Applied Surface Science 396 (2017) 249–258

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Applied Surface Science

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Full length article

Enhanced corrosion resistance and biocompatibility of AZ31 Mg alloy

using PCL/ZnO NPs via electrospinning
Jinwoo Kim a , Hamouda M. Mousa a,b , Chan Hee Park a,c,∗ , Cheol Sang Kim a,c,∗
a
Department of Bionanosystem Engineering, Graduate School, Chonbuk National University, Jeonju 561-756, Republic of Korea
b
Department of Engineering Materials and Mechanical Design, Faculty of Engineering, South Valley University, Qena 83523, Egypt
c
Division of Mechanical Design Engineering, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea

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

Article history: In the efforts to improve corrosion resistance and biocompatibility of magnesium alloys, polycarpro-
Received 21 June 2016 lactone (PCL) and zinc oxide nanoparticles (ZnO NPs) composite coatings were applied onto AZ31 Mg
Received in revised form 1 October 2016 alloys via electrospinning technique in this study. The PCL/ZnO composite coatings on Mg alloys were
Accepted 17 October 2016
characterized by using FE-SEM, EDX, XPS, and FT-IR. Moreover, coating adhesion test, electrochemical
Available online 18 October 2016
corrosion test, and biocompatibility test in vitro were performed to measure coating performance. Our
results revealed that the increase in the content of ZnO NPs in the composite coatings not only improved
Keywords:
the coating adhesion of composite coatings on Mg alloys, but also increased the corrosion resistance. Fur-
Magnesium alloy
ZnO nanoparticles (NPs)
thermore, the biocompatibility of MC3T3-E1 osteoblasts of the PCL/ZnO composite coated samples was
Polycaprolactone (PCL) superior to the biocompatibility of the bare samples. Such data suggest that applying PCL/ZnO composite
Electrospinning coating to the magnesium alloys has suitable potential in biomedical applications.
Corrosion © 2016 Published by Elsevier B.V.
Biocompatibility

1. Introduction
In recent years, magnesium (Mg) and its alloys as biodegrad-
able metallic implants have attracted a great amount of attention
for biomedical applications [1–3]. Specifically, they exhibit unique
advantages such as biodegradability, bioabsorbability, and ade-
quate mechanical/physical properties [4,5]. Case in point, it does
not require a post-surgery of removing the implant materials after
a full recovery of tissue regeneration, because it degrades in the
human body. Not only that, Mg is essential in human body for phys-
iological functions with daily consumptions of 250–500 mg/day [6].
However, on the other hand, there are main disadvantages of
the Mg implant, considering its major limitations when exposed
to the physiological media such as rapid corrosion and hydrogen
gas (H2 ) gas release [2,6,7]. These parameters induce an alkalin-
ization environment in the surrounding area of injured tissues as
well as a decrease in mechanical properties of the Mg implant.
Hence, in order to control such limitations, surface treatment is an
unavoidable process in employing Mg and its alloys as a biomedical
device, which ultimately leads to a decrease in the degradation rate
∗ Corresponding authors at: Department of Bionanosystem Engineering, Graduate
School, Chonbuk National University, Jeonju 561-756, Republic of Korea.
E-mail addresses: biochan@jbnu.ac.kr (C.H. Park), chskim@jbnu.ac.kr (C.S. Kim).
span time and maintains their mechanical properties with healing
tissues recovery [8].
To improve corrosion resistance and biocompatibility of Mg-
based biomaterials, many researchers have applied coating layers
onto Mg-based substrates using various techniques and methods
including organic/inorganic, electrodeposition, sol-gel, dipping,
and spraying and others [8–13]. Electrospinning is a versatile tech-
nique that produces micro/nanoscaled polymer fibers at a uniform
scale. The produced electrospinning nanofibers have large surface
areas to volume ratio, high porosity, and mimicked extracellular
matrix (ECM) of the cell [14]. Organic coating is one of the widely
used methods of surface modification, in which the biocompati-
ble polymer with proper structure covers the implant interface [8].
Polycarprolactone (PCL) polymer is commonly used as biomaterials
in biomedical applications such as tissue engineering scaffolds and
wound dressing because of its biocompatibility and biodegradabil-
ity [15,16].
Nanoparticles with specific properties can be added in order to
fabricate multifunctional coating layers in a nano-scaled size and
enhancing polymeric coating properties. Zinc oxide nanoparticles
(ZnO NPs) have various advantages such as antibacterial properties,
nontoxicity, and anticancer behavior [17–19]. However, controlling
the amount of ZnO NPs in polymer composite is essential for the
use in biomedical applications due to its influence on cell behavior
caused by the oxygen reactive species (ROS) [20].

http://dx.doi.org/10.1016/j.apsusc.2016.10.092
0169-4332/© 2016 Published by Elsevier B.V.
250 J. Kim et al. / Applied Surface Science 396 (2017) 249–258
Table 1
Chemical composition of AZ31 magnesium alloy (in wt.%) used in the study.
Element Al Zn Mn Si Cu
wt.% 2.5–3.5 0.7–1.3 0.2–1.0 0.05 0.01
In this study, PCL/ZnO composite coating layers were fabricated
on AZ31 Mg alloys via electrospinning with the expectation that
ZnO NPs would contribute to not only increasing coating perfor-
mance but also inducing biological activity. The goal of the study
is to explore the influence of electrospun composite nanofiber
coatings on Mg alloys (AZ31) using PCL and ZnO NPs. Different
weight percentages of ZnO NPs were incorporated with PCL poly-
mer matrix at 1 and 3 wt%. Various characterization techniques and
measurements including FE-SEM, EDX, XPS, FT-IR, coating adhe-
sion test, polarization, and EIS were carried out to investigate the
influence of PCL/ZnO composite coatings on AZ31 Mg alloys. The
biocompatibility of MC3T3-E1 osteoblasts of the coated samples
via electrospinning technique was also investigated.
2. Materials and methods
2.1. Materials
A commercial (3% Al, 1% Zn and 96% Mg) alloy (Alfa Aesar Com-
pany, South Korea) with samples size (12 mm × 12 mm × 6.35 mm)
was used, as shown in Table 1. The substrates were polished with
SiC paper up to 2000 grit. In this study, poly(ε-caprolactone) (PCL,
Mw = 70,000–90,000) and zinc oxide nanoparticles (ZnO, parti-
cle size avg. = 60 nm) were purchased from Sigma-Aldrich, USA.
Dimethylformamide (DMF) and Tetrahydrofuran (THF) were pur-
chased from Sam Chun, Korea.
2.2. Electrospinning setup and procedure
PCL solutions (10 wt%) were prepared by dissolving PCL in a sol-
vent of DMF/THF (1:1 w/w). For PCL/ZnO composite solutions, 1
and 3 wt% of zinc oxide nanoparticles from the polymer weight
were added to a PCL solution and then vigorously stirred for
12 h at room temperature. As-prepared solutions (PCL, PCL/ZnO
1 wt.%, and PCL/ZnO 3 wt%) for electrospinning coating were loaded
in 10 mL of syringes, and were connected to a metal capillary
(di = 0.51 mm, 21 G) through a plastic tube. The flow rate was set up
at 1 mL/h, and electrospinning was carried out at room conditions
(temperature = 18 ◦ C and humidity = 30%), 16 kV applied voltage
and 15 cm tip-to-collector distance. In this study, electrospinning
coating process of all samples was equally conducted for 5 min.
2.3. Characterization
Field emission scanning electron microscopy (FE-SEM, Carl
Zeiss, Supra 40VP, and Japan) was used to examine the surface
morphology of different coated Mg samples. For observing FE-
SEM images, the samples were sputter coated under argon, and
the excitation voltage was set at 2 kV. The elemental compositions
of the surface were analyzed using Energy Dispersive X-ray Spec-
troscopy (EDX), which is equipped with FE-SEM. ImageJ software
(NIH, USA) was employed in order to measure the fiber diame-
ter and analyze fiber diameter distribution. For each electrospun
coating layer, at least 120 fibers were obtained from three differ-
ent FESEM images in order to calculate the average fiber diameter
and fiber diameter distribution. Surface chemistry of the different
coatings was studied by using X-ray photoelectron spectroscopy
(XPS, AXIS-NOVA, Kratos, Inc.) with an Al K␣ irradiation source. To
determine a change in the chemical structure of the coated samples,
Fourier-transform infrared spectroscopy (FT-IR, ABB Brown MB100
spectrometer, Canada) was employed. The wettability test was con-
ducted by using a contact angle meter (GBX, Digidrop, France) to
Mg examine the surface alteration due to the composite coating on Mg
Balance alloy substrate. DI water was used in this experiment and carefully
dropped on the coated substrates. At least 3 measurements were
performed for each sample, and the average values are present in
the study.
2.4. Coating adhesion measurements
The adhesion performance between the coatings and the Mg
substrates was evaluated according to the ASTMF1044-05 stan-
dard. The coated (top) and uncoated (bottom) surfaces were fixed
with a mixture of epoxy resin and polyamide resin (1:1), and then
the prepared samples were dried at 100 ◦ C for 4 h. Coating adhesion
strength was measured by using universal tensile testing machine
(RB 301 UNITECH-M, Korea) at a test speed of 5 mm/min [21,22].
In this study, three samples for each condition were conducted to
calculate the average values for the coating adhesion properties.
2.5. Electrochemical corrosion test: polarization and EIS
Corrosion test was conducted through potentiodynamic polar-
ization technique and electrochemical impedance spectroscopy
(EIS), and the details of the test were performed according to
our previous reports [23]. Simply, the experiments were con-
ducted with three electrode systems: Mg substrate as working
electrode, platinum as the counter electrode, and reference elec-
trode as Ag/AgCl saturated in KCl solution. Polarization curves
were obtained in simulated body fluid (SBF) at the temperature of
37 ◦ C ± 0.5 ◦ C in the potential range between −2 V and 1.5 V with
scan rate 5 mV/s. Before performing the test, the samples were
immersed in the SBF solution for 1 h in order to stabilize the sam-
ples in the electrochemical cell. The specimen area exposed to the
SBF solution was 0.875 cm2 , and potential values displayed in the
study are compared to those of the counter electrode. For electro-
chemical impedance spectroscopy, the measurement was carried
out in SBF solution at 37 ± ◦ 5C with a three electrode system: Mg
substrates, platinum, and Ag/AgCl saturated in KCl as the working,
reference, and counter electrode, respectively. The corrosion test
was evaluated in the frequency range from 105 Hz to 10−1 Hz, and
the amplitude of sinusoidal potential was set at 10 mV. Before test-
ing, all samples were exposed to the solution for a while with the
intention of stabilizing the open circuit potential (OCP).
2.6. In vitro cell biocompatibility
Cell attachment on the surface of coated/uncoated Mg alloys and
proliferation were investigated according to our previous report
[24]. Pre-osteoblast cells (MC3T3-E1) were grown on a 12-well
plate containing different coated samples for 1 and 3 days at 37 ◦ C
and atmosphere conditions of 5% CO2 . The MC3T3-E1 cells were
seeded at a density of 3 × 104 cells per well and culture medium was
changed daily. Samples were immersed in culture media (alpha-
mem supplemented with 10% FBS and 1% penicillin-streptomycin).
Cell morphology and attachment on the surface of the samples were
observed through SEM. The cell viability was monitored on the first
and third days of culture using Dojindo’s cell counting kit-8 (CCK)
assay. The absorbance of CCK-8 solution was evaluated at 450 nm
using iMarkTM microplate reader. Cell-cultured Mg samples (1 and
3 days) were rinsed with PBS (phosphate buffer saline, pH 7.4) and
set in 2.5% glutaraldehyde for 1 h to observe cell proliferation. The
coated Mg samples were washed with 25%, 50%, 75%, and 100%
ethanol and dried in air overnight.
 J. Kim et al. / Applied Surface Science 396 (2017) 249–258 251

Fig. 1. Schematic illustration of applying PCL/ZnO composite coating onto AZ31 Mg substrates by the electrospinning method.

Fig. 2. FE-SEM images of electrospun coating surfaces on Mg alloys; (a) PCL coated surface, (b) PCL coating layers containing 1 wt.% and (c) 3 wt.%. (d), (e) and (f) shows EDX
(energy-dispersive X-ray spectrum) characterization. The fiber size distributions and average fiber diameters are shown in (g)–(i).

3. Results and discussion
3.1. Surface characterization
Some major concerns in using Mg and its alloys as biomedical
devices are their poor corrosion resistance and low biocompat-
ibility. In the study, to overcome these limitations of Mg-based
biomaterials, we conducted PCL/ZnO NPs composite coatings on Mg
substrates by electrospinning as shown in Fig. 1 and investigated
the effects of the composite coatings on their adhesion strength,
corrosion performance, and biocompatibility.
There is a clear indication of how the incorporation of inorganic
materials with organic nanofibers has influences in the structure
and morphology, as the conductivity, viscosity, and surface tension
of electrospinning solution was affected [25,26]. Based on prelim-
inary tests, zinc oxide nanoparticle incorporated PCL solutions of
greater than 3 wt% led to an increase in the solution viscosity, which
resulted in the production of numerous beads on the coating lay-
ers. Thus, according to the result of the preliminary experiments,
we concluded that concentrations of up to 3 wt% is optimal for elec-
trospun composite coatings because of good electrospinnability as
well as appropriate dispersion of ZnO NPs in the solution.
Fig. 2(a–c) presents that there were morphological changes on
the surfaces of Mg alloy due to the increasing the content of ZnO
NPs. All electrospun coated samples were uniformly covered by
nanofibers and highly porous. For PCL coated Mg samples, fibers on
the surface were very smooth and plain. However, individual fibers
became rougher as zinc oxide NPs (1 and 3 wt%) was incorporated
in the fibers. FE-SEM images indicate that the ZnO nanoparticles
with the composite coating impacted the nanofiber morphology
and structure, which might be attributed to the change in the solu-
tion viscosity and the agglomerates of ZnO NPs [27]. In addition,
252 J. Kim et al. / Applied Surface Science 396 (2017) 249–258
Fig. 3. Elements mapping of the coated Mg samples: Bare, PCL, PCL/ZnO 1 wt%, and PCL/ZnO 3 wt%.
it is clear from the images that electrospinning can produce nano-
structural and porous surfaces on Mg alloys.
To clarify the effect of ZnO nanoparticles on morphological
changes, EDX analysis was also employed as seen in Fig. 2(d–f)
and Table 1. The results show varying atomic percentage of PCL,
PCL-ZnO 1 wt%, and PCL-ZnO 3 wt% composite coatings on Mg sub-
strates. The EDX spectrum of PCL/ZnO NPs (1 and 3 wt%) composite
coated samples indicated the presence of the ZnO nanoparticles,
confirming the distribution of ZnO NPs in the fibers. Different chem-
ical composition (in at.%) of the electrospun coated samples also
indicates that the amount of ZnO NPs could affect porosity, pore
sizes, and roughness of nanofibrous surfaces [28]. The EDX element
mapping is present in Fig. 3, which indicates the distribution of
elements on the samples. The Zn and O elements increased as the
ZnO NP content in the PCL increased. However, the Mg elements
became lower as the ZnO concentration in the PCL fibers increased.
This result is consistent with other researches that reveal how ZnO
nanoparticles with various concentrations significantly change the
morphological changes such as fiber diameters, fiber distributions
and pore constriction [28–30].
The average fiber diameters and fiber diameter distribution of
the PCL and PCL/ZnO electrospun layers were obtained by analyz-
ing at least 120 fibers using ImageJ software as shown in Fig. 2(g–i).
Many studies have demonstrated that fiber morphology, fiber
diameters, and size distributions can be varied due to the changes in
viscosity and conductivity of electrospinning solutions [24,26,30].
Though the morphological features were, in the present study,
noticeably different at each condition, the average fiber diame-
ters showed the similar values such as 373 ± 160, 357 ± 181, and
374 ± 144 nm for the PCL layers with 0, 1, and 3 wt%, respectively.
However, the distribution of fiber diameters varied from 100 nm
to about 1 ␮m for all coated samples. The diameter distribution
clearly presented that PCL/ZnO 3 wt% layers produce more uniform
fibers than those of PCL and PCL/ZnO 1 wt%. These observations sug-
gest that the addition of ZnO NPs into the PCL nanofibers can affect
morphological properties, especially in fiber size distributions.
XPS analysis was used to examine the elemental compositions
and surface chemistry in the study. Fig. 4 presents the survey XPS
spectra of the bare and coated Mg samples. The result shows that
only carbon and oxygen atoms were present in PCL coated surfaces
of Mg alloys, indicating the successful formation of the polymeric
coating layers on Mg alloys through electrospinning process. Com-
pared to the coated samples, the bare samples shows the typical
peaks of AZ31 Mg alloys including magnesium, zinc, oxygen, car-
bon and other elements. XPS High resolution spectra for C 1s, O 1s,
Mg 2p, and Zn 2p3/2 peaks were exhibited in Fig. 5. From the C 1s
spectra, all the coated surfaces of Mg alloys showed not only the
similar curves but also the same peaks at 284.8 and 288.8 eV cor-
responding to C C and O C O components. This explains that PCL
polymeric layers were dominant in the coated samples. In the bare
 J. Kim et al. / Applied Surface Science 396 (2017) 249–258 253

Fig. 4. XPS survey spectra of PCL and PCL/ZnO composite coating layer (a), and the XPS survey including the bare AZ31 (b).

Fig. 5. High resolution XPS spectra of the bare and coated samples: (a) C1s, (b) O1s, (c) Mg2p, and (d) Zn2p3/2 . Bare samples are an AZ31 Mg alloy consisting of Mg, Al, Zn, etc.

Mg sample, clear O 1s peaks was detected at 529.8 eV that is related
to Mg oxides while the coated samples revealed that O1s peaks shift
to higher binding energies (531–533 eV) pointing at a contribution
from organic components. The intensities of O 1s peaks decreased
for the coated samples compared to the bare samples, which can
be attributed to the polymeric surface. Zn 2p3/2 peak at 1020.08 eV
was observed from the both PCL/ZnO 1 and 3 wt% composite coated
samples, which may be due to the presence of ZnO NPs in the PCL
coatings. Surface chemical compositions in Table 3 also demon-
strate that the zinc content increases as the amount of ZnO NPs in
the composite coated surfaces increases, suggesting that zinc oxide
nanoparticles were uniformly distributed in the PCL nanofibers.
FT-IR spectrum was obtained in the wavenumber range of
400–4000 cm−1 in order to determine a change in the chemical
structure from an electrospinning coating process. Fig. 6 reveals
the FT-IR spectra of PCL, PCL/ZnO 1 wt% and PCL/ZnO 3 wt% com-
posite coatings on the surface of Mg alloys. Overall, distinctive PCL
peaks dominate the FTIR spectrum of PCL/ZnO composite, and PCL
only coated Mg alloys especially in the range of 700–3100 cm−1
[31]. An intense peak is shown at 1723 cm−1 in the PCL coating,
which corresponded to the −CO stretching in the ester carbonyl
group from the PCL polymer. The C H of saturated carbons was
observed at 2867 and 2944 cm−1 [30]. The bands at 400–700 cm−1
that are attributed to the stretching of the Zn-O bond are present
in the PCL/ZnO 1 and 3 wt% coating, respectively [20].
Even though PCL polymeric layers have many advantages,
such as containing biodegradability, biocompatibility, and suitable
mechanical strength, it is well known that hydrophobicity of PCL
is one of the major obstacles that affect the low wettability and
inadequate biocompatibility for it to be used as a coating layer
254 J. Kim et al. / Applied Surface Science 396 (2017) 249–258

Fig. 6. (a) FT-IR spectra of PCL, PCL/ZnO 1 wt.%, and PCL/ZnO 3 wt.% coated Mg samples. (b) Increase in the intensity of the Zn-O bond as the ZnO content in the composite
coating increases.

Table 2
EDX analysis of surface concentrations of C, O, Zn, and Mg (in wt.%) on the coated
Mg samples.

Elements PCL PCL/ZnO 1 wt.% PCL/ZnO 3 wt.%

C 67.48 76.36 73.28

O 12.65 14.92 20.18
Zn 0.12 0.72 5.13
Mg 19.74 8.00 1.41

Table 3
Surface concentration of C, O, Zn, and Mg (in at.%) on the bare and coated samples
using XPS analysis.

Sample C O Zn Mg

Bare 13.74 51.66 0.74 30.42

PCL 77.1 22.5 0.06 0.21
PCL/ZnO 1 wt% 77 22.5 0.32 0.17
PCL/ZnO 3 wt% 76 23.5 0.53 0.13

Table 4
Fig. 7. The result of water contact angle measurement of the different coating sur-
Result of differently coated samples of potentiodynamic polarization in SBF solution
faces: Bare, PCL, PCL/ZnO 1 wt.%, and PCL/ZnO 3 wt.% (n = 3).
at 37 ◦ C.

Sample Potential (V) Current density (␮A/cm2 )

in biomedical applications [32,33]. In the study, the effect of PCL
coating layers incorporated with ZnO NPs on surface properties Bare −1.394 39.8
PCL −1.361 3.47
was investigated by using a contact angle (CA) measurement. Fig. 7
PCL/ZnO 1 wt% −1.259 2.34
shows that PCL coating layers exhibited a CA of 137.2◦ , which corre- PCL/ZnO 3 wt% −1.159 2.11
sponds to the measurement of other studies. When 1 wt% and 3 wt%
ZnO NPs were incorporated, the CA values gradually decreased,
which means they approached more hydrophilic surfaces. From the results demonstrate that the PCL/ZnO 3 wt% composite layer had
results of the CA measurement, it is expected that the modified sur- higher tensile strength and strain at failure compared to those of
face by incorporating of ZnO NPs will influence not only biological PCL and PCL/ZnO 1 wt%. This was due to the fact that good interfacial
activity but also coating adhesion. interactions were formed between the PCL/ZnO 3 wt% coating and
Mg alloys, which might be attributed to bonding force between the
3.2. Coating adhesion performance coating layer and Mg substrate as shown in Fig. 7(b) [35]. The other
reason might be the denser and compact layer formed by PCL/ZnO
Coating adhesion performance between metal substrates and 3 wt% coating due to the reduced the Mg element as you can see in
organic coatings is significant, because coating failure directly Fig. 2(f), Fig. 3, Tables 2 and 3.
deteriorates reliability, durability, and performance of bioimplant
applications [34]. Pull-off test was utilized to measure adhesion 3.3. Corrosion evaluation
strength of PCL, and PCL/ZnO (1 and 3 wt%) composite coating on
Mg alloys. Fig. 8(a) represents different stress-strain curves based 3.3.1. Polarization test
on the polymer/composite coating layers, divulging that coating Potentiodynamic polarization curves in a SBF solution was
adhesion depends on the composite materials. The adhesion stress obtained to study the corrosion behavior of different coating sam-
of PCL coated ones was about 12.2 MPa, and incorporating ZnO ples in Fig. 9, and related parameters were shown in Table 4.
NPs (1 and 3 wt%) with PCL nanofibers showed the adhesion stress Polarization curves are used to measure protective properties of
of 12.1 and 15.8 MPa against pull-off strength, respectively. The the metal surface, which can be characterized by the values of cor-
 J. Kim et al. / Applied Surface Science 396 (2017) 249–258
Fig. 8. (a) Stress-strain curves of the different coating samples for coating adhesion test and (b) schematic illustration of the interactions of ZnO NPs, PCL chains, and Mg
substrates.
Fig. 9. (a) The Potentiodynamic polarization curves obtained in SBF solution at 37 ◦ C ± 5 ◦ C from different samples: Bare, PCL, PCL/ZnO 1 wt% and PCL/ZnO 3 wt%. (b) SEM
images of the corroded surface morphology after potentiodynamic polarization test.
rosion current density and corrosion potential. It is well known
that improved corrosion resistance can be estimated as corrosion
potential increases while current density decreases [36]. Especially,
corrosion potential is one of the crucial factors in determining the
corrosion rate of the samples as well as the susceptibility of their
surface [37]. From the result, corrosion potential (Ecorr ) values of
bare and electrospinning-coated Mg alloys varied, ranging from
−1400 to −1100 mV. The bare samples showed the lowest cor-
rosion potential (Ecorr ), but all coated samples displayed higher
corrosion potential (Ecorr ). Furthermore, the coated Mg sample,
in Table 1, exhibited ten times lower corrosion current density
(A/cm2 ) than that of the bare sample. In sum, the data reveals that
electrospun layers are effective in improving corrosion resistance
as the corrosion potential values were higher than those of the
uncoated samples.
Among the coated samples, they showed no significant dif-
ferences in the current density, which suggests that polarization
measurements are not sufficient to differentiate the corrosion pro-
tection of the Mg samples, but there is additional corrosion analysis
needed such as EIS or immersion tests [38–40]. In addition, it might
be also assumed that the porous layers produced by electrospinning
have no dramatic effects on corrosion improvement [41]. However,
in the overall view of the polarization curves, it can be concluded
255
that PCL/ZnO 3 wt% composite coatings are considered better in
corrosion resistance due to their values of the highest corrosion
potential (Ecorr ) as well as the lowest current density (Icorr ). The SEM
images after the potentiodynamic polarization test for bare and
the different coated samples are present in Fig. 9(b). It was clearly
seen that the bare Mg samples were more susceptible to pitting
and cracking occurrence when compared to the coated samples,
which demonstrates that coating layers formed by the electrospin-
ning method play a significant role when it comes to corrosion
protection. Overall, the results exhibit that the presence of elec-
trospun coating layers on Mg alloys eventually slows the corrosion
and that PCL coating with optimal content of ZnO NPs enhances the
corrosion performance of Mg alloys.
3.3.2. EIS test
To further clarify the corrosion behavior of the PCL and PCL/ZnO
coated samples, EIS spectra were obtained at open circuit poten-
tial. The measurements were carried out in the simulated body
fluid at 37 ◦ C, and the results of the representative Nyquist, Bode,
and phase angle plots are present in Fig. 10(a)–(c), respectively.
Fig. 10(a) shows how all samples commonly reveal semi-circles (a
capacitive loop) but their radius varying evidently. It is well known
that the presence of the semi-circle shapes in Nyquist plots explains
256 J. Kim et al. / Applied Surface Science 396 (2017) 249–258

Fig. 10. Niquist, Bode, and Phase angle plots (a–c) of EIS analysis for uncoated and coated Mg alloy samples in the SBF at 37 ◦ C.

Fig. 11. SEM images showing cell attachment and growth on different surfaces: of (a, e) Bare, (b, f) PCL coated, (c, g) PCL/ZnO 1 wt% and (d, h) PCL/ZnO 3 wt% coated Mg
substrates at 1 day (a–d) and 3 days (e–h).

not only the resistance to charge transfer, but also the capacitance Table 5
Fitting parameters obtained from EIS spectra of bare and coated samples. EIS per-
of the double electrical layer between the electrode and electrolyte
formed in SBF solution at 37 ◦ C.
interface [42]. From Fig. 10(a), the coated samples recorded higher
global impedance values (both Zreal and Zimg) compared to the val- Samples Rs ( cm2) Rp (k cm2) Qp (␮F/cm2) n
ues of the bare samples, meaning that the coated samples have an Bare 213 1.986 16.181 0.68
increased barrier properties against corrosion occurrence. In addi- PCL 252 8.735 1.643 0.86
tion, the largest radius was produced by the PCL/ZnO 3 wt% coated PCL/ZnO 1 wt% 264 10.737 2.026 0.88
PCL/ZnO 3 wt% 211 35.309 0.755 0.84
sample, which indicates the highest corrosion resistance among the
coated samples.
Fig. 10(b) exhibits the Bode plot of EIS measurements for bare are characterized at high and mid frequencies between 105 and
and coated Mg alloy samples. According to the bode plot, it can 1, and the Mg substrates can be defined between 1 Hz–10–1 Hz
be seen that the curves of PCL and PCL/ZnO 1 wt% coated samples from Fig. 10(c). For the coated samples, the values of phase angle
were similar but only slightly different at overall frequency ranges were shifted towards more negative directions at high and medium
(10–1–105 Hz). It is well known that higher impedance modulus frequency range (105 Hz–10 Hz), indicating the higher barrier prop-
(|Z|) at low frequency regions from 10 to 1 Hz to 1 Hz represents erties than those of the bare Mg samples. However, similar curve
a better corrosion resistance of the substrates [3]. PCL/ZnO 3 wt% behaviors were revealed at low frequency regions between 1 Hz
coated samples displayed a noticeable increase in the impedance and 10−1 Hz, which suggests that the low frequency regions are
modulus. This explains that PCL/ZnO 3 wt% coating layers can pro- associated with the Mg substrate boundary. The Mg samples coated
duce more compact and denser coating layers in comparison to PCL with PCL/ZnO 3 wt% showed the most negative values in the high
and PCL/ZnO 1 wt% coatings and that it plays a significant role in frequency regions (103 Hz–105 Hz), implying that this compos-
contributing to the highest Z modulus values among the coatings. ite coating can promote a strong and effective corrosion-resistant
To understand corrosion mechanisms of the coated samples, the outer layers.
phase angle plots were examined as evident in Fig. 10(c). The sur- The numerical values of EIS parameters for the uncoated and
face state and its structural changes can be observed from the phase coated Mg samples are summarized in Table 5. Rs is the electrolyte
angle that shows a capacitive behavior over the frequency range. resistance (SBF) between the working and reference electrodes,
The trend of the four curves is distinct, showing how uncoated and and Rp , Qp and n are the fitting values obtained from the simu-
coated samples have different corrosion processes. The coating lay- lated EIS spectra. As shown in Table 5, the polarization resistance
ers or a native passivation layer on Mg samples (outer surfaces) (Rp ) of bare Mg samples presented the lowest values among the
 J. Kim et al. / Applied Surface Science 396 (2017) 249–258
Fig. 12. The osteoblast cell viability for differently coated Mg samples on 1 and
3 days. The viability of the samples was relative to the control cell set at 100%. The
data is revealed as the mean ± standard deviation (n = 3) and * indicates statistical
significance (p < 0.05), ** shows statistical significance (p < 0.01) (one way ANOVA
post hoc Turkey test).
samples, suggesting the surface of the bare Mg samples has a poor
protective ability. The polarization resistance (Rp ) for the Mg sam-
ple coated with PCL was nearly 4 times higher than that of the
bare sample while the Rp value increased more than 16 times in
the PCL/ZnO 3 wt% coated samples, indicating the PCL/ZnO 3 wt%
composite is more effective on corrosion protection. Thus, it can
be concluded that the PCL/ZnO 3 wt% composite coating formed by
electrospinning enhance the corrosion resistance of the Mg alloy.
3.4. Biocompatibility
To assess the biological response on non-coated and coated Mg
surfaces, osteoblast cells were cultured for 1 and 3 days, examining
cell attachment and growth [43]. In Fig. 11(a and e), only a few cells
were attached to the naked surface of the bare samples after 1 day
of incubation, and even fewer cells survived the 3 day mark. This
demonstrates that the surface of bare Mg samples does not pro-
vide a sufficient settlement environment for cells to survive due
to the cracks and uncontrolled hydrogen release from rapid corro-
sion occurrence [6]. On the other hand, polymeric and composite
nanofibrous coatings (Fig. 11b–d and f–h) showed drastic enhanced
cell adhesion and growth at 1 and 3 days, which might be due to the
change in surface topography via electrospinning coating process
[44].
The CCK assay was also employed to determine the biocompat-
ibility of the different coatings on Mg alloys. Fig. 12 indicates that
the cell viability of the bare Mg samples was relatively lower when
compared to that of the coated samples. This tendency remark-
able at 3 days of the untreated Mg samples showed highly inferior
cell viability to PCL, PCL/ZnO 1 wt% and 3 wt%. In contrast, the
cell viability became enhanced in the coated samples relative to
at both 1 and 3 days. The composite-coated samples (PCL/ZnO
1 and 3 wt%) exhibited a gradual proliferation pattern over the
3 days of cell culture. The SEM and cell viability results explain that
electrospun coating layer on Mg samples improves the biocompat-
ibility when compared to that of the bare sample. Furthermore,
it was revealed that 1 and 3 wt% of ZnO NPs incorporated PCL
coatings did not considerably affect cellular behavior on Mg sub-
strates but rather showed an improvement on the cell viability
result, indicating electrospun coatings containing a proper concen-
tration of ZnO NPs could be acceptable in biomedical applications
[30,44,45].
257
4. Conclusion
In the study, PCL/ZnO composite coating (1 and 3 wt%) on Mg
alloys were successfully produced using electrospinning and char-
acterized. The electrospun fabricated surfaces were seen highly
porous and nano-structural, and ZnO NPs incorporated fibers
became rougher than neat PCL fibers. EDX, XPS, and FT-IR analy-
ses confirmed the incorporation of ZnO NPs into PCL nanofibrous
coating. For coating adhesion results, PCL/ZnO 3 wt% composite Mg
samples revealed the strongest adhesive property among coated
samples, which might be due to the ZnO NPs that influence the
structure and composition of metal/polymeric interfaces. Corro-
sion test demonstrated that corrosion resistance of the electrospun
composite coated samples is superior to the resistance of bare and
only polymeric coated ones. Moreover, the electrospun composite
coated surfaces on Mg substrates showed better cell attachment
and proliferation compared to those of bare Mg surfaces. Therefore,
it is recommendable that PCL/Zinc oxide nanoparticle composite
coatings via electrospinning have promising potential for applica-
tions in magnesium-based biomaterials.
Acknowledgments
This research was supported by a grant from the Basic
Science Research Program through the National Research Foun-
dation of Korea (NRF) by Ministry of Education (Project no NRF
2015R1C1A1A02036404). And partially supported by 2015 Jeon-
Buk TechnoPark academic-industrial core technology development
and commercialization support program.
References
[1] F. Witte, The history of biodegradable magnesium implants: a review, Acta
Biomater. 6 (2010) 1680–1692.
[2] J. Kuhlmann, I. Bartsch, E. Willbold, S. Schuchardt, O. Holz, N. Hort, D. Hoche,
W.R. Heineman, F. Witte, Fast escape of hydrogen from gas cavities around
corroding magnesium implants, Acta Biomater. 9 (2013) 8714–8721.
[3] H.M. Mousa, A.P. Tiwari, J. Kim, S.P. Adhikari, C.H. Park, C.S. Kim, A novel
in situ deposition of hydroxyapatite nanoplates using
anodization/hydrothermal process onto magnesium alloy surface towards
third generation biomaterials, Mater. Lett. 164 (2016) 144–147.
[4] S. Kunjukunju, A. Roy, M. Ramanathan, B. Lee, J.E. Candiello, P.N. Kumta, A
layer-by-layer approach to natural polymer-derived bioactive coatings on
magnesium alloys, Acta Biomater. 9 (2013) 8690–8703.
[5] M.P. Staiger, A.M. Pietak, J. Huadmai, G. Dias, Magnesium and its alloys as
orthopedic biomaterials: a review, Biomaterials 27 (2006) 1728–1734.
[6] H. Hornberger, S. Virtanen, A.R. Boccaccini, Biomedical coatings on
magnesium alloys – a review, Acta Biomater. 8 (2012) 2442–2455.
[7] F. Witte, V. Kaese, H. Haferkamp, E. Switzer, A. Meyer-Lindenberg, C.J. Wirth,
H. Windhagen, In vivo corrosion of four magnesium alloys and the associated
bone response, Biomaterials 26 (2005) 3557–3563.
[8] H.M. Wong, K.W.K. Yeung, K.O. Lam, V. Tam, P.K. Chu, K.D.K. Luk, K.M.C.
Cheung, A biodegradable polymer-based coating to control the performance
of magnesium alloy orthopaedic implants, Biomaterials 31 (2010) 2084–2096.
[9] J.N. Li, P. Cao, X.N. Zhang, S.X. Zhang, Y.H. He, In vitro degradation and cell
attachment of a PLGA coated biodegradable Mg-6Zn based alloy, J. Mater. Sci.
45 (2010) 6038–6045.
[10] F. Feil, W. Furbeth, M. Schutze, Purely inorganic coatings based on
nanoparticles for magnesium alloys, Electrochim. Acta 54 (2009) 2478–2486.
[11] C.L. Wen, S.K. Guan, L. Peng, C.X. Ren, X. Wang, Z.H. Hu, Characterization and
degradation behavior of AZ31 alloy surface modified by bone-like
hydroxyapatite for implant applications, Appl. Surf. Sci. 255 (2009)
6433–6438.
[12] J.M. Seitz, K. Collier, E. Wulf, D. Bormann, F.W. Bach, Comparison of the
corrosion behavior of coated and uncoated magnesium alloys in an In vitro
corrosion environment, Adv. Eng. Mater. 13 (2011) B313–B323.
[13] B.D. Hahn, D.S. Park, J.J. Choi, J.H. Ryu, W.H. Yoon, J.H. Choi, H.E. Kim, S.G. Kim,
Aerosol deposition of hydroxyapatite-chitosan composite coatings on
biodegradable magnesium alloy, Surf. Coat. Tech. 205 (2011) 3112–3118.
[14] P. Bhattacharjee, D. Naskar, H.W. Kim, T.K. Maiti, D. Bhattacharya, S.C. Kundu,
Non-mulberry silk fibroin grafted PCL nanofibrous scaffold: promising ECM
for bone tissue engineering, Eur. Polym. J. 71 (2015) 490–509.
[15] L. Calandrelli, A. Calarco, P. Laurienzo, M. Malinconico, O. Petillo, G. Peluso,
Compatibilized polymer blends based on PDLLA and PCL for application in
bioartificial liver, Biomacromolecules 9 (2008) 1527–1534.
258 J. Kim et al. / Applied Surface Science 396 (2017) 249–258
[16] V. Romeo, G. Gorrasi, V. Vittoria, I.S. Chronakis, Encapsulation and exfoliation
of inorganic lamellar fillers into polycaprolactone by electrospinning,
Biomacromolecules 8 (2007) 3147–3152.
[17] R. Augustine, N. Kalarikkal, S. Thomas, Effect of zinc oxide nanoparticles on
the in vitro degradation of electrospun polycaprolactone membranes in
simulated body fluid, Int. J. Polym. Mater. Polym. 65 (2016) 28–37.
[18] G. Colon, B.C. Ward, T.J. Webster, Increased osteoblast and decreased
Staphylococcus epidermidis functions on nanophase ZnO and TiO2, J. Biomed.
Mater. Res. A 78a (2006) 595–604.
[19] D. Virovska, D. Paneva, N. Manolova, I. Rashkov, D. Karashanova,
Electrospinning/electrospraying vs. electrospinning: a comparative study on
the design of poly(l-lactide)/zinc oxide non-woven textile, Appl. Surf. Sci. 311
(2014) 842–850.
[20] R. Wahab, M.A. Siddiqui, Q. Saquib, S. Dwivedi, J. Ahmad, J. Musarrat, A.A.
Al-Khedhairy, H.S. Shin, ZnO nanoparticles induced oxidative stress and
apoptosis in HepG2 and MCF-7 cancer cells and their antibacterial activity,
Colloids Surf. B 117 (2014) 267–276.
[21] M.P.K. Turunen, P. Marjamaki, M. Paajanen, J. Lahtinen, J.K. Kivilahti, Pull-off
test in the assessment of adhesion at printed wiring board
metallisation/epoxy interface, Microelectron. Reliab. 44 (2004) 993–1007.
[22] D.S. Rickerby, A review of the methods for the measurement of coating
substrate adhesion, Surf. Coat. Technol. 36 (1988) 541–557.
[23] H.M. Mousa, K.H. Hussein, H.M. Woo, C.H. Park, C.S. Kim, One-step anodization
deposition of anticorrosive bioceramic compounds on AZ31B magnesium
alloy for biomedical application, Ceram. Int. 41 (2015) 10861–10870.
[24] M.K. Joshi, A.P. Tiwari, H.R. Pant, B.K. Shrestha, H.J. Kim, C.H. Park, C.S. Kim,
In situ generation of cellulose nanocrystals in polycaprolactone nanofibers:
effects on crystallinity, mechanical strength, biocompatibility, and biomimetic
mineralization, ACS Appl. Mater. Interfaces 7 (2015) 19672–19683.
[25] H.R. Pant, M.P. Bajgai, K.T. Nam, K.H. Chu, S.J. Park, H.Y. Kim, Formation of
electrospun nylon-6/methoxy poly(ethylene glycol) oligomer spider-wave
nanofibers, Mater. Lett. 64 (2010) 2087–2090.
[26] H. Rodriguez-Tobias, G. Morales, A. Ledezma, J. Romero, R. Saldivar, V.
Langlois, E. Renard, D. Grande, Electrospinning and electrospraying
techniques for designing novel antibacterial poly(3-hydroxybutyrate)/zinc
oxide nanofibrous composites, J. Mater. Sci. 51 (2016) 8593–8609.
[27] P. Wutticharoenmongkol, N. Sanchavanakit, P. Pavasant, P. Supaphol,
Preparation and characterization of novel bone scaffolds based on
electrospun polycaprolactone fibers filled with nanoparticles, Macromol.
Biosci. 6 (2006) 70–77.
[28] L.D. Tijing, Y.C. Woo, W.G. Shim, T. He, J.S. Choi, S.H. Kim, H.K. Shon,
Superhydrophobic nanofiber membrane containing carbon nanotubes for
high-performance direct contact membrane distillation, J. Membr. Sci. 502
(2016) 158–170.
[29] H. Rodriguez-Tobias, G. Morales, A. Ledezma, J. Romero, D. Grande, Novel
antibacterial electrospun mats based on poly(d,l-lactide) nanofibers and zinc
oxide nanoparticles, J. Mater. Sci. 49 (2014) 8373–8385.
[30] R. Augustine, H.N. Malik, D.K. Singhal, A. Mukherjee, D. Malakar, N. Kalarikkal,
S. Thomas, Electrospun polycaprolactone/ZnO nanocomposite membranes as
biomaterials with antibacterial and cell adhesion properties, J. Polym. Res. 21
(2014).
[31] A. Yakubu, Z. Abbas, N.A. Ibrahim, A. Fahad, Reduction of electromagnetic
interference using ZnO-PCL nanocomposites at microwave frequency, Adv.
Mater. Sci. Eng. (2015).
[32] L.L. Pan, X.B. Pei, R. He, Q.B. Wan, J. Wang, Multiwall carbon
nanotubes/polycaprolactone composites for bone tissue engineering
application, Colloids Surf. B 93 (2012) 226–234.
[33] Y.Z. Zhang, J. Venugopal, Z.M. Huang, C.T. Lim, S. Ramakrishna,
Characterization of the surface biocompatibility of the electrospun
PCL-collagen nanofibers using fibroblasts, Biomacromolecules 6 (2005)
2583–2589.
[34] R.G. Hu, S. Zhang, J.F. Bu, C.J. Lin, G.L. Song, Recent progress in corrosion
protection of magnesium alloys by organic coatings, Prog. Org. Coat. 73 (2012)
129–141.
[35] A. Abdal-hay, A.S. Hamdy, K.A. Khalil, J.H. Lim, A novel simple one-step air jet
spinning approach for deposition of poly(vinyl acetate)/hydroxyapatite
composite nanofibers on Ti implants, Mater. Sci. Eng. C: Mater. 49 (2015)
681–690.
[36] D. Ahmadkhaniha, M. Fedel, M.H. Sohi, A.Z. Hanzaki, F. Deflorian, Corrosion
behavior of magnesium and magnesium-hydroxyapatite composite
fabricated by friction stir processing in Dulbecco’s phosphate buffered saline,
Corros. Sci. 104 (2016) 319–329.
[37] H.M. Mousa, D.H. Lee, C.H. Park, C.S. Kim, A novel simple strategy for in situ
deposition of apatite layer on AZ31B magnesium alloy for bone tissue
regeneration, Appl. Surf. Sci. 351 (2015) 55–65.
[38] E. Mostaed, M. Hashempour, A. Fabrizi, D. Dellasega, M. Bestetti, F. Bonollo, M.
Vedani, Microstructure texture evolution, mechanical properties and
corrosion behavior of ECAP processed ZK60 magnesium alloy for
biodegradable applications, J. Mech. Behav. Biomed. 37 (2014) 307–322.
[39] Z.M. Shi, M. Liu, A. Atrens, Measurement of the corrosion rate of magnesium
alloys using Tafel extrapolation, Corros. Sci. 52 (2010) 579–588.
[40] G.L. Song, A. Atrens, Understanding magnesium corrosion – a framework for
improved alloy performance, Adv. Eng. Mater. 5 (2003) 837–858.
[41] A. Abdal-hay, N.A.M. Barakat, J.K. Lim, Influence of electrospinning and
dip-coating techniques on the degradation and cytocompatibility of
Mg-based alloy, Colloids Surf. A 420 (2013) 37–45.
[42] R. Arrabal, J.M. Mota, A. Criado, A. Pardo, M. Mohedano, E. Matykina,
Assessment of duplex coating combining plasma electrolytic oxidation and
polymer layer on AZ31 magnesium alloy, Surf. Coat. Technol. 206 (2012)
4692–4703.
[43] Y.Z. Zhang, J.R. Venugopal, A. El-Turki, S. Ramakrishna, B. Su, C.T. Lim,
Electrospun biomimetic nanocomposite nanofibers of
hydroxyapatite/chitosan for bone tissue engineering, Biomaterials 29 (2008)
4314–4322.
[44] R. Augustine, E.A. Dominic, I. Reju, B. Kaimal, N. Kalarikkal, S. Thomas,
Electrospun polycaprolactone membranes incorporated with ZnO
nanoparticles as skin substitutes with enhanced fibroblast proliferation and
wound healing, RSC Adv. 4 (2014) 24777–24785.
[45] H. Rodríguez-Tobías, G. Morales, D. Grande, Improvement of mechanical
properties and antibacterial activity of electrospun poly (d l-lactide)-based
mats by incorporation of ZnO-graft-poly (d,l-lactide) nanoparticles, Mater.
Chem. Phys. 182 (2016) 324–331.