TiCuO薄膜的显微结构对Cu离子释放和内皮细胞行为的影响

doi:10.15541/jim20180004

摘要/Abstract

摘要：

采用直流磁控溅射技术, 在Si片和316L SS基体上制备了不同Cu含量的TiCuO薄膜。采用X射线衍射仪(XRD)、透射电镜(TEM)、X 射线能谱仪(EDS)和X射线光电子能谱(XPS)对薄膜的显微结构和化学组成进行了分析。采用电化学腐蚀和模拟体液浸泡实验评价了薄膜的腐蚀性能和Cu离子释放特性。体外静态培养内皮细胞后, 采用细胞计数试剂盒(CCK-8)评价了TiCuO薄膜的细胞活性。研究结果表明, 未掺杂的TiO₂薄膜为金红石相, 掺入Cu后的TiCuO薄膜由非晶基体上含有Cu₂O的纳米晶粒构成。薄膜中的富Cu区引起了薄膜腐蚀。含Cu量高的TiCuO薄膜更易被腐蚀, 并释放出较多Cu离子。TiCuO薄膜释放出一定浓度的Cu离子促进了内皮细胞活性。研究表明, TiCuO薄膜的含Cu量和显微结构影响了Cu离子释放, 对其内皮细胞活性起了关键作用。

关键词: TiCuO薄膜, 磁控溅射, Cu离子释放, 内皮细胞治性

Abstract:

TiCuO films with different microstructures and Cu contents were prepared on Si and 316L SS substrates using DC magnetron sputtering. The microstructure and chemical composition of the films were analyzed by X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray energy dispersive spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS). Corrosion behavior and Cu ions release properties of the films were evaluated by electrochemical corrosion and simulated body-fluid immersion tests, respectively. Endothelial cell (EC) viability on the films was evaluated by CCK-8 assay in vitro. Un-doped TiO₂ films exhibit rutile phases. After Cu incorporation, the TiCuO films consist of nano-crystalline grains of Cu₂O on their amorphous matrix. The films with higher Cu contents are more susceptible to corrosion and release more Cu ions. The Cu-rich phases of the films result in corrosion. EC viability is enhanced by a certain concentration of Cu ions released from TiCuO films. All above results demonstrated that microstructures and Cu contents of TiCuO films play an important role in controlling copper ions release behaviors, and therefore affect the EC viability.

Key words: TiCuO films, magnetron sputtering, Cu ions release, ECs viability

中图分类号:

TG178
R318

程丹, 黄斌, 陈涛, 景凤娟, 谢东, 冷永祥, 黄楠. TiCuO薄膜的显微结构对Cu离子释放和内皮细胞行为的影响[J]. 无机材料学报, 2018, 33(10): 1089-1096.

CHENG Dan, HUANG Bin, CHEN Tao, JING Feng-Juan, XIE Dong, LENG Yong-Xiang, HUANG Nan. Microstructure of TiCuO Films on Copper Ion Release and Endothelial Cell Behavior[J]. Journal of Inorganic Materials, 2018, 33(10): 1089-1096.

图/表 11

表1 采用EDS检测TiCuO薄膜中的Ti、Cu、O元素的相对原子含量

Table 1 Atomic concentration of TiCuO films evaluated by EDS

Sample	Cu/at%	Ti/at%	O/at%
TiCuO-1	12.4	12.3	75.3
TiCuO-2	23.1	6.7	70.2

图1 TiO2和TiCuO样品的XRD图谱

Fig. 1 XRD patterns of TiO2 and TiCuO samples

图2 TiCuO-1样品的TEM照片(a), A和B区域Cu原子百分比(b), A(c)和B(d)区域的选区电子衍射图(c)及其HRTEM形貌(e)~(f)

Fig. 2 TEM image (a), Cu contents evaluated by EDS (b), the selected area electron diffraction results of A region (c) and B region (d), and their corresponding HRTEM images (e, f), respectively, of the TiCuO-1 sample

图3 TiCuO-2样品的TEM形貌照片(a), Cu、O、Ti元素的EDS 线扫描结果(b), A(c)和B(d)区域的选区电子衍射图及其HRTEM形貌图(e)~(f)

Fig. 3 TEM image(a), corresponding line-scanning of Cu, O, Ti elements (b) across a selected area (yellow line in (a)), the selected area electron diffraction results of A region (c) and B region (d), and their corresponding HRTEM images (e, f) respectively of the TiCuO-2 sample

图4 TiCuO-1和TiCuO-2的Cu (a)和Ti (b)XPS图谱

Fig. 4 Cu2p (a) and Ti2p (b) XPS spectra of TiCuO-1 and TiCuO-2 films

表2 TiO2、TiCuO-1、TiCuO-2 样品中的Ti2p和Cu2p拟合后的Ti4+、Ti3+、Ti2+在Ti2p图谱中所占面积的百分含量及Cu2+、Cu+在Cu2p图谱中的所占面积的百分含量

Table 2 The area percentages of Ti4 +, Ti3 + and Ti2 + in the Ti2p spectra and the area percentage of Cu2 + and Cu + in Cu2p spectra of TiO2, TiCuO-1 and TiCuO-2 samples

Sample	Ti2p			Cu2p
Sample	Ti⁴⁺	Ti²⁺	Ti³⁺	Cu²⁺	Cu⁺
TiCuO-2	47at%	53at%	0	30.2at%	69.8at%
TiCuO-1	39at%	43at%	18at%	6.2at%	93.8at%
TiO₂	32at%	34at%	34at%	-	-

表3 TiO2、TiCuO和SS的自腐蚀电位(Ecorr)及其自腐蚀电流密度(Icorr)

Table 3 Corrosion potential (Ecorr) and current densities (Icorr) of SS, TiO2 and TiCuO films derived from potentiodynamic polarization curves

Sample	SS	TiO₂	TiCuO-1	TiCuO-2
E_corr/V(vs.SCE)	-0.13	-0.06	-0.16	-0.17
I_corr/(μA·cm^-2)	0.49	0.03	0.62	3.89

图5 TiO2、TiCuO样品和SS的电化学腐蚀极化曲线(A)及TiCuO-2样品的腐蚀形貌图和a, b, c, d四个不同腐蚀区域的EDS结果(B)

Fig. 5 Electrochemical corrosion polarization curves of TiO2, TiCuO and 316 L SS samples (A) and morphology of the TiCuO-2 sample with its EDS results of the four different corroded areas labeled with a, b, c and d (B)

图6 TiCuO-1和TiCuO-2样品释放Cu离子累积浓度(a)和Cu离子平均释放速率(b)

Fig. 6 Accumulated Cu ions concentration (a) and release rate (b) of Cu ions released from TiCuO-1 and TiCuO-2 samples

图7 TiO2、TiCuO样品体外静态培养内皮细胞的细胞活性(a)和罗丹明荧光染色照片(b)

Fig. 7 ECs viability detected by CCK-8 assay (a) and fluorescence microscopic images stained by rhodamine (b) after being cultured on the samples of TiO2 and TiCuO

图8 TiO2、TiCuO样品体外静态培养内皮细胞的细胞活性(a)和罗丹明荧光染色照片(b)

Fig. 8 ECs viability detected by CCK-8 assay (a) and fluorescence microscopic images stained by rhodamine (b) after being cultured under condition medium with CuCl2 solution at different concentration

参考文献 37

[1]	WARD M R, STEWART D J, KUTRYK M J.Endothelial progenitor cell therapy for the treatment of coronary disease, acute MI, and pulmonary arterial hypertension: current perspectives.Catheterization and Cardiovascular Interventions, 2007, 70(7): 983-998.
[2]	NISIO D M, MIDDELDORP S, BÜLLER H R. Direct thrombin inhibitors.New England Journal of Medicine, 2005, 353(10): 1028-1040.
[3]	AKAHORI T, NIINOMI M.Fracture characteristics of fatigued Ti-6Al-4V ELI as an implant material.Materials Science & Engineering A, 1998, 243(1/2): 237-243.
[4]	LENG Y X, CHEN J Y, YANG P, et al. Structure and properties of passivating titanium oxide films fabricated by DC plasma oxidation. Surface & Coatings Technology, 2003, 166(2/3): 176-182.
[5]	TSYGANOV I A, MAITZ M F, RICHTER E,et al. Hemocompatibility of titanium-based coatings prepared by metal plasma immersion ion implantation and deposition. Nuclear Instruments & Methods in Physics Research, 2007, 257(1/2): 122-127.
[6]	KLESZCZEWSKI T, MODZELEWSKA B, BAL W,et al. Cu(II) complexation does not affect oxytocin action on pregnant human myometrium in vitro. Reproductive Toxicology, 2015, 59: 60-65.
[7]	AGGETT P J.An overview of the metabolism of copper.European Journal of Medical Research, 1999, 4(6): 214-216.
[8]	GAETKE L M, CHOW C K.Copper toxicity, oxidative stress, and antioxidant nutrients.Toxicology, 2003, 189(1/2): 147-163.
[9]	HU G.Copper stimulates proliferation of human endothelial cells under culture.Journal of Cellular Biochemistry, 1998, 69(3): 326-335.
[10]	REN L, XU L, FENG J,et al. In vitro study of role of trace amount of Cu release from Cu-bearing stainless steel targeting for reduction of in-stent restenosis. Journal of Materials Science Materials in Medicine, 2012, 23(5): 1235-1245.
[11]	SEN C K, KHANNA S, VENOJARVI M,et al. Copper-induced vascular endothelial growth factor expression and wound healing. Am. J. Physiol.-Heart Circ. Physiol., 2002, 282(5): H1821-H1827.
[12]	YIN R, LING L, XIANG Y,et al. Enhanced photocatalytic reduction of chromium (VI) by Cu-doped TiO2 under UV-A irradiation. Separation & Purification Technology, 2017, 190: 53-59.
[13]	ZONG M, LONG B, LIU Y,et al. Antibacterial ability and angiogenic activity of Cu-TiO2 nanotube arrays. Materials Science & Engineering C, 2017, 71: 93-99.
[14]	NORAMBUENA G A, PATEL R, KARAU M,et al. Antibacterial and bio compatible titanium-copper oxide coating may be a potential strategy to reduce periprosthetic infection: an in vitro study. Clinical Orthopaedics & Related Research, 2016, 475(3): 1-11.
[15]	WANG H, LI Y, BA X,et al. TiO2 thin films with rutile phase prepared by DC magnetron co-sputtering at room temperature: effect of Cu incorporation. Applied Surface Science, 2015, 345: 49-56.
[16]	TOMOLYA K, JANOVSZKY D, SYCHEVA A, et al. Peculiarities of ball-milling induced crystalline-amorphous transformation in Cu-Zr-Al-Ni-Ti alloys. Intermetallic, 2015, 65: 117-121.
[17]	NABIAŁEK M. Soft magnetic and microstructural investigation in Fe-based amorphous alloy.Journal of Alloys & Compounds, 2015, 642: 98-103.
[18]	OLEKSAK RP, DEVARAJ A, HERMAN GS.Atomic-scale structural evolution of Ta-Ni-Si amorphous metal thin films.Materials Letters, 2016, 164: 9-14.
[19]	TSENG I H, WU J C S, CHOU H Y. Effects of Sol-Gel procedures on the photocatalysis of Cu/TiO2 in CO2 photoreduction. Journal of Catalysis, 2004, 221(2): 432-440.
[20]	SUN C, ZHU J, LV Y,et al. Dispersion, reduction and catalytic performance of CuO supported on ZrO2-doped TiO2 for NO removal by CO. Applied Catalysis B: Environmental, 2011, 103(1): 206-220.
[21]	LIU J, LI X, ZHAO Q,et al. The selective catalytic reduction of NO with propene over Cu-supported Ti-Ce mixed oxide catalysts: promotional effect of ceria. Journal of Molecular Catalysis A: Chemical, 2013, 378: 115-123.
[22]	LONG R, ENGLISH N J.Electronic properties of anatase-TiO2, co-doped by cation-pairs from hybrid density functional theory calculations.Chemical Physics Letters, 2011, 513(4): 218-223.
[23]	NAKAYAMA S, KIMURA A, SHIBATA M,et al. Voltammetric characterization of oxide films formed on copper in air. Journal of the Electrochemical Society, 2001, 148(11): B467-B472.
[24]	NAKAYAMA S, NOTOYA T, OSAKAI T.Highly selective determination of copper corrosion products by voltammetric reduction in a strongly alkaline electrolyte.Analytical Sciences the International Journal of the Japan Society for Analytical Chemistry, 2012, 28(4): 323.
[25]	BELOUSOV V V.Mechanisms of accelerated oxidation of copper in the presence of molten oxides.Oxidation of Metals, 2007, 67(5/6): 235-250.
[26]	AL-MAYOUF A M, AL-SWAYIH A A, AL-MOBARAK N A,et al. Corrosion behavior of a new titanium alloy for dental implant applications in fluoride media. Materials Chemistry and Physics, 2004, 86(2/3): 320-329.
[27]	LIU H, LENG Y, HUANG N.Corrosion resistance of Ti-O film modified 316L stainless steel coronary stentsin vitro. Journal of Materials Engineering & Performance, 2012, 21(3): 424-428.
[28]	LIU H, ZHANG D, FENG S,et al. Corrosion and ion release behavior of Cu/Ti film prepared via physical vapor deposition in vitro as potential biomaterials for cardiovascular devices. Applied Surface Science, 2012, 258(19): 7286-7291.
[29]	JACEK BANAS, ANDRZEJ MAZURKIEWICZ.The effect of copper on passivity and corrosion behaviour of ferritic and ferritic- austenitic stainless steels.Materials Science and Engineering: A, 2000, 277(1/2): 183-191.
[30]	STRANAK V, WULFF H, REBL H,et al. Deposition of thin titanium copper films with antimicrobial effect by advanced magnetron sputtering methods. Materials Science & Engineering C, 2011, 31(7): 1512-1519.
[31]	SEO Y, CHO Y S, HUH Y D,et al. Copper ion from Cu2O crystal induces AMPK-mediated autophagy via superoxide in endothelial cells. Molecules & Cells, 2016, 39(3): 195-203.
[32]	LIU H, PAN C, ZHOU S,et al. Improving hemocompatibility and accelerating endothelialization of vascular stents by a copper-titanium film. Materials Science & Engineering C Materials for Biological Applications, 2016, 69: 1175-1182.
[33]	MIDANDER K, CRONHOLM P, KARLSSON H L,et al. Surface characteristics, copper release, and toxicity of nano-and micrometer- sized copper and copper(II) oxide particles: a cross-disciplinary study. Small, 2009, 5(3): 389-399.
[34]	ARIMA Y, IWATA H.Effect of wettability and surface functional groups on protein adsorption and cell adhesion using well-defined mixed self-assembled monolayers.Biomaterials, 2007, 28(20): 3074-3082.
[35]	ESHAGHI A, ESHAGHI A.Preparation and hydrophilicity of TiO2 Sol-Gel derived nanocomposite films modified with copper loaded TiO2 nanoparticles.Materials Research Bulletin, 2011, 46(12): 2342-2345.
[36]	HUANG L, NING C Q, DING D Y,et al. Wettability and in vitro bioactivity of doped TiO2 nanotubes. Journal of Inorganic Materials, 2010, 25(7): 775-779.
[37]	BRAMMER K S, OH S, GALLAGHER J O,et al. Enhanced cellular mobility guided by TiO2 nanotube surfaces. Nano Letters, 2008, 8(3): 786-793.