AlCoCrCuFeTix高熵合金细胞毒性评价及对L929细胞凋亡与Ⅰ型胶原合成的影响
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摘要
目的评价AlCoCrCuFeTix(x=0,0.25,0.5,1)系列高熵合金细胞毒性。方法对AlCoCrCuFeTix(x=0,0.25,0.5,1)系列高熵合金进行MTT毒性实验评价其细胞毒性;流式细胞术AnnexinⅤ-FITC/PI双染法评价其对L929细胞凋亡的影响;免疫荧光染色检测各组L929细胞Ⅰ型胶原的合成水平来评价该系列材料对细胞增殖的影响。结果随着Ti元素的递增,AlCoCrCuFeTix(x=0,0.25,0.5,1)系列高熵合金细胞毒性增大,凋亡率呈上升趋势,而L929细胞的Ⅰ型胶原蛋白表达水平下降。实验表明AlCoCrCuFe具有良好的细胞相容性,细胞毒性各时间点评级均为1级,且凋亡率及Ⅰ型胶原蛋白表达水平与空白对照组无明显差异。结论 AlCoCrCuFeTix系列高熵合金中的AlCoCrCuFe具有良好的细胞相容性,可初步满足医疗器械使用的基本要求。
Objective To evaluate the cytotoxicity of AlCoCrCuFeTix(x=0, 0.25, 0.5,1) series high entropy alloys and explore the effect on apoptosis and type I collagen synthesis in L929 cells by flow cytometry and immunofluorescence. Methods The AlCoCrCuFeTix(x=0, 0.25, 0.5, 1) series of high entropy alloys were tested by MTT test to evaluate the cytotoxicity of series of alloys. Annexin V-FITC/PI staining with flow cytometry was used to evaluate the effect of series of materials on apoptosis of L929 cells. Immunofluorescence staining was used to detect the level of collagen I synthesis to evaluate the effect of materials on cell proliferation. Results The cytotoxicity series of high entropy alloys showed an increasing trend with the increase of Ti elements. At the same time, the proliferation rate of L929 cells decreased, the rate of apoptosis increased and the expression level of type Ⅰ collagen in L929 cells decreased. The metal of AlCoCrCuFe had good cell compatibility, the cytotoxicity was Grade 1 at each time, and there was no statistical difference in the apoptotic rate and type Ⅰ collagen expression level compared with the blank control group. Conclusion AlCoCrCuFe alloy in the series of AlCoCrCuFeTix(x=0, 0.25, 0.5, 1) high entropy alloys has good cell compatibility and can preliminarily meet the basic requirements of the application of dental materials.
Objective To evaluate the cytotoxicity of AlCoCrCuFeTix(x=0, 0.25, 0.5,1) series high entropy alloys and explore the effect on apoptosis and type I collagen synthesis in L929 cells by flow cytometry and immunofluorescence. Methods The AlCoCrCuFeTix(x=0, 0.25, 0.5, 1) series of high entropy alloys were tested by MTT test to evaluate the cytotoxicity of series of alloys. Annexin V-FITC/PI staining with flow cytometry was used to evaluate the effect of series of materials on apoptosis of L929 cells. Immunofluorescence staining was used to detect the level of collagen I synthesis to evaluate the effect of materials on cell proliferation. Results The cytotoxicity series of high entropy alloys showed an increasing trend with the increase of Ti elements. At the same time, the proliferation rate of L929 cells decreased, the rate of apoptosis increased and the expression level of type Ⅰ collagen in L929 cells decreased. The metal of AlCoCrCuFe had good cell compatibility, the cytotoxicity was Grade 1 at each time, and there was no statistical difference in the apoptotic rate and type Ⅰ collagen expression level compared with the blank control group. Conclusion AlCoCrCuFe alloy in the series of AlCoCrCuFeTix(x=0, 0.25, 0.5, 1) high entropy alloys has good cell compatibility and can preliminarily meet the basic requirements of the application of dental materials.
引文
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[2]Tsai MH,Yeh JW.High-entropy alloys:a critical review[J].Mater Res Lett,2014,2(3):107-123.
[3]Miracle DB,Senkov ON.A critical review of high entropy alloys and related concepts[J].Acta Mater,2016,122:448-511.
[4]Pickering EJ,Jones NG.High-entropy alloys:a critical assessment of their founding principles and future prospects[J].Int Mater Rev,2016,61(3):183-202.
[5]Gludovatz B,Hohenwarter A,Thurston KVS,et al.Exceptional damage-tolerance of a medium-entropy alloy Cr Co Ni at cryogenic temperatures[J].Nat Commun,2016,7:10602.
[6]Li DY,Zhang Y.The ultrahigh charpy impact toughness of forged Alx Co Cr Fe Ni high entropy alloys at room and cryogenic temperatures[J].Intermetallics,2016,70:24-28.
[7]Qiu XW,Zhang YP,He L,et al.Microstructure and corrosion resistance of Al Cr Fe Cu Co high entropy alloy[J].J Alloy Compd,2013,549(2):195-199.
[8]Praveen S,Murty BS,Kottada RS.Alloying behavior in multicomponent Al Co Cr Cu Fe and Ni Co Cr Cu Fe high entropy alloys[J].Mat Sci Eng A,2012,534(1):83-89.
[9]Australia S.Biological evaluation of medical devices[J].Iso,2005,16(5):10-15.
[10]Hong D,Saha P,Chou D T,et al.In vitro degradation and cytotoxicity response of Mg-4%Zn-0.5%Zr(ZK40)alloy as a potential biodegradable material[J].Acta Biomater,2013,9(10):8534-8547.
[11]Fields GB.New strategies for targeting matrix metalloproteinases[J].Matrix Biol,2015,44-46:239.
[12]Gasik M.In vitro test method for implant materials:US,EP2697622 A1[P].2014-02-06.
[13]Reddy LH,Arias JL,Nicolas J,et al.Magnetic nanoparticles:design and characterization,toxicity and biocompatibility,pharmaceuticaland biomedical applications[J].Chem Rev,2012,112:5818-5878.
[14]Pogrebnyak AD,Bagdasaryan AA,Yakushchenko IV,et al.The structure and properties of high-entropy alloys and nitride coatings based on them[J].Russ Chem Rev,2014,83(11):1027-1061.
[15]He JY,Wang H,Huang HL,et al.A precipitation-hardened highentropy alloy with outstanding tensile properties[J].Acta Mater,2016,102:187-196.
[16]Li Z,Tasan CC,Springer H,et al.Interstitial atoms enable joint twinning and transformation induced plasticity in strong and ductile high-entropy alloys[J].Sci Rep,2017,7:40704.
[17]Yurchenko NY,Stepanov ND,Zherebtsov SV,et al.Structure and mechanical properties of B2 ordered refractory Al Nb Ti VZr x(x=0-1.5)high-entropy alloys[J].Mat Sci Eng A,2017,704:82-90.
[18]Dimic'I,Cvijovic'alagic'I,Hohenwarter A,et al.Electrochemical and biocompatibility examinations of high-pressure torsion processed titanium and Ti-13Nb-13Zr alloy[J].J Biomed Mater Res B,2017,3(106):1097-1107.
[19]Li Y,Wong C,Xiong J,et al.Cytotoxicity of titanium and titanium alloying elements[J].J Dent Res,2010,89(5):493.
[20]Okazaki Y,Gotoh E.Comparison of metal release from various metallic biomaterials in vitro[J].Biomaterials,2005,26(1):11-21.
[21]He Q,Shi J.Mesoporous Silicananoparticle based nano drug delivery systems:synthesis,controlled drug release and delivery,pharmacokinetics and biocompatibility[J].J Mater Chem A,2011,21(16):5845-5855.
[22]Weijia LI,Zhou J,Yuyin XU.Study of the in vitro cytotoxicity testing of medical devices[J].Biomed Rep,2015,3(5):617-620.
[23]Elshahawy WM,Watanabe I,Kramer P.In vitro cytotoxicity evaluation of elemental ions released from different prosthodontic materials[J].Dent Mater,2009,25(12):1551-1555.
[24]孟贺,韩东,战德松.五种牙科合金对小鼠成纤维细胞毒性及凋亡相关基因表达的影响[J].中华口腔医学杂志,2009,44(8):497-501.
[25]刘敏,胡平安,丁克祥.不同血清浓度对小鼠成纤维细胞L929生长增殖和胶原蛋白分泌的影响[J].细胞与分子免疫学杂志,2011,27(7):736-739.