• 四川大學華西醫(yī)院（成都，610041）1 骨科，2 生物治療國家重點實驗室?干細胞與組織工程研究室;

【摘要】　目的　 以脫細胞牛軟骨基質（acellular cartilaginous matrix，ACM）作支架體外構建組織工程軟骨，了解其作為軟骨組織工程支架的可行性。　方法　 2003年1月-2005年12月聯(lián)合應用凍干-反復凍融-酶消化法對牛軟骨基質行脫細胞處理。將體外培養(yǎng)擴增的2~5代兔軟骨細胞接種在材料上，體外培養(yǎng)3周，觀察軟骨細胞在支架材料上的生長分布情況。　結果　 軟骨細胞在制備的ACM上可較好地黏附生長，并且分泌大量Ⅱ型膠原和葡萄糖胺聚糖；但軟骨細胞不能長入ACM內部，只能在表層生長，少量軟骨細胞分布在ACM孔隙中。　結論　 ACM支架材料具有良好的細胞相容性和活性，并且能促進軟骨細胞增殖和維持軟骨細胞表型。
【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.

引用本文： 高博,孔清泉,項舟,楊志明,羅靜聰,李秀群. 以脫細胞牛軟骨基質為支架體外構建組織工程軟骨的實驗研究. 華西醫(yī)學, 2010, 25(11): 2028-2031. doi: 復制

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. 　張晨, 王者生, 高景恒. 軟骨脫細胞基質生物性的實驗研究[J]. 實用美容整形外科雜志, 2003, 14(2): 59-61.
13. 　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[J]. 中國修復重建外科雜志, 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. 　康紅軍, 盧世避, 孫明學, 等. 利用脂肪干細胞構建組織工程軟骨修復兔膝關節(jié)軟骨缺損[J]. 中華創(chuàng)傷骨科雜志, 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.
  1. 1.
  2. 2. 　Ciorba A, Martini A. Tissue engineering and cartilage regeneration for auricular reconstruction[J]. Inter J Pediatr Otorhinol, 2006, 70: 1507-1515.
  3. 3. 　Badylak SF. Xenogeneic extracellular matrix as a scaffold for tissue reconstruction[J]. Transpl Immunol, 2004, 12(3-4): 367-377.
  4. 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. 5. 　Sharma B, Elisseeff JH. Engineering structurally organized cartilage and bone tissues[J]. Ann Biomed Eng, 2004, 32(1): 148-159.
  6. 6. 　Capito RM, Spector M. Scaffold-based articular cartilage repair. IEEE Eng Med Biol Mag, 2003, 22(5): 42-50.
  7. 7. 　Kleinman HK, Philp D, Hoffman MP. Role of the extracellular matrix in morphogenesis[J]. Curr Opin Biotechnol, 2003, 14(5): 526-532.
  8. 8. 　Takezawa T. A strategy for the development of tissue engineering scaffolds that regulate cell behavior[J]. Biomaterials, 2003, 24(13): 2267-2275.
  9. 9. 　Badylak SF. The extracellular matrix as a scaffold for tissue reconstruction[J]. Semin Cell Dev Biol, 2002, 13(5): 377-383.
  10. 10. 　Abenavoli FM, Corelli R. About AlloDerm[J]. Plast Reconstr Surg, 2001, 108(7): 2175-2176.
  11. 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. 12. 　張晨, 王者生, 高景恒. 軟骨脫細胞基質生物性的實驗研究[J]. 實用美容整形外科雜志, 2003, 14(2): 59-61.
  13. 13. 　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[J]. 中國修復重建外科雜志, 2008, 22(7): 846-850.
  14. 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. 15. 　康紅軍, 盧世避, 孫明學, 等. 利用脂肪干細胞構建組織工程軟骨修復兔膝關節(jié)軟骨缺損[J]. 中華創(chuàng)傷骨科雜志, 2007, 9(4): 326-330.
  16. 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. 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.