殼聚糖復合蠶絲蛋白支架的制備及其性能研究_《中國修復重建外科雜志》

作者：

張鵬 ^1,2 , 王文良 ¹

1 武警后勤學院附屬醫院關節三科（天津，300162）;;
2 河北醫科大學研究生學院;

關鍵詞：

軟骨組織工程蠶絲蛋白殼聚糖支架材料

DOI：

10.7507/1002-1892.20130332

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目的通過調節殼聚糖（chitosan，CS）與蠶絲蛋白（silk fibroin，SF）的質量比制備不同質量比支架材料，并檢測比較其性能。方法通過冷凍干燥法按照SF∶CS質量比分別為6∶4（A組）、6∶8（B組）、6∶16（C組）制備CS-SF支架。觀察各組支架材料的結構、孔隙率、熱水溶失率、彈性模量和吸水膨脹率，掃描電鏡觀察孔徑大小及形態。采用密度梯度離心法分離4周齡SD雄性大鼠BMSCs，取第3代BMSCs接種至各組支架，MTS法檢測細胞在支架上的增殖情況。結果傅里葉紅外光譜儀檢測示，隨著CS含量增高，代表無規卷曲/α螺旋結構的吸收峰1 654 cm-1和1 540 cm-1不斷減小，而對應于1 628 cm-1和1 516 cm-1相當于β-折疊結構的吸收峰不斷增大。A、B、C組孔隙率分別為87.36% ± 2.15%、77.82% ± 1.37%、72.22% ± 1.37%，A組顯著高于B、C組（P lt; 0.05），B組顯著高于C組（P lt; 0.05）。A、B組熱水溶失率均為0，C組為3.12% ± 1.26%。各組支架均具有良好的黏彈性，A、B、C組彈性模量在人正常膝關節軟骨的彈性模量范圍（4～15 kPa）內，組間比較差異無統計學意義（F=5.523，P=0.054）。A、B、C組吸水膨脹率分別為1 528.52% ± 194.63%、1 078.22% ± 100.52%、1 320.05% ± 179.97%，A組顯著高于B組（P=0.05），A、C組間及B、C組間差異無統計學意義（P gt; 0.05）。掃描電鏡觀察示，A組支架孔徑大小較均一（50～250 μm），孔隙相通性好；B、C組結構紊亂，孔徑大小不一，孔隙相通性差。MTS檢測示，1、3、5、7 d時A組活細胞數明顯高于B、C組（P lt; 0.05）；3、5、7 d時，B、C組間差異亦有統計學意義（P lt; 0.05）。結論按SF∶CS質量比為6∶4制備的CS-SF支架適合軟骨組織工程的要求。

引用本文： 張鵬,王文良. 殼聚糖復合蠶絲蛋白支架的制備及其性能研究. 中國修復重建外科雜志, 2013, 27(12): 1517-1522. doi: 10.7507/1002-1892.20130332 復制

1.	張志成, 孫天勝. 軟骨損傷臨床治療進展. 中華外科雜志, 2006, 44(12): 862-864.
2.	姜鑫, 張益民, 李漢秀, 等. 膝關節軟骨損傷的組織工程學治療. 中國組織工程研究與臨床康復, 2007, 11(2): 328, 339.
3.	Bhardwaj N, Kundu SC. Chondrogenic differentiation of rat MSCs on porous scaffolds of silk fibroin/chitosan blends. Biomaterials, 2012, 33(10): 2848-2857.
4.	Feng XX, Zhang LL, Chen JY, et al. Preparation and characterization of novel nanocomposite films formed from silk fibroin and nano-TiO2. Int J Biol Macromol, 2007, 40(2): 105-111.
5.	Swann AC, Seedhom BB. Improved techniques for measuring the indentation and thickness of articular cartilage. Proc Insth Mech Eng H, 1989, 202(3): 143-150.
6.	Hunziker EB. Biologic repair of articular cartilage. Defect models in experimental animals and matrix requirements. Clin Orthop Relat Res, 1999, (367 Suppl): S135-146.
7.	Suh JK, Matthew HW. Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. Biomaterials, 2000, 21(24): 2589-2598.
8.	Wang L, Stegemann JP. Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering. Biomaterials, 2010, 31(14): 3976-3985.
9.	王建, 劉杰, 田華科, 等. 不同種子細胞復合殼聚糖水凝膠構建可注射組織工程髓核的比較研究. 中國修復重建外科雜志, 2010, 24(7): 806-810.
10.	Shi C, Zhu Y, Ran X, et al. Therapeutic potential of chitosan and its derivatives in regenerative medicine. J Surg Res, 2006, 133(2): 185-192.
11.	Ghaznavi AM, Kokai LE, Lovett ML, et al. Silk fibroin conduits: a cellular and functional assessment of peripheral nerve repair. Ann Plast Surg, 2011, 66(3): 273-279.
12.	Mauney JR, Cannon GM, Lovett ML, et al. Evaluation of gel spun silk-based biomaterials in a murine model of bladder augmentation. Biomaterials, 2011, 32(3): 808-818.
13.	韓寧波, 趙建寧. 軟骨組織工程常用支架制備技術. 中國組織工程研究與臨床康復, 2008, 12(19): 3694-3696.
14.	郭忠鵬, 蔣電明. 新型組織工程軟骨支架材料及其構建技術的改進. 中國組織工程研究與臨床康復, 2008, 12(36): 7076-7080.
15.	Homicz MR, Schumacher BL, Sah RL, et al. Effects of serial expansion of septal chondrocytes on tissue-engineered neocartilage composition. Otolaryngol Head Neck Surg, 2002, 127(5): 398-408.
16.	Dozin B, Malpeli M, Camardella L, et al. Response of young, aged and osteoarthritic human articular chondrocytes to inflammatory cytokines: molecular and cellular aspects. Matrix Biol, 2002, 21(5): 449-459.
17.	Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood, 2001, 98(8): 2396-2402.
18.	Lee OK, Kuo TK, Chen WM, et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood, 2004, 103(5): 1669-1675.
19.	Jiang Y, Jahagirdar BN, Reinhardt RL, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature, 2002, 418(6893): 41-49.
20.	Zhao L, Weir MD, Xu HH. An injectable calcium phosphate-alginate hydrogel-umbilical cord mesenchymal stem cell paste for bone tissue engineering. Biomaterials, 2010, 31(25): 6502-6510.
21.	Mao M, He J, Liu Y, et al. Ice-template-induced silk fibroin-chitosan scaffolds with predefined microfluidic channels and fully porous structures. Acta Biomater, 2012, 8(6): 2175-2184.
22.	Muzio G, Vernè E, Canuto RA, et al. Shock waves induce activity of human osteoblast-like cells in bioactive scaffolds. J Trauma, 2010, 68(6): 1439-1444.

1. 張志成, 孫天勝. 軟骨損傷臨床治療進展. 中華外科雜志, 2006, 44(12): 862-864.
2. 姜鑫, 張益民, 李漢秀, 等. 膝關節軟骨損傷的組織工程學治療. 中國組織工程研究與臨床康復, 2007, 11(2): 328, 339.
3. Bhardwaj N, Kundu SC. Chondrogenic differentiation of rat MSCs on porous scaffolds of silk fibroin/chitosan blends. Biomaterials, 2012, 33(10): 2848-2857.
4. Feng XX, Zhang LL, Chen JY, et al. Preparation and characterization of novel nanocomposite films formed from silk fibroin and nano-TiO2. Int J Biol Macromol, 2007, 40(2): 105-111.
5. Swann AC, Seedhom BB. Improved techniques for measuring the indentation and thickness of articular cartilage. Proc Insth Mech Eng H, 1989, 202(3): 143-150.
6. Hunziker EB. Biologic repair of articular cartilage. Defect models in experimental animals and matrix requirements. Clin Orthop Relat Res, 1999, (367 Suppl): S135-146.
7. Suh JK, Matthew HW. Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. Biomaterials, 2000, 21(24): 2589-2598.
8. Wang L, Stegemann JP. Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering. Biomaterials, 2010, 31(14): 3976-3985.
9. 王建, 劉杰, 田華科, 等. 不同種子細胞復合殼聚糖水凝膠構建可注射組織工程髓核的比較研究. 中國修復重建外科雜志, 2010, 24(7): 806-810.
10. Shi C, Zhu Y, Ran X, et al. Therapeutic potential of chitosan and its derivatives in regenerative medicine. J Surg Res, 2006, 133(2): 185-192.
11. Ghaznavi AM, Kokai LE, Lovett ML, et al. Silk fibroin conduits: a cellular and functional assessment of peripheral nerve repair. Ann Plast Surg, 2011, 66(3): 273-279.
12. Mauney JR, Cannon GM, Lovett ML, et al. Evaluation of gel spun silk-based biomaterials in a murine model of bladder augmentation. Biomaterials, 2011, 32(3): 808-818.
13. 韓寧波, 趙建寧. 軟骨組織工程常用支架制備技術. 中國組織工程研究與臨床康復, 2008, 12(19): 3694-3696.
14. 郭忠鵬, 蔣電明. 新型組織工程軟骨支架材料及其構建技術的改進. 中國組織工程研究與臨床康復, 2008, 12(36): 7076-7080.
15. Homicz MR, Schumacher BL, Sah RL, et al. Effects of serial expansion of septal chondrocytes on tissue-engineered neocartilage composition. Otolaryngol Head Neck Surg, 2002, 127(5): 398-408.
16. Dozin B, Malpeli M, Camardella L, et al. Response of young, aged and osteoarthritic human articular chondrocytes to inflammatory cytokines: molecular and cellular aspects. Matrix Biol, 2002, 21(5): 449-459.
17. Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood, 2001, 98(8): 2396-2402.
18. Lee OK, Kuo TK, Chen WM, et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood, 2004, 103(5): 1669-1675.
19. Jiang Y, Jahagirdar BN, Reinhardt RL, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature, 2002, 418(6893): 41-49.
20. Zhao L, Weir MD, Xu HH. An injectable calcium phosphate-alginate hydrogel-umbilical cord mesenchymal stem cell paste for bone tissue engineering. Biomaterials, 2010, 31(25): 6502-6510.
21. Mao M, He J, Liu Y, et al. Ice-template-induced silk fibroin-chitosan scaffolds with predefined microfluidic channels and fully porous structures. Acta Biomater, 2012, 8(6): 2175-2184.
22. Muzio G, Vernè E, Canuto RA, et al. Shock waves induce activity of human osteoblast-like cells in bioactive scaffolds. J Trauma, 2010, 68(6): 1439-1444.

《中國修復重建外科雜志》

殼聚糖復合蠶絲蛋白支架的制備及其性能研究

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《中國修復重建外科雜志》

殼聚糖復合蠶絲蛋白支架的制備及其性能研究

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