Materials Letters 268 (2020) 127631

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Materials Letters
journal homepage: www.elsevier.com/locate/mlblue

The influence of substrate electropolishing on anodization behavior,

corrosion resistance, cytocompatibility and antibacterial ability of NiTi
alloy
Yonghua Sun, Youjie Rong, Ya Zhao, Xiaohong Yao ⇑, Ruiqiang Hang ⇑
Laboratory of Biomaterial Surfaces & Interfaces, Institute of New Carbon Materials, Taiyuan University of Technology, Taiyuan 030024, China

a r t i c l e i n f o a b s t r a c t

Article history: Ni-Ti-O nanotubes (NTs) anodically grown on nearly equiatomic NiTi alloy are promising in biomedical
Received 21 December 2019 field, but mechanical polishing pretreatment of the substrate usually results in severe pitting. The present
Received in revised form 26 February 2020 work proposes to use electropolishing (EP) as pretreatment of anodization of the NiTi alloy to eliminate
Accepted 9 March 2020
the side effect of mechanical polishing. Scanning electron microscopy was used to observe the morphol-
Available online 9 March 2020
ogy of the samples. Potentiodynamic polarization test was carried out to evaluate the corrosion behavior.
Biological experiments were performed to assay their cytocomaptibility and antibacterial ability. Our
Keywords:
results show pitting can be completely avoided after substrate EP pretreatment, yielding a macro smooth
Biomaterials
Surface
surface, while micro ordered nanotubular structure is well preserved. EP pretreatment enhances the cor-
NiTi alloy rosion resistance and antibacterial ability of the NiTi alloy when compared with that of mechanically pol-
Electropolishing ished sample, and all the samples show no appreciable cytotoxicity, clearly indicating EP is a promising
Anodization pretreatment for fabricating Ni-Ti-O NTs on the NiTi alloy for biomedical applications.
Nanotubes Ó 2020 Elsevier B.V. All rights reserved.

1. Introduction It is well known that electropolishing (EP) can effectively

Nearly equiatomic NiTi alloy is preferred as orthopedic implant
material due to its unique shape memory, super–elastic property,
and other desirable properties [1,2], but in many conditions its bio-
functionality cannot satisfy clinical requirements [3,4]. Constitut-
ing Ni-Ti-O NTs via anodization has been proven to be effective
in enhancing its biofunctionality [5]. However, our previous study
showed serious substrate pitting was usually occurred during
anodization [6], which may be ascribed to stress layer generated
during mechanical polishing (MP) of the substrate. The layer con-
tains a lot of microstructural defects and these surface defects
may act as electrochemical active sites, at which oxide film is vul-
nerable to attack by fluoride (F) ions in the electrolyte, eventually
resulting in pitting formation. These macro pits on the NiTi alloy
may serve as active sites of electrochemical corrosion in vivo to
accelerate substrate corrosion. On one hand, high corrosion rate
may lead to continuous release of Ni ions at a relatively high level,
causing healthy concerns. On the other hand, accelerated corrosion
may degrade substrate mechanical properties, eventually resulting
in implant failure.
⇑ Corresponding authors.
E-mail addresses: xhyao@tyut.edu.cn (X. Yao), hangruiqiang@tyut.edu.cn
(R. Hang).
remove surface stress layer generated by MP [7]. Additionally,
external magnetic field applied during EP could further improve
surface properties [8]. We hypothesize that substrate EP pretreat-
ment may be an ideal approach to eliminate the surface stress
layer and therefore pitting corrosion, eventually yielding a defect
free surface at macro level and ordered nanostructure at micro
level. The influence of the EP on the corrosion behavior, cytocom-
patibility and antibacterial ability of the NT coated NiTi alloy was
also evaluated.
2. Materials and methods
2.1. Sample preparation and characterization
Mechanically polished NiTi alloy sheets (50.8 at.% Ni, U10 mm,
denoted as NiTi-MP) were subjected to EP in solution composed of
CH3OH (99.98 vol%) and HClO4 (72 vol%) with volume ratio of 4:1
at a constant voltage of 13 V for 5 min at 0 °C under magnetic stir-
ring. NiTi-MP and electropolished samples (denoted as NiTi-EP)
were used for anodization. NiTi-MP and NiTi-EP were anodized
in an ethylene glycol electrolyte containing 0.2 wt% NH4F and 0.5
or 1.0 vol% H2O at a fixed voltage of 25 V for 1 h and the resulting
samples were denoted as MP-NT-0.5, EP-NT-0.5, MP-NT-1.0 and

https://doi.org/10.1016/j.matlet.2020.127631
0167-577X/Ó 2020 Elsevier B.V. All rights reserved.
2 Y. Sun et al. / Materials Letters 268 (2020) 127631

EP-NT-1.0, respectively. Surface and cross-sectional morphologies 2.5. Statistical analysis

of the samples were observed using a field-emission scanning elec-
tron microscopy (FE-SEM, JSM-7001F, JEOL). Each quantitative experiment was repeated at least three times
and the data were presented as mean ± standard deviation. Statis-
tical analysis was done with one-way analysis of variance followed
2.2. Corrosion test by the Student–Newman–Keuls test. When p < 0.01, the difference
was considered to be highly significant.
Corrosion behavior of the samples was investigated by poten-
tiodynamic polarization test on an electrochemical workstation
3. Results
(CHI660E, CHI Instruments, Inc., Shanghai) in phosphate buffered
Saline (PBS, pH = 7.4) at 37 °C. After immersing in PBS for 1 h,
Fig. 1 shows the surface and cross-sectional SEM images of all
the tests were performed at the potential range of 0.8~1.0 V ver-
the samples. Low and high magnification images show the surfaces
sus saturated calomel electrode (SCE) at a constant scanning rate of
NiTi-MP and NiTi-EP are smooth. Only MP pretreatment results in
1 mV/s. The corrosion potential (Ecorr), corrosion current density
the occurrence of macro corrosion pits (MP-NT-0.5 and MP-NT-1.0),
(Icorr) and cathodic Tafel slope (bc) were analyzed by Tafel extrapo-
while defect free surface at macro level is obtained after EP pre-
lation method.
treatment. All the sample surfaces are covered with ordered Ni-
Ti-O NTs with diameter of about 60 nm, irrespective of substrate
2.3. Cytocompatibility evaluation pretreatment. Cross-sectional images reveal that EP effectively
increases the NT length when compared with that of MP.
Cytocompatibility of the samples was assessed by cell live/dead Polarization curves and the electrochemical parameters of
staining. Specifically, endothelial cells (ECs, EA. hy926) were the samples are shown in Fig. 2 and Table 1, respectively.
seeded on the sample surfaces with a density of 2  104 cells/cm2
and then cultured for 1, 3 and 5 days in Dulbecco’s modified eagle
medium (DMEM, Gibco) containing 10 vol% fetal bovine serum
(Sijiqing, China) and 1% penicillin and streptomycin. At each time
point, the samples were rinsed with PBS and then 50 lL of the
work solution (Invitrogen) was added onto sample surface and
incubated in darkness for 40 min at 37 °C. Finally, the cells were
observed by confocal laser scanning microscope (C2 Plus, Nikon).

2.4. Antibacterial assay

Antibacterial effects of the samples were evaluated by plate-

counting method using Staphylococcus aureus (S. aureus). 50 lL of
bacterial suspension with a concentration of 1.0  105 CFU/mL
was dripped on sample surface and incubated at 37 °C for 12 h.
Then the bacteria were quantified by standard serial dilution with
PBS and spread plate method. The antibacterial rate was evaluated
by the following formula: R = (B A)/B  100%. Where R is the
antibacterial rate, B and A are the mean numbers of bacteria
(CFU) on blank control and samples, respectively. Fig. 2. Potentiodynamic polarization curves of the samples.

Fig. 1. Surface and cross-sectional SEM images of the samples.

 Y. Sun et al. / Materials Letters 268 (2020) 127631 3

Table 1 Antibacterial ability of the samples against S. aureus is pre-

Corrosion potentials (Ecorr), current densities (Icorr), and cathodic Tafel slopes (bc) of sented in Fig. 4. The colony number corresponding to tested sam-
the samples.
ples is less than blank control, especially EP-NT-0.5, MP-NT-1.0,
2 1
Sample Ecorr/V vs. SCE Icorr/A cm bc/V decade and EP-NT-1.0 (Fig. 4A). Quantitative results in Fig. 4B well corrob-
NiTi-MP 0.20 4.0  10 7
0.19 orate the qualitative results.
8
NiTi-EP 0.28 1.2  10 0.15
8
MP-NT-0.5 0.61 6.3  10 0.12
EP-NT-0.5 0.36 2.1  10 8
0.13 4. Discussion
8
MP-NT-1.0 0.32 7.9  10 0.14
8
EP-NT-1.0 0.38 3.9  10 0.16 Although Ni-Ti-O NTs are promising as biomedical coatings of
the NiTi alloy, substrate MP before anodization usually results in
pitting, causing safety concerns especially for long-term implanta-
Icorr decreases by an order of magnitude after EP and anodization. It tions. Our results demonstrate that EP is an ideal alternative of MP
is also found that EP-NT-0.5 and EP-NT-1.0 are characterized by the to overcome the drawback. This may be ascribed to the surface
lower corrosion current density compared with MP-NT-0.5 and stress layer generated by MP contains a lot of active sites that
MP-NT-1.0 respectively. are sensitive to the attack of F ions during anodization, while EP
Live/dead staining results are shown in Fig. 3. No dead cells (in can effectively remove the stress layer and the active sites [9],
red) can be observed on all the sample surfaces. The number of liv- yielding a uniform surface both at macro and micro levels. In addi-
ing cells (in green) on all the sample increases with time. No obvi- tion, elimination of the pitting also extends the NT length through
ous difference in cell number can be observed between each group avoiding short–circuit current of anode dissolution, which may be
at the same culturing time. a plausible explanation of the improved corrosion resistance. As

Fig. 3. Fluorescence images of live/dead staining of ECs after culturing for 1, 3, and 5 days on the sample surfaces.

Fig. 4. (A) Optical images of bacterial plates. (B) Antibacterial rate of the samples against S. aureus. **p < 0.01.
4 Y. Sun et al. / Materials Letters 268 (2020) 127631
evidenced by our previous work [10], long NTs with high specific
surface area may also contribute to Ni2+ release to combat bacterial
infection and therefore increase the success rate of the implant.
Although excessive Ni2+ release may result in cytotoxicity
[11,12], our live/dead staining results indicate that the amount of
Ni2+ released can be well tolerated by ECs. Collectively, although
further work is required to verify its clinical applicability, the
present study indeed demonstrates that EP is a promising
pretreatment of anodization of the NiTi alloy for biomedical
applications.
5. Conclusion
Compared with mechanical polishing pretreatment, electropol-
ishing can effectively inhibit pitting corrosion of the NiTi alloy dur-
ing anodization, which may be ascribed to its effectiveness in
removing surface stress layer of mechanically polished sample.
Therefore, a uniform Ni-Ti-O NT coating is prepared on the surface
of the NiTi alloy. In comparison with the mechanically polished
samples, the NTs are longer after electropolishing, which enhances
the corrosion resistance and antibacterial ability and well pre-
serves favorable cytocompatibility.
CRediT authorship contribution statement
Yonghua Sun: Methodology, Investigation, Formal analysis,
Writing - original draft. Youjie Rong: Resources, Investigation.
Ya Zhao: Resources, Validation. Xiaohong Yao: Supervision, Fund-
ing acquisition. Ruiqiang Hang: Supervision, Writing - review &
editing.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
Acknowledgments
This work was jointly supported by the Fund for Shanxi ‘‘1331
Project” Key Innovative Research Team (PY201809) and the
Natural Science Foundation of Shanxi Province (201801D121093).
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