 3495-3498.
16.	张斌, 黄富国. 骨骼肌成肌细胞移植研究. 中国修复重建外科杂志, 2006, 20(8): 849-853.
17.	Macfelda K, Kapeller B, Wilbacher I, et al. Behavior of cardiomyocytes and skeletal muscle cells on different extracellular matrix components—relevance for cardiac tissue engineering. Artif Organs, 2007, 31(1): 4-12.
18.	Toba T, Nakamura T, Shimizu Y, et al. Regeneration of canine peroneal nerve with the use of a polyglycolic acid-collagen tube filled with laminin-soaked collagen sponge: a comparative study of collagen sponge and collagen fibers as filling materials for nerve conduits. J Biomed Mater Res, 2001, 58(6): 622-630.
19.	徐林海, 焦向阳, 季正伦. 以胶原海绵为载体培养的人表皮细胞移植. 中国修复重建外科杂志, 2001, 15(2): 118-121.
20.	Kroehne V, Heschel I, Schugner F, et al. Use of a novel collagen matrix with oriented pore structure for muscle cell differentiation in cell culture and in grafts. J Cell Mol Med, 2008, 12(5A): 1640-1648.
21.	Beier JP, Klumpp D, Rudisile M, et al. Collagen matrices from sponge to nano: new perspectives for tissue engineering of skeletal muscle. BMC Biotechnol, 2009, 9: 34.
22.	岑石强, 杨志明. 成肌细胞在基因治疗和组织工程中的应用综述. 中国修复重建外科杂志, 1999, 13(3): 173-177.
23.	Rich KM, Luszczynski JR, Osborne PA, et al. Nerve growth factor protects adult sensory neurons from cell death and atrophy caused by nerve injury. J Neurocyctol, 1987, 16(2): 261-268.
24.	Paratcha G, Ledda F. GDNF and GFRα: a versatile molecular complex for developing neurons. Trend Neurosci, 2008, 31(8): 384-391.

1. Kimata Y, Sakuraba M, Namba Y, et al. Functional reconstruction with free flaps following ablation of oropharyngeal cancer. Int J Clin Oncal, 2005, 10(4): 229-233.
2. Slaughter K, Li H, Sokoloff AJ. Neuromuscular organization of the superior longitudinalis muscle in the human tongue. Cells Tissues Organs, 2005, 181(1): 51-64.
3. Pfrieger FW. Roles of glial cells in synapse development. Cell Mol Life Sci, 2009, 66(13): 2037-2047.
4. Witzemann V. Development of the neuromuscular junction. Cell Tissue Res, 2006, 326(2): 263-271.
5. 唐休发, 温玉明, 王大章. 舌下神经颏舌肌蒂植入失神经胸大肌瓣的实验研究. 实用口腔医学杂志, 2001, 17(5): 377-380.
6. Ohishi M, Harada F, Rahman F, et al. GDNF expression in terminal schwann cells associated with the periodontal ruffini endings of the rat incisors during nerve regeneration. Anat Rec (Hoboken), 2009, 292(8): 1185-1191.
7. Zhang L, Ma Z, Smith GM, et al. GDNF-enhanced axonal regeneration and myelination following spinal cord injury is mediated by primary effects on neurons. Glia, 2009, 57(11): 1178-1191.
8. Schmidt CE, Leach JB. Neural tissue engineering: Strategies for repair and regeneration. Annu Rev Biomed Eng, 2003, 5: 293-347.
9. Wang H, Hughes I, Planer W, et al. The timing and location of glial cell line-derived neurotrophic factor expression determine enteric nervous system structure and function. J Neurosci, 2010, 30(4): 1523-1538.
10. 蔡炜嵩, 赵红宇, 袁正伟. 胶质细胞源性神经营养因子对运动神经元发育及运动神经元性疾病的作用. 中国修复重建外科杂志, 2005, 19(6): 490-493.
11. Li W, Brakefield D, Pan Y, et al. Muscle-derived but not centrally derived transgene GDNF is neuroprotective in G93A-SOD1 mouse model of ALS. Exp Neurol, 2007, 203(2): 457-471.
12. Henderson CE, Phillips HS, Pollock RA, et al. GDNF: a potent survival factor for motoneurons present in periphery nerve and muscle. Science, 1994, 266(5187): 1062-1064.
13. 李军, 张洹, 刘革修. 小鼠胎盘组织细胞移植的抗衰老作用研究. 生物医学工程学杂志, 2010, 27(6): 1312-1316.
14. Paxinos G, Waston C. 大鼠脑立体定位图谱. 诸葛启钏, 译. 3版. 北京: 人民卫生出版社, 2005: 1-26.
15. 赵硕, 张彩顺. 辣根过氧化物酶逆行示踪评价无细胞神经移植物复合骨髓间充质干细胞对坐骨神经缺损大鼠运动神经元的作用. 中国组织工程研究与临床康复, 2010, 14(19): 3495-3498.
16. 张斌, 黄富国. 骨骼肌成肌细胞移植研究. 中国修复重建外科杂志, 2006, 20(8): 849-853.
17. Macfelda K, Kapeller B, Wilbacher I, et al. Behavior of cardiomyocytes and skeletal muscle cells on different extracellular matrix components—relevance for cardiac tissue engineering. Artif Organs, 2007, 31(1): 4-12.
18. Toba T, Nakamura T, Shimizu Y, et al. Regeneration of canine peroneal nerve with the use of a polyglycolic acid-collagen tube filled with laminin-soaked collagen sponge: a comparative study of collagen sponge and collagen fibers as filling materials for nerve conduits. J Biomed Mater Res, 2001, 58(6): 622-630.
19. 徐林海, 焦向阳, 季正伦. 以胶原海绵为载体培养的人表皮细胞移植. 中国修复重建外科杂志, 2001, 15(2): 118-121.
20. Kroehne V, Heschel I, Schugner F, et al. Use of a novel collagen matrix with oriented pore structure for muscle cell differentiation in cell culture and in grafts. J Cell Mol Med, 2008, 12(5A): 1640-1648.
21. Beier JP, Klumpp D, Rudisile M, et al. Collagen matrices from sponge to nano: new perspectives for tissue engineering of skeletal muscle. BMC Biotechnol, 2009, 9: 34.
22. 岑石强, 杨志明. 成肌细胞在基因治疗和组织工程中的应用综述. 中国修复重建外科杂志, 1999, 13(3): 173-177.
23. Rich KM, Luszczynski JR, Osborne PA, et al. Nerve growth factor protects adult sensory neurons from cell death and atrophy caused by nerve injury. J Neurocyctol, 1987, 16(2): 261-268.
24. Paratcha G, Ledda F. GDNF and GFRα: a versatile molecular complex for developing neurons. Trend Neurosci, 2008, 31(8): 384-391.

Chinese Journal of Reparative and Reconstructive Surgery

IN VIVO STUDY ON TISSUE ENGINEERED SKELETAL MUSCLE WITH HYPOGLOSSAL NERVE IMPLANTATION

Abstract Full text Figures/Tables Video References Cited by

Format

Content