脱细胞小牛肌腱组织形态学和生物力学特性研究_《中国修复重建外科杂志》

作者：

江燕林 ¹ , 张忆 ¹ , 杨洁亮 ² , 罗静聪 ^1,2 , 秦廷武 ^1,2

四川大学华西医院（成都，610041）1 再生医学研究中心;;
2 生物治疗国家重点实验室·干细胞与组织工程研究室;

关键词：

脱细胞肌腱形态学组织学生物力学小牛

DOI：

10.7507/1002-1892.20130125

视频：

导出 下载 收藏 扫码 引用

摘要 全文 图表 视频 参考文献 施引文献 补充材料

目的研究反复冻融结合核酸酶处理小牛肌腱脱细胞效果以及对肌腱组织形态、结构、生化成分和力学特性的影响。方法取新鲜1日龄西门塔尔小牛跟腱48根，随机分为3组（n=16）：A组为正常对照组，B组采用液氮冷冻/37℃复温反复冻融5次，C组在反复冻融基础上结合核酸酶处理24 h。每组取2根跟腱用于扫描电镜观察，3根用于、组织学及免疫组织化学染色观察，3根用于DNA残留量检测，8根用于生物力学检测。结果扫描电镜观察示A、B组胶原纤维排列致密有序，形态完整，未见断裂；C组胶原纤维排列松散，几乎无断裂。阿利新蓝、天狼星红染色及免疫组织化学染色示C组糖胺聚糖、Ⅰ型胶原、Ⅲ型胶原、纤维连接蛋白大部分被保留；HE染色和DAPI染色示A、B组胶原纤维间可见较多完整的细胞核，而C组胶原纤维之间未见完整细胞核，但腱内膜上仍有部分细胞核残留。A、B、C组的DNA残留量分别为（0.24 ± 0.12）、（0.16 ± 0.07）、（0.05 ± 0.02）μg/mg，C组显著低于A、B组（P lt; 0.05）；A、B组间差异无统计学意义（P gt; 0.05）。生物力学检测显示，A、B、C组间拉伸强度、断裂应变、刚度和弹性模量差异均无统计学意义（P gt; 0.05）。结论反复冻融结合核酸酶处理小牛肌腱，可有效去除细胞成分，同时基本保留肌腱原始胶原纤维结构、形态、大部分细胞外基质成分和力学特性。

引用本文： 江燕林,张忆,杨洁亮,罗静聪,秦廷武. 脱细胞小牛肌腱组织形态学和生物力学特性研究. 中国修复重建外科杂志, 2013, 27(5): 565-570. doi: 10.7507/1002-1892.20130125 复制

1.	Carey JL, Dunn WR, Dahm DL, et al. A systematic review of anterior cruciate ligament reconstruction with autograft compared with allograft. J Bone Joint Surg (Am), 2009, 91(9): 2242-2250.
2.	Whitlock PW, Smith TL, Poehling GG, et al. A naturally derived, cytocompatible, and architecturally optimized scaffold for tendon and ligament regeneration. Biomaterials, 2007, 28(29): 4321-4329.
3.	Stone KR, Walgenbach AW, Turek TJ, et al. Anterior cruciate ligament reconstruction a porcinexenograft: a serologic, histologic, and biomechanical study in primates. Arthroscopy, 2007, 23(4): 411-419.
4.	Stone KR, Abdel-Motal UM, Walgenbach AW, et al. Replacement of human anterior cruciate ligaments with pig ligaments: a model for anti-non-gal antibody response in long-term xenotransplantation. Transplantation, 2007, 83(2): 211-219.
5.	Randall KL, Booth BA, Miller AJ, et al. Use of an acellular regenerative tissue matrix in combination with vacuumassisted closure therapy for treatment of a diabetic foot wound. J Foot Ankle Surg, 2008, 47(5): 430-433.
6.	Pridgen BC, Woon CY, Kim K, et al. Flexor tendon tissue engineering: acellularization of human flexor tendons with preservation of biomechanical properties and biocompatibility. Tissue Eng Part C Methods, 2011, 17(8): 819-828.
7.	Harrison RD, Gratzer PF. Effect of extraction protocols and epidermal growth factor on the cellular repopulation of decellularized anterior cruciate ligament allografts. J Biomed Mater Res A, 2005, 75(4): 841-854.
8.	Ning LJ, Zhang Y, Chen XH, et al. Preparation and characterization of decellularized tendon slices for tendon tissue engineering. J Biomed Mater Res A, 2012, 100(6): 1448-1456.
9.	Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: Structure and function. Acta Biomater, 2009, 5(1): 1-13.
10.	Azuma C, Tohyama H, Nakamura H, et al. Antibody neutralization of TGF-beta enhances the deterioration of collagen fascicles in a tissue-cultured tendon matrix with ex vivo fibroblast infiltration. J Biomech, 2007, 40(10): 2184-2190.
11.	Ponce Márquez S, Martínez VS, McIntosh Ambrose W, et al. Decellularization of bovine corneas for tissue engineering applications. Acta Biomater, 2009, 5(6): 1839-1847.
12.	Aurora A, McCarron J, Iannotti JP, et al. Commercially available extracellular matrix materials for rotator cuff repairs: state of the art and future trends. J Shoulder Elbow Surg, 2007, 16(5 Suppl): S171-178.
13.	Yang JJ, Jang EC, Lee JS, et al. The Effect of amniotic membrane transplantation on tendon-healing in a rabbit Achilles tendon model. Tissue Engineering and Regenerative Medicine, 2010, 7(3): 323-329.
14.	Gilbert TW, Stewart AM, Simmons BA, et al. Degradation and remodeling of small intestinal submucosa in canine Achilles tendon repair. J Bone Joint Surg (Am), 2007, 89(6): 621-630.
15.	Androgens C, Spragg RK, Derwin KA. Mechanical conditioning of cell-seeded small intestine submucosa: a potential tissue-engineering strategy for tendon repair. Tissue Eng, 2007, 13(2): 233-240.
16.	Derwin KA, Badylak SF, Steinmann SP, et al. Extracellular matrix scaffold devices for rotator cuff repair. J Shoulder Elbow Surg, 2010, 19(3): 467-476.
17.	Vavken P, Joshi S, Murray MM. Triton-X is most effective among three decellularization agents for ACL tissue engineering. J Orthop Res, 2009, 27(12): 1612-1618.
18.	Cartmell JS, Dunn MG. Effect of chemical treatments on tendon cellularity and mechanical properties. J Biomed Mater Res, 2000, 49(1): 134-140.
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.	Minami A, Ishii S, Ogino T, et al. Effect of the immunological antigenicity of the allogenaic tendon grafting. Hand, 1982, 14(2): 111-119.
21.	Qin TW, Chen QS, Sun YL, et al. Mechanical characteristics of native tendon slices for tissue engineering scaffold. J Biomed Mater Res B, 2012, 100(3): 752-758.

1. Carey JL, Dunn WR, Dahm DL, et al. A systematic review of anterior cruciate ligament reconstruction with autograft compared with allograft. J Bone Joint Surg (Am), 2009, 91(9): 2242-2250.
2. Whitlock PW, Smith TL, Poehling GG, et al. A naturally derived, cytocompatible, and architecturally optimized scaffold for tendon and ligament regeneration. Biomaterials, 2007, 28(29): 4321-4329.
3. Stone KR, Walgenbach AW, Turek TJ, et al. Anterior cruciate ligament reconstruction a porcinexenograft: a serologic, histologic, and biomechanical study in primates. Arthroscopy, 2007, 23(4): 411-419.
4. Stone KR, Abdel-Motal UM, Walgenbach AW, et al. Replacement of human anterior cruciate ligaments with pig ligaments: a model for anti-non-gal antibody response in long-term xenotransplantation. Transplantation, 2007, 83(2): 211-219.
5. Randall KL, Booth BA, Miller AJ, et al. Use of an acellular regenerative tissue matrix in combination with vacuumassisted closure therapy for treatment of a diabetic foot wound. J Foot Ankle Surg, 2008, 47(5): 430-433.
6. Pridgen BC, Woon CY, Kim K, et al. Flexor tendon tissue engineering: acellularization of human flexor tendons with preservation of biomechanical properties and biocompatibility. Tissue Eng Part C Methods, 2011, 17(8): 819-828.
7. Harrison RD, Gratzer PF. Effect of extraction protocols and epidermal growth factor on the cellular repopulation of decellularized anterior cruciate ligament allografts. J Biomed Mater Res A, 2005, 75(4): 841-854.
8. Ning LJ, Zhang Y, Chen XH, et al. Preparation and characterization of decellularized tendon slices for tendon tissue engineering. J Biomed Mater Res A, 2012, 100(6): 1448-1456.
9. Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: Structure and function. Acta Biomater, 2009, 5(1): 1-13.
10. Azuma C, Tohyama H, Nakamura H, et al. Antibody neutralization of TGF-beta enhances the deterioration of collagen fascicles in a tissue-cultured tendon matrix with ex vivo fibroblast infiltration. J Biomech, 2007, 40(10): 2184-2190.
11. Ponce Márquez S, Martínez VS, McIntosh Ambrose W, et al. Decellularization of bovine corneas for tissue engineering applications. Acta Biomater, 2009, 5(6): 1839-1847.
12. Aurora A, McCarron J, Iannotti JP, et al. Commercially available extracellular matrix materials for rotator cuff repairs: state of the art and future trends. J Shoulder Elbow Surg, 2007, 16(5 Suppl): S171-178.
13. Yang JJ, Jang EC, Lee JS, et al. The Effect of amniotic membrane transplantation on tendon-healing in a rabbit Achilles tendon model. Tissue Engineering and Regenerative Medicine, 2010, 7(3): 323-329.
14. Gilbert TW, Stewart AM, Simmons BA, et al. Degradation and remodeling of small intestinal submucosa in canine Achilles tendon repair. J Bone Joint Surg (Am), 2007, 89(6): 621-630.
15. Androgens C, Spragg RK, Derwin KA. Mechanical conditioning of cell-seeded small intestine submucosa: a potential tissue-engineering strategy for tendon repair. Tissue Eng, 2007, 13(2): 233-240.
16. Derwin KA, Badylak SF, Steinmann SP, et al. Extracellular matrix scaffold devices for rotator cuff repair. J Shoulder Elbow Surg, 2010, 19(3): 467-476.
17. Vavken P, Joshi S, Murray MM. Triton-X is most effective among three decellularization agents for ACL tissue engineering. J Orthop Res, 2009, 27(12): 1612-1618.
18. Cartmell JS, Dunn MG. Effect of chemical treatments on tendon cellularity and mechanical properties. J Biomed Mater Res, 2000, 49(1): 134-140.
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. Minami A, Ishii S, Ogino T, et al. Effect of the immunological antigenicity of the allogenaic tendon grafting. Hand, 1982, 14(2): 111-119.
21. Qin TW, Chen QS, Sun YL, et al. Mechanical characteristics of native tendon slices for tissue engineering scaffold. J Biomed Mater Res B, 2012, 100(3): 752-758.

上一篇
人孤雌胚胎干细胞无饲养层培养体系的建立
下一篇
MSCs修复烧伤创面的研究进展

《中国修复重建外科杂志》

脱细胞小牛肌腱组织形态学和生物力学特性研究

摘要 全文 图表 视频 参考文献 施引文献 补充材料

上一篇

下一篇

Format

Content

《中国修复重建外科杂志》

脱细胞小牛肌腱组织形态学和生物力学特性研究

摘要 全文 图表 视频 参考文献 施引文献 补充材料

上一篇

下一篇

Format

Content

摘要全文图表视频参考文献施引文献补充材料