Materials Today Communications 33 (2022) 104563

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Materials Today Communications

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Ion implantation of 109Ag stable isotope as a tracer in SS316L biomedical

implant for failure detection
Bharti Malvi a, Ramesh Chaudhari b, Balasubramanian C c, d, Ashutosh Kumar b, Asokan K e, g,
Swagat Das f, Manas Paliwal f, Superb K. Misra a, *
a
Materials Engineering, Indian Institute of Technology Gandhinagar, Gujarat, India
b
School of Arts and Sciences, Ahmedabad University, Gujarat, India
c
Institute for Plasma Research, Gandhinagar, Gujarat, India
d
Homi Bhabha National Institute, Mumbai, India
e
Material Science Division, Inter University Accelerator Centre, New Delhi, India
f
Indian Institute of Technology Kharagpur, West Bengal, India
g
Department of Physics & Centre for Interdisciplinary Research, University of Petroleum and Energy Studies-Dehradun, 248007, India

A R T I C L E I N F O A B S T R A C T

Keywords: Stainless steel alloys are widely used for biomedical applications ranging from surgical tools to biomedical
Ion implantation implant prostheses. Apart from surgical failure, surface degradation (wear and corrosion) leading to poor
Tracers osteointegration of materials are a major cause of the failure of these bioimplants. Detection of the implant
Biological implants
failure, once it is placed inside the body, is very difficult and not so accurate. In this research, high-energy ion
Failure detection
Stable isotopes
implantation is used to develop a tracer embedded alloy. In this novel approach, a stable isotope of silver (109Ag)
SS316L is embedded as a tracer in SS316L to detect and monitor its degradation. Accelerated and normal static im­
mersion tests were performed in 30 % H2SO4 and simulated body fluid (SBF) to represent the degradation of the
implant. These results show a correlation between the isotopic tracer releases with material degradation. Elec­
trochemical corrosion tests demonstrate the ion-implanted specimens exhibit similar corrosion resistance as the
bare specimens. In-vitro cytotoxicity assays indicated no significant difference between the cell viability on
SS316L and 109Ag ion-implanted samples. The MG-63 osteoblast cells cultured on 109Ag ion-implanted SS316L
show a significant enhancement in osteocalcin production compared to SS316L.

1. Introduction routine check-ups and radiographic evaluation for any infections or

Medical grade austenitic stainless steel SS316L (ASTM A240 grade
316L) is commonly used for manufacturing biomedical implants (pros­
thesis and/or trauma implants) and surgical tools due to their corrosion
resistance, reasonable mechanical strength, and low cost [1]. Metallic
biomedical implants are load-bearing implants and are thus subjected to
cyclic loading in a corrosive biological environment. This condition
leads to fretting wear, sliding wear, and fatigue resulting in crack
initiation, propagation, and early implant failure. Considering the crit­
icality of the implants, it is very important to determine their working
life. Over time, the release of metal ions and wear debris from the
implant causes infections, implant loosening, irritation in adjacent tis­
sues, and ultimately leads to implant failure [2]. Monitoring the con­
dition of implants after surgery is crucial and difficult. It includes
wear-related particles for years [3]. Early failure detection of the
implant failure would save a lot of pain and medication.
Unger et al. [4] used non-invasive resonance frequency monitoring
for early detection of the implant healing process. In this technique, the
lower initial output is gained from a poorly fitted implant and a
well-fitted implant shows higher resonance frequency (RF) output and
shorter simulated healing period. This technique might have a high
signal-to-noise ratio in-vivo [4]. A study by Chiva et al. [5] suggested
implant failure detection by electrophoresis of joint fluid proteins, but
this is an invasive technique and needs to be correlated with radio­
graphic evaluation. The use of macromolecular imaging agents to detect
the inflammation induced by polymethyl methacrylate (PMMA) parti­
cles in a murine peri-implant osteolysis model has also been performed
[6]. It can detect particle-induced inflammation before the development

* Corresponding author.
E-mail address: smisra@iitgn.ac.in (S.K. Misra).

https://doi.org/10.1016/j.mtcomm.2022.104563
Received 9 June 2022; Received in revised form 31 August 2022; Accepted 26 September 2022
Available online 28 September 2022
2352-4928/© 2022 Elsevier Ltd. All rights reserved.
B. Malvi et al.
Fig. 1. Schematic of tracer inducing and releasing mechanism. The biomedical implant surface is ion implanted with high energy 109Ag ions on the top surface.109Ag
ions is embedded in the sub surface of the implant material and gets released upon implant material degradation.
of anatomically detectable bone loss. The challenge in this technique is
that it is an imaging technique and involves constant monitoring of dye
in the system. The limitations of these early failure detection techniques
lead to the requirement of developing a non-invasive, safe, and viable
early failure detection technique for biomedical implants. The concen­
tration of metal ions in biological fluids could also be utilized as an in­
dicator of the performance of biomedical implants. Nicolli et al. [7]
measured the concentration of Co, Cr, As, Ni, Mn Cd, Cu, Be, Bi, Pb, Hg,
Se, Tl, Zn, and V in the blood and urine samples of people with a hip
prosthesis. Samples were taken after an average of 7.6 years of hip
replacement surgery. A significant increase of the urinary concentra­
tions were confirmed for Co, Cr, and As, while the other metals tested
showed concentrations within the SIVR (Italian Society of Reference
Values) recommended range [7,8]. A correlation between the change in
the inclination angle of the acetabular cup and metal ion release is
established [8]. The released Cr and Ni metal ions from metallic im­
plants were quantified by atomic absorption spectroscopy to correlate
with implant failure [9]. This approach lacks a strong diagnostic accu­
racy, and these metal ions are already present in the body as trace
amounts. Bauer and Shanbhag [10] studied how high metal ion levels
may help recognize patients with unexpected wear of metal-on-metal
inserts, but, wear measurements by existing methods were not accu­
rate to provide a correlation between metal ion release levels with the
implant failure. In any case, the challenge is to distinguish markers
explicitly released by wear that are not raised by different conditions in
patients. Therefore, a marker/tracer is needed to detect the implant
failure at the preliminary stage and quantify the implant failure rate not
only on the mating surfaces of the implant but also in the bulk.
In this study, a stable isotopic tracer of silver (109Ag) is introduced in
SS316L alloy by high energy ion implantation to detect the surface as
well as bulk failure. In the adult human body, the background total silver
concentration in blood is less than 1 µg L− 1 [11] For all forms of silver,
the permissible exposure limit (PEL) according to Occupational Safety
and Health Administration, the Mine Safety and Health Administration,
as well as the National Institute for Occupational Safety and Health,
suggest a permissible exposure limit (PEL) of 0.01 mg/m3 [12]. Fig. 1
shows the mechanism of tracer inducing and releasing upon implant
failure. The biomedical implant surface is ion implanted with high en­
ergy 109Ag ions on the top surface. The 109Ag ions are embedded in the
sub-surface of the implant material and expected to release when the
implant material degrades as shown in Fig. 1. Stable isotopes have been
extensively used as tracers in various environmental and geological
applications in the last couple of decades [13–15]. The natural abun­
dance of 109Ag is 48.16 % [16]. Silver is known to have low toxicity on
human cells and has antimicrobial properties at lower concentrations
[17–19]. Due to these qualities, silver has been used for artificial heart
valves and urinary tract catheters in the human body [20]. Sun et al.
[21] concluded that ion-implanted Ag+ enhances cell proliferation,
antibacterial and mechanical properties of Ti6Al4V alloy. The ion im­
plantation method of inserting tracers provides better depth control,
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Materials Today Communications 33 (2022) 104563
Table 1
109
Ion implantation parameters used to embed Ag on SS316L
substrate.
Ion Beam Energy 100 MeV
Ion Beam Current 1 pn A
Ion Fluence 1 × 1013 ionscm-2
Ion Chamber Pressure 9 × 10− 6 mbar
Ion charge state Ag+7
Temperature 25 ◦ C
allows better diffusivity, facilitates co-implantation of two or more ion
species, and can be done post manufacturing of the implant [22]. The
energy of ions is decided by the mass of an impacting ion and acceler­
ating voltage [23].
A coating containing the tracers will not serve the purpose of implant
material failure detection. For example, if the tracer is embedded in the
coating, the tracer release upon coating degradation will only signify the
failure of the coating; the SS316L implant will remain intact. Therefore,
a deeper layer of ion-implanted 109Ag ions in SS316L will facilitate 109Ag
ions-controlled release as a tracer. When the 109Ag ion-implanted
SS316L (109Ag-SS316L) alloy starts to degrade, the 109Ag ions will be
released with other host metal ions (viz. Fe, Cr, Ni, Mn, and Mo), indi­
cating failure (degradation) of the 109Ag-SS316L material. Silver (109Ag)
concentration in the blood, urine or synovial fluid will be a direct
indication of implant degradation. In this research, an immersion test of
109
Ag ion-implanted SS316L alloy is carried out in two different me­
diums to demonstrate the implant degradation. Static immersion tests
are done in 30 % H2SO4 and in vitro static immersion tests are performed
in simulated body fluid (SBF).
2. Experimental details
2.1. Tracer deposition through ion implantation
Medical grade SS316L samples of 1 cm × 1 cm × 0.2 cm were me­
chanically polished using silicon carbide emery papers of grade 220,
400, 600, 800, 1000, and 1200 on one side (area 1 cm × 1 cm). Final
polishing was done using alumina suspension to obtain a mirror finish
and after that, samples were cleaned with acetone in an ultrasonic bath
for 15 min. The polished surface of the samples was ion-implanted with
a stable isotope of silver, 109Ag ions at the 15UD Tandem Accelerator
facility at Inter-University Accelerator Centre (IUAC, New Delhi). The
ion implantation parameters are listed in Table 1. For ion implantation,
the SS316L samples were mounted on the copper ladder equipped with a
stepper motor for vertical movement. The focused ion beam of ~ 1 mm
spot diameter was used to continuously scan the complete area (1 cm2)
of the sample. The simulation of ion implantation depth and ion distri­
bution profile of 109Ag in SS was calculated by using the SRIM/TRIM-
2008 software [24].
B. Malvi et al.
2.2. Characterization
The surface morphology of SS316L and 109Ag-SS316L samples were
studied using SEM-EDS (Carl Zeiss, Germany). XRD analysis (Bruker D8,
Cu-Kα, 40 kV, 30 mA, 2θ = 20–100◦ ) was performed and the diffraction
pattern was matched with the ICSD database. The chemical composition
was measured with the help of inductively coupled plasma optical
emission spectroscopy (ICP-OES) after complete digestion using a mi­
crowave sample preparation system (Perkin-Elmer, Titan MPS). XPS
measurements were performed on the cross-section of both samples to
confirm the presence of implanted 109Ag ions. The microhardness of the
samples was measured using Vickers hardness testing equipment (500 gf
and 15 s). At least 20 indentations were made for each sample at random
positions. (n = 3). The roughness of the samples was measured using
Bruker Dektak XT stylus profilometer using a stylus radius of 2 µm,
5 mm scan length, and 3 mgf for 120 s. A minimum of five scans was
done for each sample (n = 3). The water contact angle and surface en­
ergy measurements were performed on the samples using contact Angle
(Kruss, DSA 25) under ambient conditions using the sessile drop method.
Deionized water and di-iodomethane were used to measure the surface
energy. Water contact angle measurements were conducted on three
different sites of three different specimens (n = 3) for statistical
accountability.
109
2.3. Ag tracer release studies
To demonstrate the concept of tracer release as a function of implant
material degradation, the samples were kept in 30 % H2SO4 for 72 h (at
room temperature). The accelerated failure of the implants in presence
of the acid was correlated with the release of 109Ag ions. Aliquots were
taken at specific time points, and the released metal ion concentrations
were measured using ICP-OES (PerkinElmer Avio 200). Tracer release
measurements were also performed in simulated body fluid (SBF). SBF is
a commonly used media for implant testing due to its ionic composition
being similar to blood plasma [25]. The samples were ultrasonically
cleaned, weighed, and immersed in freshly prepared 50 mL SBF. The
samples were kept in a shaker incubator at 37 ◦ C for seventy days. Ali­
quots were taken at various time points and analyzed using ICP-MS
(PerkinElmer NexION 2000) for the estimation of tracer (109Ag) and
metal ions released. SS316L and 109Ag-SS316L samples were retrieved
after 70 days and washed with deionized water to remove any salt
deposition. The samples were then analyzed in SEM and EDX. For the
limit of detection (LOD) and lower limit of quantitation (LLOQ) deter­
mination, standards and samples were prepared in the respective matrix.
Calibration curves were linear meeting the acceptance criteria of R2
values greater than 0.997 [26].
2.4. Electrochemical corrosion test
Electrochemical impedance spectroscopy (EIS) and potentiodynamic
polarization (PDP) tests were performed on SS316L and 109Ag-SS316L
samples in 50 mL SBF as an electrolyte at ambient temperature (Met­
rohm Autolab, Nova 2.1 analyst software). The pH of the electrolyte was
maintained at 7.4. A three-electrode corrosion experiment setup was
used for corrosion testing where samples were used as a working elec­
trode with only the top area (1 cm2) exposed for reaction. A saturated
calomel electrode (SCE) was used as a reference electrode and platinum
mesh was used as a counter electrode. To maintain the stability of the
electrochemical system, the open-circuit potential (OCP) was measured
for 14 h, after a stable potential was achieved for at least 30 min
(± 0.002 V). The EIS spectra were measured for a frequency range of
100 kHz to 0.01 Hz with 10 points per decade and at the amplitude of
0.01 V. A potentiodynamic polarization test was performed at the scan
rate of 0.005 V s− 1. The electrode potential, also known as the scanning
potential, was increased from − 0.5 V to 1.2 V vs OCP. In all cases, the
contact area was 1 cm2. The corrosion current density (icorr), corrosion
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Materials Today Communications 33 (2022) 104563
potentials (Ecorr), and corrosion rate were estimated using linear fit and
Tafel extrapolation to the cathodic region of the polarization curve after
the corrosion tests were completed for both SS316L and 109Ag-SS316L.
Three samples were analyzed to ensure the repeatability of the results.
The retrieved samples were analyzed in SEM for pitting corrosion
morphology after rinsing with deionized water and air drying.
2.5. Cell culture studies
Osteoblast-like MG-63 cells were obtained from National Centre for
Cell Science (NCCS), India. MG-63 cells were maintained in minimal
essential medium (MEM, Hi-Media) without phenol red supplemented
with 10 % fetal bovine serum (FBS, Gibco) and 1 % penicillin/strepto­
mycin (Hi-Media) under the humidified condition at 37 ◦ C and 5 % CO2.
At confluency, cells were harvested using trypsin-EDTA solution (Hi-
Media) and passaged at a 1:4 split ratio.
2.5.1. Cell viability assay
The propidium iodide (PI) uptake assay was carried out according to
the method described by Crowley et al. [27]. Briefly, 105 cells per well in
a 12 well plate were seeded along with samples and incubated at 37 ◦ C
and 5% CO2 for 24 h and 48 h. After the exposure, the supernatant was
collected and cells were harvested using trypsin–EDTA solution. Cells
were resuspended in PI solution (10 μg mL− 1) and incubated in dark for
30 min. Cell death was measured using a flow cytometer (BD FACS
Calibur™). The cytotoxic effect of the sample was also measured by
colorimetric assay, which determines the ability of the live cell to reduce
the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetreazoliym bromide
(MTT) dye. Briefly, 3 × 104 cells were seeded per well in 24 well plates,
and samples were placed in wells and incubated at 37 ◦ C and 5 % CO2
for 24 h and 48 h. Once the treatment was over, supernatant was
removed and replaced with 0.5 mg mL− 1 MTT solution in MEM media
without phenol red and incubated for 4 h. Formulated formazan crystals
were solubilized in DMSO and absorbance was measured at 570 nm
using a UV-Vis microplate reader (Synergy-HT, BioTek).
2.5.2. Alkaline phosphatase (ALP) activity
MG-63 cells were cultured on samples in 48 well culture plates at
1 × 104 cells per well for 7 days. Further, the cell culture medium was
replaced by a cell culture medium containing 1 × 10− 7 mM vitamin D.
After 48 h of stimulation, the cells were harvested and the pellet was
washed twice with PBS. Cell lysates were prepared by vortexing 500 μL
MQ water and 25 μL 1 % Triton X-100, and homogenised via sonifica­
tion [28]. The total protein content of cells was determined using
Pierce™ BCA protein assay kit (Thermoscientific). ALP activity was
assayed using an alkaline phosphatase detection kit (Sigma) as per the
manufacturer’s protocol depending on the conversion of p-nitrophenyl
phosphate to p-nitrophenol (coloured product). The colour change was
measured spectrophotometrically at 405 nm using a UV-Vis microplate
reader (Synergy-HT, BioTek). ALP level was normalized to the total
protein content of cells.
2.5.3. Osteocalcin production
MG-63 cells were cultured on samples in 48 well culture plates at
1 × 104 cells per well for 7 days. Thereafter, the cell culture medium was
replaced by a cell culture medium (MEM) containing 1 × 10− 7 mM
Vitamin D [28]. After 48 h stimulation, cell culture media was collected
and osteocalcin production was measured using the human osteocalcin
instant ELISA (Sigma) as per the manufacturer’s protocol.
2.6. Statistical analysis
All analyses were done in triplicates (n = 3) for statistical account­
ability. Statistical differences were calculated by unpaired t-test and
two-way ANOVA using GraphPad PRISM. In all cases the ‘p’ value or
probability value * = p < 0.05, were considered significant, and the
B. Malvi et al. Materials Today Communications 33 (2022) 104563

Fig. 2. SRIM plots generated after 109Ag ion implantation on SS316L substrates showing (a) 109Ag depth profile, (b) 109Ag ion distribution, (c) collision events, and
(d) vacancies generated by 109Ag ion implantation.

Fig. 3. (a) SS316L and 109Ag-SS316L samples, (b) XRD pattern for SS316L and 109
Ag-SS316L samples, SEM images of (c) SS316L (d) 109
Ag-SS316L, with the
respective water contact angle in inset. (Scale bar: 200 nm).

4
B. Malvi et al.
Fig. 4. (a) Vickers hardness (VHN) and (b) Water contact angle comparison for SS316L and 109Ag-SS316L samples. The box represents the interquartile range of the
data (25–75 %), with the median represented by the horizontal line in the box. The lower whiskers represent the minimum value and the upper whisker represents
the maximum value, excluding the outliers. (**p ≤ 0.01, ****p ≤ 0.0001).
results were expressed as mean ± standard deviation (SD), where
** = p ≤ 0.01, *** = p ≤ 0.001, and **** = p ≤ 0.0001.
3. Results and discussion
3.1. Tracer implantation
Ion implantation was used in this study to embed a stable isotopic
109
Ag tracer into the SS316L substrate. The enriched stable isotopes are
very costly and not easily available; therefore, a very small amount of
tracer is embedded. Table 1 shows the ion implantation parameters. At a
small dose of 1 × 1013 ionscm-2, the stoichiometry of the subsurface is
not much affected. However, the 109Ag is still detectable using ICP-MS
when it comes out of the system upon implant failure. Silver ions are
found to enhance cell adhesion and antibacterial effects when embedded
via ion implantation, therefore, 109Ag ions are used in this study [21].
Although the natural abundance of 109Ag (48.18 %) is similar to the
natural abundance of 107Ag (51.82 %) isotope, 109Ag is still differen­
tiable over the background silver present in the human body at trace
level using ICP-MS. In the adult human body, the background total silver
concentration in blood is less than 1 µg L− 1 [11]. At such lower back­
ground concentration, the 109Ag ions released from the 109Ag-SS316L
can be detected and tracked using ICP-MS and the advantage of using a
particular isotope is to get an absolute idea of the implant degradation.
High energy ion implantation of 109Ag ions was carried out to embed the
ions beneath the top surface of SS316L. Depth profile and ion distribu­
tion plot of 109Ag ions in 109Ag-SS316L was created using the SRIM
software and are shown in Fig. 2 [29]. Owing to the high ion beam
energy (100 MeV), the 109Ag ions were embedded deep into the sample,
with an ion stopping range of 6.54 µm from the surface. Fig. 2b indicates
the maximum 109Ag ion concentration to be accumulated at a depth of
4–7 µm from the irradiated surface. The ion irradiation collision event
and the number of vacancies generated due to ion implantation are
shown in Fig. 2c–d. These defects have been shown to contribute to­
wards increasing the hardness after ion implantation [30]. The ions
implantation at a shallow depth of a few hundred nanometers may not
be able to represent the actual implant failure. As illustrated in Fig. 1, a
tracer implanted at shallow depth will only suggest the surface wear and
will fail to trace the crack initiation and propagation in depth. Therefore,
a range of depth of about 4–7 µm is chosen for this study. 109Ag ion
implantation causes the solid surface to be highly concentrated with
non-equilibrium defects and radiation-induced vacancies in the sub­
surface layers [31].
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Materials Today Communications 33 (2022) 104563
3.2. Characterization of tracer implanted substrates
The composition of SS316L was confirmed using EDS and ICP-OES
(Supplementary Table S1). Fig. 3a shows the images of SS316L sub­
strate and 109Ag-SS316L samples. XRD pattern of SS316L and 109Ag-
SS316L is shown in Fig. 3b and matched with the ICDD database for γ
austenite stainless steel (ICDD:00-031-0619) and silver (ICDD:00-004-
0783). Additional Ag peaks were not found in the XRD pattern of 109Ag-
SS316L sample, due to the low concentration of silver. No peak shift and
peak profile change for 109Ag-SS316L was observed after 109Ag im­
plantation (Supplementary Table S2, Supplementary Fig. S1). To
confirm the presence of implanted 109Ag ions, XPS analysis was per­
formed on the cross section of implanted sample. The surface was
sputtered up to 5 µm depth and the signals were recorded. (Supple­
mentary Fig. S7) The intensity of 109Ag signal was very weak as the
number of implanted 109Ag ions are very small (1 × 1013 ionscm-2).
Surface morphology of SS316L and 109Ag-SS316L samples was observed
using SEM and is shown in Fig. 3c–d along with their respective water
contact angle in the inset. The polished surfaces show the scratches from
the polishing paper and the inclusion defects on the surface. These de­
fects and inclusions act as corrosion initiating sites and are prone to
rapid dissolution. Fig. 4a–b shows the change in Vickers hardness (VHN)
and water contact angle of SS316L before and after 109Ag ion implan­
tation, respectively. There was a significant (****p < 0.001) increase in
the mean VHN value of SS316L after ion implantation, wherein the VHN
value increased from 149 ± 6 to 154 ± 7. The increase in hardness after
ion implantation for SS316L could be attributed due to lattice defor­
mation and residual compressive stresses generated during ion implan­
tation [21,32]. The number of vacancies caused due to high energy ion
bombardment into the SS target is shown in Fig. 2d. The generation of
displacements and vacancies may lead to point defects and dislocations.
High energy ion implantation has been shown to induce dislocation into
the target material [33]. The surface roughness of SS316L decreased
from 282 nm to 184 nm after ion implantation. It has been shown that
the surface roughness of a smooth surface increases, and for a rough
surface, the roughness decreases after ion implantation (ion fluence
2 × 1017 ions cm− 2) [23]. The ion implantation generated dislocations
and inclusions in the polycrystalline microstructure of SS316L and
increased the surface roughness [34]. The water contact angle of the
surface of bare SS316L was 57.6◦ ± 5.6◦ and there was a 21.8 % increase
in water contact angle after ion implantation. The increase in contact
angle can be attributed to decreased interfacial tension between water
and sample surface leading to lower surface energy. The trend reported
in this study is consistent with the literature [35,36] Different surface
oxides before and after Ag implantation could change water contact
angles. Typically, metal surfaces have high surface energy due to solid
B. Malvi et al. Materials Today Communications 33 (2022) 104563

Table 2 the literature for SS316L [39]. The total silver present in 109Ag-SS316L is
Surface characteristics for SS316L and 109Ag-SS316L implants. (Data presented as measured to be 0.101 ± 0.017 % by weight. Since only 109Ag isotope is
mean ± standard deviation and n = 3). embedded in SS316L, the concentration of Ag measured in ICP-OES is
Material Water Surface Vickers Average the concentration of 109Ag isotope. The limit of detection (LOD) for Ag in
contact energy (mN/ hardness (HV) roughness (nm) ICP-OES is determined as 0.02 mg L− 1 (ppm). The lowest limit of
angle m) quantification (LLOQ) is determined as 0.05 mg L− 1 (ppm) for Ag in the
SS316L 57.6◦ ± 5.6◦ 51.0 ± 4.0 148.9 ± 6.0 282.3 acid matrix. The Iso-corrosion curve for SS316L in mildly concentrated
109
Ag- 70.2◦ ± 6.1◦ 43.8 ± 3.2 154.5 ± 6.8 184.4 H2SO4 (> 20 % v/v) suggests a uniform attack of acid on SS316L at
SS316L
room temperature. Therefore, an acidic environment was used to test the
hypothesis of this study, i.e. using tracer release as a measure of material
bonding. Surface free energy, γ, is written as a component of surface failure. Fig. 5 shows the release of 109Ag with SS316L degradation in
internal energy, US, temperature, T, and surface entropy, SS 30 % H2SO4 for 72 h. The acid attack on the surface was instant and
uniform on SS316L, which resulted in exposing the implanted silver
γ = Us − TSs (1) from the sub-surface. Thus, causing silver concentration in the medium
to increase with the degradation of the 109Ag-SS316L implant. After 2 h,
where, degree of disorder in the surface (SS), increases with the
0.0015 ± 0.0004 mg of 109Ag was released in the medium (corre­
embedding of other elements in the surface. According to Eq. (1), if the
sponding to 0.024 mg L− 1), whereas for Fe the concentration was
ion implantation energy is higher than the interatomic bond energy
13.99 mg L− 1 corresponding to 0.84 ± 0.13 mg Fe release. Fig. 5b
(US), the surface energy will decrease after ion implantation [37].
shows that for almost all the elements, very less amount (~ 1.3 %) of
Change in surface roughness is a product of the changes induced in the
degradation occurred for SS316L during the first 6 h, which increased to
surface chemistry of the target and the atomic mass of the ion. The
25 % in 12 h, 80 % in 24 h and 95 % in the 36 h. The percentage
change in surface roughness and surface energy in the target is due to
degradation of SS316L and 109Ag-SS316L samples followed a similar
high energy (100 MeV in this case) and the higher mass of the impacting
109 trend increasing % release of Fe correlating with 109Ag. A rise in the
Ag ions. The mean surface roughness, surface energy, water contact
concentration of 109Ag is correlated with the dissolution or degradation
angle, and hardness for SS316L and 109Ag-SS316L are presented in
of total implant material. No significant difference is observed in the
Table 2. The cell adhesion and proliferation are enhanced with increase
degradation rate of SS316L and 109Ag-SS samples in 30 % H2SO4, with a
in water contact angle and decreased surface energy [38].
95 % confidence interval using an unpaired t-test. After 72 h in 30%
H2SO4, the samples were completely dissolved. For non-ion implanted
3.3. Tracer release study under accelerated acidic environment SS316L samples, no silver was detected. At the end of 72 h, 0.22
± 0.02 mg of 109Ag was released from the ion-implanted samples, cor­
The chemical composition of SS316L alloy measured using ICP-OES responding to 100 % of silver release and representing 0.09 % of the
and SEM-EDS is listed in Supplementary information Table S1. The total weight of the sample. In comparison, the Fe released in the acid
measured concentration of elements was within the range reported in

Fig. 5. (a) Metal and tracer release in mg, (b) % metal ion release in the percentage of total metal present in SS316L and 109Ag-SS316L samples in 30 % H2SO4 with
time (Paired t-test for individual elements indicate no significant difference in the mass of metal ion release except for 109Ag (***p = 0.0002).

6
B. Malvi et al. Materials Today Communications 33 (2022) 104563

Fig. 6. SEM images of alloy surfaces after corrosion for (a and b) SS316L and (c and d)109Ag-SS316L.

Fig. 7. (a) Metal and tracer release in µg (b) metal ion release in the percentage of total metal present in SS316L and 109Ag-SS316L samples in SBF for 70 days.
(Paired t-test for individual elements indicate no significant difference in the mass of metal ion release at each time point).

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B. Malvi et al. Materials Today Communications 33 (2022) 104563

Fig. 8. (a) Variation in open circuit potential with time, (b) Equivalent circuit diagram, (c) Nyquist plot, (d) Potentiodynamic polarization curve for SS316L and
109
Ag-SS316L in SBF, SEM morphology of (e) SS316L surface (f) 109Ag-SS316L surface with pits after potentiodynamic polarization.

medium was 150.7 ± 6.31 mg, which is 65.8 % of the total weight of the
samples or 100 % of total Fe present in the samples.
Fig. 6 shows the surface morphology of corroded SS316L and 109Ag-
SS316L samples after 6 h of immersion in 30 % H2SO4. The surface
elemental composition is analyzed by the point scan EDS of corroded
surfaces at different points. (Supplementary Fig. S6) SEM images show
the aggressive uniform attack of H2SO4 acid on the sample surface. Se­
vere pitting corrosion and intergranular corrosion are visible, as shown
in Fig. 6c–d. The corrosion damage on the individual grain is evident
since the grain boundaries are etched preferentially due to the carbides
present on the grain boundaries [40]. The presence of defects and in­
clusions on the surface acted as corrosion starting sites. These inclusions
are prone to rapid dissolution due to the presence of iron-rich phases
present in the inclusions. A study by Arwati and Sianipar [41] suggests
that the uniform corrosion rate increases with time for the immersion
time of five days in sulfuric acid and typical corroded surface
morphology contains local blasting of the metal surface, indicating the
local defects as the first corrosion starting sites. Once the corrosion re­
action starts, SS316L demonstrates general uniform corrosion instead of
localized corrosion [42]. However, the change in surface chemistry
during ion implantation includes the generation of defects and disloca­
tion [43]. Therefore, the crystallinity near the surface is compromised.
Fig. 6c–d shows the corroded 109Ag-SS316L surfaces, and the micro­
cracks generated. The presence of cracks indicates a brittle or amor­
phous phase, which is consistent with the literature [34].

8
B. Malvi et al.
3.4. Tracer release study under simulated body fluid (SBF)
The number of metal ions released during immersion of SS316L and
109
Ag-SS316L alloys is shown in Fig. 7. In general, SBF does not react/
dissolve metal/alloy, but it can be seen from Fig. 7 that a minimal
quantity is degrading in the form of released metal ions. The amount of
metal ion release is more in ion-implanted alloy samples as compared to
bare samples. The reason could be the rupture of passive oxide film due
to ion implantation. Since the ion implantation simulation demonstrated
that the maximum 109Ag ion distribution is about 4–7 µm inside the
surface, there is no silver present on the surface and that is why the silver
release from the ion-implanted samples is negligible, as shown in
Fig. 7a. However, the mass of metal ions released at particular time
points for SS316L and 109Ag-SS316L show no significant difference
(p > 0.05). Although the tracer release was detected in the acidic
environment, for the benign SBF media even after 70 days in SBF, the
tracer release was minimal. The percentage release of all the metal ions
(Fe, Cr, Ni, Mn, Mo) is very small. In Fig. 7b, it is clear that only 0.02 %
Fe was released from bare SS and 0.03 % Fe was released from 109Ag-
SS316L, which corresponds to 61.8 ± 20.4 µg and 63.3 ± 11.6 µg of Fe
released in SBF respectively. The limit of detection (LOD) for 109Ag in
ICP-MS is determined as 0.01 µg L− 1. The lowest limit of quantification
(LLOQ) is determined as 1 µg L− 1 for 109Ag in the SBF matrix. SEM
images of the samples after 70 days confirm the presence of nanosized
pits formed due to static immersion. (Supplementary Fig. S2). Silver
stable isotope (109Ag) embedded SS316L failure is detectable by quan­
tifying the 109Ag ion release. It has been shown in our previous studies,
that stable isotopes are detectable in concentrations as low as 1 µg mL− 1
by using ICP-MS techniques [15]. An absolute remark on implant failure
is drawn based on; negligible quantity of 109Ag tracer in the human body
and, the only source of 109Ag is the biomedical implant in which 109Ag is
embedded as a tracer.
3.5. Electrochemical corrosion behavior
Electrochemical impedance spectroscopy (EIS) was used to under­
take a quantitative assessment of uniform corrosion. EISanalysis is
performed to know the frequency response of SS316L and 109Ag-SS316L
using the Nyquist plot in the measured range of frequency. OCP was
measured for 14 h (Fig. 8a).
It is evident from Fig. 8a that the OCP of 109Ag-SS316L is slightly
shifted towards a positive potential as compared to SS316L but even­
tually stabilized as somewhat the same potential as SS316L. In terms of
the OCP fluctuations, the OCP of bare SS316L is comparatively smooth
and stabiles after 6 h with no potential spikes beyond that. It means a
stable oxide layer is formed on the surface. In 109Ag-SS316L, several
distinct potential spikes are visible in OCP, marked from G to N. This can
be linked to the deterioration and re-passivation of the oxide layer. The
extent of potential drop is most likely associated with the dissolution of
metal ions due to the deterioration of passive oxide layer from ion im­
plantation. Once the oxide film breaks, the potential decreases drasti­
cally and the upward shift in potential indicates re-passivation of the
oxide film [44].
After the OCP is stabilized, the frequency response of the sample is
measured. In the Nyquist plot, the real and imaginary part of the
impedance is plotted. Fig. 8b represents a semi-circular pattern of the
Nyquist plot which corresponds to the charge transfer-controlled
mechanism whereas a straight line at a 45◦ angle represents a
diffusion-controlled reaction mechanism. The diameter of the semi­
circular Nyquist plot is a measure of corrosion resistance of the material
[45]. In this case, the Nyquist plots show a corrosion reaction controlled
by a charge transfer mechanism. Nyquist plots for both the samples show
a similar pattern, however, the larger diameter of the semicircle of the
Nyquist plot for SS316L indicates high corrosion resistance and better
stability of passive film in SS316L as compared to 109Ag-SS316L [45].
The interface of electrode/electrolyte is simulated by employing an
9
Materials Today Communications 33 (2022) 104563
Table 3
Electrochemical parameters determined from the potentiodynamic polarization
test and EIS fitting (all potential values are with respect to saturated calomel
electrode (SCE); n = 3).
Sample SS316L 109
Ag-SS316L
Corrosion parameters from EIS measurements
Solution resistance Rs (Ω) 31.11 ± 0.94 38.14 ± 1.62
Polarization resistance Rp (Ω) 607.83 × 103 71.45 × 103
Corrosion parameters from potentiodynamic polarization
OCP (V) -0.02 -0.02
Ecorr (V) -0.25 -0.26
icorr (µAcm-2) 2.41 2.64
βa (mV) 10.45 2.00
βc (mV) 0.16 0.16
Polarization resistance Rp (Ω) 28.69 × 103 24.38 × 103
Epit (V) 0.44 0.43
Corrosion rate (mm/y) 0.028 0.030
equivalent electrical circuit model (EEC) (Randle Circuit) to quantify the
charge transfer and charge distribution within the double layer using
Nova 2.1 software. The observed values were inserted to fit the curves.
The behavior of the high-frequency impedance is related to the electrical
resistance of the electrolyte and is represented by “Rs” in the EEC. The
low and medium frequency impedance responses are related to the
charge distribution of the double layer (non-Faraday) and the Faraday
processes occurring at the electrode-electrolyte interface. In CEE, these
processes are quantified as constant phase element “CPE" and charge
transfer/polarization resistance "Rp" respectively. The CPE is due to the
non-uniform distribution of the charges at the oxide layer/electrolyte
interface and is related to the dielectric properties of the oxide and the
properties of the ionic species present at the oxide/electrolyte interface.
The stability of the oxide film is called "Rp" and exhibits resistance to
charge transport across the oxide film and oxide/electrolyte surfaces.
The values of Rs and Rp are listed in Table 3 and it is clear that
109
Ag-SS316L has lower polarization resistance resulting in higher
corrosion due to charge transfer.
Biomedical implants generally fails due to localized (both pitting and
crevice) corrosion, intergranular corrosion, dealloying, galvanic corro­
sion, stress corrosion cracking, corrosion fatigue and fretting corrosion
[46]. Potentiodynamic polarization (PDP) tests along with Tafel analysis
is a well-known electrochemical technique and is indicative of localized
corrosion behavior [47,48]. In Tafel analysis, a sample is subjected to a
typical potential scan of ± 25 mV with respect to the open circuit
voltage (OCP), and the current value produced is measured. The
potentio-dynamic polarization curves of both 109Ag ion implanted and
bare SS316L samples in SBF at a pH of 7.4 at room temperature are
shown in Fig. 8c. Table 3 shows all of the essential parameters measured.
The values for the polarization resistance are not same as the ones ob­
tained from EIS technique as represented in Table 3. This might be
because of the inherent distinction in both the techniques, and consis­
tent with the literature [49]. However, both the EIS and PDP method
confirmed that SS exhibit better polarization resistance as compared to
109
Ag-SS. Fig. 8c shows a typical anodic polarization curve for SS316L
(marked as P in Fig. 8c) and 109Ag-SS316L (marked as Q in Fig. 8c).
Anodic polarization curve is consisting of two different regions; a
cathodic (P1–P2 and Q1–Q2 in Fig. 8c) and an anodic region (P2 and Q2
onwards). The three distinct areas of the anodic part of the curve are, the
active region (P2–P3 and Q2–Q3), passive region (P3–P4 and Q3–Q4), and
transpassive region (P4–P5 and Q4–Q5). Corrosion potential (Ecorr) is the
potential at which the current density is the lowest, i.e., at points P2 and
Q2. At P2, SS316L shows a corrosion potential of − 0.256 V vs SCE, and
at point Q2 109Ag-SS316L exhibits the corrosion potential of − 0.268 V
vs SCE. In the active region (P2–P3 and Q2–Q3), the dissolution of both
the samples starts, and corrosion current density increases. At passiv­
ation potential, points P3 and Q3, the dissolution became kinetically
B. Malvi et al. Materials Today Communications 33 (2022) 104563

Fig. 9. Influence of 109Ag ion implantation on osteoblast like MG-63 cells survival and differentiation (a) A histogram of PI uptake for control, SS316L and 109Ag-
SS316L for 24 h and 48 h (b) Bar graph representing percent PI uptake (c) Graph representing percent cell viability after 24 h and 48 h of incubation with different
samples. (d) Alkaline Phosphatase is an early marker for osteoblast differentiation which is expressed in the bar graph as ALP activity per microgram of protein and
(e) Osteocalcin production presented in ng mL− 1 for control, SS316L and 109Ag- SS316L.

confined and the increase in anodic current was slowed down with the
potential displaying a passive behavior. This potential is called passive
potential. Finally, there is a transpassive second area (P4–P5 and Q4–Q5)
that starts at a critical potential, where the current value rapidly in­
creases owing to the breakdown of the passive film. This phenomenon is
known as pitting corrosion and the potential is called pitting (Epit) or
breakdown potential [50]. After ion implantation, the shift in corrosion
potential towards slight negative indicates the presence of unstable
passive film in the ion implanted sample. At lower applied potentials,
109
Ag ion implantation creates a non-equilibrium surface with a
near-amorphous surface layer that is prone to attract the corrosive
species to damage the protecting passive layer and damage the surface.

10
B. Malvi et al.
With Tafel analysis, corrosion current density (icorr) for SS316L and
109
Ag-SS316L is calculated as 2.416 µA cm− 2 and 2.649 µA cm− 2,
respectively. Corrosion current density is a representation of uniform
corrosion and an increased value of icorr suggests deterioration in the
overall corrosion resistance after 109Ag ion implantation. This is in
agreement with the metal ion release profiles in Section 3.3. From the
SEM images of corroded samples in Fig. 8 d–e, it is clear that the density
of corrosion pits is more in 109Ag-SS316L as compared to bare SS316L.
The typical diameter of pits in 109Ag-SS316L is about 65.3 ± 21.5 µm
whereas in SS316L it is 58.7 ± 9.8 µm. The localized or pitting attack on
both samples can be studied by the pitting potential (Epit) which is
decreased from 0.442 V vs SCE to 0.430 V vs SCE after ion implantation.
This decrease in pitting potential is attributed to the breakdown of the
passive film. Pitting potential is a measure of localized corrosion and the
fluctuations in the transpassive region are caused by the appearance of
metastable pitting and its subsequent repassivation. The unexpected
depassivation might cause metastable pitting and the increase in the
current, however, the subsequent repassivation of the metastable pitting
decreased the current, resulting in current fluctuations [51]. In the
transpassive region, after points P5 and Q5, the sample surface becomes
highly reactive and a constant current flows through the system causing
severe pitting corrosion.
3.6. In-vitro cytocompatibility studies
Propidium iodide (PI) dye is impermeable to the living cells and can
only be taken up by cells with compromised cell membranes or dead
cells. No significant increase in PI-positive population was observed in
MG-63 cells incubated with 109Ag-SS316L and SS136L compared to
control after 24 and 48 h (Fig. 9a–b). However, in case of cells incubated
with 109Ag-SS316L for 48 h shows less PI positive population (7.84 %) as
compared to 24 h (9.74 %) which indicates that 109Ag ion implanted
SS316L is well tolerated by MG-63 cells. MTT assay also indicates
maximum cell viability in the control group followed by SS316L and
109
Ag-SS316L while the difference between SS316L and 109Ag-SS316L is
not statistically significant. Both the viability assays exhibit a similar
trend where the cells are more viable after 48 h of exposure as compared
to 24 h (Fig. 9c).
It is well known that extracellular matrix production and differen­
tiation of osteoblast are sensitive to properties of implant material sur­
face [28,52,53]. According to the properties of the implant surface (viz.
roughness, surface energy, hydrophilicity), osteoblast shows the
increased activity of ALP and osteocalcin. Osteoblast with higher ALP
activity indicates mineralization of their matrix as ALP is an early
marker for osteogenic differentiation [42]. MG-63 cells grown on the
109
Ag ion implanted SS316L indicated the highest ALP production
compared to SS316L (Fig. 9d). Osteocalcin is a non-collagenous matrix
protein that is most abundant in bone and associated with bone minerals
[28,54]. We observed the osteocalcin production data in line with ALP
activity. MG-63 cells cultured on 109Ag ion implanted SS316L showed
significantly increased osteocalcin production compared to SS316L
(Fig. 9e). This could be due to the changes in the surface of the material
post ion implantation and not because of the presence of 109Ag ions.
4. Conclusions
Predicting the lifetime and failure of the metallic biomedical im­
plants is challenging due to the various complexities involved in joint
replacement and trauma surgeries. A new method to detect biomedical
implant failure and degradation using silver stable isotope (109Ag) as
tracer, is demonstrated in this research. Ion implantation is one of the
best methods to introduce the stable isotopic tracer in biomedical
implant material. Although stable isotopes can be costly, but they do
have exceptional uniqueness in their labelling strategy. Stable isotopes
have exceptional tracing ability due to their abundance and their
detection in ultra-low level. The isotopic tracer once in the system can be
11
Materials Today Communications 33 (2022) 104563
tracked irrespective of its form (particles, ions, complexes). Ion im­
plantation ensured that the isotopic tracer is not on the surface of the
implants but rather sub-surface of the implant. High energy ion im­
plantation facilitated the slow release of implanted 109Ag ions simulta­
neously with implant material degradation. After 109Ag ion
implantation, the hardness is increased, surface roughness and surface
energy are decreased. Electrochemical corrosion test indicated the
damage of corrosion-resistant oxide film due to ion implantation but the
effect of ion implantation on the electrochemical corrosion resistance of
SS316L is found to be insignificant. The cell viability and osteocalcin
production of MG-63 cells have improved after ion implantation. The
degradation of SS316L and 109Ag-SS316L samples in 30 % H2SO4 fol­
lowed a similar trend increasing percentage release of Fe, Cr, Ni, Mo,
and Mn correlating with 109Ag. A rise in the concentration of 109Ag is
correlated with the dissolution or degradation of total implant material.
This study demonstrates that metallic biomaterial failure can be detec­
ted using 109Ag isotope tracer. The ion implantation of stable isotopes
can be applied industrially in the manufacturing process for
manufacturing tracer embedded biomedical implants.
CRediT authorship contribution statement
Bharti Malvi: Conceptualization, Methodology, Writing - original
draft, Writing - review & editing. Ramesh Chaudhari: Investigation,
Methodology, Writing - original draft. Balasubramanian C: Writing –
review & editing. Ashutosh Kumar: Writing – original draft, Writing –
review & editing. Swagat Das: Conceptualization, Writing – review &
editing. Manas Paliwal: Conceptualization, Writing – review & editing.
Superb K. Misra: Conceptualization, Methodology.
Declaration of Competing Interest
The authors declare the following financial interests/personal re­
lationships which may be considered as potential competing interests:
Superb Misra reports financial support was provided by Ministry of
Education.
Data Availability
Data will be made available on request.
Acknowledgments
The authors are grateful to IUAC New Delhi for facilitating 109Ag ion
implantation. We acknowledge the Department of Science and Tech­
nology (SERB), Government of India (CRG/2019/006165), and
IMPRINT (Project no. 6408) research grant for funding this project.
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.mtcomm.2022.104563.
References
[1] J. Wilson, Metallic biomaterials, in: Fundam. Biomater. Met., Elsevier, 2018,
pp. 1–33, https://doi.org/10.1016/B978-0-08-102205-4.00001-5.
[2] Q. Chen, G.A. Thouas, Metallic implant biomaterials, Mater. Sci. Eng. R Rep. 87
(2015) 1–57, https://doi.org/10.1016/j.mser.2014.10.001.
[3] J.K. Lord, D.J. Langton, A.V.F. Nargol, T.J. Joyce, Volumetric wear assessment of
failed metal-on-metal hip resurfacing prostheses, Wear 272 (2011) 79–87, https://
doi.org/10.1016/j.wear.2011.07.009.
[4] A. Unger, H. Cabrera-Palacios, A. Schulz, C. Jürgens, A. Paech, Acoustic monitoring
(RFM) of total hip arthroplasty results of a cadaver study, Eur. J. Med. Res. 14
(2009) 264–271, https://doi.org/10.1186/2047-783X-14-6-264.
[5] A. Chiva, Biochemical markers in total joint arthroplasty: electrophoresis of joint
fluid proteins as a new diagnostic tool for prosthetic performance, Eur. J. Orthop.
Surg. Traumatol. 21 (2011) 545–551, https://doi.org/10.1007/s00590-011-0766-
1.
B. Malvi et al.
[6] K. Ren, A. Dusad, Y. Zhang, P.E. Purdue, E.V. Fehringer, K.L. Garvin, S.R. Goldring,
D. Wang, Early diagnosis of orthopedic implant failure using macromolecular
imaging agents, Pharmaceut. Res. 31 (2014) 2086–2094, https://doi.org/10.1007/
s11095-014-1310-x.
[7] A. Nicolli, A. Trevisan, I. Bortoletti, A. Pozzuoli, P. Ruggieri, A. Martinelli,
A. Gambalunga, M. Carrieri, Biological monitoring of metal ions released from hip
prostheses, Int. J. Environ. Res. Public. Health 17 (2020) 3223, https://doi.org/
10.3390/ijerph17093223.
[8] A. Nicolli, G. Bisinella, G. Padovani, A. Vitella, F. Chiara, A. Trevisan, Predictivity
and fate of metal ion release from metal-on-metal total hip prostheses,
J. Arthroplast. 29 (2014) 1763–1767, https://doi.org/10.1016/j.arth.2014.04.041.
[9] L. Savarino, G.S. Maci, M. Greco, N. Baldini, A. Giunti, Metal ion release from
fracture fixation devices: a potential marker of implant failure, J. Biomed. Mater.
Res. B Appl. Biomater. 86 (2008) 389–395, https://doi.org/10.1002/jbm.b.31032.
[10] T.W. Bauer, A.S. Shanbhag, Are there biological markers of wear? JAAOS - J. Am.
Acad. Orthop. Surg. 16 (2008) S68.
[11] Z. Ferdous, A. Nemmar, Health impact of silver nanoparticles: a review of the
biodistribution and toxicity following various routes of exposure, Int. J. Mol. Sci.
21 (2020) 2375, https://doi.org/10.3390/ijms21072375.
[12] P.L. Drake, K.J. Hazelwood, Exposure-related health effects of silver and silver
compounds: a review, Ann. Occup. Hyg. 49 (2005) 575–585, https://doi.org/
10.1093/annhyg/mei019.
[13] W. Li, W. Gou, W. Li, T. Zhang, B. Yu, Q. Liu, J. Shi, Environmental applications of
metal stable isotopes: silver, mercury and zinc, Environ. Pollut. 252 (2019)
1344–1356, https://doi.org/10.1016/j.envpol.2019.06.037.
[14] I. Rodushkin, E. Engstrom, D.C. Baxter, Isotopic analyses by ICP-MS in clinical
samples, Anal. Bioanal. Chem. 405 (2013) 2785–2797, https://doi.org/10.1007/
s00216-012-6457-x.
[15] S.K. Misra, A. Dybowska, D. Berhanu, M.N. Croteau, S.N. Luoma, A.R. Boccaccini,
E. Valsami-Jones, Isotopically modified nanoparticles for enhanced detection in
bioaccumulation studies, Environ. Sci. Technol. 46 (2012) 1216–1222, https://doi.
org/10.1021/es2039757.
[16] W.R. Shields, D.N. Craig, V.H. Dibeler, Absolute isotopic abundance ratio and the
atomic weight of silver, J. Am. Chem. Soc. 82 (1960) 5033–5036, https://doi.org/
10.1021/ja01504a005.
[17] N.K. Manninen, S. Calderon, I. Carvalho, M. Henriques, A. Cavaleiro, S. Carvalho,
Antibacterial Ag/a-C nanocomposite coatings: the influence of nano-galvanic a-C
and Ag couples on Ag ionization rates, Appl. Surf. Sci. 377 (2016) 283–291,
https://doi.org/10.1016/j.apsusc.2016.03.113.
[18] R. Rebelo, N.K. Manninen, L. Fialho, M. Henriques, S. Carvalho, Morphology and
oxygen incorporation effect on antimicrobial activity of silver thin films, Appl.
Surf. Sci. 371 (2016) 1–8, https://doi.org/10.1016/j.apsusc.2016.02.148.
[19] J.M. Schierholz, L.J. Lucas, A. Rump, G. Pulverer, Efficacy of silver-coated medical
devices, J. Hosp. Infect. 40 (1998) 257–262, https://doi.org/10.1016/S0195-6701
(98)90301-2.
[20] G. Cook, J.W. Costerton, R.O. Darouiche, Direct confocal microscopy studies of the
bacterial colonization in vitro of a silver-coated heart valve sewing cuff, Int. J.
Antimicrob. Agents 13 (2000) 169–173, https://doi.org/10.1016/S0924-8579(99)
00120-X.
[21] X. Sun, H. Gong, D. Li, L. Dong, M. Zhao, R. Wan, H. Gu, Ag+ implantation induces
mechanical properties, cell adhesion and antibacterial effects of TiN/Ag
multilayers in vitro, Nanomedicine 12 (2017) 2257–2268, https://doi.org/
10.2217/nnm-2017-0087.
[22] J. Wood, Ion Implantation, in: K.H.J. Buschow, R.W. Cahn, M.C. Flemings,
B. Ilschner, E.J. Kramer, S. Mahajan, P. Veyssière (Eds.), Encycl. Mater. Sci.
Technol., Elsevier, Oxford, 2001, pp. 4284–4286, https://doi.org/10.1016/B0-08-
043152-6/00751-8.
[23] L.L. Meisner, A.I. Lotkov, V.A. Matveeva, L.V. Artemieva, S.N. Meisner, A.
L. Matveev, Effect of silicon, titanium, and zirconium ion implantation on NiTi
biocompatibility, Adv. Mater. Sci. Eng. 2012 (2012), e706094, https://doi.org/
10.1155/2012/706094.
[24] J.F. Ziegler, M.D. Ziegler, J.P. Biersack, SRIM – the stopping and range of ions in
matter (2010), Nucl. Instrum. Methods Phys. Res. B 268 (2010) 1818–1823,
https://doi.org/10.1016/j.nimb.2010.02.091.
[25] M.R.C. Marques, R. Loebenberg, M. Almukainzi, Simulated biological fluids with
possible application in dissolution testing, Dissolut. Technol. 18 (2011) 15–28,
https://doi.org/10.14227/DT180311P15.
[26] J. Van Loco, M. Elskens, C. Croux, H. Beernaert, Linearity of calibration curves: use
and misuse of the correlation coefficient, Kathol. Univ. Leuven Open Access Publ.
Kathol. Univ. Leuven 7 (2002), https://doi.org/10.1007/s00769-002-0487-6.
[27] L.C. Crowley, A.P. Scott, B.J. Marfell, J.A. Boughaba, G. Chojnowski, N.
J. Waterhouse, Measuring cell death by propidium iodide uptake and flow
cytometry, Cold Spring Harb. Protoc. (2016) (2016), https://doi.org/10.1101/pdb.
prot087163.
[28] X. Rausch-fan, Z. Qu, M. Wieland, M. Matejka, A. Schedle, Differentiation and
cytokine synthesis of human alveolar osteoblasts compared to osteoblast-like cells
(MG63) in response to titanium surfaces, Dent. Mater. 24 (2008) 102–110, https://
doi.org/10.1016/j.dental.2007.03.001.
[29] S.H.H.H. Gazestani, M.G. Shahraki, M.R. Hantehzadeh, M. Ghoranneviss,
A. Shokouhy, Comparison of experimental and theoretical results of nitrogen
implantation in AISI 304 stainless steel, Appl. Surf. Sci. 237 (2004) 332–335,
https://doi.org/10.1016/j.apsusc.2004.06.107.
12
Materials Today Communications 33 (2022) 104563
[30] V. Muthukumaran, V. Selladurai, S. Nandhakumar, S. Mouleeswaran, Experimental
investigation on corrosion and hardness of ion implanted AISI 316L stainless steel,
Mater. Des. 31 (2010) 2813–2817, https://doi.org/10.1016/j.
matdes.2010.01.007.
[31] H. Wollenberger, Phase transformations under irradiation, J. Nucl. Mater. 216
(1994) 63–77, https://doi.org/10.1016/0022-3115(94)90007-8.
[32] F. Wang, C. Zhou, L. Zheng, H. Zhang, Improvement of the corrosion and
tribological properties of CSS-42L aerospace bearing steel using carbon ion
implantation, Appl. Surf. Sci. 392 (2017) 305–311, https://doi.org/10.1016/j.
apsusc.2016.09.068.
[33] N. Kumar, S. Kataria, S. Dash, S.K. Srivastava, C.R. Das, P. Chandramohan, A.
K. Tyagi, K.G.M. Nair, B. Raj, Tribological properties of nitrogen ion implanted
steel, Wear 274–275 (2012) 60–67, https://doi.org/10.1016/j.wear.2011.08.017.
[34] K. Feng, X. Cai, Z. Li, P.K. Chu, Improved corrosion resistance of stainless steel
316L by Ti ion implantation, Mater. Lett. 68 (2012) 450–452, https://doi.org/
10.1016/j.matlet.2011.11.014.
[35] X. Chen, X. Yin, J. Jin, A study on the wettability of ion-implanted stainless and
bearing steels, Metals 9 (2019) 208, https://doi.org/10.3390/met9020208.
[36] Q. Zhao, Y. Liu, C. Wang, S. Wang, N. Peng, C. Jeynes, Reduction of bacterial
adhesion on ion-implanted stainless steel surfaces, Med. Eng. Phys. 30 (2008)
341–349, https://doi.org/10.1016/j.medengphy.2007.04.004.
[37] H. Müller-Steinhagen, Q. Zhao, Investigation of low fouling surface alloys made by
ion implantation technology, Chem. Eng. Sci. 52 (1997) 3321–3332, https://doi.
org/10.1016/S0009-2509(97)00162-0.
[38] S.B. Kennedy, N.R. Washburn, C.G. Simon, E.J. Amis, Combinatorial screen of the
effect of surface energy on fibronectin-mediated osteoblast adhesion, spreading
and proliferation, Biomaterials 27 (2006) 3817–3824, https://doi.org/10.1016/j.
biomaterials.2006.02.044.
[39] Y. Yang, L. Guo, H. Liu, Effect of fluoride ions on corrosion behavior of SS316L in
simulated proton exchange membrane fuel cell (PEMFC) cathode environments,
J. Power Sources 195 (2010) 5651–5659, https://doi.org/10.1016/j.
jpowsour.2010.03.064.
[40] M. Terada, D.M. Escriba, I. Costa, E. Materna-Morris, A.F. Padilha, Investigation on
the intergranular corrosion resistance of the AISI 316L(N) stainless steel after long
time creep testing at 600 ◦ C, Mater. Charact. 59 (2008) 663–668, https://doi.org/
10.1016/j.matchar.2007.05.017.
[41] A. Arwati, S. Sianipar, Corrosion study of SS316L in environment sulphur acid
using weight loss method, SINERGI 22 (2018) 24–28, https://doi.org/10.22441/
sinergi.2018.1.005.
[42] M.P. Ryan, D.E. Williams, R.J. Chater, B.M. Hutton, D.S. McPhail, Why stainless
steel corrodes, Nature 415 (2002) 770–774, https://doi.org/10.1038/415770a.
[43] F. Riffard, H. Buscail, E. Caudron, R. Cueff, C. Issartel, S. Perrier, The influence of
implanted yttrium on the cyclic oxidation behaviour of 304 stainless steel, Appl.
Surf. Sci. 252 (2006) 3697–3706, https://doi.org/10.1016/j.apsusc.2005.05.054.
[44] L. Neelakantan, J.K. Zglinski, M. Frotscher, G. Eggeler, Design and fabrication of a
bending rotation fatigue test rig for in situ electrochemical analysis during fatigue
testing of NiTi shape memory alloy wires, Rev. Sci. Instrum. 84 (2013), 035102,
https://doi.org/10.1063/1.4793488.
[45] V.P. Mahesh, A. Gumaste, N. Meena, J. Alphonsa, A. Arora, Corrosion behavior of
aluminum surface composites with metallic, ceramic, and hybrid reinforcements
using friction stir processing, Metall. Mater. Trans. B (2020), https://doi.org/
10.1007/s11663-020-01932-7.
[46] N. Eliaz, Corrosion of metallic biomaterials: a review, Materials 12 (2019) 407,
https://doi.org/10.3390/ma12030407.
[47] D. Li, C. Lin, C. Batchelor-McAuley, L. Chen, R.G. Compton, Tafel analysis in
practice, J. Electroanal. Chem. 826 (2018) 117–124, https://doi.org/10.1016/j.
jelechem.2018.08.018.
[48] E. McCafferty, Validation of corrosion rates measured by the Tafel extrapolation
method, Corros. Sci. 47 (2005) 3202–3215, https://doi.org/10.1016/j.
corsci.2005.05.046.
[49] C. Yang, J. Wang, X. Xie, S. Wang, Z. Mao, H. Wang, Electrochemical behavior of
surface treated metal bipolar plates used in passive direct methanol fuel cell, Int. J.
Hydrog. Energy 37 (2012) 867–872, https://doi.org/10.1016/j.
ijhydene.2011.04.033.
[50] L. Gil, S. Brühl, L. Jiménez, O. Leon, R. Guevara, M.H. Staia, Corrosion
performance of the plasma nitrided 316L stainless steel, Surf. Coat. Technol. 201
(2006) 4424–4429, https://doi.org/10.1016/j.surfcoat.2006.08.081.
[51] L. Tan, Z. Wang, Y. Ma, Tribocorrosion behavior and degradation mechanism of
316L stainless steel in typical corrosive media, Acta Metall. Sin. Engl. Lett. (2021),
https://doi.org/10.1007/s40195-020-01182-1.
[52] I. Degasne, M.F. Baslé, V. Demais, G. Huré, M. Lesourd, B. Grolleau, L. Mercier,
D. Chappard, Effects of roughness, fibronectin and vitronectin on attachment,
spreading, and proliferation of human osteoblast-like cells (Saos-2) on titanium
surfaces, Calcif. Tissue Int. 64 (1999) 499–507, https://doi.org/10.1007/
s002239900640.
[53] J.L. Ong, E.G. Bess, K. Bessho, Osteoblast progenitor cell responses to characterized
titanium surfaces in the presence of bone morphogenetic protein–atelopeptide type
I collagen IN VITRO, J. Oral Implantol. 25 (1999) 95–100, https://doi.org/
10.1563/1548-1336(1999)025<0095:OPCRTC>2.3.CO;2.
[54] A.L. Boskey, S. Gadaleta, C. Gundberg, S.B. Doty, P. Ducy, G. Karsenty, Fourier
transform infrared microspectroscopic analysis of bones of osteocalcin-deficient
mice provides insight into the function of osteocalcin, Bone 23 (1998) 187–196,
https://doi.org/10.1016/S8756-3282(98)00092-1.