• 中山大學(xué)附屬第一醫(yī)院（廣州，510080）1顯微創(chuàng)傷骨科，2骨科研究所，3黃埔院區(qū)上肢骨科;

目的觀察去細胞異種神經(jīng)復(fù)合同種異體脂肪干細胞修復(fù)獼猴周圍神經(jīng)缺損后的全身及局部免疫排斥反應(yīng)，評價該修復(fù)材料的安全性。 方法健康成年雄性長白豬1只，體重48 kg，取脛神經(jīng)制備去細胞異種神經(jīng)；健康成年雄性獼猴1只，體重4.5 kg，分離培養(yǎng)脂肪干細胞；健康成年雌性獼猴10只，體重3～5 kg，制備25 mm長橈神經(jīng)缺損動物模型，隨機分為復(fù)合細胞組和無細胞組（n=5），前者采用去細胞異種神經(jīng)復(fù)合第3代脂肪干細胞移植修復(fù)，后者采用去細胞異種神經(jīng)移植修復(fù)。于術(shù)前及術(shù)后14、60、90 d抽取外周靜脈血行淋巴細胞分析；術(shù)后5個月取材觀察移植物組織免疫反應(yīng)和神經(jīng)再生情況，并與自體神經(jīng)移植組作比較。 結(jié)果復(fù)合細胞組與無細胞組術(shù)前及術(shù)后各時間點外周靜脈血淋巴細胞數(shù)量、T淋巴細胞百分率及其數(shù)量、CD8+ T淋巴細胞百分率、CD4+/CD8+ T淋巴細胞比值間比較，差異均無統(tǒng)計學(xué)意義（P  gt; 0.05）；術(shù)后14 d，復(fù)合細胞組CD4+ T淋巴細胞百分率低于其余時間點， 差異有統(tǒng)計學(xué)意義（P  lt; 0.05）；同一時間點兩組間比較，除術(shù)后14 d復(fù)合細胞組CD4+ T淋巴細胞百分率低于無細胞組，差異有統(tǒng)計學(xué)意義（P  lt; 0.05）外，其余各時間點各指標(biāo)組間比較差異均無統(tǒng)計學(xué)意義（P  gt; 0.05）。術(shù)后5個月，去細胞異種神經(jīng)與周圍組織有輕度粘連，神經(jīng)外膜較自體神經(jīng)厚，未見組織壞死、纖維瘢痕形成，移植物內(nèi)見再生神經(jīng)纖維，復(fù)合細胞組、無細胞組均有稀疏的CD3+、CD4+、CD8+、CD68+、CD163+ T淋巴細胞散在分布，細胞浸潤情況與自體神經(jīng)移植組類似。 結(jié)論去細胞異種神經(jīng)移植修復(fù)獼猴周圍神經(jīng)缺損后未產(chǎn)生全身及局部免疫排斥反應(yīng)；復(fù)合同種異體脂肪干細胞移植后也未發(fā)現(xiàn)免疫排斥反應(yīng)，且術(shù)后早期可能抑制CD4+ T淋巴細胞增殖。

引用本文： 黃喜軍,朱慶棠,江麗,鄭燦鑌,朱昭煒,路慶森,許銀峰,顧立強,劉小林. 去細胞異種神經(jīng)復(fù)合同種異體脂肪干細胞修復(fù)獼猴周圍神經(jīng)缺損的免疫反應(yīng)研究. 中國修復(fù)重建外科雜志, 2012, 26(8): 993-1000. doi: 復(fù)制

1. Siemionow M, Bozkurt M, Zor F. Regeneration and repair of peripheral nerves with different biomaterials: review. Microsurgery, 2010, 30(7): 574-588.
2. 范偉杰, 丁志清. 不同方法制備的脫細胞神經(jīng)組織學(xué)及組織相容性研究. 中國修復(fù)重建外科雜志, 2011, 25(2): 176-180.
3. 王冠軍, 盧世璧, 匡正達, 等. 化學(xué)去細胞異體神經(jīng)移植促神經(jīng)趨化性再生實驗研究. 中國修復(fù)重建外科雜志, 2010, 24(11): 1288-1292.
4. 趙喆, 王玉, 彭江, 等. 化學(xué)去細胞異體神經(jīng)周圍復(fù)合BMSCs生物蛋白膠復(fù)合物促周圍神經(jīng)缺損修復(fù). 中國修復(fù)重建外科雜志, 2011, 25(4): 488-493.
5. Moore AM, Macewan M, Santosa KB, et al. Acellular nerve allografts in peripheral nerve regeneration: a comparative study. Muscle Nerve, 2011, 44(2): 221-234.
6. Johnson PJ, Newton P, Hunter DA, et al. Nerve endoneurial microstructure facilitates uniform distribution of regenerative fibers: a post hoc comparison of midgraft nerve fiber densities. J Reconstr Microsurg, 2011, 27(2): 83-90.
7. 丁小珩, 劉小林, 劉育杰, 等. 去細胞同種異體神經(jīng)修復(fù)材料臨床應(yīng)用初步報告. 中華顯微外科雜志, 2009, 32(6): 448-450.
8. 郭義柱, 王巖, 劉相成, 等. 同種異體神經(jīng)移植治療銳性臂叢神經(jīng)缺損5例臨床研究. 軍醫(yī)進修學(xué)院學(xué)報, 2011, 32(4): 317-318.
9. Brooks DN, Weber RV, Chao JD, et al. Processed nerve allografts for peripheral nerve reconstruction: A multicenter study of utilization and outcomes in sensory, mixed, and motor nerve reconstructions. Microsurgery, 2012, 32(1): 1-14.
10. 何彩鳳, 朱慶棠, 江麗, 等. 豬與人去細胞神經(jīng)結(jié)構(gòu)與成分的對比研究. 實用手外科雜志, 2011, 25(1): 39-42, 46.
11. 趙喆, 趙斌, 王玉, 等. 化學(xué)去細胞異體神經(jīng)添加不同組織來源雪旺細胞對周圍神經(jīng)損傷修復(fù)的功能評價. 中國修復(fù)重建外科雜志, 2010, 24(11): 1281-1287.
12. di Summa PG, Kingham PJ, Raffoul W, et al. Adipose-derived stem cells enhance peripheral nerve regeneration. J Plast Reconstr Aesthet Surg, 2010, 63(9): 1544-1552.
13. Liu G, Cheng Y, Guo S, et al. Transplantation of adipose-derived stem cells for peripheral nerve repair. Int J Mol Med, 2011, 28(4): 565-572.
14. 朱慶棠, 朱家愷, 賴英榮, 等. 去細胞組織工程化神經(jīng)支架的制備與形態(tài)學(xué)研究. 中華顯微外科雜志, 2004, 27(1): 35-37.
15. Wang D, Liu XL, Zhu JK, et al. Repairing large radial nerve defects by acellular nerve allografts seeded with autologous bone marrow stromal cells in a monkey model. J Neurotrauma, 2010, 27(10): 1935-1943.
16. 金婷, 林星石, 楊若佳, 等. 小鼠同種異體及異種神經(jīng)移植后脾臟T淋巴細胞亞群的變化. 實驗動物科學(xué), 2007, 24(4): 1-4.
17. Rovak JM, Bishop DK, Boxer LK, et al. Peripheral nerve transplantation: the role of chemical acellularization in eliminating allograft antigenicity. J Reconstr Microsurg, 2005, 21(3): 207-213.
18. Kvist M, Sondell M, Kanje M, et al. Regeneration in, and properties of, extracted peripheral nerve allografts and xenografts. J Plast Surg Hand Surg, 2011, 45(3): 122-128.
19. Zhang Y, Luo H, Zhang Z, et al. A nerve graft constructed with xenogeneic acellular nerve matrix and autologous adipose-derived mesenchymal stem cells. Biomaterials, 2010, 31(20): 5312-5324.
20. Tse KH, Sun M, Mantovani C, et al. In vitro evaluation of polyester-based scaffolds seeded with adipose derived stem cells for peripheral nerve regeneration. J Biomed Mater Res A, 2010, 95(3): 701-708.
21. Sun F, Zhou K, Mi WJ, et al. Combined use of decellularized allogeneic artery conduits with autologous transdifferentiated adipose-derived stem cells for facial nerve regeneration in rats. Biomaterials, 2011, 32(32): 8118-8128.
22. Klyushnenkova E, Mosca JD, Zernetkina V, et al. T cell responses to allogeneic human mesenchymal stem cells: immunogenicity, tolerance, and suppression. J Biomed Sci, 2005, 12(1): 47-57.
23. Sioud M, Mobergslien A, Boudabous A, et al. Evidence for the involvement of galectin-3 in mesenchymal stem cell suppression of allogeneic T-cell proliferation. Scand J Immunol, 2010, 71(4): 267-274.
24. Comoli P, Ginevri F, Maccario R, et al. Human mesenchymal stem cells inhibit antibody production induced in vitro by allostimulation. Nephrol Dial Transplant, 2008, 23(4): 1196-1202.
25. Sotiropoulou PA, Perez SA, Gritzapis AD, et al. Interactions between human mesenchymal stem cells and natural killer cells. Stem Cells, 2006, 24(1): 74-85.
26. Kuo YR, Chen CC, Goto S, et al. Modulation of immune response and T-cell regulation by donor adipose-derived stem cells in a rodent hind-limb allotransplant model. Plast Reconstr Surg, 2011, 128(6): 661e-672e.
  1. 1. Siemionow M, Bozkurt M, Zor F. Regeneration and repair of peripheral nerves with different biomaterials: review. Microsurgery, 2010, 30(7): 574-588.
  2. 2. 范偉杰, 丁志清. 不同方法制備的脫細胞神經(jīng)組織學(xué)及組織相容性研究. 中國修復(fù)重建外科雜志, 2011, 25(2): 176-180.
  3. 3. 王冠軍, 盧世璧, 匡正達, 等. 化學(xué)去細胞異體神經(jīng)移植促神經(jīng)趨化性再生實驗研究. 中國修復(fù)重建外科雜志, 2010, 24(11): 1288-1292.
  4. 4. 趙喆, 王玉, 彭江, 等. 化學(xué)去細胞異體神經(jīng)周圍復(fù)合BMSCs生物蛋白膠復(fù)合物促周圍神經(jīng)缺損修復(fù). 中國修復(fù)重建外科雜志, 2011, 25(4): 488-493.
  5. 5. Moore AM, Macewan M, Santosa KB, et al. Acellular nerve allografts in peripheral nerve regeneration: a comparative study. Muscle Nerve, 2011, 44(2): 221-234.
  6. 6. Johnson PJ, Newton P, Hunter DA, et al. Nerve endoneurial microstructure facilitates uniform distribution of regenerative fibers: a post hoc comparison of midgraft nerve fiber densities. J Reconstr Microsurg, 2011, 27(2): 83-90.
  7. 7. 丁小珩, 劉小林, 劉育杰, 等. 去細胞同種異體神經(jīng)修復(fù)材料臨床應(yīng)用初步報告. 中華顯微外科雜志, 2009, 32(6): 448-450.
  8. 8. 郭義柱, 王巖, 劉相成, 等. 同種異體神經(jīng)移植治療銳性臂叢神經(jīng)缺損5例臨床研究. 軍醫(yī)進修學(xué)院學(xué)報, 2011, 32(4): 317-318.
  9. 9. Brooks DN, Weber RV, Chao JD, et al. Processed nerve allografts for peripheral nerve reconstruction: A multicenter study of utilization and outcomes in sensory, mixed, and motor nerve reconstructions. Microsurgery, 2012, 32(1): 1-14.
  10. 10. 何彩鳳, 朱慶棠, 江麗, 等. 豬與人去細胞神經(jīng)結(jié)構(gòu)與成分的對比研究. 實用手外科雜志, 2011, 25(1): 39-42, 46.
  11. 11. 趙喆, 趙斌, 王玉, 等. 化學(xué)去細胞異體神經(jīng)添加不同組織來源雪旺細胞對周圍神經(jīng)損傷修復(fù)的功能評價. 中國修復(fù)重建外科雜志, 2010, 24(11): 1281-1287.
  12. 12. di Summa PG, Kingham PJ, Raffoul W, et al. Adipose-derived stem cells enhance peripheral nerve regeneration. J Plast Reconstr Aesthet Surg, 2010, 63(9): 1544-1552.
  13. 13. Liu G, Cheng Y, Guo S, et al. Transplantation of adipose-derived stem cells for peripheral nerve repair. Int J Mol Med, 2011, 28(4): 565-572.
  14. 14. 朱慶棠, 朱家愷, 賴英榮, 等. 去細胞組織工程化神經(jīng)支架的制備與形態(tài)學(xué)研究. 中華顯微外科雜志, 2004, 27(1): 35-37.
  15. 15. Wang D, Liu XL, Zhu JK, et al. Repairing large radial nerve defects by acellular nerve allografts seeded with autologous bone marrow stromal cells in a monkey model. J Neurotrauma, 2010, 27(10): 1935-1943.
  16. 16. 金婷, 林星石, 楊若佳, 等. 小鼠同種異體及異種神經(jīng)移植后脾臟T淋巴細胞亞群的變化. 實驗動物科學(xué), 2007, 24(4): 1-4.
  17. 17. Rovak JM, Bishop DK, Boxer LK, et al. Peripheral nerve transplantation: the role of chemical acellularization in eliminating allograft antigenicity. J Reconstr Microsurg, 2005, 21(3): 207-213.
  18. 18. Kvist M, Sondell M, Kanje M, et al. Regeneration in, and properties of, extracted peripheral nerve allografts and xenografts. J Plast Surg Hand Surg, 2011, 45(3): 122-128.
  19. 19. Zhang Y, Luo H, Zhang Z, et al. A nerve graft constructed with xenogeneic acellular nerve matrix and autologous adipose-derived mesenchymal stem cells. Biomaterials, 2010, 31(20): 5312-5324.
  20. 20. Tse KH, Sun M, Mantovani C, et al. In vitro evaluation of polyester-based scaffolds seeded with adipose derived stem cells for peripheral nerve regeneration. J Biomed Mater Res A, 2010, 95(3): 701-708.
  21. 21. Sun F, Zhou K, Mi WJ, et al. Combined use of decellularized allogeneic artery conduits with autologous transdifferentiated adipose-derived stem cells for facial nerve regeneration in rats. Biomaterials, 2011, 32(32): 8118-8128.
  22. 22. Klyushnenkova E, Mosca JD, Zernetkina V, et al. T cell responses to allogeneic human mesenchymal stem cells: immunogenicity, tolerance, and suppression. J Biomed Sci, 2005, 12(1): 47-57.
  23. 23. Sioud M, Mobergslien A, Boudabous A, et al. Evidence for the involvement of galectin-3 in mesenchymal stem cell suppression of allogeneic T-cell proliferation. Scand J Immunol, 2010, 71(4): 267-274.
  24. 24. Comoli P, Ginevri F, Maccario R, et al. Human mesenchymal stem cells inhibit antibody production induced in vitro by allostimulation. Nephrol Dial Transplant, 2008, 23(4): 1196-1202.
  25. 25. Sotiropoulou PA, Perez SA, Gritzapis AD, et al. Interactions between human mesenchymal stem cells and natural killer cells. Stem Cells, 2006, 24(1): 74-85.
  26. 26. Kuo YR, Chen CC, Goto S, et al. Modulation of immune response and T-cell regulation by donor adipose-derived stem cells in a rodent hind-limb allotransplant model. Plast Reconstr Surg, 2011, 128(6): 661e-672e.