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Short Communication

Enhanced biocompatibility of a Ni–Cr alloy

prepared by selective laser melting: a preliminary
in vitro study

Min-Ho Hong a , Takao Hanawa b , Si Hoon Song c , Bong Ki Min d , Tae-Yub Kwon a,e,∗
a Institute for Biomaterials Research & Development, Kyungpook National University, Daegu, Republic of Korea
b Department of Metallic Biomaterials, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan
c Department of Medical & Biological Engineering, Graduate School, Kyungpook National University, Daegu, Republic of Korea
d Center for Research Facilities, Yeungnam University, Gyeongsan, Republic of Korea
e Department of Dental Biomaterials, School of Dentistry, Kyungpook National University, Daegu, Republic of Korea

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

Article history: Against a general consensus regarding the high toxicity and low biocompatibility of cast
Received 3 February 2018 nickel–chromium (Ni–Cr) alloys, the in vitro biocompatibility of selective laser melting (SLM)-
Accepted 22 August 2018 fabricated Ni–Cr alloys was investigated and compared with that of cast alloy. Using a single
Available online xxx Ni–Cr alloy powder, two SLM alloys with high (SLM a) and low (SML b) porosity were prepared
by adjusting the laser process parameters in addition to a cast alloy. The alloys were studied
Keywords: in terms of their microstructure, metal ion release, and cell response. All three alloys were
Metallurgy composed of only the  (face-centered cubic) phase. However, both SLM alloys showed more
Ni–Cr alloy homogeneous dispersion of Ni, Cr, and molybdenum elements and finer grain formation
Selective laser melting than the cast alloy. Immersion test results indicated that both SLM alloys (in particular,
Microstructure SML b) exhibited a significantly lower Ni ion release than the cast alloy (P < 0.001). A water-
Biocompatibility soluble tetrazolium salt-8 assay also showed that the viability of L929 mouse fibroblasts was
significantly higher for both SLM groups (in particular, SML b) than for the cast alloy group
(P < 0.001). Thus, the combined results of this in vitro study suggest that the SLM-processed
Ni–Cr alloys exhibited greatly enhanced biocompatibility in comparison with the cast alloy.
© 2018 Brazilian Metallurgical, Materials and Mining Association. Published by Elsevier
Editora Ltda. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).

∗
Corresponding author.
E-mail: tykwon@knu.ac.kr (T. Kwon).
https://doi.org/10.1016/j.jmrt.2018.08.004
2238-7854/© 2018 Brazilian Metallurgical, Materials and Mining Association. Published by Elsevier Editora Ltda. This is an open access
article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Please cite this article in press as: Hong M, et al. Enhanced biocompatibility of a Ni–Cr alloy prepared by selective laser melting: a preliminary
in vitro study. J Mater Res Technol. 2018. https://doi.org/10.1016/j.jmrt.2018.08.004
JMRTEC-478; No. of Pages 6
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SLM b) were prepared by adjusting two laser process parame-

1. Introduction ters (scan speed and laser power). An SLM device (MetalSys150,
Winforsys Co., Ltd., Korea) with an ytterbium fiber laser beam
In dentistry, metal–ceramic restorations are still a popular
(IPG YLR-200 CW) was operated under a nitrogen gas atmo-
choice for fixed partial dentures because of their excel-
sphere (flow rate: 5 L/mm). The scan speeds for the SLM a
lent esthetic and mechanical properties [1,2]. Traditionally,
and SLM b alloys were 600 and 1000 mm/s, respectively. The
the metal frameworks are manufactured by lost-wax cast-
laser power levels for the SLM a and SLM b alloys were 125
ing, and then the veneering ceramic is fused to the metal.
and 175 W, respectively. For both SLM alloys, the scan-line
Nickel–chromium (Ni–Cr) and cobalt–chromium (Co–Cr) alloys
spacing, layer thickness, and laser spot size were 0.10 mm,
are the most commonly employed dental cast base-metal
35 ␮m, and 0.08 mm, respectively. The building direction was
alloys for such use [3].
perpendicular to the radial direction of the disk specimens.
Although Ni–Cr alloys have advantages, including high
The densities of the two SLM alloys were measured by the
acid/alkali resistance as well as great toughness and strength
Archimedes method.
at high temperatures [4], they pose higher biologic risk than
The microstructures of the alloys were investigated using
other alloys because of the common hypersensitivity to Ni
optical microscopy (MM-40/2U, Nikon) and X-ray diffractome-
[5]. Moreover, Ni–Cr–beryllium (Be) alloys corrode far more
try (XRD, X’Pert PRO, PANalytical). The surfaces were polished
than their non-Be counterparts [5], although the addition of
with silicon carbide papers and then diamond suspensions
Be improves casting manipulation and enhances porcelain
until a 0.05-␮m surface finish was achieved. Phase iden-
bonding [3,6]. Therefore, Co–Cr alloys are the most common
tification was carried out by XRD, using Cu K␣ radiation
base-metal alternative for patients who are allergic to Ni [3,5].
( = 0.1541 nm) at an accelerating voltage of 40 kV, a beam cur-
However, Co–Cr alloys have the highest melting ranges of the
rent of 30 mA, a 2-angle scan range of 20–100◦ , a scanning
casting alloys, with the exception of titanium-based alloys,
speed of 2◦ /min, a sampling pitch of 0.02◦ , and a preset time
making laboratory manipulation (casting, finishing, and pol-
of 0.6 s.
ishing) of these alloys difficult [3,5,7]. In addition, the surface
To analyze the elemental composition, the specimens for
oxide of Co–Cr alloys is difficult to mask, and the thermal
each alloy were examined using field emission-scanning elec-
expansion coefficients are often not as compatible with the
tron microscopy (FE-SEM, Merlin, Carl Zeiss AG) with energy
porcelains as Ni–Cr alloys [5,8]. Co is also the second-most
dispersive X-ray spectroscopy (EDS, Oxford Instruments)
common metal allergen [5].
under an accelerating voltage of 20 kV. To determine the crys-
Selective laser melting (SLM), an additive manufacturing
tallographic orientation, electron backscattered diffraction
technique, has attracted increasing attention as an effec-
(EBSD) scans were performed on the FE-SEM equipped with
tive alternative to traditional casting. In comparison with
a Nordlys Nano EBSD detector (Oxford Instruments) under an
traditional casting, SLM can avoid errors made by techni-
accelerating voltage of 20 kV.
cians, prevent casting defects, and reduce the extent of
To evaluate the amounts of Ni, Cr, and Mo ions released
impurities [9]. In addition, SLM alters the microstructures
from the alloys, the specimens were immersed in either
which determine the characteristics of an alloy in biologi-
Roswell Park Memorial Institute (RPMI) 1640 culture medium
cal environments where there is direct contact with body
or acidic (pH 2.3) artificial saliva. The experimental protocols
fluids [9,10]. Xin et al. [9] demonstrated that a Co–Cr alloy
have been described elsewhere [9,11]. Briefly, each specimen
prepared by SLM showed lower ion release and better cell
was immersed in 7.5 mL of solution and stored in an oven at
response than cast alloy because of the homogeneous and
37 ◦ C. After 7-day immersion, the solutions (n = 5) were tested
compact microstructure formation. A similar tendency can
using inductively coupled plasma mass spectroscopy (NexIon
also be expected for SLM-fabricated Ni–Cr alloys. To the best
300X, PerkinElmer), based on matrix-matched standards. The
of our knowledge, however, little research has been con-
concentrations of Ni, Cr, and Mo ions were converted to units
ducted to investigate the biocompatibility of Ni–Cr alloys
of ␮g/cm2 .
fabricated via SLM. The purpose of this preliminary in vitro
The effect of ion release on the viability of L929 mouse
study was to evaluate metal ion release from, and the
fibroblasts was evaluated by a water-soluble tetrazolium salt-
cell response to, Ni–Cr alloys prepared by either SLM or
8 (WST-8) assay. The experimental protocols have also been
casting.
described elsewhere [9,11]. Briefly, the extracts were prepared
by immersing the specimens in the RPMI 1640 culture medium
2. Experimental at a ratio of 3:1 volume of solution to specimen surface area for
7 days. L929 mouse fibroblasts were seeded in 96-well plates
A non-Be-containing SLM alloy powder (Metal Player Co., Ltd.), at 4 × 104 cell/mL and incubated for 24 h [12]. The medium in
with a composition of Ni 68 wt%, Cr 22 wt%, and molybde- each well was then replaced with each extract. After 1, 3, and
num (Mo) 9 wt%, was used to prepare disk-shaped specimens 7 days of culture, the absorbance of each solution was mea-
(10 mm in diameter and 3 mm in thickness). The average par- sured using a spectrophotometer at 450 nm (n = 9), according
ticle size of the powder was less than 40 ␮m. In the CAST the WST-8 protocol. The medium itself and a 10% dimethyl
group, wax patterns were prepared, embedded in an invest- sulfoxide-containing medium were used as negative and pos-
ment material (BC-VEST P-1000, Bukwang), and finally cast itive controls, respectively [11,12]. The cell morphology at 1 day
using the Ni–Cr alloy in a casting apparatus (Centrifico, Kerr of culture was also examined using a phase-contrast micro-
Corp.). In the SLM groups, two different specimens (SLM a and scope after Giemsa staining [11].

Please cite this article in press as: Hong M, et al. Enhanced biocompatibility of a Ni–Cr alloy prepared by selective laser melting: a preliminary
in vitro study. J Mater Res Technol. 2018. https://doi.org/10.1016/j.jmrt.2018.08.004
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CAST SLM_a SLM_b

10 µm 10 µm 10 µm

(111)
Ni-based y (fcc) Ni-based y (fcc) Ni-based y (fcc)

(111)

(111)
Intensity (arbitrary unit)

Intensity (arbitrary unit)

Intensity (arbitrary unit)

(200)

(200)
(200)

(311)
(220)

(311)
(220)
(311)
(222)

(222)
(220)

(222)
20 30 40 50 60 70 80 90 100 20 30 40 50 60 70 80 90 100 20 30 40 50 60 70 80 90 100
Diffraction angle (2 theta) Diffraction angle (2 theta) Diffraction angle (2 theta)

Fig. 1 – Optical microscopy images and corresponding XRD patterns of the CAST, SLM a, and SLM b alloy specimens. For all
the three groups, only the Ni-based  (face-centered cubic, fcc) matrix phase was identified (JCPDS card no.: 87-0712).

The results were analyzed using one-way analysis of vari- different manufacturing processes (one casting and two SLM)
ance, followed by Tukey’s post hoc test to determine whether to prepare three different alloys.
there were significant differences among groups (˛ = 0.05). If Fig. 1 shows the optical microscopy images and XRD pat-
appropriate, the data were transformed before analysis using a terns of the alloys produced by casting or SLM processes. The
square root transformation to meet homogeneity of variance. CAST and SLM b specimens showed less pore formation on
the surface than the SLM a specimen. The XRD patterns indi-
cate that the CAST, SLM a, and SLM b alloys were composed of
3. Results and discussion only  (face-centered cubic, fcc) phase, which presents a solid
solution of Ni, without any precipitates.
Ni–Cr alloys with Cr contents of over 20 wt% have superior cor- Fig. 2 shows the SEM, EDS, and EBSD images of the three
rosion resistance qualities [5,13]. In addition, Mo is an effective prepared alloys. The EDS maps of both SLM groups show a
hardening element for Ni–Cr alloys and has the added bene- more homogeneous dispersion of the metal elements than
fit of influencing the coefficient of thermal expansion [3]. The those of the CAST group, indicating superior corrosion resis-
pitting corrosion resistance of Ni–Cr alloys in an acid medium tance of the SLM alloys [14]. The EBSD images show that the
is dramatically improved with increased Mo content [13]. In samples of the SLM groups were composed of finer grains,
this study, a single Mo-containing Ni–Cr alloy, composed of Ni probably due to rapid solidification by a very high cooling rate
68 wt%, Cr 22 wt%, and Mo 9 wt%, was subjected to the three [8], than the CAST group. Such small grains of the SLM groups

EDS map
SEM EBSD
Ni cr Mo Band contrast map IPF map
CAST
SLM_a
SLM_b

Fig. 2 – SEM, EDS and EBSD images of the CAST, SLM a, and SLM b specimens (250×, scale bar = 100 ␮m). The CAST
specimen showed a compositional segregation of the Cr and Mo elements.

Please cite this article in press as: Hong M, et al. Enhanced biocompatibility of a Ni–Cr alloy prepared by selective laser melting: a preliminary
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Table 1 – Metal ion release (␮g/cm2 ) in the immersion solutions.

Immersion solution Group Ni Cr Mo

Culture medium CAST 0.663 ± 0.019 a

0.009 ± 0.002a
0.101 ± 0.010a
SLM a 0.199 ± 0.014b 0.008 ± 0.001a 0.064 ± 0.009b
SLM b 0.018 ± 0.001c 0.008 ± 0.001a 0.006 ± 0.001c

Artificial saliva (pH 2.3) CAST 0.932 ± 0.117a 0.118 ± 0.023a 0.270 ± 0.030a
SLM a 0.361 ± 0.025b 0.126 ± 0.022a 0.102 ± 0.015b
SLM b 0.175 ± 0.029c 0.117 ± 0.011a 0.022 ± 0.004c

For each solution, the same superscripted lowercase letters within each column indicate statistically similar means (P > 0.05).

NC PC

CAST SLM_a SLM_b

Fig. 3 – Phase-contrast microscopy images of L929 fibroblasts after 1 day of culture (200×, stained with Giemsa) (NC,
negative control; PC, positive control).

also could potentially enhance corrosion resistance under
acidic condition [11,15]. In addition, the CAST alloy showed a
compositional segregation of the Cr and Mo elements during
solidification [16]. The measured density of the CAST alloy was
8.31 g/cm3 . Within the SLM groups, the SLM b alloy showed a
more dense and compact structure, with less porosity than the
SLM a alloy (measured densities 8.52 and 7.11 g/cm3 , respec-
tively) (Figs. 1 and 2). Such porosity may increase susceptibility
to corrosion, such as crevice corrosion and pitting corrosion
[17].
Table 1 shows the results of the Ni, Cr, and Mo ions released
from the alloy specimens. In both immersion solutions, a
greater release of Ni ions was detected in comparison with
that of the other ions. The concentrations of Ni and Mo ions
released from the SLM alloys (in particular, in SLM b) were
significantly lower than those from the CAST alloy (P < 0.001).
Acidic artificial saliva was also used as an immersion solu-
tion because the corrosion of Ni–Cr alloys may be particularly
evident in acidic environments [5,11]. Overall, the acidic arti-
ficial saliva aggravated the toxic metal ion release from the
alloy specimens in comparison to the culture medium [11].
In particular, the release of Cr ions was greater in the acidic
saliva than in the culture medium. It was assumed that rela-
tively low proportion of Cr (22 wt%) in the alloys caused slight
breakdown and repassivation of the surface in the saliva and,
as a result, an acceleration of Cr ion release [18].
The phase-contrast microscopy and cell viability results
(Figs. 3 and 4) were paralleled by the metal ion release results
(Table 1). The SLM groups exhibited greater cell density and
viability than the CAST group (P < 0.05). Moreover, the SLM b
group exhibited superior cell viability (approaching that of
the negative control) in comparison to the other two groups
(P < 0.05). The findings of this in vitro biocompatibility test sug-
gest that the Ni–Cr alloy prepared via an optimally controlled
SLM process (SLM b) potentially possesses much lower bio-
logical risk than cast Ni–Cr alloy. Even the SLM a alloy, which
had much higher porosity than the cast alloy (Fig. 1), yielded
more favorable results than the cast alloy. Thus, the combined
results of the present study suggest that the application of the
SLM technique for Ni–Cr alloys may have potential to over-
come the toxicity problem of cast Ni–Cr alloy. However, further
comprehensive studies are required to confirm the enhanced
in vitro and in vivo biocompatibility of SLM-fabricated Ni–Cr
alloys.

Please cite this article in press as: Hong M, et al. Enhanced biocompatibility of a Ni–Cr alloy prepared by selective laser melting: a preliminary
in vitro study. J Mater Res Technol. 2018. https://doi.org/10.1016/j.jmrt.2018.08.004
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NC PC CAST SLM_a SLM_b

120
*** *** **
*** *** ***
*** *** **
*** *** ***
100

***
*** ***
Cell viability (%/NC)
80

*** ***
60 ***

40

20

0
Day 1 Day 3 Day 7
Time

Fig. 4 – Cell viability of L929 fibroblasts after 1, 3, and 7 days of culture (**P < 0.01; ***P < 0.001) (NC, negative control; PC,
positive control).

restorations prepared by different methods. J Prosthet Dent

4. Conclusions
The results of this in vitro study suggest that the Ni–Cr alloys
fabricated by SLM were superior to the traditional-cast Ni–Cr
alloy in terms of biocompatibility, mainly due to the more
homogeneous dispersion of Ni, Cr, and Mo and finer grain for-
mation within the alloys. Moreover, the SLM b alloy, which
had a highly dense structure with less porosity, resulted in
enhanced cell viability in comparison to the cast alloy due to
further reduction in the amount of toxic metal ions released.
The SLM technique thus merits consideration for the fabrica-
tion of Ni–Cr dental prostheses with low health risks.
Conflicts of interest
The authors declare no conflicts of interest.
Acknowledgements
This research was supported by Basic Science Research
Program through the National Research Foundation of
Korea (NRF) funded by the Ministry of Education (NRF-
2017R1A6A3A11036498).
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in vitro study. J Mater Res Technol. 2018. https://doi.org/10.1016/j.jmrt.2018.08.004
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