纯镁超声微弧氧化载黄芪提取物膜层动物实验研究
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摘要
目的纯镁种植体经超声微弧氧后,以植酸为偶联剂载入黄芪甲苷,研究其在动物体内的降解情况及骨结合情况。方法以纯镁超声微弧氧化为A组,在A组基础上以植酸偶联剂封孔为B组,在B组基础上加入黄芪提取物(浓度0. 0025 g/mL)为C组,将不同植体植入家兔单侧下颌骨中,采用SEM观察植体表面形态,三维锥形束CT(CBCT)观察植入8周后植体表面骨结合及植体降解情况,HE染色观察2周、4周、8周植体与骨组织界面成骨情况,免疫组化测定2周、4周BMP-2灰度值。结果纯镁种植体经微弧氧化处理,再经植酸和黄芪提取物复合,种植体表面膜层空隙明显缩小实现封孔,CBCT观察第2周、4周、8周植表面骨结合情况及植体降解能力情况均为C组>B组>A组,HE及免疫组化染色法显示植体周围成骨情况C组>B组>A组。结论纯镁种植体超声微弧氧化以植酸为偶联剂载黄芪提取物黄芪甲苷复合膜层达到了封孔效果,植入动物体内后可减缓植体的降解速率,且具有更好地促进骨结合和早期骨生长的能力。
Objective The degradation and osseointegration of astragaloside of pure magnesium implants in animals were studied by using phytic acid as coupling agent after ultrasonic micro-arc oxidation. Methods Ultrasonic micro-arc oxidation of pure magnesium was used as group A. On the basis of group A,the phytic acid coupling agent was used to seal the hole for group B. On the basis of group B,the extract of astragalus(concentration: 0.0025 g/ml) was added for group C. Different implants were implanted into the unilateral mandible of rabbits. The surface morphology of the implants was observed by SEM. Threedimensional cone beam computed tomography(CBCT) was used to observe the osseointegration and implant degradation of the implants after 2,4,and 8 weeks of implantation. HE staining was used to observe the osteogenicity of the interface. The gray value of BMP-2 was measured by immunohistochemistry for 2 and 4 weeks. Results The pure magnesium implants were treated by micro-arc oxidation,and then phytic acid and Astragalus membranaceus extracts were combined. The gaps on the surface of the implant were significantly reduced to achieve sealing. The surface bone bonding of the implants at 2,4 and 8 weeks from CBCT observation and the implant degradation ability were group C > group B > group A. HE and immunohistochemical staining showed that the osteogenesis around the implant in group C > group B >group A. Conclusions Ultrasonic micro-arc oxidation of pure magnesium implants with phytic acid as a coupling agent loaded with Astragalus membranaceus extract and Astragaloside IV complex membrane layer achieved sealing effect. It can slow the degradation rate of implantsand has better ability to promote osseointegration and early bone growth.
Objective The degradation and osseointegration of astragaloside of pure magnesium implants in animals were studied by using phytic acid as coupling agent after ultrasonic micro-arc oxidation. Methods Ultrasonic micro-arc oxidation of pure magnesium was used as group A. On the basis of group A,the phytic acid coupling agent was used to seal the hole for group B. On the basis of group B,the extract of astragalus(concentration: 0.0025 g/ml) was added for group C. Different implants were implanted into the unilateral mandible of rabbits. The surface morphology of the implants was observed by SEM. Threedimensional cone beam computed tomography(CBCT) was used to observe the osseointegration and implant degradation of the implants after 2,4,and 8 weeks of implantation. HE staining was used to observe the osteogenicity of the interface. The gray value of BMP-2 was measured by immunohistochemistry for 2 and 4 weeks. Results The pure magnesium implants were treated by micro-arc oxidation,and then phytic acid and Astragalus membranaceus extracts were combined. The gaps on the surface of the implant were significantly reduced to achieve sealing. The surface bone bonding of the implants at 2,4 and 8 weeks from CBCT observation and the implant degradation ability were group C > group B > group A. HE and immunohistochemical staining showed that the osteogenesis around the implant in group C > group B >group A. Conclusions Ultrasonic micro-arc oxidation of pure magnesium implants with phytic acid as a coupling agent loaded with Astragalus membranaceus extract and Astragaloside IV complex membrane layer achieved sealing effect. It can slow the degradation rate of implantsand has better ability to promote osseointegration and early bone growth.
引文
[1] Sandro Carrara. Integrated Bio/Nano/CMOS interfaces for electrochemical molecular sensing[J]. IEEE Transactions on Electrical and Electronic Engineering,2018,13(11):1534-1539.
[2] Uhrig B A,Clements I P,Boerckel J D,et al. Characterization of a composite injury model of severe lower limb bone and nerve trauma[J]. Journal of Tissue Engineering and Regenerative Medicine,2014,8(6):432-441.
[3] Razavi M,Fathi M,Savabi O,et al. Coating of biodegradable magnesium alloy bone implants using nanostructured diopside(CaMgSi2O6)[J]. Applied Surface Science,2014,288:130-137.
[4]Staiger M P,Pietak M A,Huadmai J,et al. Magnesium and its alloys as orthopedic biomaterials:A review[J].Biomaterials,2006,27(9):1728-1734.
[5]王建利,马斌,张楠,等.生物可降解医用镁合金研究进展[J].热加工工艺,2015,44(10):5-11.Wang Jianli,Ma Bin,Zhang Nan,et al. Advances in research on biodegradable medical magnesium alloys[J].Thermal Processing,2015,44(10):5-11.
[6] Farraro K F,Kim K E,Woo S L,et al.Revolutionizing orthopedic biomaterials:The potential of biodegradable and bioresorbable magnesium based materials for functional tissueengineering[J]. Journal of Biomechanics,2014,47(9):1979-1986.
[7]张永强,赵建宁,包倪荣.医用镁合金表面处理的研究与应用[J].中国组织工程研究,2018,22(22):3589-3594.Zhang Yongqiang,Zhao Jianning,Bao Nirong. Research and application of surface treatment of medical magnesium alloys[J]. China Tissue Engineering Research,2018,22(22):3589-3594.
[8]Wang B J,Wang S D,Xu D K,et al. Recent progress in fatigue behavior of Mg alloys in air and aqueous media:Areview[J]. Journal of Materials Science and Technology,2017,33(10):1075-1086.
[9]Song Yingwei,Shan Dayong,Han Enhou. Pitting corrosion of a Rare Earth Mg alloy GW93[J]. Journal of Materials Science&Technology,2017,33(9):954-960.
[10]罗军明,吴小红,徐吉林.电解液组分对Ti CP/Ti6Al4V复合材料微弧氧化膜耐蚀性及耐磨性影响[J].无机材料学报,2017,32(4):418-424.Luo Junming,Wu Xiaohong,Xu Jilin. Electrolytic composition has an effect on corrosion resistance and wear resistance of Ti CP/Ti6Al4V composite microarc oxidation film[J]. Journal of Inorganic Materials,2017,32(4):418-424.
[11]曲立杰,李慕勤,张二林,等.超声微弧氧化处理镁合金模拟体液中的耐蚀性[J].材料热处理学报,2013,34(3):130-135.Qu Lijie,Li Muqin,Zhang Erlin,et al. Ultrasonic microarc oxidation to simulate corrosion resistance in magnesium alloys in body fluids[J]. Journal of Material Heat Treatment,2013,34(3):130-135.
[12]韩旭,刘继光,李慕勤,等.纯镁微弧氧化载绿原酸复合膜层的家兔骨结合能力研究[J].中国体视学与图像分析,2018,23(2):198-205.Han Xu,Liu Jiguang,Li Muqin,et al. The study of rabbit bone binding ability of the chlorogenic acid composite film layer[J]. Chinese Journal of Stereology and Image Analysis,2018,23(2):198-205.
[13]Duan H,Du K,Yan C,et al. Electrochemical corrosion behavior of composite coatings of sealed MAO film on magnesium alloy AZ91D[J]. Electrochimica Acta,2006,51(14):2898-2908.
[14]刘继光,王娅涵,李慕勤,等.纯镁超声微弧氧化-植酸-丝素复合处理种植体的体内降解特性[J].中国体视学与图像分析,2016,21(2):197-205.Liu Jiguang,Wang Yahan,Li Muqin,et al. Pure magnesium ultrasound microarc oxidation phytate silk compound treatment of implants in vivo degradation characteristics[J]. Chinese Journal of Stereology and Image Analysis,2016,21(2):197-205.
[15]Zhang Xuan,Chen Yingqi,Lei Huili,et al. Phytic acid layer template-assisted deposition of Ti O2film on titanium:Surface electronic properties, superhydrophilicity and bending strength[J]. Materials&Design,2016,89:476-484.
[16]林红强,杨娜,王涵,等.黄芪的化学成分、药理活性及临床应用研究进展[J].世界最新医学信息文摘,2018,18(38):45-47,+49.Lin Hongqiang,Yang Na,Wang Han,et al. Advances in the study of chemical constituents,pharmacological activities and clinical application of Astragalus membranaceus[J]. Abstracts of the Latest Medical Information in the World,2018,18(38):45-47,+49.
[17]王青,赵林华,邸莎.黄芪的临床应用及其用量探究[J].吉林中医药,2018,38(12):1450-1454.Wang Qing,Zhao Linhua,Di Sha. Exploration of clinical application and dosage of Astragalus membranaceus[J].Jilin Traditional Chinese Medicine, 2018, 38(12):1450-1454.
[18]魏方红,李慕勤,郭小娟,等.植酸对纯镁超声微弧氧化层防护性能的影响[J].中国表面工程,2015,28(2):78-83.Wei Fanghong,Li Muqin,Guo Xiaojuan,et al. Effect of phytic acid on protective properties of pure magnesium ultrasonic micro-arc oxide layer[J]. China Surface Engineering,2015,28(2):78-83.
[19]Reznikov N,Shahar R,Weiner S. Bone hierarchical structure in three dimensions[J]. Acta Biomaterialia,2014,10(9):3815-3826.
[20]白燕,王士斌,陈爱政,等. BMP-2及FGF-2对骨髓间充质干细胞成骨分化的影响[J].重庆医科大学学报,2017,42(12):1575-1581.Bai Yan,Wang Shibin,Chen Aizheng,et al. BMP-2 and FGF-2 on bone marrow mesenchymal stem cell osteogenesis of effects[J]. Journal of the ChongqingMedical University,2017,42(12):1575-1581.
[21]赵华桥,朱红,王家洪,等.黄芪甲苷抑制COUP-TFII表达对成骨细胞分化的影响及相关分子机制研究[J].四川中医,2018,36(5):54-58.Zhao Huaqiao,Zhu Hong,Wang Jiahong,et al. Huangqi inhibit the effect of COUP-TFII expression on osteoblasts differentiation and related molecular mechanisms[J].Sichuan Traditional Chinese Medicine,2018,36(5):54-58.
[2] Uhrig B A,Clements I P,Boerckel J D,et al. Characterization of a composite injury model of severe lower limb bone and nerve trauma[J]. Journal of Tissue Engineering and Regenerative Medicine,2014,8(6):432-441.
[3] Razavi M,Fathi M,Savabi O,et al. Coating of biodegradable magnesium alloy bone implants using nanostructured diopside(CaMgSi2O6)[J]. Applied Surface Science,2014,288:130-137.
[4]Staiger M P,Pietak M A,Huadmai J,et al. Magnesium and its alloys as orthopedic biomaterials:A review[J].Biomaterials,2006,27(9):1728-1734.
[5]王建利,马斌,张楠,等.生物可降解医用镁合金研究进展[J].热加工工艺,2015,44(10):5-11.Wang Jianli,Ma Bin,Zhang Nan,et al. Advances in research on biodegradable medical magnesium alloys[J].Thermal Processing,2015,44(10):5-11.
[6] Farraro K F,Kim K E,Woo S L,et al.Revolutionizing orthopedic biomaterials:The potential of biodegradable and bioresorbable magnesium based materials for functional tissueengineering[J]. Journal of Biomechanics,2014,47(9):1979-1986.
[7]张永强,赵建宁,包倪荣.医用镁合金表面处理的研究与应用[J].中国组织工程研究,2018,22(22):3589-3594.Zhang Yongqiang,Zhao Jianning,Bao Nirong. Research and application of surface treatment of medical magnesium alloys[J]. China Tissue Engineering Research,2018,22(22):3589-3594.
[8]Wang B J,Wang S D,Xu D K,et al. Recent progress in fatigue behavior of Mg alloys in air and aqueous media:Areview[J]. Journal of Materials Science and Technology,2017,33(10):1075-1086.
[9]Song Yingwei,Shan Dayong,Han Enhou. Pitting corrosion of a Rare Earth Mg alloy GW93[J]. Journal of Materials Science&Technology,2017,33(9):954-960.
[10]罗军明,吴小红,徐吉林.电解液组分对Ti CP/Ti6Al4V复合材料微弧氧化膜耐蚀性及耐磨性影响[J].无机材料学报,2017,32(4):418-424.Luo Junming,Wu Xiaohong,Xu Jilin. Electrolytic composition has an effect on corrosion resistance and wear resistance of Ti CP/Ti6Al4V composite microarc oxidation film[J]. Journal of Inorganic Materials,2017,32(4):418-424.
[11]曲立杰,李慕勤,张二林,等.超声微弧氧化处理镁合金模拟体液中的耐蚀性[J].材料热处理学报,2013,34(3):130-135.Qu Lijie,Li Muqin,Zhang Erlin,et al. Ultrasonic microarc oxidation to simulate corrosion resistance in magnesium alloys in body fluids[J]. Journal of Material Heat Treatment,2013,34(3):130-135.
[12]韩旭,刘继光,李慕勤,等.纯镁微弧氧化载绿原酸复合膜层的家兔骨结合能力研究[J].中国体视学与图像分析,2018,23(2):198-205.Han Xu,Liu Jiguang,Li Muqin,et al. The study of rabbit bone binding ability of the chlorogenic acid composite film layer[J]. Chinese Journal of Stereology and Image Analysis,2018,23(2):198-205.
[13]Duan H,Du K,Yan C,et al. Electrochemical corrosion behavior of composite coatings of sealed MAO film on magnesium alloy AZ91D[J]. Electrochimica Acta,2006,51(14):2898-2908.
[14]刘继光,王娅涵,李慕勤,等.纯镁超声微弧氧化-植酸-丝素复合处理种植体的体内降解特性[J].中国体视学与图像分析,2016,21(2):197-205.Liu Jiguang,Wang Yahan,Li Muqin,et al. Pure magnesium ultrasound microarc oxidation phytate silk compound treatment of implants in vivo degradation characteristics[J]. Chinese Journal of Stereology and Image Analysis,2016,21(2):197-205.
[15]Zhang Xuan,Chen Yingqi,Lei Huili,et al. Phytic acid layer template-assisted deposition of Ti O2film on titanium:Surface electronic properties, superhydrophilicity and bending strength[J]. Materials&Design,2016,89:476-484.
[16]林红强,杨娜,王涵,等.黄芪的化学成分、药理活性及临床应用研究进展[J].世界最新医学信息文摘,2018,18(38):45-47,+49.Lin Hongqiang,Yang Na,Wang Han,et al. Advances in the study of chemical constituents,pharmacological activities and clinical application of Astragalus membranaceus[J]. Abstracts of the Latest Medical Information in the World,2018,18(38):45-47,+49.
[17]王青,赵林华,邸莎.黄芪的临床应用及其用量探究[J].吉林中医药,2018,38(12):1450-1454.Wang Qing,Zhao Linhua,Di Sha. Exploration of clinical application and dosage of Astragalus membranaceus[J].Jilin Traditional Chinese Medicine, 2018, 38(12):1450-1454.
[18]魏方红,李慕勤,郭小娟,等.植酸对纯镁超声微弧氧化层防护性能的影响[J].中国表面工程,2015,28(2):78-83.Wei Fanghong,Li Muqin,Guo Xiaojuan,et al. Effect of phytic acid on protective properties of pure magnesium ultrasonic micro-arc oxide layer[J]. China Surface Engineering,2015,28(2):78-83.
[19]Reznikov N,Shahar R,Weiner S. Bone hierarchical structure in three dimensions[J]. Acta Biomaterialia,2014,10(9):3815-3826.
[20]白燕,王士斌,陈爱政,等. BMP-2及FGF-2对骨髓间充质干细胞成骨分化的影响[J].重庆医科大学学报,2017,42(12):1575-1581.Bai Yan,Wang Shibin,Chen Aizheng,et al. BMP-2 and FGF-2 on bone marrow mesenchymal stem cell osteogenesis of effects[J]. Journal of the ChongqingMedical University,2017,42(12):1575-1581.
[21]赵华桥,朱红,王家洪,等.黄芪甲苷抑制COUP-TFII表达对成骨细胞分化的影响及相关分子机制研究[J].四川中医,2018,36(5):54-58.Zhao Huaqiao,Zhu Hong,Wang Jiahong,et al. Huangqi inhibit the effect of COUP-TFII expression on osteoblasts differentiation and related molecular mechanisms[J].Sichuan Traditional Chinese Medicine,2018,36(5):54-58.