Acellular Cartilaginous Matrix as A Scaffold for Cartilage Reconstruction in vitro_West China Medical Journal

Authors：

GAO Bo ¹ , KONG Qingquan ¹ ,  XIANG Zhou ^1,2 , YANG Zhiming ² , LUO Jingcong ² , LI Xiuqun ²

1. Department of Orthopaedic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China; 2. Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy;

Corresponding author：

XIANG Zhou, Email: xiangzhou@hotmail.com

Keywords：

组织工程软骨; 脱细胞牛软骨基质; 软骨细胞; 体外构建

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【摘要】　目的　以脱细胞牛软骨基质（acellular cartilaginous matrix，ACM）作支架体外构建组织工程软骨，了解其作为软骨组织工程支架的可行性。　方法　 2003年1月-2005年12月联合应用冻干-反复冻融-酶消化法对牛软骨基质行脱细胞处理。将体外培养扩增的2~5代兔软骨细胞接种在材料上，体外培养3周，观察软骨细胞在支架材料上的生长分布情况。　结果　软骨细胞在制备的ACM上可较好地黏附生长，并且分泌大量Ⅱ型胶原和葡萄糖胺聚糖；但软骨细胞不能长入ACM内部，只能在表层生长，少量软骨细胞分布在ACM孔隙中。　结论　 ACM支架材料具有良好的细胞相容性和活性，并且能促进软骨细胞增殖和维持软骨细胞表型。
【Abstract】　Objective　 To investigate the feasibility of applying the acellular cartilaginous matrix (ACM) as the scaffold in constructing tissue engineering cartilage in vitro.　Methods　 We processed the bovine cartilaginous matrix by combination of lyophilization (freeze dry) and repeated freeze-thaw (12 cycles) and enzyme digestion in order to remove the cell component. The 2-5 passage chondrocytes were seeded onto the ACM, and had been cultured for 3 weeks. The cell growth, attachment and distribution were detected by histochemical stain, immunohistochemical stain, and scan electron microscope.　Results　 The chondrocytes could adhere and grow well on the matrix surface, and synthesize plenty of glycosaminoglycanand type Ⅱ collagen. However, the chondrocytes grew only on the surface and superficial layer of the scaffold, and they did not move into the inner part of the scaffold.　Conclusion　 ACM has good cellular compatibility without cytotoxicity and provide temporary substrate to which these anchorage-dependent cells can adhere, and stimulate the chondrocytes anchored on the scaffold to proliferate and keep differentiated phenotype.

Citation： GAO Bo,KONG Qingquan,XIANG Zhou,YANG Zhiming,LUO Jingcong,LI Xiuqun. Acellular Cartilaginous Matrix as A Scaffold for Cartilage Reconstruction in vitro. West China Medical Journal, 2010, 25(11): 2028-2031. doi: Copy

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15.	康红军, 卢世避, 孙明学, 等. 利用脂肪干细胞构建组织工程软骨修复兔膝关节软骨缺损[J]. 中华创伤骨科杂志, 2007, 9(4): 326-330.
16.	Kelley TF, Sutton FM, Wallace, et al. Chondrocyte repopulation of allograft cartilage: a preliminary investigation and strategy for developing cartilage matrices for reconstruction[J]. Otolaryngol Head Neck Surg, 2002, 127(4): 265-270.
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1.
2. 　Ciorba A, Martini A. Tissue engineering and cartilage regeneration for auricular reconstruction[J]. Inter J Pediatr Otorhinol, 2006, 70: 1507-1515.
3. 　Badylak SF. Xenogeneic extracellular matrix as a scaffold for tissue reconstruction[J]. Transpl Immunol, 2004, 12(3-4): 367-377.
4. 　Schlote N, Wefer J, Sievert KD. Acellular matrix for functional reconstruction of the urogenital tract. Special form of "tissue engineering"[J]? Urologe A, 2004, 43(10): 1209-1212.
5. 　Sharma B, Elisseeff JH. Engineering structurally organized cartilage and bone tissues[J]. Ann Biomed Eng, 2004, 32(1): 148-159.
6. 　Capito RM, Spector M. Scaffold-based articular cartilage repair. IEEE Eng Med Biol Mag, 2003, 22(5): 42-50.
7. 　Kleinman HK, Philp D, Hoffman MP. Role of the extracellular matrix in morphogenesis[J]. Curr Opin Biotechnol, 2003, 14(5): 526-532.
8. 　Takezawa T. A strategy for the development of tissue engineering scaffolds that regulate cell behavior[J]. Biomaterials, 2003, 24(13): 2267-2275.
9. 　Badylak SF. The extracellular matrix as a scaffold for tissue reconstruction[J]. Semin Cell Dev Biol, 2002, 13(5): 377-383.
10. 　Abenavoli FM, Corelli R. About AlloDerm[J]. Plast Reconstr Surg, 2001, 108(7): 2175-2176.
11. 　Sclafani AP, Romo T 3rd, Jacono AA, et al. Evaluation of acellular dermal graft in sheet (AlloDerm) and injectable (micronized AlloDerm) forms for soft tissue augmentation. Clinical observations and histological analysis[J]. Arch Facial Plast Surg, 2000, 2(2): 130-136.
12. 　张晨, 王者生, 高景恒. 软骨脱细胞基质生物性的实验研究[J]. 实用美容整形外科杂志, 2003, 14(2): 59-61.
13. 　张晨, 景士兵, 杨琨, 等. 软骨脱细胞基质支架材料的软骨组织工程实验研究[J]. 中国修复重建外科杂志, 2008, 22(7): 846-850.
14. 　Peretti GM, Randolph MA, Villa MT, et al. Cell-based tissue-engineered allogeneic implant for cartilage repair[J]. Tissue Eng, 2000, 6(5): 567-576.
15. 　康红军, 卢世避, 孙明学, 等. 利用脂肪干细胞构建组织工程软骨修复兔膝关节软骨缺损[J]. 中华创伤骨科杂志, 2007, 9(4): 326-330.
16. 　Kelley TF, Sutton FM, Wallace, et al. Chondrocyte repopulation of allograft cartilage: a preliminary investigation and strategy for developing cartilage matrices for reconstruction[J]. Otolaryngol Head Neck Surg, 2002, 127(4): 265-270.
17. 　Zaleske D, Peretti G, Allemann F, et al. Engineering a joint: a chimeric construct with bovine chondrocytes in a devitalized chick knee[J]. Tissue Eng, 2003, 9(5): 949-956.

West China Medical Journal

Acellular Cartilaginous Matrix as A Scaffold for Cartilage Reconstruction in vitro

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