表面含硅微弧氧化涂层镁合金ZK60体外与成骨细胞生物相容性的研究_《中国修复重建外科杂志》

作者：

杨小明 ^1,2 , 尹庆水 ² , 张余 ² , 李梅 ² , 蓝国波 ² , 林潇 ³ , 谭丽丽 ³ , 杨柯 ³

1 南方医科大学研究生学院（广州，510515）;;
2 广州军区总医院骨科医院骨科实验室;;
3 中国科学院金属研究所;

关键词：

硅微弧氧化涂层镁合金成骨细胞生物相容性

DOI：

10.7507/1002-1892.20130135

视频：

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摘要 全文 图表 视频 参考文献 施引文献 补充材料

目的研究表面含硅微弧氧化（micro-arc oxidation，MAO）涂层镁合金ZK60体外与成骨细胞的生物相容性。方法扫描电镜观察表面含硅MAO涂层镁合金ZK60的表面形貌，并用能谱分析仪检测涂层元素组成。实验分为表面含硅MAO涂层镁合金ZK60组（A组）、无涂层镁合金ZK60组（B组）、医用钛合金组（C组）及空白对照组（D组）。取A、B、C组对应材料按照ISO 10993-12标准（试样表面积/浸体介质= 1.25 cm2/mL）分别制备材料浸提液并培养小鼠前成骨细胞MC3T3-El，D组采用含10%FBS的α-MEM培养基进行培养。倒置相差显微镜观察细胞形态，MTT法检测细胞增殖活力，并检测成骨细胞ALP表达活性。扫描电镜观察A、B、C组材料表面细胞黏附形态，检测涂层表面的蛋白吸附能力，采用DAPI观察细胞早期黏附情况，钙黄绿素/乙锭均二聚物1（calcein-AM/ethidium homodimer 1，calcein- AM/ EthD- 1）双重染色观察细胞生长状态。结果扫描电镜下见表面含硅MAO涂层镁合金ZK60呈粗糙、多孔表面形态，涂层的主要组成元素为Mg、O和Si。材料浸提液培养后各组细胞轮廓清晰、形态良好，其中A组细胞伸展性更好。培养5 d时，MTT检测示A组细胞增殖活力明显高于其他组（P lt; 0.05）。扫描电镜观察见A、C组材料表面细胞伸展性良好，优于B组，且A组细胞与材料表面黏附更紧密。A组材料表面蛋白吸附量为（152.7 ± 6.3）µg/mL，明显高于B、C组的（96.3 ± 3.9）、（96.1 ± 8.7） µg/ mL（P lt; 0.05）；各时间点A组细胞黏附数量均明显高于B、C组（P lt; 0.05）。calcein-AM/EthD-1双染观察见A、C组材料表面细胞生长状态优于B组。 A、B组ALP表达分别为15.55 ± 0.29和13.75 ± 0.44，明显高于C、D组的10.43 ± 0.79和10.73 ± 0.47（P lt; 0.05），且A组高于B组（P lt; 0.05）。结论表面含硅MAO涂层镁合金ZK60可促进成骨细胞增殖、黏附及分化，具有良好生物相容性，是一种较理想的镁合金表面改性方法。

引用本文： 杨小明,尹庆水,张余,李梅,蓝国波,林潇,谭丽丽,杨柯. 表面含硅微弧氧化涂层镁合金ZK60体外与成骨细胞生物相容性的研究. 中国修复重建外科杂志, 2013, 27(5): 612-618. doi: 10.7507/1002-1892.20130135 复制

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2.	Li Z, Gu X, Lou S, et al. The development of binary Mg-Ca alloys for use as biodegradable materials within bone. Biomaterials, 2008, 29(10): 1329-1344.
3.	Zhang S, Zhang X, Zhao C, et al. Research on an Mg-Zn alloy as a degradable biomaterial. Acta Biomater, 2010, 6(2): 626-640.
4.	Song GL. Control of biodegradation of biocompatible magnesium alloys. Corrosion Science, 2007, 49: 1696-1701.
5.	Staiger MP, Pietak AM, Huadmai J, et al. Magnesium and its alloy as orthopaedic biomaterials: a review. Biomater, 2006, 27(9): 1728-1734.
6.	Wong HM, Yeung KW, Lam KO, et al. A biodegradable polymer-based coating to control the performance of magnesium alloy orthopaedic implants. Biomaterials, 2010, 31(8): 2084-2096.
7.	Xin Y, Jiang J, Huo K, et al. Corrosion resistance and cytocompatibility of biodegradable surgical magnesium alloy coated with hydrogenated amorphous silicon. J Biomed Mater Res A, 2009, 89(3): 717-726.
8.	Gray JE, Luan B. Protective coatings on magnesium and its alloys—a critical review. J Alloys Compounds, 2002, 336(1-2): 88-113.
9.	Cai J, Cao F, Chang L, et al. The preparation and corrosion behaviors of MAO coating on AZ91D with rare earth conversion precursor film. Applied Surface Science, 2011, 257 (8): 3804-3811.
10.	Guo HF, An MZ. Growth of ceramic coatings on AZ91D magnesium alloys by micro-arc oxidation in aluminate-fluoride solutions and evaluation of corrosion resistance. Applied Surface Science, 2005, 246(1-3): 229-238.
11.	Guo HF, An MZ, Huo HB, et al. Microstructure characteristic of ceramic coatings fabricated on magnesium alloys by micro-arc oxidation in alkaline silicate solutions. Applied Surface Science, 2006, 252(22): 7911-7916.
12.	Liang J, Hu L, Hao JC. Improvement of corrosion properties of microarc oxidation coating on magnesium alloy by optimizing current density parameters. Applied Surface Science, 2007, 253(16): 6939-6945.
13.	Jugdaohsingh R. Silicon and bone health. J Nutr Health Aging, 2007, 11(2): 99-110.
14.	Ni S, Chou L, Chang J. Preparation and characterization of forsterite (Mg2SiO4) bioceramics. Ceram Inter, 2007, 33: 83-88.
15.	Xie Y, Zhai W, Chen L, et al. Preparation and in vitro evaluation of plasma-sprayed Mg2SiO4 coating on titanium alloy. Acta Biomater, 2009, 5(6): 2331-2337.
16.	Lin X, Tan L, Zhang Q, et al. The in vitro degradation process and biocompatibility of a ZK60 magnesium alloy with forsterite-containing micro-arc oxidation coating. Acta Biomaterialia, 2012. [Epub ahead of print].
17.	ISO 10993-12. Biological evaluation of medical devices—Part 12: Sample preparation and reference materials. Geneva: ISO copyright office, 2007: 1-17.
18.	Chiu KY, Wong MH, Cheng FT, et al. Characterization and corrosion studies of fiuoride conversion coating on degradable Mg implants. Surf Coat Tech, 2007, 202(3): 590-598.
19.	邹静恂, 白进发, 叶玲. 高分子材料在组织工程中的应用. 首都医科大学学报, 2002, 23(4): 364-366.
20.	Sun J, Li J, Liu X, et al. Proliferation and gene expression of osteoblasts cultured in DMEM containing the ionic products of dicalcium silicate coating. Biomed Pharmacother, 2009, 63(9): 650-657.
21.	左林, 柏树令, 潘锋, 等. 成骨细胞与镁合金表面硅酸盐-磷酸盐复合涂层的体外相容性. 解剖学报, 2011, 42(1): 65-69.
22.	Zhu X, Chen J, Scheideler L, et al. Effects of topography and composition of titanium surface oxides on osteoblast responses. Biomaterials, 2004, 25(18): 4087-4103.
23.	Paul W, Sharma CP. Nanoceramic matrices: biomedical applications. Am J Biochem Biotechnol, 2006, 2(2): 41-48.
24.	Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol, 2005, 23(1): 47-55.
25.	Paital SR, Dahotre NB. Laser surface treatment for porous and textured Ca-P bio-ceramic coating on Ti-6Al-4V. Biomed Mater, 2007, 2(4): 274-281.
26.	Assender H, Bliznyuk V, Porfyrakis K. How surface topography relates to materials’ properties. Science, 2002, 297(5583): 973-976.
27.	Webster TJ, Ejiofor JU. Increased osteoblast adhesion on nanophase metals: Ti, Ti6Al4V, and CoCrMo. Biomaterials, 2004, 25(19): 4731-4739.
28.	Johnson I, Perchy D, Liu H. In vitro evaluation of the surface effects on magnesium-yttrium alloy degradation and mesenchymal stem cell adhesion. J Biomed Mater Res Part A, 2011, 176(20): 1778-1784.
29.	Gu XN, Li N, Zheng YF, et al. In vitro degradation performance and biological response of a Mg-Zn-Zr alloy. Materials Science and Engineering: B, 2011, 176(20): 1178-1784.
30.	Revell PA, Damien E, Zhang XS, et al. The effect of magnesium ions on bone bonding to hydroxyapatite coating on titanium alloy implants. Key Engineering Materials, 2004, 254-256: 447-450.
31.	Valerio P, Pereira MM, Goes AM, et al. The effect of ionic products from bioactive glass dissolution on osteoblast proliferation and collagen production. Biomaterials, 2004, 25(15): 2941-2948.
32.	Obata A, Kasuga T. Cellular compatibility of bone-like apatite containing silicon species. J Biomed Res, 2007, 85(1): 140-144.

1. 刘振东, 范清宇. 应力遮挡效应——寻找丢失的钥匙. 中华创伤骨科杂志, 2002, 4(1): 62-64.
2. Li Z, Gu X, Lou S, et al. The development of binary Mg-Ca alloys for use as biodegradable materials within bone. Biomaterials, 2008, 29(10): 1329-1344.
3. Zhang S, Zhang X, Zhao C, et al. Research on an Mg-Zn alloy as a degradable biomaterial. Acta Biomater, 2010, 6(2): 626-640.
4. Song GL. Control of biodegradation of biocompatible magnesium alloys. Corrosion Science, 2007, 49: 1696-1701.
5. Staiger MP, Pietak AM, Huadmai J, et al. Magnesium and its alloy as orthopaedic biomaterials: a review. Biomater, 2006, 27(9): 1728-1734.
6. Wong HM, Yeung KW, Lam KO, et al. A biodegradable polymer-based coating to control the performance of magnesium alloy orthopaedic implants. Biomaterials, 2010, 31(8): 2084-2096.
7. Xin Y, Jiang J, Huo K, et al. Corrosion resistance and cytocompatibility of biodegradable surgical magnesium alloy coated with hydrogenated amorphous silicon. J Biomed Mater Res A, 2009, 89(3): 717-726.
8. Gray JE, Luan B. Protective coatings on magnesium and its alloys—a critical review. J Alloys Compounds, 2002, 336(1-2): 88-113.
9. Cai J, Cao F, Chang L, et al. The preparation and corrosion behaviors of MAO coating on AZ91D with rare earth conversion precursor film. Applied Surface Science, 2011, 257 (8): 3804-3811.
10. Guo HF, An MZ. Growth of ceramic coatings on AZ91D magnesium alloys by micro-arc oxidation in aluminate-fluoride solutions and evaluation of corrosion resistance. Applied Surface Science, 2005, 246(1-3): 229-238.
11. Guo HF, An MZ, Huo HB, et al. Microstructure characteristic of ceramic coatings fabricated on magnesium alloys by micro-arc oxidation in alkaline silicate solutions. Applied Surface Science, 2006, 252(22): 7911-7916.
12. Liang J, Hu L, Hao JC. Improvement of corrosion properties of microarc oxidation coating on magnesium alloy by optimizing current density parameters. Applied Surface Science, 2007, 253(16): 6939-6945.
13. Jugdaohsingh R. Silicon and bone health. J Nutr Health Aging, 2007, 11(2): 99-110.
14. Ni S, Chou L, Chang J. Preparation and characterization of forsterite (Mg2SiO4) bioceramics. Ceram Inter, 2007, 33: 83-88.
15. Xie Y, Zhai W, Chen L, et al. Preparation and in vitro evaluation of plasma-sprayed Mg2SiO4 coating on titanium alloy. Acta Biomater, 2009, 5(6): 2331-2337.
16. Lin X, Tan L, Zhang Q, et al. The in vitro degradation process and biocompatibility of a ZK60 magnesium alloy with forsterite-containing micro-arc oxidation coating. Acta Biomaterialia, 2012. [Epub ahead of print].
17. ISO 10993-12. Biological evaluation of medical devices—Part 12: Sample preparation and reference materials. Geneva: ISO copyright office, 2007: 1-17.
18. Chiu KY, Wong MH, Cheng FT, et al. Characterization and corrosion studies of fiuoride conversion coating on degradable Mg implants. Surf Coat Tech, 2007, 202(3): 590-598.
19. 邹静恂, 白进发, 叶玲. 高分子材料在组织工程中的应用. 首都医科大学学报, 2002, 23(4): 364-366.
20. Sun J, Li J, Liu X, et al. Proliferation and gene expression of osteoblasts cultured in DMEM containing the ionic products of dicalcium silicate coating. Biomed Pharmacother, 2009, 63(9): 650-657.
21. 左林, 柏树令, 潘锋, 等. 成骨细胞与镁合金表面硅酸盐-磷酸盐复合涂层的体外相容性. 解剖学报, 2011, 42(1): 65-69.
22. Zhu X, Chen J, Scheideler L, et al. Effects of topography and composition of titanium surface oxides on osteoblast responses. Biomaterials, 2004, 25(18): 4087-4103.
23. Paul W, Sharma CP. Nanoceramic matrices: biomedical applications. Am J Biochem Biotechnol, 2006, 2(2): 41-48.
24. Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol, 2005, 23(1): 47-55.
25. Paital SR, Dahotre NB. Laser surface treatment for porous and textured Ca-P bio-ceramic coating on Ti-6Al-4V. Biomed Mater, 2007, 2(4): 274-281.
26. Assender H, Bliznyuk V, Porfyrakis K. How surface topography relates to materials’ properties. Science, 2002, 297(5583): 973-976.
27. Webster TJ, Ejiofor JU. Increased osteoblast adhesion on nanophase metals: Ti, Ti6Al4V, and CoCrMo. Biomaterials, 2004, 25(19): 4731-4739.
28. Johnson I, Perchy D, Liu H. In vitro evaluation of the surface effects on magnesium-yttrium alloy degradation and mesenchymal stem cell adhesion. J Biomed Mater Res Part A, 2011, 176(20): 1778-1784.
29. Gu XN, Li N, Zheng YF, et al. In vitro degradation performance and biological response of a Mg-Zn-Zr alloy. Materials Science and Engineering: B, 2011, 176(20): 1178-1784.
30. Revell PA, Damien E, Zhang XS, et al. The effect of magnesium ions on bone bonding to hydroxyapatite coating on titanium alloy implants. Key Engineering Materials, 2004, 254-256: 447-450.
31. Valerio P, Pereira MM, Goes AM, et al. The effect of ionic products from bioactive glass dissolution on osteoblast proliferation and collagen production. Biomaterials, 2004, 25(15): 2941-2948.
32. Obata A, Kasuga T. Cellular compatibility of bone-like apatite containing silicon species. J Biomed Res, 2007, 85(1): 140-144.

《中国修复重建外科杂志》

表面含硅微弧氧化涂层镁合金ZK60体外与成骨细胞生物相容性的研究

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《中国修复重建外科杂志》

表面含硅微弧氧化涂层镁合金ZK60体外与成骨细胞生物相容性的研究

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