脫鈣皮質骨基質的拉伸力學性能研究_《中國修復重建外科雜志》

作者：

劉國明 ¹ , 張憶 ^2,3 , 江燕林 ³ , 黃富國 ¹ , 秦廷武 ^2,3

四川大學華西醫院（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;;
3 再生醫學研究中心;

關鍵詞：

脫鈣皮質骨基質力學性能組織工程支架

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目的探討脫鈣皮質骨基質厚度與拉伸力學性能的關系，為將其作為組織工程支架材料奠定實驗基礎。方法取市售新鮮小牛脛骨，常規快速脫鈣制備脫鈣皮質骨基質，大體觀察其顏色、質地等物理性狀，并進行脫鈣檢測。將脫鈣皮質骨基質沿徑向縱切成不同厚度的片材，并根據厚度分為4 組（n=16），分別為A 組100 ～ 300 μm，B 組300 ～ 500 μm，C 組500 ～ 700 μm，D 組700 ～ 1 000 μm；對每組標本進行拉伸性能和組織學觀測。結果大體觀察顯示脫鈣皮質骨基質經H2O2 處理后呈乳白色，柔韌性好，富有彈性。脫鈣皮質骨基質的脫鈣率達97.6%。A 組強度及彈性模量明顯小于B、C、D 組（P lt; 0.05），B、C、D 組間比較差異均無統計學意義（P gt; 0.05）；脫鈣皮質骨基質剛度隨厚度增加呈逐漸增大趨勢，A 組剛度顯著低于B、C、D 組（P lt; 0.05），B、C 組低于D 組（P lt; 0.05），B、C 組間差異無統計學意義（P gt; 0.05）；各組極限應變差異均無統計學意義（P gt; 0.05）。組織學觀察各組均可見典型骨單位結構，其最大直徑范圍為102 ～ 325 μm，平均最大直徑為182 μm。結論骨單位的完整性對脫鈣皮質骨基質的力學性能有重要影響；脫鈣皮質骨基質作為組織工程支架材料，在厚度 gt; 300 μm 時可保持其拉伸力學性能。

引用本文： 劉國明 ,張憶,江燕林,黃富國,秦廷武. 脫鈣皮質骨基質的拉伸力學性能研究. 中國修復重建外科雜志, 2012, 26(4): 501-505. doi: 復制

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2.	Swadener JG, Rho JY, Pharr GM. Effects of anisotropy on elastic moduli measured by nanoindentation in human tibial cortical bone. J Biomed Mater Res, 2001, 57(1): 108-112.
3.	Novitskaya E, Chen PY, Lee S, et al. Anisotropy in the compressive mechanical properties of bovine cortical bone and the mineral and protein constituents. Acta Biomater, 2011, 7(8): 3170-3177.
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5.	Urist MR, Dawson E. Intertransverse process fusion with the aid of chemosterilized autolyzed antigen-extracted allogeneic (AAA) bone. Clin Orthop Relat Res, 1981, (154): 97-113.
6.	Urist MR, Silverman BF, Büring K, et al. The bone induction principle. Clin Orthop Relat Res, 1967, (53): 243-283.
7.	Bigham AS, Dehghani SN, Shafiei Z, et al. Xenogenic demineralized bone matrix and fresh autogenous cortical bone effects on experimental bone healing: radiological, histopathological and biomechanical evaluation. J Orthop Traumatol, 2008, 9(2): 73-80.
8.	Mistry AS, Mikos AG. Tissue engineering strategies for bone regeneration. Adv Biochem Eng Biotechnol, 2005, 94: 1-22.
9.	Sasso RC, LeHuec JC, Shaffrey C, et al. Iliac crest bone graft donor site pain after anterior lumbar interbody fusion: a prospective patient satisfaction outcome assessment. J Spinal Disord Tech, 2005, 18(Suppl): S77-81.
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11.	Buck BE, Resnick L, Shah SM, et al. Human immunodeficiency virus cultured from bone. Implications for transplantation. Clin Orthop Relat Res, 1990, (251): 249-253.
12.	Honsawek S, Bumrungpanichthaworn P, Thanakit V. Osteoinductive potential of small intestinal submucosa/demineralized bone matrix as composite scaffolds for bone tissue engineering. Asian Biomed, 2010, 4 (6): 913-922.
13.	Guizzardi S, Di Silvestre M, Scandroglio R, et al. Implants of heterologous demineralized bone matrix for induction of posterior spinal fusion in rats. Spine (Phlia Pa 1976), 1992, 17(6): 701-707.
14.	Pietrzak WS, Perns SV, Keyes J, et al. Demineralized bone matrix graft: a scientific and clinical case study assessment. J Foot Ankle Surg, 2005, 44(5): 345-353.
15.	Honsawek S, Dhitiseith D, Phupong V. Gene expression characteristics of osteoblast differentiation in human umbilical cord mesenchymal stem cells induced by demineralized bone matrix. Asian Biomed, 2007, 1(4): 383-391.
16.	Kasten P, Luginbühl R, van Griensven M, et al. Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix. Biomaterials, 2003, 24(15): 2593-603.
17.	Xie H, Yang F, Deng L, et al. The performance of a bone-derived scaffold material in the repair of critical bone defects in a rhesus monkey model. Biomaterials, 2007, 28(22): 3314-3324.
18.	Honsawek S, Bumrungpanichthaworn P, Thitiset T, et al. Gene expression analysis of demineralized bone matrix-induced osteogenesis in human periosteal cells using cDNA array technology. Genet Mol Res, 2011, 10(3): 2093-2103.
19.	Jackson DW, Simon TM, Lowery W, et al. Biologic remodeling after anterior cruciate ligament reconstruction using a collagen matrix derived from demineralized bone. An experimental study in the goat model. Am J Sports Med, 1996, 24(4): 405-414.
20.	Yamada T. Bone-demineralized bone-bone graft for ligament reconstruction in rats. J Med Dent Sci, 2004, 51(1): 45-52.
21.	Burstein AH, Zika JM, Heiple KG, et al. Contribution of collagen and mineral to the elastic-plastic properties of bone. J Bone Joint Surg (Am), 1975, 57(7): 956-961.
22.	Paterson CR. Collagen chemistry and the brittle bone diseases. Endeavour, 1988, 12(2): 56-59.
23.	Wang X, Bank RA, Tekoppele JM, et al. The role of collagen in determining bone mechanical properties. J Orthop Res, 2001, 19(6): 1021-1026.
24.	侯振德, 高瑞亭, 周欣竹. 干牛骨拉伸彈性模量沿徑向的分布. 中國生物醫學工程學報, 1995, 14(2): 149-154.
25.	Bowman SM, Zeind J, Gibson LJ, et al. The tensile behavior of demineralized bovine cortical bone. J Biomech, 1996, 29(11): 1497-501.
26.	Catanese J 3rd, Iverson EP, Ng RK, et al. Heterogeneity of the mechanical properties of demineralized bone. J Biomech, 1999, 32(12): 1365-1369.
27.	Omae H, Zhao C, Sun YL, et al. Multilayer tendon slices seeded with bone marrow stromal cells: a novel composite for tendon engineering. J Orthop Res, 2009, 27(7): 937-942.
28.	Mauney JR, Sjostorm S, Blumberg J, et al. Mechanical stimulation promotes osteogenic differentiation of human bone marrow stromal cells on 3-D partially demineralized bone scaffolds in vitro. Calcif Tissue Int, 2004, 74(5): 458-468.
29.	向強, 鄧聰穎, 張遠, 等. 成骨細胞特異性鈣黏蛋白涂布脫鈣骨基質材料對BMSCs 黏附及成骨分化能力的影響. 中國修復重建外科雜志, 2009, 23(5): 602-606.
30.	Hou T, Li Q, Luo F, et al. Controlled dynamization to enhance reconstruction capacity of tissue-engineered bone in healing critically sized bone defects: an in vivo study in goats. Tissue Eng Part A, 2010, 16(1): 201-212.
31.	侯天勇, 羅飛, 劉杰, 等. 凍干組織工程骨與組織工程骨成骨活性比較研究. 中國修復重建外科雜志, 2010, 24(7): 779-784.
32.	楊軍, 周振東, 李建軍, 等. 動態壓力促進骨基質支架中微血管形成的實驗研究. 中華骨科雜志, 2011, 31(4): 372-378.
33.	Pan Y, Dong SW, Hao Y, et al. Demineralized bone matrix gelatin as scaffold for tissue engineering. Afr J Microbiol Res, 2010, 4(9): 865-870.
34.	Summitt MC, Reisinger KD. Characterization of the mechanical properties of demineralized bone. J Biomed Mater Res A, 2003, 67(3): 742-750.

1. Rho JY, Kuhn-Spearing L, Zioupos P. Mechanical properties and the hierarchical structure of bone. Med Eng Phys, 1998, 20(2): 92-102.
2. Swadener JG, Rho JY, Pharr GM. Effects of anisotropy on elastic moduli measured by nanoindentation in human tibial cortical bone. J Biomed Mater Res, 2001, 57(1): 108-112.
3. Novitskaya E, Chen PY, Lee S, et al. Anisotropy in the compressive mechanical properties of bovine cortical bone and the mineral and protein constituents. Acta Biomater, 2011, 7(8): 3170-3177.
4. Martin RB, Ishida J. The relative effects of collagen fiber orientation, porosity, density, and mineralization on bone strength. J Biomech, 1989, 22(5): 419-426.
5. Urist MR, Dawson E. Intertransverse process fusion with the aid of chemosterilized autolyzed antigen-extracted allogeneic (AAA) bone. Clin Orthop Relat Res, 1981, (154): 97-113.
6. Urist MR, Silverman BF, Büring K, et al. The bone induction principle. Clin Orthop Relat Res, 1967, (53): 243-283.
7. Bigham AS, Dehghani SN, Shafiei Z, et al. Xenogenic demineralized bone matrix and fresh autogenous cortical bone effects on experimental bone healing: radiological, histopathological and biomechanical evaluation. J Orthop Traumatol, 2008, 9(2): 73-80.
8. Mistry AS, Mikos AG. Tissue engineering strategies for bone regeneration. Adv Biochem Eng Biotechnol, 2005, 94: 1-22.
9. Sasso RC, LeHuec JC, Shaffrey C, et al. Iliac crest bone graft donor site pain after anterior lumbar interbody fusion: a prospective patient satisfaction outcome assessment. J Spinal Disord Tech, 2005, 18(Suppl): S77-81.
10. Silber JS, Anderson DG, Daffner SD, et al. Donor site morbidity after anterior iliac crest bone harvest for single-level anterior cervical discectomy and fusion. Spine (Phila Pa 1976), 2003, 28(2): 134-139.
11. Buck BE, Resnick L, Shah SM, et al. Human immunodeficiency virus cultured from bone. Implications for transplantation. Clin Orthop Relat Res, 1990, (251): 249-253.
12. Honsawek S, Bumrungpanichthaworn P, Thanakit V. Osteoinductive potential of small intestinal submucosa/demineralized bone matrix as composite scaffolds for bone tissue engineering. Asian Biomed, 2010, 4 (6): 913-922.
13. Guizzardi S, Di Silvestre M, Scandroglio R, et al. Implants of heterologous demineralized bone matrix for induction of posterior spinal fusion in rats. Spine (Phlia Pa 1976), 1992, 17(6): 701-707.
14. Pietrzak WS, Perns SV, Keyes J, et al. Demineralized bone matrix graft: a scientific and clinical case study assessment. J Foot Ankle Surg, 2005, 44(5): 345-353.
15. Honsawek S, Dhitiseith D, Phupong V. Gene expression characteristics of osteoblast differentiation in human umbilical cord mesenchymal stem cells induced by demineralized bone matrix. Asian Biomed, 2007, 1(4): 383-391.
16. Kasten P, Luginbühl R, van Griensven M, et al. Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix. Biomaterials, 2003, 24(15): 2593-603.
17. Xie H, Yang F, Deng L, et al. The performance of a bone-derived scaffold material in the repair of critical bone defects in a rhesus monkey model. Biomaterials, 2007, 28(22): 3314-3324.
18. Honsawek S, Bumrungpanichthaworn P, Thitiset T, et al. Gene expression analysis of demineralized bone matrix-induced osteogenesis in human periosteal cells using cDNA array technology. Genet Mol Res, 2011, 10(3): 2093-2103.
19. Jackson DW, Simon TM, Lowery W, et al. Biologic remodeling after anterior cruciate ligament reconstruction using a collagen matrix derived from demineralized bone. An experimental study in the goat model. Am J Sports Med, 1996, 24(4): 405-414.
20. Yamada T. Bone-demineralized bone-bone graft for ligament reconstruction in rats. J Med Dent Sci, 2004, 51(1): 45-52.
21. Burstein AH, Zika JM, Heiple KG, et al. Contribution of collagen and mineral to the elastic-plastic properties of bone. J Bone Joint Surg (Am), 1975, 57(7): 956-961.
22. Paterson CR. Collagen chemistry and the brittle bone diseases. Endeavour, 1988, 12(2): 56-59.
23. Wang X, Bank RA, Tekoppele JM, et al. The role of collagen in determining bone mechanical properties. J Orthop Res, 2001, 19(6): 1021-1026.
24. 侯振德, 高瑞亭, 周欣竹. 干牛骨拉伸彈性模量沿徑向的分布. 中國生物醫學工程學報, 1995, 14(2): 149-154.
25. Bowman SM, Zeind J, Gibson LJ, et al. The tensile behavior of demineralized bovine cortical bone. J Biomech, 1996, 29(11): 1497-501.
26. Catanese J 3rd, Iverson EP, Ng RK, et al. Heterogeneity of the mechanical properties of demineralized bone. J Biomech, 1999, 32(12): 1365-1369.
27. Omae H, Zhao C, Sun YL, et al. Multilayer tendon slices seeded with bone marrow stromal cells: a novel composite for tendon engineering. J Orthop Res, 2009, 27(7): 937-942.
28. Mauney JR, Sjostorm S, Blumberg J, et al. Mechanical stimulation promotes osteogenic differentiation of human bone marrow stromal cells on 3-D partially demineralized bone scaffolds in vitro. Calcif Tissue Int, 2004, 74(5): 458-468.
29. 向強, 鄧聰穎, 張遠, 等. 成骨細胞特異性鈣黏蛋白涂布脫鈣骨基質材料對BMSCs 黏附及成骨分化能力的影響. 中國修復重建外科雜志, 2009, 23(5): 602-606.
30. Hou T, Li Q, Luo F, et al. Controlled dynamization to enhance reconstruction capacity of tissue-engineered bone in healing critically sized bone defects: an in vivo study in goats. Tissue Eng Part A, 2010, 16(1): 201-212.
31. 侯天勇, 羅飛, 劉杰, 等. 凍干組織工程骨與組織工程骨成骨活性比較研究. 中國修復重建外科雜志, 2010, 24(7): 779-784.
32. 楊軍, 周振東, 李建軍, 等. 動態壓力促進骨基質支架中微血管形成的實驗研究. 中華骨科雜志, 2011, 31(4): 372-378.
33. Pan Y, Dong SW, Hao Y, et al. Demineralized bone matrix gelatin as scaffold for tissue engineering. Afr J Microbiol Res, 2010, 4(9): 865-870.
34. Summitt MC, Reisinger KD. Characterization of the mechanical properties of demineralized bone. J Biomed Mater Res A, 2003, 67(3): 742-750.

《中國修復重建外科雜志》

脫鈣皮質骨基質的拉伸力學性能研究

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《中國修復重建外科雜志》

脫鈣皮質骨基質的拉伸力學性能研究

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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