3D打印医用钛合金的抗菌性能和体外生物相容性
|
摘要
应用选择性激光熔融技术(SLM)制备出3D打印医用钛合金Ti-6Al-4V和Ti-6Al-4V-5Cu,用平板共培养法研究测定其抗菌性能,用CCK8细胞增殖测定法、鬼笔环肽细胞骨架染色法和Annexin-V/PI流式细胞术研究了这种合金的抗菌性能和对小鼠胚胎成骨前体细胞(MC3T3-E1)的体外生物相容性影响。结果表明,3D打印Ti-6Al-4V-5Cu合金具有较高的抗菌性能,对金黄色葡萄球菌的抗菌率达到(57.03±1.55)%。在CCK8细胞增殖毒性测定、细胞骨架鬼笔环肽染色实验和Annexin-V/PI双标记法流式分析三种研究中Ti-6Al-4V-5Cu表现的优越,具有更好的体外生物相容性。
The 3 D printing medical titanium alloys Ti-6 Al-4 V and Ti-6 Al-4 V-5 Cu were prepared by selective laser melting technology(SLM), and their antibacterial properties were assessed by plate co-culture method. The in vitro biocompatibility with the mouse embryonic osteogenic precursor cells(MC3 T3-E1) of the prepared alloys was systematically investigated by means of methods of CCK8 cell proliferation assay, phalloidin cytoskeleton staining and Annexin-V/PI flow cytometry. The results show that the 3 D printing Ti-6 Al-4 V-5 Cu alloy has high antibacterial property and the antibacterial rate against Staphylococcus aureus is 57.03%. The alloy Ti-6 Al-4 V-5 Cu performed well with better in vitro biocompatibility during the three assessments,namely,CCK8 cell proliferation toxicity assay, cytoskeleton phalloidin staining experiment and Annexin-V/PI double labeling flow analysis.
The 3 D printing medical titanium alloys Ti-6 Al-4 V and Ti-6 Al-4 V-5 Cu were prepared by selective laser melting technology(SLM), and their antibacterial properties were assessed by plate co-culture method. The in vitro biocompatibility with the mouse embryonic osteogenic precursor cells(MC3 T3-E1) of the prepared alloys was systematically investigated by means of methods of CCK8 cell proliferation assay, phalloidin cytoskeleton staining and Annexin-V/PI flow cytometry. The results show that the 3 D printing Ti-6 Al-4 V-5 Cu alloy has high antibacterial property and the antibacterial rate against Staphylococcus aureus is 57.03%. The alloy Ti-6 Al-4 V-5 Cu performed well with better in vitro biocompatibility during the three assessments,namely,CCK8 cell proliferation toxicity assay, cytoskeleton phalloidin staining experiment and Annexin-V/PI double labeling flow analysis.
引文
[1]Yan T F.The current situation and developmental trend of biomedical materials[J].China Med.Dev.Inf.,2006,12(5):1(奚廷斐.生物医用材料现状和发展趋势[J].中国医疗器械信息,2006,12(5):1)
[2]Li Y,Liu L N,Wan P,et al.Biodegradable Mg-Cu alloy implants with antibacterial activity for the treatment of osteomyelitis:In vitro and in vivo evaluations[J].Biomaterials,2016,106:250
[3]Mombelli A,Müller N,Cionca N.The epidemiology of peri-implantitis[J].Clin.Oral.Implants Res.,2012,23 Suppl6:67
[4]Liu R,Memarzadeh K,Chang B,et al.Antibacterial effect of copper-bearing titanium alloy(Ti-Cu)against Streptococcus mutans and Porphyromonas gingivalis[J].Sci.Rep.,2016,6:29985
[5]Cacciotti I,Bianco A.High thermally stable Mg-substituted tricalcium phosphate via precipitation[J].Ceram.Int.,2011,37:127
[6]Kalita S J,Bhatt H A.Nanocrystalline hydroxyapatite doped with magnesium and zinc:Synthesis and characterization[J].Mater.Sci.Eng.,2007,27C:837
[7]Wu C T,Ramaswamy Y,Kwik D,et al.The effect of strontium incorporation into CaSiO3ceramics on their physical and biological properties[J].Biomaterials,2007,28:3171
[8]Rensing C,Grass G.Escherichia coli mechanisms of copper homeostasis in a changing environment[J].FEMS Microbiol.Rev.,2003,27:197
[9]Ma Z,Ren L,Yang K,et al.Effect of heat treatment on Cu distribution,antibacterial performance and cytotoxicity of Ti-6Al-4V-5Cu alloy[J].J.Mater.Sci.Technol.,2015,31:723
[10]Ren L,Li M,Zhang Y,et al.Antibacterial properties of Ti-6Al-4V-x Cu alloys[J].J.Mater.Sci.Technol.,2014,30:699
[11]Burghardt I,Lüthen F,Prinz C,et al.A dual function of copper in designing regenerative implants[J].Biomaterials,2015,44:36
[12]Zhang B,Huang Q R,Gao Y,et al.Preliminary study on some properties of Co-Cr dental alloy formed by selective laser melting technique[J].J.Wuhan Univ.Technol.Mater.,2012,27:665
[13]Huang W Y,Jiang M Z,Zhan D S.Single crown restoration with 3shape Trios scanner and 3D printing technology[J].Chin.J.Pract.Stomatol.,2017,10:279(黄婉怡,姜慕舟,战德松.口内扫描仪结合3D打印技术单冠固定修复临床研究[J].中国实用口腔科杂志,2017,10:279)
[14]Habijan T,Haberland C,Meier H,et al.The biocompatibility of dense and porous Nickel-Titanium produced by selective laser melting[J].Mater.Sci.Eng.,2013,33C:419
[15]Figliuzzi M,Mangano F,Mangano C.A novel root analogue dental implant using CT scan and CAD/CAM:selective laser melting technology[J].Int.J.Oral Maxill.Surg.,2012,41:858
[16]Zhao B,Xu D K,Sun Z Q,et al.In vitro biocompatibility and antibacterial property of a novel magnesium phosphate whisker[J].Chin.J.Mater.Res.,2016,30:220(赵冰,徐大可,孙子晴等.新型磷镁晶须的体外生物相容性和抗菌性能[J].材料研究学报,2016,30:220)
[17]Foreman A,Boase S,Psaltis A,et al.Role of bacterial and fungal biofilms in chronic rhinosinusitis[J].Curr.Allergy Asthma Rep.,2012,12:127
[18]Rensing C,Grass G.Escherichia coli mechanisms of copper homeostasis in a changing environment[J].FEMS Microbiol.Rev.,2003,27:197
[19]Kang M K,Moon S K,Kwon J S,et al.Antibacterial effect of sand blasted,large-grit,acid-etched treated Ti-Ag alloys[J].Mater.Res.Bull.,2012,47:2952
[20]Mei S L,Wang H Y,Wang W,et al.Antibacterial effects and biocompatibility of titanium surfaces with graded silver incorporation in titania nanotubes[J].Biomaterials,2014,35:4255
[21]Zheng Y H,Li J B,Liu X Y,et al.Antimicrobial and osteogenic effect of Ag-implanted titanium with a nanostructured surface[J].Int J Nanomedicine,2012,7:875
[22]Holmes C J,Faict D.Peritoneal dialysis solution biocompatibility:definitions and evaluation strategies[J].Kidney Int.,2003,64(Suppl.88):S50
[23]Scheiber I F,Mercer J F B,Dringen R.Metabolism and functions of copper in brain[J].Progr.Neurobiol.,2014,116:33
[24]Szymański P,Fraczek T,Markowicz M,et al.Development of copper based drugs,radiopharmaceuticals and medical materials[J].Biometals,2012,25:1089
[25]Qi X H,Jiang H W.Matrix vesicles and their relationship with cytoskeleton-associated proteins[J].Int.J.Stomatol.,2018,45:204(冼雪红,蒋宏伟.基质小泡及其与细胞骨架蛋白的关系[J].国际口腔医学杂志,2018,45:204))
[26]Yu T H,Zhang N,Zhan D S.Evaluation of biocompatibilities of three dental alloys by flow cytometry[J].Chin.J.Mater.Res.,2013,27:652(毓天昊,张宁,战德松.使用流式细胞仪评价3种牙科合金材料的生物相容性[J].材料研究学报,2013,27:652))
[27]Jin S J,Qi X,Wang T M,et al.In vitro study of stimulation effect on endothelialization by a copper bearing cobalt alloy[J].J.Biomed.Mater.Res.,2018,106A:561
[28]Ren L,Wong H M,Yan C H,et al.Osteogenic ability of Cu-bearing stainless steel[J].J.Biomed.Mater.Res,2015,103B:1433
[29]Kemp M G.Crosstalk between apoptosis and autophagy:environmental Genotoxins,infection,and innate immunity[J].J.Cell Death,2017,10,doi:10.1177/1179670716685085.
[30]Jivan R,Damelin L H,Birkhead M,et al.Disulfiram/copper-disulfiram damages multiple protein degradation and turnover pathways and cytotoxicity is enhanced by metformin in oesophageal squamous cell carcinoma cell lines[J].J Cell Biochem,2015,116:2334
[31]Wu X,Xue X,Wang W J,et al.Suppressing autophagy enhances disulfiram/copper-induced apoptosis in non-small cell lung cancer[J].Eur.J.Pharmacol.,2018,827:1
[2]Li Y,Liu L N,Wan P,et al.Biodegradable Mg-Cu alloy implants with antibacterial activity for the treatment of osteomyelitis:In vitro and in vivo evaluations[J].Biomaterials,2016,106:250
[3]Mombelli A,Müller N,Cionca N.The epidemiology of peri-implantitis[J].Clin.Oral.Implants Res.,2012,23 Suppl6:67
[4]Liu R,Memarzadeh K,Chang B,et al.Antibacterial effect of copper-bearing titanium alloy(Ti-Cu)against Streptococcus mutans and Porphyromonas gingivalis[J].Sci.Rep.,2016,6:29985
[5]Cacciotti I,Bianco A.High thermally stable Mg-substituted tricalcium phosphate via precipitation[J].Ceram.Int.,2011,37:127
[6]Kalita S J,Bhatt H A.Nanocrystalline hydroxyapatite doped with magnesium and zinc:Synthesis and characterization[J].Mater.Sci.Eng.,2007,27C:837
[7]Wu C T,Ramaswamy Y,Kwik D,et al.The effect of strontium incorporation into CaSiO3ceramics on their physical and biological properties[J].Biomaterials,2007,28:3171
[8]Rensing C,Grass G.Escherichia coli mechanisms of copper homeostasis in a changing environment[J].FEMS Microbiol.Rev.,2003,27:197
[9]Ma Z,Ren L,Yang K,et al.Effect of heat treatment on Cu distribution,antibacterial performance and cytotoxicity of Ti-6Al-4V-5Cu alloy[J].J.Mater.Sci.Technol.,2015,31:723
[10]Ren L,Li M,Zhang Y,et al.Antibacterial properties of Ti-6Al-4V-x Cu alloys[J].J.Mater.Sci.Technol.,2014,30:699
[11]Burghardt I,Lüthen F,Prinz C,et al.A dual function of copper in designing regenerative implants[J].Biomaterials,2015,44:36
[12]Zhang B,Huang Q R,Gao Y,et al.Preliminary study on some properties of Co-Cr dental alloy formed by selective laser melting technique[J].J.Wuhan Univ.Technol.Mater.,2012,27:665
[13]Huang W Y,Jiang M Z,Zhan D S.Single crown restoration with 3shape Trios scanner and 3D printing technology[J].Chin.J.Pract.Stomatol.,2017,10:279(黄婉怡,姜慕舟,战德松.口内扫描仪结合3D打印技术单冠固定修复临床研究[J].中国实用口腔科杂志,2017,10:279)
[14]Habijan T,Haberland C,Meier H,et al.The biocompatibility of dense and porous Nickel-Titanium produced by selective laser melting[J].Mater.Sci.Eng.,2013,33C:419
[15]Figliuzzi M,Mangano F,Mangano C.A novel root analogue dental implant using CT scan and CAD/CAM:selective laser melting technology[J].Int.J.Oral Maxill.Surg.,2012,41:858
[16]Zhao B,Xu D K,Sun Z Q,et al.In vitro biocompatibility and antibacterial property of a novel magnesium phosphate whisker[J].Chin.J.Mater.Res.,2016,30:220(赵冰,徐大可,孙子晴等.新型磷镁晶须的体外生物相容性和抗菌性能[J].材料研究学报,2016,30:220)
[17]Foreman A,Boase S,Psaltis A,et al.Role of bacterial and fungal biofilms in chronic rhinosinusitis[J].Curr.Allergy Asthma Rep.,2012,12:127
[18]Rensing C,Grass G.Escherichia coli mechanisms of copper homeostasis in a changing environment[J].FEMS Microbiol.Rev.,2003,27:197
[19]Kang M K,Moon S K,Kwon J S,et al.Antibacterial effect of sand blasted,large-grit,acid-etched treated Ti-Ag alloys[J].Mater.Res.Bull.,2012,47:2952
[20]Mei S L,Wang H Y,Wang W,et al.Antibacterial effects and biocompatibility of titanium surfaces with graded silver incorporation in titania nanotubes[J].Biomaterials,2014,35:4255
[21]Zheng Y H,Li J B,Liu X Y,et al.Antimicrobial and osteogenic effect of Ag-implanted titanium with a nanostructured surface[J].Int J Nanomedicine,2012,7:875
[22]Holmes C J,Faict D.Peritoneal dialysis solution biocompatibility:definitions and evaluation strategies[J].Kidney Int.,2003,64(Suppl.88):S50
[23]Scheiber I F,Mercer J F B,Dringen R.Metabolism and functions of copper in brain[J].Progr.Neurobiol.,2014,116:33
[24]Szymański P,Fraczek T,Markowicz M,et al.Development of copper based drugs,radiopharmaceuticals and medical materials[J].Biometals,2012,25:1089
[25]Qi X H,Jiang H W.Matrix vesicles and their relationship with cytoskeleton-associated proteins[J].Int.J.Stomatol.,2018,45:204(冼雪红,蒋宏伟.基质小泡及其与细胞骨架蛋白的关系[J].国际口腔医学杂志,2018,45:204))
[26]Yu T H,Zhang N,Zhan D S.Evaluation of biocompatibilities of three dental alloys by flow cytometry[J].Chin.J.Mater.Res.,2013,27:652(毓天昊,张宁,战德松.使用流式细胞仪评价3种牙科合金材料的生物相容性[J].材料研究学报,2013,27:652))
[27]Jin S J,Qi X,Wang T M,et al.In vitro study of stimulation effect on endothelialization by a copper bearing cobalt alloy[J].J.Biomed.Mater.Res.,2018,106A:561
[28]Ren L,Wong H M,Yan C H,et al.Osteogenic ability of Cu-bearing stainless steel[J].J.Biomed.Mater.Res,2015,103B:1433
[29]Kemp M G.Crosstalk between apoptosis and autophagy:environmental Genotoxins,infection,and innate immunity[J].J.Cell Death,2017,10,doi:10.1177/1179670716685085.
[30]Jivan R,Damelin L H,Birkhead M,et al.Disulfiram/copper-disulfiram damages multiple protein degradation and turnover pathways and cytotoxicity is enhanced by metformin in oesophageal squamous cell carcinoma cell lines[J].J Cell Biochem,2015,116:2334
[31]Wu X,Xue X,Wang W J,et al.Suppressing autophagy enhances disulfiram/copper-induced apoptosis in non-small cell lung cancer[J].Eur.J.Pharmacol.,2018,827:1
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |