新型微弧氧化涂层镁-锌-钙合金支架/自体颗粒骨修复兔临界性骨缺损的研究
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摘要
目的探讨一种新型微弧氧化(micro-arc oxidation,MAO)涂层镁(Mg)-锌(Zn)-钙(Ca)合金支架/自体颗粒骨修复兔临界性骨缺损(critical size bone defect,CSD)的效果,以及该支架在体内的耐腐蚀性和生物相容性。方法将72只新西兰白兔随机分为3组(n=24),A组为无涂层Mg-Zn-Ca合金支架组;B组为10μm厚MAO涂层Mg-Zn-Ca合金支架组;C组为单纯自体颗粒骨植骨组。所有动物制备双侧尺骨15 mm长CSD模型,A、B组将截取的尺骨制成颗粒骨后填充至支架内修复尺骨缺损,C组采用自体颗粒骨修复。术后2、4、8、12周,行大体观察并记录局部皮下积气量;X线片和Van Gieson染色观察骨缺损愈合情况,根据Lane-Sandhu标准行X线片评分;Micro-CT扫描观察并计算支架降解丢失体积百分比(ΔV)及降解速度(corrosion rate,CR);监测实验过程中血清Mg~(2+)、Ca~(2+)浓度变化,并于术后12周收集肝、脑、肾和脾组织行病理学观察。结果术后2、4、8周B组皮下积气量少于A组,其中2、4周两组间比较差异有统计学意义(P<0.05);但术后12周时B组显著多于A组(P<0.05)。术后4、8周C组X线片评分显著高于A、B组(P<0.05),术后8周时B组显著高于A组(P<0.05);术后12周B、C组显著高于A组(P<0.05),但B、C组间差异无统计学意义(P>0.05),同时B组骨缺损部位新骨塑形明显优于A组。Micro-CT示术后4、8周,B组CR及ΔV均显著低于A组(P<0.05)。Van Gieson染色示B组较A组具有更好的生物相容性和促成骨性;血清离子监测结果显示术后各时间点3组血清Mg~(2+)、Ca~(2+)浓度比较,差异均无统计学意义(P>0.05);术后12周,3组实验动物肝、脑、肾及脾组织HE染色均未见明显病理改变。结论新型MAO涂层Mg-Zn-Ca合金支架/自体颗粒骨能够有效修复CSD;同时,10μm厚MAO涂层能有效改良Mg-Zn-Ca合金支架的骨修复效果、耐腐蚀性及生物相容性。
Objective To evaluate the effect of a novel micro-arc oxidation(MAO) coated magnesium-zinccalcium(Mg-Zn-Ca) alloy scaffold/autologous bone particles to repair critical size bone defect(CSD) in rabbit and explore the novel scaffold in vivo corrosion resistance and biocompatibility. Methods Seventy-two New Zealand white rabbits were randomly divided into 3 groups(n=24), group A was uncoated Mg-Zn-Ca alloy scaffold group, group B was 10 μm MAO coated Mg-Zn-Ca alloy scaffold group, and group C was control group with only autologous bone graft. The animals were operated to obtain bilateral ulnar CSD(15 mm in length) models. The bone fragment was removed and minced into small particles and were filled into the scaffolds of groups A and B. Then, the scaffolds or autologous bone particles were replanted into the defects. The animals were sacrificed at 2, 4, 8, and 12 weeks after surgery(6 rabbits each group). The local subcutaneous pneumatosis was observed and recorded. The ulna defect healing was evaluated by X-ray image and Van Gieson staining. The X-ray images were assessed and scored by Lane-Sandhu criteria. The percentage of the lost volume of the scaffold(ΔV) and corrosion rate(CR) were calculated by the Micro-CT. The Mg~(2+) and Ca~(2+) concentrations were monitored during experiment and the rabbit liver, brain, kidney, and spleen were obtained to process HE staining at 12 weeks after surgery. Results The local subcutaneous pneumatosis in group B was less than that in group A at 2, 4, and8 weeks after surgery, showing significant differences between 2 groups at 2 and 4 weeks after surgery(P<0.05); and the local subcutaneous pneumatosis was significantly higher in group B than that in group A at 12 weeks after surgery(P<0.05). The X-ray result showed that the score of group C was significantly higher than those of groups A and B at 4 and 8 weeks after surgery(P<0.05), and the score of group B was significantly higher than that of group A at 8 weeks(P<0.05).At 12 weeks after surgery, the scores of groups B and C were significantly higher than that of group A(P<0.05).Meanwhile, the renew bone moulding of group B was better than that in group A at 12 weeks after surgery. Micro-CT showed that ΔV and CR in group B were significantly lower than those in group A(P<0.05). Van Gieson staining showed that group B had better biocompatibility and osteanagenesis than group A. The Mg~(2+) and Ca~(2+) concentrations in serum showed no significant difference between groups during experiments(P>0.05). And there was no obvious pathological changes in the liver, brain, kidney, and spleen of the 3 groups with HE staining at 12 weeks. Conclusion The MAO coated Mg-Zn-Ca alloy scaffold/autologous bone particles could be used to repair CSD effectively. At the same time,10 μm MAO coating can effectively improve the osteanagenesis, corrosion resistance, and biocompatibility of Mg-Zn-Ca alloy scaffold.
Objective To evaluate the effect of a novel micro-arc oxidation(MAO) coated magnesium-zinccalcium(Mg-Zn-Ca) alloy scaffold/autologous bone particles to repair critical size bone defect(CSD) in rabbit and explore the novel scaffold in vivo corrosion resistance and biocompatibility. Methods Seventy-two New Zealand white rabbits were randomly divided into 3 groups(n=24), group A was uncoated Mg-Zn-Ca alloy scaffold group, group B was 10 μm MAO coated Mg-Zn-Ca alloy scaffold group, and group C was control group with only autologous bone graft. The animals were operated to obtain bilateral ulnar CSD(15 mm in length) models. The bone fragment was removed and minced into small particles and were filled into the scaffolds of groups A and B. Then, the scaffolds or autologous bone particles were replanted into the defects. The animals were sacrificed at 2, 4, 8, and 12 weeks after surgery(6 rabbits each group). The local subcutaneous pneumatosis was observed and recorded. The ulna defect healing was evaluated by X-ray image and Van Gieson staining. The X-ray images were assessed and scored by Lane-Sandhu criteria. The percentage of the lost volume of the scaffold(ΔV) and corrosion rate(CR) were calculated by the Micro-CT. The Mg~(2+) and Ca~(2+) concentrations were monitored during experiment and the rabbit liver, brain, kidney, and spleen were obtained to process HE staining at 12 weeks after surgery. Results The local subcutaneous pneumatosis in group B was less than that in group A at 2, 4, and8 weeks after surgery, showing significant differences between 2 groups at 2 and 4 weeks after surgery(P<0.05); and the local subcutaneous pneumatosis was significantly higher in group B than that in group A at 12 weeks after surgery(P<0.05). The X-ray result showed that the score of group C was significantly higher than those of groups A and B at 4 and 8 weeks after surgery(P<0.05), and the score of group B was significantly higher than that of group A at 8 weeks(P<0.05).At 12 weeks after surgery, the scores of groups B and C were significantly higher than that of group A(P<0.05).Meanwhile, the renew bone moulding of group B was better than that in group A at 12 weeks after surgery. Micro-CT showed that ΔV and CR in group B were significantly lower than those in group A(P<0.05). Van Gieson staining showed that group B had better biocompatibility and osteanagenesis than group A. The Mg~(2+) and Ca~(2+) concentrations in serum showed no significant difference between groups during experiments(P>0.05). And there was no obvious pathological changes in the liver, brain, kidney, and spleen of the 3 groups with HE staining at 12 weeks. Conclusion The MAO coated Mg-Zn-Ca alloy scaffold/autologous bone particles could be used to repair CSD effectively. At the same time,10 μm MAO coating can effectively improve the osteanagenesis, corrosion resistance, and biocompatibility of Mg-Zn-Ca alloy scaffold.
引文
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2Clemens MW,Chang EI,Selber JC,et al.Composite extremity and trunk reconstruction with vascularized fibula flap in postoncologic bone defects:a 10-year experience.Plast Reconstr Surg,2012,129(1):170-178.
3杨运发,张光明,徐中和.下肢创伤后大段感染性骨缺损的分型及修复.中华创伤骨科杂志,2010,12(5):417-420.
4Brandoff JF,Silber JS,Vaccaro AR.Contemporary alternatives to synthetic bone grafts for spine surgery.Am J Orthop(Belle Mead NJ),2008,37(8):410-414.
5Lindsey RW,Gugala Z,Milne E,et al.The efficacy of cylindrical titanium mesh cage for the reconstruction of a critical-size canine segmental femoral diaphyseal defect.J Orthop Res,2006,24(7):1438-1453.
6Staiger MP,Pietak AM,Huadmai J,et al.Magnesium and its alloys as orthopedic biomaterials:a review.Biomaterials,2006,27(9):1728-1734.
7Kim WJ,Chung SW,Chung CS,et al.Superplasticity in thin magnesium alloy sheets and deformation mechanism maps for magnesium alloys at elevated temperatures.Acta Materialia,2001,49(16):3337-3345.
8Flatman PW.Magnesium transport across cell membranes.J Membr Biol,1984,80(1):1-14.
9Wu L,Luthringer BJ,Feyerabend F,et al.Effects of extracellular magnesium on the differentiation and function of human osteoclasts.Acta Biomater,2014,10(6):2843-2854.
10Arnaud MJ.Update on the assessment of magnesium status.Br J Nutri,2008,99(Suppl 3):S24-36.
11Elin RJ.Assessment of magnesium status.Clin Chem,1987,33(11):1965-1970.
12Gibson IR,Huang J,Best SM,et al.Enhanced in vitro cell activity and surface apatite layer formation on novel silicon substituted hydroxyapatites.Bioceramics,1999,12:191-194.
13Park JW,Kim YJ,Jang JH,et al.Osteoblast response to magnesium ion-incorporated nanoporous titanium oxide surfaces.Clin Oral Implants Res,2010,21(11):1278-1287.
14Xu L,Yu G,Zhang E,et al.In vivo corrosion behavior of Mg-MnZn alloy for bone implant application.J Biomed Mater Res,2007,83(3):703-711.
15Smith MR,Atkinson P,White D,et al.Design and assessment of a wrapped cylindrical Ca-P AZ31 Mg alloy for critical-size ulna defect repair.J Biomed Mater Res B Appl Biomater,2012,100(1):206-216.
16葛野,李世慧,李建军,等.镁锶合金修复兔桡骨骨缺损的实验研究.生物骨科材料与临床研究,2016,13(5):1-5.
17Guo JW,Sun SY,Wang YM,et al.Hydrothermal biomimetic modification of micro-arc oxidized magnesium alloy for enhanced corrosion resistance and deposition behaviors in SBF.Surface and Coatings Technology,2015,269(15):183-190.
18Lane J,Sandhu H.Current approaches to experimental bone grafting.Orthop Clin North Am,1987,18(2):213-225.
19Fazel Anvari-Yazdi A,Tahermanesh K,Hadavi SM,et al.Cytotoxicity assessment of adipose-derived mesenchymal stem cells on synthesized biodegradable Mg-Zn-Ca alloys.Mater Sci Eng C Mater Biol Appl,2016,69:584-597.
20Zhao J,CHEN JL,YU K,et al.Effects of chitosan coating on biocompatibility of Mg-6%Zn-10%Ca3(PO4)2 implant.Trans Nonferrous Met Soc.China,2015,3:824-831.
21Jang Y,Tan Z,Jurey C,et al.Understanding corrosion behavior of Mg-Zn-Ca alloys from subcutaneous mouse model:effect of Zn element concentration and plasma electrolytic oxidation.Mater Sci Eng C Mater Biol Appl,2015,48:28-40.
22Zeng RC,Cui LY,Jiang K,et al.In vitro corrosion and cytocompatibility of a microarc oxidation coating and poly(Llactic acid)composite coating on Mg-1Li-1Ca alloy for orthopedic implants.ACS Appl Mater Interfaces,2016,8(15):10014-10028.
23Wu YF,Wang YM,Jing YB,et al.In vivo study of microarc oxidation coated biodegradable magnesium plate to heal bone fracture defect of 3mm width.Colloids Surf B Biointerfaces,2017,158:147-156.
24Chen S,Guan SK,Li W,et al.In vivo degradation and bone response of a composite coating on Mg-Zn-Ca alloy prepared by microarc oxidation and electrochemical deposition.J Biomed Mater Res B Appl Biomater,2012,100(2):533-543.
2Clemens MW,Chang EI,Selber JC,et al.Composite extremity and trunk reconstruction with vascularized fibula flap in postoncologic bone defects:a 10-year experience.Plast Reconstr Surg,2012,129(1):170-178.
3杨运发,张光明,徐中和.下肢创伤后大段感染性骨缺损的分型及修复.中华创伤骨科杂志,2010,12(5):417-420.
4Brandoff JF,Silber JS,Vaccaro AR.Contemporary alternatives to synthetic bone grafts for spine surgery.Am J Orthop(Belle Mead NJ),2008,37(8):410-414.
5Lindsey RW,Gugala Z,Milne E,et al.The efficacy of cylindrical titanium mesh cage for the reconstruction of a critical-size canine segmental femoral diaphyseal defect.J Orthop Res,2006,24(7):1438-1453.
6Staiger MP,Pietak AM,Huadmai J,et al.Magnesium and its alloys as orthopedic biomaterials:a review.Biomaterials,2006,27(9):1728-1734.
7Kim WJ,Chung SW,Chung CS,et al.Superplasticity in thin magnesium alloy sheets and deformation mechanism maps for magnesium alloys at elevated temperatures.Acta Materialia,2001,49(16):3337-3345.
8Flatman PW.Magnesium transport across cell membranes.J Membr Biol,1984,80(1):1-14.
9Wu L,Luthringer BJ,Feyerabend F,et al.Effects of extracellular magnesium on the differentiation and function of human osteoclasts.Acta Biomater,2014,10(6):2843-2854.
10Arnaud MJ.Update on the assessment of magnesium status.Br J Nutri,2008,99(Suppl 3):S24-36.
11Elin RJ.Assessment of magnesium status.Clin Chem,1987,33(11):1965-1970.
12Gibson IR,Huang J,Best SM,et al.Enhanced in vitro cell activity and surface apatite layer formation on novel silicon substituted hydroxyapatites.Bioceramics,1999,12:191-194.
13Park JW,Kim YJ,Jang JH,et al.Osteoblast response to magnesium ion-incorporated nanoporous titanium oxide surfaces.Clin Oral Implants Res,2010,21(11):1278-1287.
14Xu L,Yu G,Zhang E,et al.In vivo corrosion behavior of Mg-MnZn alloy for bone implant application.J Biomed Mater Res,2007,83(3):703-711.
15Smith MR,Atkinson P,White D,et al.Design and assessment of a wrapped cylindrical Ca-P AZ31 Mg alloy for critical-size ulna defect repair.J Biomed Mater Res B Appl Biomater,2012,100(1):206-216.
16葛野,李世慧,李建军,等.镁锶合金修复兔桡骨骨缺损的实验研究.生物骨科材料与临床研究,2016,13(5):1-5.
17Guo JW,Sun SY,Wang YM,et al.Hydrothermal biomimetic modification of micro-arc oxidized magnesium alloy for enhanced corrosion resistance and deposition behaviors in SBF.Surface and Coatings Technology,2015,269(15):183-190.
18Lane J,Sandhu H.Current approaches to experimental bone grafting.Orthop Clin North Am,1987,18(2):213-225.
19Fazel Anvari-Yazdi A,Tahermanesh K,Hadavi SM,et al.Cytotoxicity assessment of adipose-derived mesenchymal stem cells on synthesized biodegradable Mg-Zn-Ca alloys.Mater Sci Eng C Mater Biol Appl,2016,69:584-597.
20Zhao J,CHEN JL,YU K,et al.Effects of chitosan coating on biocompatibility of Mg-6%Zn-10%Ca3(PO4)2 implant.Trans Nonferrous Met Soc.China,2015,3:824-831.
21Jang Y,Tan Z,Jurey C,et al.Understanding corrosion behavior of Mg-Zn-Ca alloys from subcutaneous mouse model:effect of Zn element concentration and plasma electrolytic oxidation.Mater Sci Eng C Mater Biol Appl,2015,48:28-40.
22Zeng RC,Cui LY,Jiang K,et al.In vitro corrosion and cytocompatibility of a microarc oxidation coating and poly(Llactic acid)composite coating on Mg-1Li-1Ca alloy for orthopedic implants.ACS Appl Mater Interfaces,2016,8(15):10014-10028.
23Wu YF,Wang YM,Jing YB,et al.In vivo study of microarc oxidation coated biodegradable magnesium plate to heal bone fracture defect of 3mm width.Colloids Surf B Biointerfaces,2017,158:147-156.
24Chen S,Guan SK,Li W,et al.In vivo degradation and bone response of a composite coating on Mg-Zn-Ca alloy prepared by microarc oxidation and electrochemical deposition.J Biomed Mater Res B Appl Biomater,2012,100(2):533-543.