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344 Surface Modification of Magnesium and its Alloys for Biomedical Applications
These are all well studied and commonly used in industrial applications, and an
excellent review on these methods was written by Gray and Luan (2002). While
such methods may protect magnesium, the use of bare magnesium is not ideal for
biomedical application in general, or neural application specifically. Instead, this chapter will concentrate on methods to apply polymeric coatings.
12.7.1 Pretreatment of magnesium surface
It should be noted that pretreatment through use of the surface-altering methods listed
above has been found to improve the adhesion strength of subsequent polymeric
coating (Hornberger et al., 2012; Ostrowski et al., 2013). Some refer to the use of
such pretreatment as a “two-step” coating method and application of coating to
bare, unaltered Mg surfaces is a “one-st ep” method. No studies are currently published
in which neural cells have been grown on Mg substrates coated through a two-step process, but it seems reasonable to assume that this method might increase surface roughness of this applied polymer coating. It has been reported that nano-rough gold
surfaces cause high rates of cell death by necrosis in neurons (Brunetti et al., 2010).
Therefore, the question remains whether the increase in adhesion strength achieved
through pretreatments is an optimal strategy for creating neural biomaterials.
12.7.2 Dip-coating
Dip-coating is one of the most facile coating methods. It involves dissolving monom ers
in a solvent and then dipping the magnesium substrate. Coating thickness can be
controlled by the viscosity of the monomer solution; the number of dipping cycles;
and most importantly, the dipping rate. This type of coating can be used to apply thermoplastic polymers as well as polymers that need to be dissolved in organic solvents.
Although this method is simple, it offers less exact control over coating thickness and
uniformity. This method is well suited for thicker coatings, tens of microns to the submillimeter range. However, film thickness per dipping cycle varies widely with the viscosity of the dipping solution and rate of dipping. Zomorodian et al. (2013) reported
films of less than 5 mm thickness (see Figure 12.5) when coating magnesium with
PEI dispersed with nano-sized hydroxyapatite (HA). In addition, they showed that
agglomeration of their nanoparticles resulted in defects in the uniformity of the coating,
which can be seen in Figure 12.5, panel C. Others have also reported difficulties in
creating uniform, nonporous coatings through the dipping process (Xu & Yamamoto,
2012). In the field of neural biomaterials, this may not necessarily be a drawback, as a
certain degree of porosity is desirable for many neural applications. However, being
able to exert strict control over the features of polymer coatings on magnesium is important since the degradation of magnesium is highly unpredictable. Even with the reported
defects, magnesium samples with the coating in the study were stable in DMEM solution for 3 months.
Application of polymer coatings onto magnesium substrates through a dipping process is the most reported method, most likely because it requires no special equipment.
This method also has value as an experimental tool because thickness parameters can
Surface modification of magnesium by functional polymer coatings for neural applications 345
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(a) (b)
(c)
Figure 12.5 a) SEM image showing 4e5 mm coating achieved through dipping. (b) High-
magnification image showing un-agglomerated nano HA. (c) Agglomerated HA and associated
defect in surrounding polymer film. (d) Agglomerated HA particles as seen from the surface of
the coated Mg substrate.
Reproduced from Zomorodian et al. (2013).
(d)
be easily changed by addition of dipping cycles or altering the speed of dipping. Such
simple optimization methods could be especially valuable when designing neural guidance conduits, where wall thickness is a key parameter (Kokai, Lin, Oyster, & Marra,
2009).
12.7.3 Spin-coating
Spin-coating is an ideal method for applying a uniform thin coating onto a flat substrate. The substrate is secured to a spinning stage, and then the coating material is
applied to the center of the substrate and allowed to spread evenly over the surface
through centripetal acceleration forces.
Spin-coating is a good method for spreading uniform thin films of polymers that are
extremely sticky or hydrophobic and suitable for coating flat, axi-symmetrical substrates.
It is also a good researchtool for screening potentialcoating materials because this method
is compatible with a wide variety of coating materials including solvent-dissolved solutions,thermoplastics, and coatings that require additionalprocessingsuch as curingor sintering.However, it is less effective for coating cylindrical or oddly shapedobjects and may
present edge effects if it is used to coat non-axi-symmetric shapes (Boellaard, Pham,
Sarro, & Burghartz, 2002,pp.81e86). In addition, if the substrate to be coated has
346 Surface Modification of Magnesium and its Alloys for Biomedical Applications
any depressions of features, this method is not optimal as there is a tendency for polymer to
collect in features, which results in nonuniform coating thickness (Boellaard et al., 2002,
pp. 81e86). This has the potential to be a strong negative effect when coating conducting
objects such as electrodes, where charge density at the surface must be closely controlled.
Altering the spinning speed changes the thickness and surface roughness of the
resulting coatings. In addition, when using solvent casting methods, the choice of solvent may be important, as differences in vapor pressure of the solvent will result in
different rates of vaporization speed and ultimately alter the morphology and characteristics of the resulting film (Kamanyi, Ngwa, Luo, & Grill, 2008).
12.7.4 Spray-coating
Spray-coating is similar to dip-coating, but requi res additional equipment to create a
continuous, controlled spray. Spray-coating is appropriate for polymeric material
that can exist in a liquid or colloid form, which includes most of the popular synthetic
biodegradable polymers. The coatings created by spraying are inherently porous in nature (Gray & Luan, 2002), which may be beneficial in neural applications, where a
degree of porosity is ideal, as previously discussed. Jo et al. recently used spraycoating of PCL in combination with aerosol deposition of bioactive nanoparticles
(Jo, Li, Kim, Kim, & Koh, 2013). This combination coating method allowed the researchers to create a flexible, uniform coating of biodegradable polymer to temper
the corrosion processes of the magnesium substrate. Spray-coating is an excellent option when trying to coat objects that are three dimensional, nonsymmetrical, or with
topographical features. Generally, spray-coating will result in a layer of more uniform
thickness than can be achieved with spin-coating (Boellaard et al., 2002, pp. 81e86).
There are a variety of sub-technologies that come under the heading of spraycoating. The previous paragraph concentrated on the spraying of colloids, but there
is also thermal spray-coating, which allows for the coating of a surface in any material
that is meltable (Gray & Luan, 2002). Another variation is electrostatic spray-coating
(Liu, Jiang, & Malshe, 2009). In this solvent-free process, powders of solid charged
particles or atomized liquids are accelerated toward a surface by the application of
an electrical field. For such a process to work, both the particles and the item to be
coated must be conductive and able to carry charge. This process is advantageous in
that it does not require solvents, which are often not biocompatible and need to be
removed carefully, and it is more efficient, reducing the waste of coating material
due to overspray, which is common with traditional spray techniques.
Wong et al. (2010) created what they called polymer membranes of PCL on mag-
nesium through layers of spray-coating. As seen in Figure 12.6, pores of different sizes
can be created by changing the concentration of PCL used in the spraying apparatus.
Small implants covered in these membranes were implanted into the greater trochanter
of rabbits for 2 months, and an interesting finding in this study was that no evidence of
excessive hydrogen gas was found, which is very different from previous studies.
Based on these findings, Wong et al. hypothesizes that implantation in different areas
of the body as well as the protection provided by the coating limited the evolution of
hydrogen gas to a level that was able to be absorbed easily into the body.
Surface modification of magnesium by functional polymer coatings for neural applications 347
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(a) (b)
Figure 12.6 Porous membranes of PCL produced through spray-coating onto magnesium. The
size and density of resulting pores was easily controlled by changing the density of PCL used in
the spraying apparatus. LMP, low-porosity membrane; HPM, high-porosity membrane. PCL,
poly-ε-caprolactone.
12.7.5 Electrospinning
In electrospinning, a liquid-phase polymer is released from a syringe into an electric
field. The field disrupts the surface tension of the polymer, stretching the droplets
until they form a thin, continuous stream from the syringe ti p. The thin fibers are
collected at a certain distance from the syringe tip where they are generated. Fibers
produced through electrospinning can range from microns to nanometers in diameter
and can be collected on a stationary plate, resulting in random alignment; o r the collecting p late can be rotated, which will result in controlled alignment of fibers.
Electrospinning is the most promising coating method for supporting neural cell
growth and adhesion (Aurand, Lampr, & Bjugstad, 2012; Subramanian et al., 2009),
as it can be used to create highly porous scaffolds, such as the surface shown in
Figure 12.7, which allow flow of nutrients and waste products. The interconnected
network structure of the pores created from this method makes it more suitable than
the isolate d pores created throug h other methods such as salt-leaching. In addition,
Figure 12.7 Example of an electrospun scaffold for neural tissue engineering, created from
PLGA-PANi composite fibers.
Reproduced from Subramanian et al. (2009).
348 Surface Modification of Magnesium and its Alloys for Biomedical Applications
control of alignment of fibers creates micro and nano-patterning, and such aniso-
tropic topographical cues have been shown to influence man y factors of regeneration
including neurite alignment, neurite outgrowth, formation of synaptic connections,
and cell differentiation (Nectow et al., 2012). This method creates porous coatings,
which might present a problem when coating magnesium substrates that need to be
protected from premature degradation in the physiological environment. Possible solutions to that problem would be to use dual coating methods: one “inner coating”
that is not as porous to protect the magnesium from corrosion followed by a second
“outer coating” that supports active cellular adhesion, proliferation, and migration.
12.7.6 Electrophoretic deposition
Electrophoretic deposition is a two-step process by which the particles suspended in a
colloid solution are collected onto a substrate. Deposition takes place in an electrochemical cell in which one of the electrodes is the substrate to be coated. When the potential is
applied, the suspended coating particles are polarized and pulled toward the substrate
(electrophoresis) to form a loose coating. The coating at this point is a powder agglomeration and must undergo additional processing such as sintering to secure the bonds
between the particles and to the substrate (Vandeperre & Van der Beist, 1999). Polymers
that can be used in electrophoretic deposition must be polarizable or possess the ability to
sustain induced dipoles in order for the electrophoresis to take place. Advantages of this
method include that it tends to produce uniform coatings on three-dimensional
substrates, and the entire exposed surface is easily coated, making it simple to coat inner
and outer surfaces of cylindrical objects (Boellaard et al., 2002, pp. 81e86).
12.7.7 Electrochemical deposition
Electrochemical deposition is similar to electrophoretic deposition in that it also takes
place in an electrochemical cell, but in this case, a chemical bonding process occurs, as
opposed to electrophoretic deposition. In electrochemical deposition, a monomer solution is loaded into an electrochemical cell and the magnesium substrate serves as
one of the electrodes. When voltage is applied, the monomers undergo a chemical
polymerization onto the magnesium. An example of the electrochemical cell used
by Sebaa et al. (2013) is shown in Figure 12.8. Advantages of electrochemical deposition include its relatively low cost and improved interfacial bonding between the
coating material and the substrate before heat treatment or sintering. However, the uniformity and morphology of the coating depends on the polymerization pattern or the
polymer used. For example, PEDOT coating on magnesium substrates (see
Figure 12.9) appeared particulate. Luo et al. explored direct electrodeposition of
conductive polymer coating onto magnesium substrates using magnesium as a working electrode in the electrochemical cell and an ionic liquid solution (Luo & Cui,
2011). Sebaa et al. compared various electrochemical deposition parameters to pro-
duce films of uniform thickness and reduce the cost of the electrochemical deposition
(Sebaa et al., 2013). The electrochemically deposited PEDOT on magnesium substrates was thoroughly characterized by Sebaa et al.
Surface modification of magnesium by functional polymer coatings for neural applications 349
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Magnesium (working)
Platinum or stainless steel (counter)
Silver/silver chloride or copper (reference)
Potentiostat
10 mm
5 mm
Figure 12.8 Schematic of electrochemical cell for the direct electrodeposition of conductive
polymer onto magnesium working electrode.
Figure 12.9 Representative image of morphology of PEDOT deposited on magnesium
substrate by electrochemical deposition (Sebaa et al., 2013).
12.8 Evaluation of coating
Coating of the magnesium substrates will have an effect on the biological, chemical,
and mechanical performance of the material. It is extremely important to evaluate how
the coating changes the substrate properties, as well as the stability and strength of the
coating itself. Despite this being crucial for proper evaluation of coatings and preclinical or clinical adoption of technologies, there is little consensus around which tests
should be used.
12.8.1 Characterization of microstructure
The topography and surface roughness of objects plays an important role in determining cell adhesion properties and more specifically the neuroconductive and
350 Surface Modification of Magnesium and its Alloys for Biomedical Applications
neuroinductive abilities of materials in vivo. Many experimental NGCs have incorporated micropatterning in an attempt to provide directional growth cues for regener ating
neuritis.
To evaluate structures and features at the submicron a nd nanometer scales,
higher resolving power is needed than can be provided through optical microscopy. Therefore, character izing micro- and nanostructure of polymeric coatings
must be done through more sensitive methods such as electron microscopy, the
most widely used of which is scanning electron microscopy (SEM). If the coating
polymer is not conductive, it is necessary to prepare the samples with a sputter co at
or to use an environmental SEM (ESEM), an excellent method for analyzing biological samples as it ca n look at “wet” and un-sputter-coat ed samples. However,
the resolution of ESEM is usually not as good as that provided by a standar d
SEM set-up. Polymeric samples are delicate, and even sputter-coated samples
may be damaged if the acceleration voltage used is too high.
12.8.2 Adhesion strength (ASTM tape test, ASTM
microscratch test)
While many in the literature agreed that adhesion strength is essential for a good
coating material, as Hornberger et al. (2012)andXu and Yamamoto (2012) have
previously pointed out, there is a dearth of concrete data being reported. Some authors give detailed mechanisms of how polymers and magnesium might interact
(Zomorodian et al., 2013), but in most papers no physical testing results are reported. In addition, adhesion properties that are reported are often given for tests
performed in dry conditions, which may or may not be relevant for neural and other
biomedical applications.
The American Society for Testing and Materials (ASTM) is a valuab le resource
for information about accepted and proper testing of the mechanical properties of
materials. Strong, uniform adhesion is desirable to create mag nesium with stable
coating and to predict its properties in living systems. Therefore, it is nece ssary to
test the adhesion strength of the coating to the material. A simple tes t of adhesion
strength is the tape test. In this test, a grid pattern is scored into the coating and
then tape is applied to the scored grid and removed at a set angle an d speed. Strength
of adhesion is ranked on a five-point scale depend ing on the amount of coating that
has detached from the underlying magnesium and become stuck to the tape instead.
Examples of how to evaluate adhesion strength with the tape tests are shown i n
Figure 12.10.
The microscratch and closely related nanoscratch tests are techniques from which
one can glean a range of information, including coating material hardness, yield
strength, and Young’s modulus. This test is often done with a specialized piece of
equipment, which can apply a constant and controlled force directed at an angle normal
to the coating surface. This technique can be used to evaluate organic or inorganic thin
film coatings (less than 5 mm in thickness for the microscratch test and less than
800 nm for the nanoscratch test).
Surface modification of magnesium by functional polymer coatings for neural applications 351
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Classification
5B None
4B
3B
2B
1B
0B Greater than 65%
Surface appearance
Figure 12.10 ASTM 3359 tape test classifications.
Reproduced from Sebaa et al. (2013).
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