JOM, Vol. 68, No.

4, 2016
DOI: 10.1007/s11837-015-1775-z
Ó 2016 The Minerals, Metals & Materials Society

Monitoring Biodegradation of Magnesium Implants with Sensors

DAOLI ZHAO,1 TINGTING WANG,1 XUEFEI GUO,1 JULIA KUHLMANN,1

AMOS DOEPKE,1 ZHONGYUN DONG,2 VESSELIN N. SHANOV,3
and WILLIAM R. HEINEMAN1,4

1.—Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221, USA. 2.—Depart-

ment of Internal Medicine, University of Cincinnati, Cincinnati, OH 45267, USA. 3.—Department
of Biomedical, Chemical and Environmental Engineering, University of Cincinnati, Cincinnati,
OH 45221, USA. 4.—e-mail: william.heineman@uc.edu

Magnesium and its alloys exhibit properties such as high strength, light
weight, and in vivo corrosion that make them promising candidates for the
development of biodegradable metallic implant materials for bone repair,
stents and other medical applications. Sensors have been used to monitor the
corrosion of magnesium and its alloys by measuring the concentrations of the
following corrosion products: magnesium ions, hydroxyl ions and hydrogen
gas. The corrosion characterization system with home-made capillary pH and
Mg2+ microsensors has been developed for real-time detection of magnesium
corrosion in vitro. A hydrogen gas sensor was used to monitor the corrosion of
magnesium by measuring the concentration of the hydrogen gas reaction
product in vivo. The high permeability of hydrogen through skin allows
transdermal monitoring of the biodegradation of a magnesium alloy implanted
beneath the skin by detecting hydrogen gas at the skin surface. The sensor
was used to map hydrogen concentration in the vicinity of an implanted
magnesium alloy.

presence in bone tissues, magnesium has stimula-

INTRODUCTION
Magnesium and its alloys have recently been a
focus for biomedical implants due to their biocom-
patibility and suitable mechanical properties. Mag-
nesium alloys are lightweight: the density of
magnesium and its alloys is about 1.74 g/cm3 at
20°C, which is 1.6, 3 and 4.5 times less dense than
permanent aluminum- and titanium-based alloys
and stainless steel, respectively. The density of Mg
is close to that of bone, which is in the range of
1.8–2.1 g/cm3.1–3 Magnesium alloys have a modu-
lus elasticity of 45 GPa, which is also very close to
that of bone (15–40 GPa).4–6 This property mini-
mizes the mismatch between the mechanical prop-
erties of conventional metals such as cobalt-
chrome, titanium and stainless steel and the
surrounding natural bone tissue.2 Magnesium pos-
sesses good biocompatibility and its biodegradation
does not release toxic metal ions into the body.
Relatively large amounts of magnesium are toler-
ated by the body without ill effect; the adult body
stores about 30 g of magnesium in muscle and bone
tissue.7 Moreover, due to the functional roles and
tory effects on the growth of new bone tissue.3,8,9
Importantly, magnesium alloys used as screws,
needles, and plates for fracture fixtures do not need
to be removed by a second surgical procedure after
the tissue heals sufficiently, reducing the cost to the
health care system and additional stress to the
patient.4
Magnesium degrades in aqueous environments,
generating Mg2+ and OH ions, as well as H2 gas
(Eq. 1).
Mg þ 2H2 O ! Mg2þ þ 2OH þ H2 ð1Þ
Many methods have been developed to monitor
the degradation of Mg and its alloys: weight loss
methods,10,11 collection of the evolving hydrogen,12,13
pH measurement,14,15 electrochemical impedance
spectroscopy (EIS),16 microscopy and spectroscopy.17,18
However, only one or two measurements are carried
out at a time with the above methods, followed by
different analyses at the termination of the immer-
sion test. In addition, those methods do not monitor
the corrosion of Mg in real time.

1204 (Published online January 7, 2016)

Monitoring Biodegradation of Magnesium Implants with Sensors 1205

Recently, we have developed a corrosion charac-
terization system (CCS) for real-time monitoring of
Mg corrosion.19 The CCS provides more insight into
the time-dependent corrosion behavior of Mg and its
alloys and the influence of the immersion solution.
The unique feature of the CCS is the simultaneous
use of multiple electrochemical sensors to measure
the solution-soluble corrosion products of the oxida-
tion of Mg in water. In this system, a glass electrode
pH sensor was used to monitor OH, a water
hardness ion-selective electrode (ISE) was employed
to monitor Mg2+, and a Pt disc sensor was applied
for monitoring dissolved H2 in the solution.
To reduce dimensions and volume requirements
for other applications, we developed a new genera-
tion CCS with home-made capillary pH and Mg2+
microsensors.20 We have also explored the use of a
hydrogen microsensor for in vivo corrosion monitor-
ing.21–25 As seen from Eq. 1, one molecule of H2 is
liberated for every atom of Mg that biodegrades.
Thus, measuring evolved hydrogen allows a direct
measurement of Mg corrosion. This method has
opened the way to using non-invasive H2 sensing as
an effective way to track biodegradation rates of Mg
and its alloys in vivo.
EXPERIMENTAL
Reagents
Sodium chloride, magnesium chloride, and sodium
hydroxide were purchased from Fisher Scientific
(Pittsburgh, PA, USA). 4-(2-hydroxyethyl) piper-
azine-1-ethanesulfonic acid (HEPES), magnesium
ionophore II-ETH5214, 2-nitrophenyl octyl ether,
and potassium tetrakis(4-chlorophenyl)borate were
obtained from Sigma Aldrich (St. Louis, MO, USA).
Magnesium 99.9% from Goodfellow (Oakdale, PA,
USA) was cut from stock material with a lathe to form
disks 7.5 mm diameter 9 2–3 mm thick. Epo-Thin
epoxy used to mount the disks was from Buehler
(Lake Bluff, IL, USA). Magnesium single crystals
were kindly provided by Dr. Shanov’s group at the
University of Cincinnati.
Instruments
Potentiometric sensors and electrochemical impe-
dance spectroscopy measurements have been previ-
ously described.19,26 Amperometric H2 measurements
were performed using a H2 microsensor (50 lm tip
diameter) connected to a multimeter (both from
Unisense, Aarhus, Denmark) polarized at +1000 mV
for at least 1 h. After a stable current in the low
picoampere range was established, the amperometric
sensor was ready for use.
Corrosion Study
The preparation and treatment of Mg samples for
corrosion studies was done by a previously described
method.19 The developed CCS and pH microsensor
based on capillary tubes were used to study the
corrosion of Mg 99.9% in 0.9% NaCl solutions at
37°C. The pH sensor was calibrated with standard
buffer solutions at pH 5, 7, 8 and 10. The pH
microsensor was about 2 cm from Mg metal. The pH
monitoring period was 4 h for Mg corrosion in
stirred 0.9% NaCl solutions at 37°C.
Electrochemical Impedance Spectroscopy
(EIS)
Impedance spectra were recorded using a Gamry
REF 600 potentiostat (Gamry Instruments, Warmin-
ster, PA, USA) with the Mg sample as the working
electrode in 0.9% NaCl solution. A Ag/AgCl electrode
filled with 3.0 M KCl (BASi, Lafayette, IN, USA) was
used as the reference electrode and a coiled Pt wire
(BASi) was used as the auxiliary electrode. The
frequency range of the impedance spectra was 0.1 Hz
to 1 MHz at open circuit potential with a 10 mV

Fig. 1. The corrosion characterization system (CCS) instrumentation and cell configuration. (1) pH electrode, (2) magnesium ion selective
electrode, (3) hydrogen electrochemical sensor, (4) temperature probe, (5) Mg sample, (6) platinum auxiliary electrode and (7) silver–silver
chloride reference electrode.
1206 Zhao, Wang, Guo, Kuhlmann, Doepke, Dong, Shanov, and Heineman

alternating current. The measurements were made RESULTS AND DISCUSSION

from 0 to 20 h. Echem Analyst from Gamry was used
Set Up and Instrumentation for the CCS
to analyze the impedance spectra.
The experimental setup including electrochemi-
cal/corrosion cell and associated instrumentation for
the CCS is shown in Fig. 1. The cell consisted of a 1000-
mL water-jacketed glass beaker as the corrosion cell
and a water-jacketed 65-mL calibration cell with
conical sides and a flat bottom.19 A thermostatic
recirculating water bath maintained the temperature
of the solution in the cells at 37°C. The solution was
constantly stirred using a magnetic stirring plate. A
custom-made PVC lid was used to reduce evaporation
of the solution from the corrosion cell and to consis-
tently position the Mg sample and sensors. The height
of the lid could be adjusted in the corrosion cell to
maintain a constant headspace above the corrosion
solution when the volume of the solution was changed.
The CCS integrates the potentiometric electrochemi-
cal sensors (pH sensor, Mg-ISE and electrochemical H2
sensor) and EIS immersion test for evaluating the
corrosion of Mg and its alloys. A double-junction
reference electrode Ag/AgCl was used to ensure that
the potentials from all three potentiometric sensors
were based on the same reference potential and
Fig. 2. (a) pH and Mg2+ ion selective microelectrode and (b) pH
microelectrodes.

Fig. 3. Potentiometric response of pH microelectrode based on glass capillaries to (a) standard buffer solutions and (b) corrosion of 99.9% Mg in
0.9% NaCl. (c) Nyquist plots for EIS of 99.9% Mg in 0.9% NaCl.
Monitoring Biodegradation of Magnesium Implants with Sensors
junction potential drift. This CCS was demonstrated
by comparing the corrosion of Mg (99.9%) in 0.9%
NaCl, HEPES buffer, and HEPES buffer with NaCl.19
pH and Mg2+ Microsensors
The microsensors for pH and Mg2+ were developed
based on small diameter capillary tubing that was
drawn to a small point at one end after melting as
shown in Fig. 2. The microsensor capillary contained
an AgCl-coated Ag wire immersed in a solution of
MgCl2 as the internal reference electrode and a plug of
liquid ionophore membrane in the tip (Fig. 2a) to
provide selective response for the targeted ion. A
photograph of three pH electrodes is shown in Fig. 2b.
The tips of the microelectrodes based on glass capil-
laries are micrometers in diameter (Fig. 2b) and thus
have the potential to provide spatial information
about corrosion at the surface of a Mg alloy.
Representative results are shown for the pH
microelectrode. The pH microelectrode gave a good
response to pH over a range of 5–10 as shown by the
Fig. 4. H2 measurement by H2 microsensor on a gas cavity, adjacent
to the cavity and far from the cavity on a mouse with Mg single crystal
implanted subcutaneously. Vertical axis is raw output of sensor in mV.
1207
calibration in Fig. 3a. The correlation equation is
y = 51.3x + 474.7 (R2 = 0.99) where y is the poten-
tial in mV versus the Ag/AgCl reference electrode,
and x is the pH of the buffer. The main advantages of
the pH microelectrode, its microscopic size and
biocompatibility, make it potentially useful for cell
biology applications and other experiments in small
volumes. The attainable tip size (50 lm) is compa-
rable to or smaller than that of the existing commer-
cial pH sensors. It may also be used as a scanning
probe for pH microscopy, where its small size can
provide good spatial resolution. To accommodate the
monitoring of pH in the Mg corrosion process, MgCl2
was used as the internal reference electrode solution
instead of the conventional KCl or NaCl solution. To
improve selectivity, the ionophore membrane was
used for the fabrication process. Microelectrodes
based on glass capillaries for pH and Mg2+ were
fabricated and were able to sense the changes in pH
and Mg2+ accompanying the corrosion of 99.9% Mg in
0.9% NaCl. The change in pH is shown in Fig. 3b. The
pH increases rapidly from the baseline pH of the 0.9%
NaCl solution when the Mg sample is immersed at
700 s. At this point, corrosion of Mg commences,
generating OH as in Eq. 1. When the solution
becomes sufficiently basic (ca. pH 9), OH reacts
with the Mg2+ that is also being generated to form
insoluble Mg(OH)2, which precipitates on the sample
surface, slowing the corrosion rate.19
Electrochemical Impedance Spectroscopy
EIS was used to monitor changes in the resistance
of Mg samples during corrosion. The benefit of EIS
combined with a sensor such as pH is the ability to
monitor corrosion in real time during immersion
tests without needing to remove solutions for off-line
analysis. The Nyquist plots of Mg corrosion display
the typical impedance responses in 0.9% NaCl solu-
tion (Fig. 3c). Mg alloys show two capacitance loops,
one at high frequency and the other one at medium
frequency. The diameters of the capacitive loops at
high frequency and medium frequency are directly
proportional to the charge transfer resistance (Rct)
and surface film resistance (Rf) for mass transfer,
respectively.16 During immersion in NaCl solution
for 2 h, the loop at low frequency disappears. This
observation has been reported for AZ31 alloy in
simulated body fluid.26 The diameter of the loop at
high frequency increases with longer immersion
time. Increments of the impedance modulus with
exposure time demonstrate that the corrosion prod-
ucts play an important role in the corrosion protec-
tion of metal. The increment of the resistance is
primarily due to the precipitation of Mg(OH)2 and
other compounds on the alloy surface.
Electrochemical Hydrogen Microsensor
An electrochemical hydrogen microsensor can be
used for monitoring corrosion in vivo for Mg alloys
tested in the mouse model.21 We found that the
1208 Zhao, Wang, Guo, Kuhlmann, Doepke, Dong, Shanov, and Heineman
concentration of H2 in gas cavities associated with
rapidly corroding Mg implants was actually very
low, even shortly after formation of the cavities.
Measurements made noninvasively by just pressing
the sensor tip against the skin covering the implant
are very similar to those made invasively by insert-
ing the sensor tip inside the cavity. This observation
points to extraordinarily fast transport of H2
through skin. So, we have used the H2 microsensor
to transdermally measure the H2 generated by a Mg
disk implanted subcutaneously in a mouse.21–25 The
measurements are made rapidly and noninvasively
by just pressing the sensor tip against the skin
covering the implant. This simple method opens the
way to developing H2 sensing as an effective way to
track biodegradation rates of Mg and its alloys
in vivo and noninvasively.
The H2 microsensor was used to map out the
hydrogen permeating through the skin. Measure-
ments made on the skin on top of the gas cavity, and
close to the gas cavity and far away from the gas
cavity are shown in Fig. 4. As seen, the H2
microsensor response is very fast, less than 1 min.
Some variability in H2 was found depending on
exactly where the tip was placed on the cavity.
When the sensor tip was moved off the cavity, H2
was still detected from the skin immediately adja-
cent to the cavity, but at considerably lower
concentrations.
CONCLUSION
A new generation of CCS has been developed,
which allows for the real-time monitoring of pH,
Mg2+ and dissolved H2, as well as surface changes
by EIS during corrosion of Mg and its alloys.
Capillary microsensors for pH and Mg2+ have been
successfully fabricated and used for Mg corrosion
real-time measurement. These microsensors have
the potential to provide spatial information for
corrosion at the surface of Mg alloys.
An electrochemical H2 sensor has been used to
transdermally measure the concentration of H2 in
subcutaneous gas cavities generated by Mg alloy
biodegradation. This method is simple and nonin-
vasive and makes the monitoring process of Mg and
its alloy biodegradation in vivo easier, with no
requirement for expensive and large equipment
such as x-ray and Micro-CT.
ACKNOWLEDGEMENT
The authors thank the National Science Founda-
tion (NSF ERC 0812348) for financial support.
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