The novel toxic free titanium-based amorphous alloy for biomedical application

Abstract

Ti based amorphous alloy exhibits excellent properties for biomedical applications. In general, it has high strength, low elastic modulus, good corrosion resistance and satisfactory biocompatibility. This work reported on a systematic study of a novel Ti44Zr10Pd10Cu6Co23Ta7, Ti44Zr10Pd10Cu10Co19Ta7 and Ti44Zr10Pd10Cu14Co15Ta7 metallic glass. Cylindrical rod samples with diameter of 5mm and 20mm length were fabricated by induction melting and casting into copper mold. The cast rod was then used as plasma cathode in filtered cathodic vacuum arc (FCVA) deposition chamber. The Ti-based metallic glass (MG) thin film was produced and tested for subsequent cell culture investigation to understand the biocompatibility nature of the new alloy. X-ray photoelectron spectrometry (XPS) was employed to characterize the surface chemistry. The Ti-6Al-4V alloy was studied in parallel as a control material. This novel Ti-based MG composition has shown promising osteoblast biocompatible characteristics and no cytotoxicity on human osteoblast-like cells (SaOS-2). Moreover, cells on Ti-based MG thin film exhibited greater levels of calcium deposition using Alizarin red staining technique to those of the control. All results point out that the novel Ti-based amorphous alloy has potential for using as a new coating for biomedical application and deserve further study.

Full text

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www.jmrt.com.br
Available
online
at
www.sciencedirect.com
Original
Article
The
novel
toxic
free
titanium-based
amorphous
alloy
for
biomedical
application
Saran
Tantavisuta,
Boonrat
Lohwongwatanab,∗,
Atchara
Khamkongkaeob,
Aree
Tanavaleea,
Pairat
Tangpornprasertc,
Pibul
Ittiravivonga
aDepartment
of
Orthopaedic,
Chulalongkorn
University,
Bangkok
10330,
Thailand
bDepartment
of
Metallurgical
Engineer,
Chulalongkorn
University,
Bangkok
10330,
Thailand
cDepartment
of
Mechanical
Engineer,
Chulalongkorn
University,
Bangkok
10330,
Thailand
a
r
t
i
c
l
e
i
n
f
o
Article
history:
Received
8
March
2017
Accepted
19
August
2017
Available
online
6
December
2017
Keywords:
Ti-based
amorphous
alloy
Metallic
glass
Toxic
free
Filtered
cathodic
vacuum
arc
X-ray
photoelectron
spectrometry
Biocompatibility
a
b
s
t
r
a
c
t
Ti
based
amorphous
alloy
exhibits
excellent
properties
for
biomedical
applications.
In
general,
it
has
high
strength,
low
elastic
modulus,
good
corrosion
resistance
and
satisfactory
biocompatibility.
This
work
reported
on
a
systematic
study
of
a
novel
Ti44 Zr10Pd10 Cu6Co23Ta7,
Ti44 Zr10Pd10 Cu10Co19Ta 7and
Ti44 Zr10Pd10 Cu14Co15Ta 7metallic
glass.
Cylindrical
rod
samples
with
diameter
of
5
mm
and
20
mm
length
were
fabricated
by
induc-
tion
melting
and
casting
into
copper
mold.
The
cast
rod
was
then
used
as
plasma
cathode
in
filtered
cathodic
vacuum
arc
(FCVA)
deposition
chamber.
The
Ti-based
metallic
glass
(MG)
thin
film
was
produced
and
tested
for
subsequent
cell
culture
investigation
to
understand
the
biocompatibility
nature
of
the
new
alloy.
X-ray
photoelectron
spectrometry
(XPS)
was
employed
to
characterize
the
surface
chemistry.
The
Ti–6Al–4V
alloy
was
studied
in
parallel
as
a
control
material.
This
novel
Ti-based
MG
composition
has
shown
promising
osteoblast
biocompatible
characteristics
and
no
cytotoxicity
on
human
osteoblast-like
cells
(SaOS-2).
Moreover,
cells
on
Ti-based
MG
thin
film
exhibited
greater
levels
of
calcium
deposition
using
Alizarin
red
staining
technique
to
those
of
the
control.
All
results
point
out
that
the
novel
Ti-
based
amorphous
alloy
has
potential
for
using
as
a
new
coating
for
biomedical
application
and
deserve
further
study.
©
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/).
1.
Introduction
Metallic
glass
or
amorphous
alloys
are
metal
with
an
amor-
phous
microstructure.
The
component
atoms
of
amorphous
alloys
are
randomly
packed,
instead
of
arranged
in
usual
crys-
talline
structures.
This
microstructure
leads
to
many
excellent
∗Corresponding
author.
E-mail:
boonrat@gmail.com
(B.
Lohwongwatana).
mechanical
properties
and
high
processability,
which
has
inspired
great
interest
in
their
biomedical
applications
[1–3].
The
unique
properties
of
amorphous
alloys
have
a
potential
to
solve
problems
encountered
by
current
biomedical
materi-
als
for
example
elastic
moduli
closer
to
bone
which
leading
to
reduction
in
stress
shielding
problem,
better
hardness
and
corrosion
resistance
[4,5].
Amorphous
alloy
can
be
manu-
factured
in
a
form
of
metal
foam
to
gain
further
Young’s
modulus
matching
to
human
bone
[6,7].
Amorphous
alloy
gen-
erally
demonstrates
excellent
wear
resistance
comparing
with
https://doi.org/10.1016/j.jmrt.2017.08.007
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/).

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249
Target
10cm
Solenoid
filter
Source
Fig.
1
–
Setting
for
filtered
cathodic
vacuum
arc
coating.
traditional
alloy
[8],
then
it
has
potential
to
prevent
wear
debris
and
its
sequelae.
Without
crystalline
structure,
the
amor-
phous
alloy
is
considered
free
of
crystalline
defects.
These
lead
to
superior
corrosion
resistance
to
crystalline
alloys
in
previous
reports
[9,10].
Finally,
the
high
processability
of
the
amorphous
alloy
is
a
huge
advantage
to
their
clinical
applica-
tion
potential.
Many
of
current
amorphous
alloys
contain
toxic
elements
like
Beryllium,
Aluminum
or
Nickel
that
had
been
reported
about
causing
cancer
or
allergy.
These
toxic
ions
limit
the
use
of
amorphous
alloy
as
biomaterial.
To
create
a
new
type
of
biomedical
amorphous
alloy,
the
safety
of
use
is
the
first
concerning
issue.
The
purposes
of
this
work
are
to
synthesize
the
new
type
of
amorphous
alloys
for
with-
out
toxic
elements
for
using
as
a
new
coating
for
biomedical
application.
2.
Materials
and
methods
Ti-based
alloy
ingots
with
a
nominal
composition
of
the
Ti44 Zr10Pd10 Cu6+xCo23−xTa7(x
=
0,
4,
8)
was
synthesized
using
arc-melting
6
elements
of
99.9%
or
better
purity
in
a
titanium-
gettered
argon
atmosphere.
The
Ti–6Al–4V
alloy
was
used
as
a
reference
material.
Cylindrical
rod
samples
with
a
diame-
ter
of
5
mm
and
length
of
20
mm
were
fabricated
by
copper
mold
casting
technique.
The
cylindrical
rods
of
3
new
alloy
formula
and
Ti6-Al-4V
as
a
reference
material
were
then
used
as
plasma
cathode
in
filtered
cathodic
vacuum
arc
(FCVA)
deposition
technique
to
make
Ti-based
metallic
glass
thin
film
(Fig.
1).
The
round
glass
substrate
with
a
diameter
of
1.5
cm
and
thickness
of
0.5
mm
were
attached
to
the
aluminum
plate
with
an
aluminum
tape
then
posted
at
10
cm
away
from
the
solenoid
filter
as
a
target
for
FCVA
coating
(Fig.
1).
The
param-
eter
for
FCVA
is
shown
in
Table
1.
The
Ti-based
thin
film
on
glass
substrate
was
further
employed
for
alloy
characteriza-
tion
tests
and
biocompatibility
tests.
Table
1
–
Parameters
for
filtered
cathodic
vacuum
arc
(FCVA)
deposition.
Parameters
for
FCVA
Resistivity <2
k
Distance
from
solenoid
to
target 10
cm
Time
60
min
Pressure
<5.0
×
10−5
Pulse
2.1
Hz
Voltage
600
V
Bias
target
1
kV
2.1.
Thin
film
characterization
2.1.1.
X-ray
photoelectron
spectrometer
(XPS)
X-ray
photoelectron
spectroscopy
(XPS)
is
an
essentially
non-destructive
technique
that
used
to
analytical
chemical
bonding,
element
composition,
chemical
and
electronic
state
of
every
element
in
the
material.
In
this
work,
the
XPS
spectra
were
studied
at
Synchrotron
Light
Research
Institute
(Public
Organization,
Thailand)
BL5.2:
SUT-NANOTEC-SLRI
by
using
a
ULVAC-PHI
Versa-Probe
II
XPS
at
energy
step
0.05
eV,
pass
energy
46.95.
2.2.
Biocompatibility
testing
We
performed
biocompatibility
test
with
osteoblast
like
cell
(SaOS-2)
In
Vitro.
Before
every
test,
the
coated
discs
were
ster-
ilized
by
autoclaving.
2.2.1.
Culture
condition
Osteoblast
cell
line,
osteoblast-like
cell
(SaOS-2),
was
used
for
biocompatibility
test
in
the
present
study.
The
SaOS-
2
was
cultured
in
Dulbecco’s
modified
Eagle’s
medium
(DMEM).
The
medium
was
supplemented
with
10%
fetal
bovine
serum,
2
mM
l-glutamine,
100
unit
mL−1penicillin,
100
␮g
mL−1streptomycin
and
0.25
␮g
mL−1amphoteracin
B.
Cells
were
maintained
at
37 ◦C
in
100%
humidity
and
5%
CO2.
Confluent
cells
were
detached
using
0.25%
trypsin
with
ethylene
diamine
tetraacetic
acid
and
re-suspended
in
fresh
culture
medium.
The
media
were
changed
every
2–3
days.
2.2.2.
Cell
proliferation
Cell
proliferation
was
determined
by
Methylthiazol
Tetra-
zolium
(MTT)
assay.
Cells
were
seeds
on
triplicate
samples
discs
(n
=
3)
with
a
concentration
of
50,000
cells/well
in
a
24-
well
plate.
The
assay
was
performed
at
3,
5
and
7
days.
After
each
culture
period,
the
media
was
gently
removed
and
the
specimens
were
rinsed
with
phosphate-buffered
saline
(PBS)
to
remove
unattached
cells
and
to
avoid
the
effects
of
media
on
the
biochemical
assays.
Then
MTT
solution
(300
␮L;
0.5
mg/mL
3-(4,
5-dimethylthiazol-2-yl)-2,
5-diphenyltetrazolium
bro-
mide
in
culture
medium
without
phenol
red)
was
added.
After
30
min
of
incubation,
MTT
solution
was
discarded
and
then
the
formazan
crystals
were
dissolved
in
dimethylsulfoxide
(DMSO)
(900
␮L/well)
and
glycine
buffer
(pH
=
10)
(125
␮L/well).
The
absorbance
was
read
at
a
wavelength
of
570
nm
by
Ther-
mospectronic
Genesis
10
UV-vis
spectrometer.

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2.2.3.
Cell
differentiation
For
the
osteoblast
differentiation,
cells
were
grown
on
triplicate
sample
disc
(n
=
3)
with
a
concentration
of
60,000
cells/well
in
a
24-well
plate.
After
2
days
of
cultured,
the
media
was
changed
to
differentiation
medium
(normal
culture
medium
supplemented
with
50
␮g
mL−1ascorbic
acid,
5
mM
␤-glycerophosphate
and
250
nM
Dexamethasone).
Cells
were
cultured
in
differentiation
medium
for
1,
5
and
10
days.
Cell
differentiation
behavior
was
characterized
by
Alizarin
red
staining
for
calcium
deposition
analysis.
2.2.3.1.
Alizarin
red
staining.
Alizarin
is
an
organic
compound
that
could
react
with
calcium
ions.
Alizarin
red
S,
a
dye
that
stains
calcium
salts
selectively
and
is
widely
used
for
mineral
histochemistry
of
calcium,
served
to
analyze
the
mineral-
ization
level
of
cells.
In
this
study,
SaOS-2
100,000
cells
were
seeded
onto
the
disc.
The
osteogenic
inductions
were
induced
after
24
h.
On
day
28,
the
medium
was
discarded
then
the
cells
on
the
discs
were
fixed
with
95%
ethanol
for
10
min
then
rinsed
several
times
with
distilled
water.
0.1%
Alizarin
red
was
added
onto
the
disc
then
incubated
at
37 ◦C
for
30
min
and
rinse
several
times
with
distilled
water
before
proceeded
to
light
microscope
evaluation
under
10×
magnification.
2.2.3.2.
Alkaline
phosphatase.
The
differentiation
of
osteoblasts
cells
was
evaluated
via
a
function
of
alkaline
phosphatase
activity.
After
each
culture
period,
the
media
were
gently
removed
and
rinsed
with
PBS.
The
extract
buffer
(100
␮L)
was
added
to
each
well.
The
aliquot
was
incubated
with
the
substrate
of
p-Nitrophenyl
Phosphate
(pNPP;
110
␮L)
containing
0.1
M
aminopropanol.
After
incubating
at
37 ◦C
for
15
min,
NaOH
(0.1
M;
900
␮L)
was
added
to
stop
the
reac-
tion.
ALP
activity
was
measured
by
monitoring
the
color
change
of
pNPP.
The
absorbance
was
measured
at
410
nm
wavelength
using
Thermospectronic
Genesis
10
UV-vis
spec-
trometer.
Total
protein
was
determined
using
BCA
assay.
ALP
enzymatic
activity
was
normalized
to
total
protein.
3.
Results
3.1.
Thin
film
characterization
results
3.1.1.
X-ray
photoelectron
spectrometer
(XPS)
We
found
that
the
binding
energies
of
Ti
2p,
O
1s,
Cu
2p,
Co
2p,
Pd
3d
and
Ta
4d
shift
from
the
reference
[11].
Figs.
2
and
3
rep-
resent
the
XPS
spectrum
of
Ti
2p
at
binding
energy
457.55
eV
and
463.23
eV
and
O
1s
at
528.81
eV
corresponds
to
binding
energies
for
TiO2[12,13].
3.2.
Biocompatibility
test
results
After
the
SaOS-2
cells
were
thawed
from
cryopreserve
vial,
the
cells
were
resuspended
in
fresh
medium
then
incubated
in
the
cell
culture
incubator
supplied
with
5%
CO2,
95%
O2at
37 ◦C.
At
5
days
after
thawing
process,
the
photo
was
taking
at
con-
fluency
around
90%.
It
is
shown
epithelial
like
cell
morphology
with
adherent
on
plastic
surface
(culture
container)
(Fig.
4).
Ti 2p1/2
Ti44Zr10Pd10Cu14Co15
Ti44Zr10Pd10Cu14Co19
Ti44Zr10Pd10Cu6Co23
Ti-6AI-4V
470
465
460
Binding energy (eV)
Intensity (a.u.)
455
450
Ti 2p3/2
Fig.
2
–
XPS
spectrum
of
Ti
2p
at
binding
energy
at
457.55
eV
and
463.23
eV.
O 1s
Ti44Zr10Pd10Cu14Co15
Ti44Zr10Pd10Cu10Co19
Ti44Zr10Pd10Cu6Co23
Ti-6AI-4V
440
435
430
Binding energy (eV)
Intensity (a.u.)
425
Fig.
3
–
XPS
spectrum
of
O
1s
at
528.81
eV.
3.2.1.
Cell
proliferation
The
MTT
assay
is
a
colorimetric
assay
technique
utilizing
the
ability
of
metabolically
active
cells
that
reduce
a
yellow
tetrazolium-based
compound
to
a
purple
formazan
product.
The
number
of
living
cells
in
the
culture
is
directly
related
to
the
quantity
of
formazan
product,
which
is
measured
by
the
absorbance
at
570
nm
of
wavelength.
After
cells
were
exposed
to
Ti-based
MG
thin
film
for
3
days,
5
days
and
7
days,
there
was
no
significant
difference
compared
to
the
glass
substrate

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251
Fig.
4
–
The
SaOS-2
cells
morphology
and
adherent
at
5
days
after
thawing
process.
MTT assay
Ti-6AI-4V
Ti44Zr10Pd10Cu6Co23Ta
7
Ti44Zr10Pd10Cu10Co19Ta7
Ti44Zr10Pd10Cu14Co15Ta7
Cell number
300000
200000
100000
0
Day 3
Day 5
Day 7
Fig.
5
–
The
MTT
assay
of
SaOS-2
cells
on
Ti-based
MG
thin
film.
Fig.
6
–
The
Alizarin
red
staining
result
in
negative
control
samples.
and
Ti–6Al–4V
control,
suggesting
that
Ti-based
MG
thin
film
was
not
toxic
to
SaOS-2
(Fig.
5).
3.2.2.
Cell
differentiation
3.2.2.1.
Alizarin
red
staining.
The
Alizarin
red
staining
result
in
negative
control
samples
clearly
revealed
that
no
calcium
mineralization
had
been
detected
(Fig.
6).
Alizarin
red
staining
in
Ti–6Al–4V
and
3
novel
metallic
glass
alloys,
the
results
are
shown
in
Fig.
7.
The
alkaline
phos-
phatase
study
results
are
shown
in
Fig.
8.
Alizarin
red
staining
results
was
analyzed
using
Image
J
program
(set
color
threshold
as
RGB:
200,
120,
80)
to
quantify
the
calcium
mineralization.
The
results
are
demon-
strated
in
Tables
2
and
3.
The
Ti44 Zr10Pd10 Cu6Co23Ta7and
Ti44 Zr10Pd10 Cu6Co23Ta7shown
significantly
more
Ca
deposit
area
and
also
the
bigger
size
of
calcium
deposit
comparing
with
Ti–6Al–4V.
4.
Discussion
In
the
previous
report,
the
new
Ti-based
amorphous
alloy
composite
without
toxic
elements
has
been
synthesized
in
Ti–Zr–Cu–Pd
alloy
system
such
as
Ti40 Zr10Cu36 Pd14,
which
exhibit
high
corrosion
resistance
and
good
combination
of
strength
and
ductility,
implying
a
high
potential
as
bioma-
terials
[14].
We
developed
our
metallic
glass
based
on
this
combination.
We
decided
to
decrease
Copper
due
to
the
reported
cytotoxicity
of
Cu
as
released
from
Zr-based
BMG
dur-
ing
3T3
fibroblast
cell
line
tests
[15].
We
had
decided
on
many
potential
element
to
replace
Cu.
Finally,
the
additional
ele-
ments
that
we
chose
were
Ta
and
Co.
We
selected
Ta
because
it
is
believed
to
be
a
more
effective
element
to
increase
corrosion
resistance
and
biocompatibility
of
metal
materials.
Qin
et
al.
added
Ta
into
Ti–Zr–Cu–Pd
alloy
and
found
higher
strength
and
better
plastic
deformation
comparing
with
Ti–Zr–Cu–Pd
base
alloy
[16].
Another
element
that
we
chose
is
Cobalt.
Louzguine
et
al.
described
the
effect
of
the
addition
of
5
at.%
Cobalt
to
replace
Copper
on
the
mechanical
properties
and
stability
of
the
supercooled
liquid
of
Cu60Zr30 Ti10 amorphous
alloy.
They
concluded
that
the
addition
of
Copper
to
Cu-based
metallic
glass
improved
the
mechanical
properties,
stabilizes
the
supercooled
liquid,
increased
Young’s
modulus
and
com-
pressive
strength
of
the
alloy
[17].
XPS
spectra
suggested
that
on
the
surface
of
all
samples
(Ti–6Al–4V
and
series
of
novel
amorphous
alloy)
demonstrated
oxide
of
Ti.
The
oxidation
of
each
element
depends
on
“the
Standard
Electrode
Potential
(SEP),
which
is
evaluated
relative
to
the
standard
oxidation
of
hydrogen
gas,
measures
the
ability
of
the
metal
atoms
to
get
oxidized”
[18].
A
lower
SEP
corresponds
to
an
easier
oxidization
reaction.
The
mechanism
for
oxidation
is
still
being
explored
in
the
field
of
bulk
metallic
glass
due
to
the
fact
that
the
oxidation
behaviors
are
combinations
of
various
factors.
For
instance,
Oh
et
al.
[19]
reported
the
phase
separation
that
occurred
in
Cu43Zr43 Al7Ag7bulk
metallic
glass
in
which
gold
and
silver
formed
nanometer
level
localized
cluster.
The
copper
cluster
would
readily
oxidize
in
ambient
according
to
SEP.
However,
the
standard
potential
could
not
directly
relate
to
the
oxide
observation
that
was
created
during
the
synthesis
because
of
elevated
temperature
and
environ-
ment.
In
this
report,
the
detection
of
oxides
could
not
be
directly
related
to
SEP.
The
thin
oxidized
layer
was
created
at
high
temperatures
during
the
synthesis,
and
consequently
the
film
became
protective
film
at
room
temperature
after
the
syn-
thesis.
When
Ti–6Al–4V
and
Ti44 Zr10Pd10 Cu6+xCo23−xTa7(x
=
0,
4,
8)
were
exposed
to
air
and
investment
mold
material
dur-
ing
casting,
oxidation
layer
formed.
Similar
XPS
results
were

252
j
m
a
t
e
r
r
e
s
t
e
c
h
n
o
l
.
2
0
1
8;7(3):248–253
Fig.
7
–
The
Alizarin
red
staining
results
of
SaOS-2
cells
on
Ti-based
MG
thin
film.
Alkaline phosphatase study
Ti-AI-4V
Ti44Zr10Pd10Cu6Co23Ta
7
Ti44Zr10Pd10Cu10Co19Ta7
Ti44Zr10Pd10Cu14Co15Ta7
Normalize ALP activity (%)
Time
600
400
200
0
Day 1
Day 5
Day 10
Fig.
8
–
The
alkaline
phosphatase
study
results.
previously
reported
on
Ti–Zr–Pd–Cu–Sn
BMG
systems
[20].
The
XPS
also
confirmed
that
the
thin
film
created
by
FCAV
tech-
nique
contained
all
elements
the
same
as
their
novel
alloy
ingots.
Copper
containing
crystalline
alloy
and
amorphous
alloy
has
been
reported
about
cell
toxicity.
Buzzi
et
al.
[15]
reported
cytotoxicity
to
fibroblast
cell
line
from
Zr
based
amorphous
alloy
containing
Cu
(Zr58Cu22 Fe8Al12).
Elshahawy
et
al.
[21]
stated
that
Cu
released
from
gold
alloys,
which
are
commonly
used
as
fixed
prosthodontic
restorations,
show
evidence
of
a
high
cytotoxic
effect
on
fibroblast
cells.
In
contrast,
some
pre-
vious
research
did
not
demonstrated
negative
effects
of
copper
containing
alloys
to
the
cells
[22]
and
there
were
compatible
with
results
of
our
study.
The
explanation
of
this
issue
is
the
formation
of
TiO2that
developed
on
the
surface
of
novel
Ti
based
amorphous
alloy
which
contain
Ti
44
at.%.
The
TiO2has
been
reported
about
ability
to
provide
good
biocompatibility
and
bactericidal
effect
[23].
It
may
conceal
the
copper
from
direct
contact
to
the
cells
or
decrease
the
copper
ion
release
into
the
cell
growth
medium
to
the
optimum
level.
In
addi-
tion,
our
novel
alloy
compositions
contain
lower
amount
of
copper
than
the
alloy
previously
reported,
then
may
lead
to
less
toxicity
from
copper.
Some
of
previous
study
reported
about
biocompatibility
and
mechanical
property
of
the
toxic
free
amorphous
alloy.
Zhu
et
al.
[24]
have
developed
amorphous
alloy
with
com-
ponent
of
Ti40 Zr10Cu40−xPd10+x(with
x
=
0,
2,
4,
6,
8
and
10).
They
found
good
glass
forming
ability,
compressive
strength,
Young’s
modulus
and
an
elastic
elongation.
They
suggested
that
their
series
of
new
alloy
were
suitable
for
application
to
biomaterials.
Oak
conducted
human
cell
test
(SaOS-2)
on
Ti45 Zr10Pd10 Cu31Sn4.
They
found
results
of
good
biocompat-
ibility
and
glass
forming
ability
[25,26].
However,
due
to
the
Table
2
–
Quantitative
analyzation
of
Alizarin
red
staining
using
Image
J
program.
Uncoated
cover
glass
Ti–6Al–4V
Ti44 Zr10Pd10 –Cu6Co23Ta7Ti44Zr10 Pd10–Cu10 Co19Ta 7Ti44Zr10 Pd10–Cu14 Co15Ta 7
Total
area
(pixels)
0
41,533
±
25,726
63,936
±
12,569
78,721
±
23,853
188,047
±
34,563
Average
size
(pixels)
0
96.3
±
18.6
103.7
±
5.3
310.5
±
27.6
398.9
±
106.5

j
m
a
t
e
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r
e
s
t
e
c
h
n
o
l
.
2
0
1
8;7(3):248–253
253
Table
3
–
The
comparison
of
area
and
size
of
calcium
deposit
between
Ti–6Al–4V
and
the
novel
alloys.
Ti44 Zr10Pd10 –Cu6Co23Ta7Ti44Zr10 Pd10–Cu10 Co19Ta 7Ti44Zr10 Pd10–Cu14 Co15Ta 7
Ti–6Al–4V
Total
area
P
0.03
P
0.006*P
<
0.0001*
Average
size
P
0.27
P
0.0001*P
<
0.0001*
∗Significant,
p
value
<
0.05.
nature
of
novel
material,
it
is
impossible
to
find
a
previously
match
amorphous
alloy
study
to
compare
with
current
study
results.
In
our
study,
after
cells
were
exposed
to
all
novel
amor-
phous
alloy
samples
for
3
days,
5
days
and
7
days,
there
was
no
significant
difference
compared
to
the
glass
substrate
and
Ti–6Al–4V
control,
suggesting
that
Ti-based
MG
thin
film
was
not
toxic
to
SaOS-2.
5.
Conclusion
A
novel
Ti-based
amorphous
alloy
Ti44 Zr10Pd10 Cu6+xCo23−xTa7
(x
=
0,
4,
8)
demonstrated
biocompatible
characteristic
to
osteoblast
like
cells
(SaOS-2).
No
cytotoxic
was
found
during
the
cell
culture
and
proliferation.
A
novel
amorphous
alloy
thin
film
shown
supporting
of
the
osteogenic
differentiation
process
of
SaOS-2
cells
and
the
differentiation
was
even
bet-
ter
than
control
Ti–6Al–4V.
All
results
point
out
that
the
novel
Ti
based
amorphous
alloy
has
potential
for
using
as
a
new
coating
for
biomedical
application.
Conflicts
of
interest
The
authors
declare
no
conflicts
of
interest.
r
e
f
e
r
e
n
c
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