脫細胞技術及其在組織工程中的應用研究進展_《中國修復重建外科雜志》

作者：

趙宇 ^1,2 , 于淼 ² , 柏樹令 ²

1中國醫科大學附屬盛京醫院整形美容外科（沈陽，110004）;;
2中國醫科大學基礎醫學院組織工程教研室;

關鍵詞：

組織工程支架細胞外基質脫細胞滅菌

DOI：

10.7507/1002-1892.20130207

視頻：

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目的對獲取組織細胞外基質（extracellular matrix，ECM）的脫細胞方法及脫細胞ECM支架在組織工程領域的應用進行綜述。方法查閱近年國內外相關文獻，對脫細胞方法進行歸納和分類，分析滅菌方法對脫細胞支架的影響，提出脫細胞程度的評價標準，總結脫細胞ECM支架在不同組織器官中的應用。結果脫細胞方法主要有物理法、化學試劑法和生物制劑法，不同的脫細胞方法對細胞去除程度及ECM成分的影響不同，需要根據組織和脫細胞方法的特點選擇最佳脫細胞方法，以達到理想效果。結論選擇合適的脫細胞方法對制備組織工程所需的生物材料非常重要。

引用本文： 趙宇,于淼,柏樹令. 脫細胞技術及其在組織工程中的應用研究進展. 中國修復重建外科雜志, 2013, 27(8): 950-954. doi: 10.7507/1002-1892.20130207 復制

1.	Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol, 2005, 23(1): 47-55.
2.	Weber B, Emmert MY, Schoenauer R, et al. Tissue engineering on matrix: future of autologous tissue replacement. Semin Immunopathol, 2011, 33(3): 307-315.
3.	Flynn LE. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials, 2010, 31(17): 4715-4724.
4.	Francis MP, Sachs PC, Madurantakam PA, et al. Electrospinning adipose tissue-derived extracellular matrix for adipose stem cell culture. J Biomed Mater Res A, 2012, 100(7): 1716-1724.
5.	江燕林, 張憶, 楊潔亮, 等. 脫細胞小牛肌腱組織形態學和生物力學特性研究. 中國修復重建外科雜志, 2013, 27(5): 552-553.
6.	Hopkinson A, Shanmuganathan VA, Gray T, et al. Optimization of amniotic membrane (AM) denuding for tissue engineering. Tissue Eng Part C Methods, 2008, 14(4): 371-381.
7.	Funamoto S, Nam K, Kimura T, et al. The use of high-hydrostatic pressure treatment to decellularize blood vessels. Biomaterials, 2010, 31(13): 3590-3595.
8.	Negishi J, Funamoto S, Kimura T, et al. Porcine radial artery decellularization by high hydrostatic pressure. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
9.	Phillips M, Maor E, Rubinsky B. Nonthermal irreversible electroporation for tissue decellularization. J Biomech Eng, 2010, 132(9): 091003.
10.	Alhamdani MS, Schroder C, Werner J, et al. Single-step procedure for the isolation of proteins at near-native conditions from mammalian tissue for proteomic analysis on antibody microarrays. J Proteome Res, 2010, 9(2): 963-971.
11.	Du L, Wu X, Pang K, et al. Histological evaluation and biomechanical characterisation of an acellular porcine cornea scaffold. Br J Ophthalmol, 2011, 95(3): 410-414.
12.	Sicari BM, Johnson SA, Siu BF, et al. The effect of source animal age upon the in vivo remodeling characteristics of an extracellular matrix scaffold. Biomaterials, 2012, 33(22): 5524-5533.
13.	Ren H, Shi X, Tao L, et al. Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold. Liver Int, 2013, 33(3): 448-458.
14.	Sullivan DC, Mirmalek-Sani SH, Deegan DB, et al. Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials, 2012, 33(31): 7756-7764.
15.	Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
16.	Hudson TW, Liu SY, Schmidt CE. Engineering an improved acellular nerve graft via optimized chemical processing. Tissue Eng, 2004, 10(9-10): 1346-1358.
17.	Cebotari S, Tudorache I, Jaekel T, et al. Detergent decellularization of heart valves for tissue engineering: toxicological effects of residual detergents on human endothelial cells. Artif Organs, 2010, 34(3): 206-210.
18.	Meyer SR, Chiu B, Churchill TA, et al. Comparison of aortic valve allograft decellularization techniques in the rat. J Biomed Mater Res A, 2006, 79(2): 254-262.
19.	Deeken CR, White AK, Bachman SL, et al. Method of preparing a decellularized porcine tendon using tributyl phosphate. J Biomed Mater Res B Appl Biomater, 2011, 96(2): 199-206.
20.	Dong X, Wei X, Yi W, et al. RGD-modified acellular bovine pericardium as a bioprosthetic scaffold for tissue engineering. J Mater Sci Mater Med, 2009, 20(11): 2327-2336.
21.	Reing JE, Brown BN, Daly KA, et al. The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds. Biomaterials, 2010, 31(33): 8626-8633.
22.	Brown BN, Freund JM, Han L, et al. Comparison of three methods for the derivation of a biologic scaffold composed of adipose tissue extracellular matrix. Tissue Eng Part C Methods, 2011, 17(4): 411-421.
23.	Rice RD, Ayubi FS, Shaub ZJ, et al. Comparison of Surgisis, AlloDerm, and Vicryl Woven Mesh grafts for abdominal wall defect repair in an animal model. Aesthetic Plast Surg, 2010, 34(3): 290-296.
24.	Ota T, Taketani S, Iwai S, et al. Novel method of decellularization of porcine valves using polyethylene glycol and gamma irradiation. Ann Thorac Surg, 2007, 83(4): 1501-1507.
25.	Mangold S, Schrammel S, Huber G, et al. Evaluation of decellularized human umbilical vein (HUV) for vascular tissue engineering-comparison with endothelium-denuded HUV. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
26.	Schenke-Layland K, Vasilevski O, Opitz F, et al. Impact of decellularization of xenogeneic tissue on extracellular matrix integrity for tissue engineering of heart valves. J Struct Biol, 2003, 143(3): 201-208.
27.	Nam J, Choi SY, Sung SC, et al. Changes of the structural and biomechanical properties of the bovine pericardium after the removal of alpha-Gal epitopes by decellularization and alpha-Galactosidase treatment. Korean J Thorac Cardiovasc Surg, 2012, 45(6): 380-389.
28.	Fishman JM, Ansari T, Sibbons P, et al. Decellularized rabbit cricoarytenoid dorsalis muscle for laryngeal regeneration. Ann Otol Rhinol Laryngol, 2012, 121(2): 129-138.
29.	Gui L, Chan SA, Breuer CK, et al. Novel utilization of serum in tissue decellularization. Tissue Eng Part C Methods, 2010, 16(2): 173-184.
30.	Freytes DO, Stoner RM, Badylak SF. Uniaxial and biaxial properties of terminally sterilized porcine urinary bladder matrix scaffolds. J Biomed Mater Res B Appl Biomater, 2008, 84(2): 408-414.
31.	Kajbafzadeh AM, Javan-Farazmand N, Monajemzadeh M, et al. Determining the optimal decellularization and sterilization protocol for preparing a tissue scaffold of a human-sized liver tissue. Tissue Eng Part C Methods, 2013. [Epub ahead of print].
32.	Russell NA, Rives A, Pelletier MH, et al. The effect of sterilization on the mechanical properties of intact rabbit humeri in three-point bending, four-point bending and torsion. Cell Tissue Bank, 2012. [Epub ahead of print].
33.	Gilbert TW. Strategies for tissue and organ decellularization. J Cell Biochem, 2012, 113(7): 2217-2222.
34.	Uzarski JS, Van De Walle AB, Mcfetridge PS. Preimplantation processing of ex vivo-derived vascular biomaterials: effects on peripheral cell adhesion. J Biomed Mater Res A, 2013, 101(1): 123-131.
35.	Bondioli E, Fini M, Veronesi F, et al. Development and evaluation of a decellularized membrane from human dermis. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
36.	Choi JS, Kim BS, Kim JY, et al. Decellularized extracellular matrix derived from human adipose tissue as a potential scaffold for allograft tissue engineering. J Biomed Mater Res A, 2011, 97(3): 292-299.
37.	Wang JQ, Fan J, Gao JH, et al. Comparison of in vivo adipogenic capabilities of two different extracellular matrix microparticle scaffolds. Plast Reconstr Surg, 2013, 131(2): 174e-187e.
38.	Wolf MT, Daly KA, Reing JE, et al. Biologic scaffold composed of skeletal muscle extracellular matrix. Biomaterials, 2012, 33(10): 2916-2925.
39.	Wang L, Johnson JA, Chang DW, et al. Decellularized musculofascial extracellular matrix for tissue engineering. Biomaterials, 2013, 34(11): 2641-2654.
40.	張晨, 王者生, 高景恒. 軟骨脫細胞基質生物性的實驗研究. 實用美容整形外科雜志, 2003, 14(2): 59-61.
41.	張晨, 景士兵, 楊琨, 等. 軟骨脫細胞基質支架材料的軟骨組織工程實驗研究. 中國修復重建外科雜志, 2008, 22(7): 846-850.
42.	王壽宇, 呂德成, 張晨, 等. 軟骨脫細胞基質的生物力學性質研究. 組織工程與重建外科雜志, 2005, 1(5): 265-267.
43.	Crapo PM, Medberry CJ, Reing JE, et al. Biologic scaffolds composed of central nervous system extracellular matrix. Biomaterials, 2012, 33(13): 3539-3547.
44.	鞠曉軍, 鄭玲, 柏樹令, 等. 人羊膜與人脫細胞羊膜形態結構的對比觀察. 現代生物醫學進展, 2013, 13(2): 249-252.
45.	鞠曉軍, 潘鋒, 柏樹令, 等. 人脫細胞羊膜復合脂肪源性干細胞修復大鼠全層皮膚缺損的實驗研究. 中國修復重建外科雜志, 2010, 24(2): 143-149.
46.	Santi PA, Johnson SB. Decellularized ear tissues as scaffolds for stem cell differentiation. J Assoc Res Otolaryngol, 2013, 14(1): 3-15.
47.	Ma R, Li M, Luo J, et al. Structural integrity, ECM components and immunogenicity of decellularized laryngeal scaffold with preserved cartilage. Biomaterials, 2013, 34(7): 1790-1798.
48.	Totonelli G, Maghsoudlou P, Georgiades F, et al. Detergent enzymatic treatment for the development of a natural acellular matrix for oesophageal regeneration. Pediatr Surg Int, 2013, 29(1): 87-95.

1. Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol, 2005, 23(1): 47-55.
2. Weber B, Emmert MY, Schoenauer R, et al. Tissue engineering on matrix: future of autologous tissue replacement. Semin Immunopathol, 2011, 33(3): 307-315.
3. Flynn LE. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials, 2010, 31(17): 4715-4724.
4. Francis MP, Sachs PC, Madurantakam PA, et al. Electrospinning adipose tissue-derived extracellular matrix for adipose stem cell culture. J Biomed Mater Res A, 2012, 100(7): 1716-1724.
5. 江燕林, 張憶, 楊潔亮, 等. 脫細胞小牛肌腱組織形態學和生物力學特性研究. 中國修復重建外科雜志, 2013, 27(5): 552-553.
6. Hopkinson A, Shanmuganathan VA, Gray T, et al. Optimization of amniotic membrane (AM) denuding for tissue engineering. Tissue Eng Part C Methods, 2008, 14(4): 371-381.
7. Funamoto S, Nam K, Kimura T, et al. The use of high-hydrostatic pressure treatment to decellularize blood vessels. Biomaterials, 2010, 31(13): 3590-3595.
8. Negishi J, Funamoto S, Kimura T, et al. Porcine radial artery decellularization by high hydrostatic pressure. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
9. Phillips M, Maor E, Rubinsky B. Nonthermal irreversible electroporation for tissue decellularization. J Biomech Eng, 2010, 132(9): 091003.
10. Alhamdani MS, Schroder C, Werner J, et al. Single-step procedure for the isolation of proteins at near-native conditions from mammalian tissue for proteomic analysis on antibody microarrays. J Proteome Res, 2010, 9(2): 963-971.
11. Du L, Wu X, Pang K, et al. Histological evaluation and biomechanical characterisation of an acellular porcine cornea scaffold. Br J Ophthalmol, 2011, 95(3): 410-414.
12. Sicari BM, Johnson SA, Siu BF, et al. The effect of source animal age upon the in vivo remodeling characteristics of an extracellular matrix scaffold. Biomaterials, 2012, 33(22): 5524-5533.
13. Ren H, Shi X, Tao L, et al. Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold. Liver Int, 2013, 33(3): 448-458.
14. Sullivan DC, Mirmalek-Sani SH, Deegan DB, et al. Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials, 2012, 33(31): 7756-7764.
15. Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
16. Hudson TW, Liu SY, Schmidt CE. Engineering an improved acellular nerve graft via optimized chemical processing. Tissue Eng, 2004, 10(9-10): 1346-1358.
17. Cebotari S, Tudorache I, Jaekel T, et al. Detergent decellularization of heart valves for tissue engineering: toxicological effects of residual detergents on human endothelial cells. Artif Organs, 2010, 34(3): 206-210.
18. Meyer SR, Chiu B, Churchill TA, et al. Comparison of aortic valve allograft decellularization techniques in the rat. J Biomed Mater Res A, 2006, 79(2): 254-262.
19. Deeken CR, White AK, Bachman SL, et al. Method of preparing a decellularized porcine tendon using tributyl phosphate. J Biomed Mater Res B Appl Biomater, 2011, 96(2): 199-206.
20. Dong X, Wei X, Yi W, et al. RGD-modified acellular bovine pericardium as a bioprosthetic scaffold for tissue engineering. J Mater Sci Mater Med, 2009, 20(11): 2327-2336.
21. Reing JE, Brown BN, Daly KA, et al. The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds. Biomaterials, 2010, 31(33): 8626-8633.
22. Brown BN, Freund JM, Han L, et al. Comparison of three methods for the derivation of a biologic scaffold composed of adipose tissue extracellular matrix. Tissue Eng Part C Methods, 2011, 17(4): 411-421.
23. Rice RD, Ayubi FS, Shaub ZJ, et al. Comparison of Surgisis, AlloDerm, and Vicryl Woven Mesh grafts for abdominal wall defect repair in an animal model. Aesthetic Plast Surg, 2010, 34(3): 290-296.
24. Ota T, Taketani S, Iwai S, et al. Novel method of decellularization of porcine valves using polyethylene glycol and gamma irradiation. Ann Thorac Surg, 2007, 83(4): 1501-1507.
25. Mangold S, Schrammel S, Huber G, et al. Evaluation of decellularized human umbilical vein (HUV) for vascular tissue engineering-comparison with endothelium-denuded HUV. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
26. Schenke-Layland K, Vasilevski O, Opitz F, et al. Impact of decellularization of xenogeneic tissue on extracellular matrix integrity for tissue engineering of heart valves. J Struct Biol, 2003, 143(3): 201-208.
27. Nam J, Choi SY, Sung SC, et al. Changes of the structural and biomechanical properties of the bovine pericardium after the removal of alpha-Gal epitopes by decellularization and alpha-Galactosidase treatment. Korean J Thorac Cardiovasc Surg, 2012, 45(6): 380-389.
28. Fishman JM, Ansari T, Sibbons P, et al. Decellularized rabbit cricoarytenoid dorsalis muscle for laryngeal regeneration. Ann Otol Rhinol Laryngol, 2012, 121(2): 129-138.
29. Gui L, Chan SA, Breuer CK, et al. Novel utilization of serum in tissue decellularization. Tissue Eng Part C Methods, 2010, 16(2): 173-184.
30. Freytes DO, Stoner RM, Badylak SF. Uniaxial and biaxial properties of terminally sterilized porcine urinary bladder matrix scaffolds. J Biomed Mater Res B Appl Biomater, 2008, 84(2): 408-414.
31. Kajbafzadeh AM, Javan-Farazmand N, Monajemzadeh M, et al. Determining the optimal decellularization and sterilization protocol for preparing a tissue scaffold of a human-sized liver tissue. Tissue Eng Part C Methods, 2013. [Epub ahead of print].
32. Russell NA, Rives A, Pelletier MH, et al. The effect of sterilization on the mechanical properties of intact rabbit humeri in three-point bending, four-point bending and torsion. Cell Tissue Bank, 2012. [Epub ahead of print].
33. Gilbert TW. Strategies for tissue and organ decellularization. J Cell Biochem, 2012, 113(7): 2217-2222.
34. Uzarski JS, Van De Walle AB, Mcfetridge PS. Preimplantation processing of ex vivo-derived vascular biomaterials: effects on peripheral cell adhesion. J Biomed Mater Res A, 2013, 101(1): 123-131.
35. Bondioli E, Fini M, Veronesi F, et al. Development and evaluation of a decellularized membrane from human dermis. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
36. Choi JS, Kim BS, Kim JY, et al. Decellularized extracellular matrix derived from human adipose tissue as a potential scaffold for allograft tissue engineering. J Biomed Mater Res A, 2011, 97(3): 292-299.
37. Wang JQ, Fan J, Gao JH, et al. Comparison of in vivo adipogenic capabilities of two different extracellular matrix microparticle scaffolds. Plast Reconstr Surg, 2013, 131(2): 174e-187e.
38. Wolf MT, Daly KA, Reing JE, et al. Biologic scaffold composed of skeletal muscle extracellular matrix. Biomaterials, 2012, 33(10): 2916-2925.
39. Wang L, Johnson JA, Chang DW, et al. Decellularized musculofascial extracellular matrix for tissue engineering. Biomaterials, 2013, 34(11): 2641-2654.
40. 張晨, 王者生, 高景恒. 軟骨脫細胞基質生物性的實驗研究. 實用美容整形外科雜志, 2003, 14(2): 59-61.
41. 張晨, 景士兵, 楊琨, 等. 軟骨脫細胞基質支架材料的軟骨組織工程實驗研究. 中國修復重建外科雜志, 2008, 22(7): 846-850.
42. 王壽宇, 呂德成, 張晨, 等. 軟骨脫細胞基質的生物力學性質研究. 組織工程與重建外科雜志, 2005, 1(5): 265-267.
43. Crapo PM, Medberry CJ, Reing JE, et al. Biologic scaffolds composed of central nervous system extracellular matrix. Biomaterials, 2012, 33(13): 3539-3547.
44. 鞠曉軍, 鄭玲, 柏樹令, 等. 人羊膜與人脫細胞羊膜形態結構的對比觀察. 現代生物醫學進展, 2013, 13(2): 249-252.
45. 鞠曉軍, 潘鋒, 柏樹令, 等. 人脫細胞羊膜復合脂肪源性干細胞修復大鼠全層皮膚缺損的實驗研究. 中國修復重建外科雜志, 2010, 24(2): 143-149.
46. Santi PA, Johnson SB. Decellularized ear tissues as scaffolds for stem cell differentiation. J Assoc Res Otolaryngol, 2013, 14(1): 3-15.
47. Ma R, Li M, Luo J, et al. Structural integrity, ECM components and immunogenicity of decellularized laryngeal scaffold with preserved cartilage. Biomaterials, 2013, 34(7): 1790-1798.
48. Totonelli G, Maghsoudlou P, Georgiades F, et al. Detergent enzymatic treatment for the development of a natural acellular matrix for oesophageal regeneration. Pediatr Surg Int, 2013, 29(1): 87-95.

《中國修復重建外科雜志》

脫細胞技術及其在組織工程中的應用研究進展

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《中國修復重建外科雜志》

脫細胞技術及其在組織工程中的應用研究進展

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