脫細胞跟腱復合人成纖維細胞共培養的實驗研究_《中國修復重建外科雜志》

作者：

王直兵 , 張峽 , 郭新宇 , 秦川

第三軍醫大學附屬新橋醫院骨科（重慶，400037）;

關鍵詞：

組織工程韌帶脫細胞跟腱人成纖維細胞細胞相容性兔

DOI：

10.7507/1002-1892.20130177

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目的探討脫細胞跟腱的制備及其與人成纖維細胞復合培養效果，為組織工程韌帶的構建提供一種支架材料。方法取10只5月齡雄性新西蘭大白兔（體重4～5 kg）雙后肢跟腱，經胰蛋白酶、Triton X-100、SDS脫細胞處理后，行大體、組織學及掃描電鏡觀察。將脫細胞跟腱與人成纖維細胞復合培養，于培養1、3、5、7、9 d行細胞相容性檢測，并于4周后取材行組織學、掃描電鏡觀察以及生物力學檢測，并與脫細胞前、后跟腱進行比較。結果跟腱經脫細胞處理后腱膜完整，質柔軟、韌性好，體積較處理前跟腱顯著增大；跟腱腱束間未見疏松結締組織及腱細胞，膠原纖維排列疏松，呈三維網狀分布，腱束間殘留較多孔隙；且細胞相容性良好。復合培養4周后，細胞遷移至纖維束間的孔隙內，三維網狀結構消失。生物力學測試示，與脫細胞前跟腱組及細胞-支架復合組比較，脫細胞跟腱組拉伸強度及楊氏彈性模量均顯著降低（P lt; 0.05），細胞-支架復合組與脫細胞前跟腱組比較差異無統計學意義（P gt; 0.05）；各組間斷裂伸長率比較，差異均無統計學意義（P gt; 0.05）。結論兔脫細胞跟腱與人成纖維細胞生物相容性良好，可作為支架材料用于組織工程韌帶構建。

引用本文： 王直兵,張峽,郭新宇,秦川. 脫細胞跟腱復合人成纖維細胞共培養的實驗研究. 中國修復重建外科雜志, 2013, 27(7): 805-809. doi: 10.7507/1002-1892.20130177 復制

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11.	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.
12.	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.
13.	Feil G, Christ-Adler M, Maurer S, et al. Investigations of urothelial cells seeded on commercially available small intestine submucosa. Eur Urol, 2006, 50(6): 1330-1337.
14.	Elder BD, Eleswarapu SV, Athanasiou KA. Extraction techniques for the decellularization of tissue engineered articular cartilage constructs. Biomaterials, 2009, 30(22): 3749-3756．.
15.	Ott HC, Matthiesen TS, Goh SK, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med, 2008, 14(2): 213-221.
16.	Uygun BE, Soto-Gutierrez A, Yagi H, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010, 16(7): 814-820.
17.	Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
18.	Nakayama KH, Batchelder CA, Lee CI, et al. Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng Part A, 2010, 16(7): 2207-2216.
19.	Ni M, Rui YF, Tan Q, et al. Engineered scaffold-free tendon tissue produced by tendon-derived stem cells. Biomaterials, 2013, 34(8): 2024-2037.
20.	Zhang J, Li B, Wang JH. The role of engineered tendon matrix in the stemness of tendon stem cells in vitro and the promotion of tendon-like tissue formation in vivo. Biomaterials, 2011, 32(29): 6972-6981.
21.	Hashimoto T, Sun YL, An KN, et al. The effect of tendon surface treatment on cell attachment for potential enhancement of tendon graft healing: an ex vivo model. Med Eng Phys, 2012, 34(10): 1387-1393.
22.	Kew SJ, Gwynne JH, Enea D, et al. Regeneration and repair of tendon and ligament tissue using collagen fibre biomaterials. Acta Biomater, 2011, 7(9): 3237-3247.
23.	Ikeda J, Zhao C, Chen Q, et al. Compressive properties of cd-HA-gelatin modified intrasynovial tendon allograft in canine model in vivo. Biomechanics, 2011, 44(9): 1793-1796.
24.	Wang JH, Guo Q, Li B. Tendon biomechanics and mechanobiology—a minireview of basic concepts and recent advancements. J Hand Ther, 2012, 25(2): 133-141.
25.	Garvin J, Qi J, Maloney M, et al. Novel system for engineering bioartificial tendons and application of mechanical load. Tissue Eng, 2003, 9(5): 967-979.

1. Liu W, Chen B, Deng D, et al. Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model. Tissue Eng, 2006, 12(4): 775-788.
2. Cao D, Liu W, Wei X, et al. In vitro tendon engineering with avian tenocytes and polyglycolic acids: a preliminary report. Tissue Eng, 2006, 12(5): 1369-1377.
3. 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.
4. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials, 2011, 32(12): 3233-3243.
5. 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.
6. 解傳飚, 林月秋, 阮默, 等. EDC 交聯改性的脫細胞異種(豬)肌腱生物學特性研究. 中國臨床解剖學雜志, 2010, 28(4): 425-429.
7. 冷元曦, 林月秋, 阮默, 等. 肌腱移植材料的研究進展. 中國矯形外科雜志, 2009, 17(22): 1704-1706.
8. Yang B, Zhang Y, Zhou L, et al. Development of a porcine bladder acellular matrix with well-preserved extracellular bioactive factors for tissue engineering. Tissue Eng Part C Methods, 2010, 16(5): 1201-1211.
9. 胡成棟, 劉曦, 張伯勛, 等. 脫細胞雞肌腱的制備與體外生物力學測定的實驗研究. 河北醫藥, 2011, 33(18): 2761-2763.
10. 梁黎明, 柴家科, 楊紅明, 等. 脫細胞肌腱制備的實驗研究. 中國美容醫學, 2006, 15(3): 239-240.
11. 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.
12. 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.
13. Feil G, Christ-Adler M, Maurer S, et al. Investigations of urothelial cells seeded on commercially available small intestine submucosa. Eur Urol, 2006, 50(6): 1330-1337.
14. Elder BD, Eleswarapu SV, Athanasiou KA. Extraction techniques for the decellularization of tissue engineered articular cartilage constructs. Biomaterials, 2009, 30(22): 3749-3756．.
15. Ott HC, Matthiesen TS, Goh SK, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med, 2008, 14(2): 213-221.
16. Uygun BE, Soto-Gutierrez A, Yagi H, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010, 16(7): 814-820.
17. Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
18. Nakayama KH, Batchelder CA, Lee CI, et al. Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng Part A, 2010, 16(7): 2207-2216.
19. Ni M, Rui YF, Tan Q, et al. Engineered scaffold-free tendon tissue produced by tendon-derived stem cells. Biomaterials, 2013, 34(8): 2024-2037.
20. Zhang J, Li B, Wang JH. The role of engineered tendon matrix in the stemness of tendon stem cells in vitro and the promotion of tendon-like tissue formation in vivo. Biomaterials, 2011, 32(29): 6972-6981.
21. Hashimoto T, Sun YL, An KN, et al. The effect of tendon surface treatment on cell attachment for potential enhancement of tendon graft healing: an ex vivo model. Med Eng Phys, 2012, 34(10): 1387-1393.
22. Kew SJ, Gwynne JH, Enea D, et al. Regeneration and repair of tendon and ligament tissue using collagen fibre biomaterials. Acta Biomater, 2011, 7(9): 3237-3247.
23. Ikeda J, Zhao C, Chen Q, et al. Compressive properties of cd-HA-gelatin modified intrasynovial tendon allograft in canine model in vivo. Biomechanics, 2011, 44(9): 1793-1796.
24. Wang JH, Guo Q, Li B. Tendon biomechanics and mechanobiology—a minireview of basic concepts and recent advancements. J Hand Ther, 2012, 25(2): 133-141.
25. Garvin J, Qi J, Maloney M, et al. Novel system for engineering bioartificial tendons and application of mechanical load. Tissue Eng, 2003, 9(5): 967-979.

《中國修復重建外科雜志》

脫細胞跟腱復合人成纖維細胞共培養的實驗研究

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脫細胞跟腱復合人成纖維細胞共培養的實驗研究

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