Mechanical characterisation of hydrogel

materials
M. L. Oyen*
Interest in hydrogel materials is growing rapidly, due to the potential for hydrogel use in tissue
engineering and drug delivery applications, and as coatings on medical devices. However, a key
limitation with the use of hydrogel materials in many applications is their relatively poor
mechanical properties compared with those of (less biocompatible) solid polymers. In this review,
basic chemistry, microstructure and processing routes for common natural and synthetic
hydrogel materials are explored first. Underlying structure–properties relationships for hydrogels
are considered. A series of mechanical testing modalities suitable for hydrogel characterisation
are next considered, including emerging test modalities, such as nanoindentation and atomic
force microscopy (AFM) indentation. As the data analysis depends in part on the material’s
constitutive behaviour, a series of increasingly complex constitutive models will be examined,
including elastic, viscoelastic and theories that explicitly treat the multiphasic poroelastic nature of
hydrogel materials. Results from the existing literature on agar and polyacrylamide mechanical
properties are compiled and compared, highlighting the challenges and uncertainties inherent in
the process of gel mechanical characterisation.
Keywords: Hydrogel, Mechanical characterisation, Viscoelasticity, Poroelasticity, Rubber elasticity

Introduction Because a substantial fraction of hydrogel materials is

Hydrogels are network polymeric materials in which the
polymer chains are very hydrophilic, such that they can
associate with large quantities of water without dissolving.
The water can be tightly bound to the polymer network or
free to move within the polymer network.1 Because of the
large water content, hydrogels are quite biocompatible,
and can be made with water contents quite similar to those
of biological tissues (y70%) or much greater (up to and
beyond 99% water). They were first used for soft contact
lenses in the 1960s2 and have since been adopted for use as
catheter coatings,3 wound dressings,4 and in drug delivery
applications.5,6 Current research for future uses of
hydrogels indicates that they have great promise in tissue
engineering, cell encapsulation, nanoparticle coatings, and
in diagnostic microdevices such as microfluidics and
MEMS devices.1,7–10 Hydrogels have also increasingly
been used in basic science biological studies investigating
cell–material interactions,8 including those in which gel
mechanical properties are varied systematically in order to
study mechanical influences on stem cell differentiation.11–14
The mechanical behaviour of hydrogels has long been
recognised as fundamentally important, and previous
reviews have highlighted different aspects of hydrogel
mechanics without addressing all of the topics that will be
considered here.12,15,16
University of Cambridge, Cambridge, UK
*Corresponding author, email mlo29@cam.ac.uk
water, a limiting factor in the widespread adoption of
hydrogels for biomedical applications has been their
poor mechanical properties.7 This is particularly the case
when hydrogels are compared with traditional engineer-
ing materials in the context of mechanical behaviour. As
these are materials that have relatively small elastic
modulus values and which exhibit neither the behaviour
of something solid nor something liquid, both the
measurement and interpretation of mechanical data
presents significant challenges for the researcher. First, it
can be difficult to ‘grip’ the samples for testing. Second,
the elastic modulus of most hydrogels is on the order of
kiloPascals (kPa), while most mechanical testing equip-
ment is optimised for the range of MegaPascals (MPa)
to GigaPascals (GPa). Further, since hydrogels are
multi-phase materials consisting of a porous ‘solid’ with
a liquid (water) solvent phase, full understanding of the
mechanical response does not result from simple
treatments established for solid polymers, and more
complex analysis is required. All of these issues must be
addressed for researchers working to develop more
mechanically robust hydrogel materials, and all will be
addressed in the course of this review.
Hydrogel chemistry, preparation and
microstructure
Hydrogels can be categorised according to the nature of
the polymer network, including the origin of polymeric
material, basic polymer chemistry, details of preparation

ß 2014 Institute of Materials, Minerals and Mining and ASM International

Published by Maney for the Institute and ASM International
44 DOI 10.1179/1743280413Y.0000000022 International Materials Reviews 2014 VOL 59 NO 1

a poly(ethylene glycol), also poly(ethylene oxide) (PEG/PEO); b polyHEMA; c poly(vinyl alcohol) (PVA); d poly(acrylamide)
(PAAm); e an amino acid, the repeating unit of a protein; f a disaccharide, the repeating unit of a polysaccharide
1 Chemical structures for some common hydrogels
and types of chemical bonding present in the final
network.17,18 Hydrogel polymer network origins fall into
three main categories, those of synthetic polymer, those
of natural polymer and hybrid materials with both
synthetic and natural elements.10,17 Synthetic polymers
(Figure 1a–d) that have been commonly employed in
hydrogel studies include include poly(ethylene oxide) [PEO
although sometimes poly(ethylene glycol), abbreviated
PEG, when the molecular mass is below 20 000 g mol21],
poly(2-hydroxyethyl methacrylate) (polyHEMA), poly(-
vinyl alcohol) (PVA) and poly(acrylamide) (PAAm).19–28
Natural polymer (Figure 1, e-f) hydrogels include those
of proteins, such as collagen29 and silk,30 denatured
proteins, such as gelatin,31–35 and polysaccharides, such
as agar36–42 and alginate.43,44 Both synthetic and natural
gels can be multicomponent, either as grafted block
copolymers or as interpenetrating networks of two or
more independent polymers. Degradable linkers have
been incorporated into a chemically stable gel for
controlled release or regenerative medicine applica-
tions.6 Another mechanism of generating hybrid multi-
component materials is the grafting of a biological
recognition motif, such as the RGD (arginine–glycine–
aspartic acid) cell recognition sequence found in many
cell adhesive proteins such as integrins,45 to a synthetic
polymer in order to improve cellular biocompatibility
with the gel network.
Oyen Mechanical characterisation of hydrogel materials
In terms of determining the physical properties of gels,
the most important distinction is that between polymer
networks that are chemically bonded and those that are
networks of physical entanglements between polymer
chains. Figure 2 illustrates a series of chemical (Fig. 2a–
c) and physical (Fig. 2d–f) hydrogels. For chemical gels,
there are covalent cross-links at intersection points of
polymer chains, and in which the network can be (1)
ideally chemically bonded; (2) non-ideally chemically-
bonded, with polymer chain self-loops and free ends or
(3) a double network gel, in which there are two distinct
networks, each only covalently linked to ‘like’ chains
and forming an interpenetrating network structure. For
physical gels, there are effective cross-link points which
can have a range of sizes of the region connected by non-
covalent bonds. In the simplest case, chains loop around
other chains and form physical entanglements (Fig. 2d).
In both Fig. 2e and f, the regions of polymer chain
engagement at the effective cross-link points are larger
than at a single covalent point or a single chain
entanglement; in Fig. 2e, the formation of helices is
shown, and Fig. 2f represents the algae based gel
alginate, in which divalent cations (most commonly
Ca2z) form complexes joining the guluronic acid
domains in the polysaccharide chains.44 The nature
and definition of the cross-link points is fundamentally
distinct when comparing chemical and physical gels,
International Materials Reviews 2014 VOL 59 NO 1 45
Oyen Mechanical characterisation of hydrogel materials
a ideal cross-linked network with all tetra-functional linkages; b non-ideal cross-linked network including molecular ends
and loops; c ideal cross-linked double network gel; d physically entangled network; e physically entangled network with
helix formation; f alginate-like network with divalent Calcium ions forming local bridges between adjacent chains
2 Schematic microstructures of gels to define types of cross-linking
related to the relative ease of shifting these cross-link
points under applied mechanical loading. Chemical
cross-links are relatively immobile compared with
physical cross-links. Chemical and physical gels can be
produced from the same base monomer chain, as has
been shown in comparing gelatin32 or alginate44 gels
with different bondings.
Hydrogels can be considered as molecular scale
porous materials in which the pore space is occupied
by water.1 In a strict definition of a hydrogel, first the
chemical network is formed and then the network is
swollen in water,5 although sometimes the network
formation also takes place in an aqueous environment.
For chemical gels, cross-linking methods include chemi-
cal reaction and photopolymerisation with ultraviolet
light or by irradiation; physical cross-linking methods
include thermal, as in setting from a melt, or from a
reversible ‘bridging’ reaction such as in the divalent
Ca2z bridges in alginate gels, which are not covalently
linked (Fig. 2f).1
Swelling behaviour in hydrogels is important both
dynamically and at equilibrium. (poroelastic relaxation
experiments, described later in the section on ‘Poroelastic’,
have at times been called deswelling experiments, as they
involve the movement of water out of the polymer
network under physical stress.) A volume swelling
coefficient (Q) is defined as
Vswollen
Q~ (1)
Vdry
46 International Materials Reviews 2014 VOL 59 NO 1
The swelling coefficient is of fundamental importance
since it defines many of the properties that are critical in
hydrogels, including, but not limited to, the mechanical
and transport properties. The swelling coefficient differs
based on the polymer backbone chemistry and can vary
widely between different polymers; polyHEMA is a
moderate to poor swelling hydrogel compared with
highly swollen gels such as PEG and PVA. The swelling
coefficient also relates to the polymer volume fraction in
the swollen (equilibrium) state v2,s
1 Vpolymer
v2,s ~ ~ (2)
Q Vpolymer zVwater,initial zVwater,imbibed
Thus there can be two contributions to the overall water
content, the initial water in the system and the
additional volume of water imbibed during the swelling
phase. It is worth noting that Vpolymer in this formalism
is the same as Vdry above when network formation
occurs under dry conditions.
Another key parameter in understanding fundamental
scaling laws in the context of hydrogels is the (number)
average molecular weight between two adjacent cross-
linking points M- c . This is a complicated function of v2,s
(or Q) although a simple approximation46 for large
swelling ratios (Q.10) is
- c3=5
Q~bM (3)
where b is a constant related to the specific volume of the
polymer, the molar volume of water, and x12, the Flory
polymer–solvent interaction parameter.47,48 The swelling

3 Definition of hydrogel mesh size j, the distance
between effective cross-linking points
ratio is further related to the network mesh size (pore
size) j, the distance between cross-linking points in the
network10
 1=2
j~Q1=3 r- 20 (4)
 2 1=2
where r- 0 is the root mean square distance between
adjacent cross-links,46 which can be written as
 2 1=2
r- 0 ~l ðCn Nb Þ1=2 (5)
where l is the bond length, Nb the number of bonds and
Cn the Flory characteristic ratio, essentially a factor on
the order of 5–10 that describes the deviation from an
ideal freely jointed chain due to the restrictions of real
bonds in polymer chains (limited bond angles and steric
hindrance).46 A reasonable approximation for the value
of Cn for long polymer chains where nR‘ is
1zcosh
C? &
 (6)
1{cosh
where h is the bond angle between adjacent segments.
The mesh size j (Fig. 3) can be influenced by the gel
chemistry, including the cross-linking density, but also
by the external environment including the temperature
and solvent chemistry (pH and ionic strength).46 An
example of a fine porous structure with reasonably
uniform mesh size is shown for gelatine in Fig. 4.
Any of the structural parameters introduced here (v2,s,
Q, M - c , j in particular), which determine the behaviour
of the hydrogel network, are of fundamental importance
in gel function. While the swelling ratios can be
determined relatively simply, the determination of the
remainder of these parameters can be made challenging
by the fact that the pore sizes of interest for typical
hydrogels are in the 1–100 nm range. This is further
complicated by the fact that dehydrating the highly
swollen network to enable direct measurements in
vacuum, such as by electron microscopy (Fig. 4) or
porosimetry, could potentially change the pore size and
structure. Indirect methods of mesh size determination
include by light scattering, and most important for
Oyen Mechanical characterisation of hydrogel materials
4 A scanning electron microscope (SEM) image of a 15%
gelatine mixture quenched from molten in liquid pro-
pane and freeze dried prior to imaging. The specimen
preparation is unlikely to result in an identical micro-
structure to the one that results from the slow cooling
of the hydrated molten mixture as in typical gelation.
The scale bar is 2 mm in length
this review, by inference from measured mechanical
properties.5 Thus, in addition to being useful for the
obvious reason, measurements of the physical properties
of gels constitutes one of the most important mechan-
isms for inferring gel structure, thanks to the existence of
well established structure–properties relationships. Since
the mechanical properties of gels are relatively poor (kPa
elastic stiffnesses), thus limiting their applications for
biomedical use where structural properties are impor-
tant, there is great need for the development of robust
methods of measuring the mechanical properties in their
own right. Next, we will consider how the mechanical
properties of gels can be measured before returning to
fundamental relationships between mesh size and
mechanical properties.
Hydrogel mechanical characterisation
techniques
The basic mechanical testing techniques and apparatuses
used for polymeric materials more generally are also
used for mechanical characterisation of hydrogels. As
with most polymers, hydrogels exhibit time dependent
mechanical behaviour due to intrinsic viscoelasticity of
the polymer network, but with an additional time
dependent deformation mechanism due to fluid flow.
Thus, time factors heavily in the planning and execution
of mechanical experiments on hydrogels, which can be
characterised in either the time or frequency domain.
Here we will consider the six most common testing
techniques (Fig. 5): tension, compression, which can be
either unconfined or confined, local indentation with a
probe and frequency based tests such as shear rheometry
or dynamic mechanical analysis (DMA). Finally, there
are some non-contact mechanisms used to infer mechan-
ical properties.
Universal Test-frame
The most common tool used for mechanical character-
isation of materials is a Universal Test-frame, which is
capable of carrying out a wide variety of experimental
tests. Most frames are uniaxial, in that motion is
actuated and force measured along a single axis,
although biaxial instruments have become increasingly
common in recent years.49 The mechanism of motion
actuation can vary, including servohydraulic systems,
electromechanical systems (both very common) and
International Materials Reviews 2014 VOL 59 NO 1 47
Oyen Mechanical characterisation of hydrogel materials
a tension; b compression; c confined compression; d indentation; e shear rheometry; f dynamic mechanical analysis
(DMA), illustrated here for three point bending
5 Schematics of different types of mechanical testing set-ups used for testing hydrogel mechanical properties
systems driven by speaker coils (less common). The same
systems can be used for tension and compression testing,
with different hardware at the point of sample contact.
For tensile testing, key to execution of effective tests is
good sample gripping, something that is particularly
difficult when the specimens are compliant and
hydrated, as in the case of hydrogels. Strategies for
addressing this challenge include the use of cardboard
tabs, double sided tape and glue to assist in the gripping
the hydrogels.39 For compressive testing, the sample can
either be unconfined (Fig. 5b) and compressed between
two non-porous platens,26,39,40,50 or confined (Fig. 5c) to
a container and compressed with a single porous
platen.51 The latter case is a testing modality unique to
multiphase materials, as the motion of fluid out of the
sample, through the porous platen, is expected (this will
be discussed further in the section on ‘Poroelastic’).
Universal Test-frames range in size from extremely large
(meganewton, MN) scale machines used in testing civil
engineering components to small modern machines
optimised for soft biological materials or biomedical
components including hydrogels. A typical load cell for
such testing would have a maximum load capacity of
only 5 or 10 N.
For both tension and unconfined compression testing,
the load–displacement (F–x) data are converted to
stress–strain (s–e) data using simple geometrical rela-
tionships52 and the Young’s modulus E and failure
48 International Materials Reviews 2014 VOL 59 NO
strength sf are reported. Typical tests are done at fixed
rate until specimen failure, and due to the time
dependent nature of hydrogels, testing at a variety of
strain rates may be required (Fig. 6a).53 Especially given
the nature of the time dependent response of hydrogels,
in addition to simple tests at fixed rate, the specimen
may be subjected to creep (extension at fixed load or
fixed stress) or relaxation (load decrease at fixed
deformation or strain) experiments where variation in
response with time is of interest and the loading is fixed
(Fig. 6b).
Indentation
In indentation testing, a probe of known geometry is
brought into contact with a material surface, pressed
into the material, and retracted again. Historically,
indentation testing was the gold standard for examining
the hardness of metals,54 and indenters were dead weight
loading devices coupled with optical microscopes for
examination of the residual indent impression. However,
the growth in recent years has been in depth sensing
indentation, in which the full indentation load–depth–
time (P–h–t) profile is recorded and analysed for
material property deconvolution. Thus, the indentation
test is a local version of a compression test, where the
sample is compressed in a small region instead of across
the entire surface. The most commonly reported
parameter associated with indentation testing is the
1

6 Mechanical test types: imposed displacement–time (x–t) or force–time (F–t) for a fixed rate for varying rates, b static
creep or relaxation at fixed force or displacement and c sinusoidal loading at varying frequency
reduced modulus ER, which for compliant materials is
equivalent to the plane strain modulus
E
ER ~ (7)
1{n2
where v is the Poisson’s ratio.52
Minimal sample preparation is required for indenta-
tion testing. As a result, indentation has been growing in
popularity for testing materials, such as hydrogels,
which would be difficult to ‘machine’ into a regular
specimen geometry. Hydrated, compliant specimens can
also be difficult to ‘grip’ for tensile or compressive
mechanical testing, which is not required in probe based
testing. The relatively small volumes of material
required for quantitative indentation testing means that
valuable samples can be examined, and high throughput
screening methods may prove useful for materials
selection. Further, inhomogeneous graded or patterned
gels can be probed locally for quantification of the local
inhomogeneity, using a mapping technique across the
sample surface.55 Finally, it is straightforward to keep
hydrogel samples hydrated throughout the duration of
an indentation test.
Indentation testing has been used to study hydrogels
at large (mm)40,56,57 and at small (nm to mm) length
scales.13,19,22,27,41,58,59 The technique is highly adaptable
with regard to length scale, as the probe geometry and
contact area can be tuned to optimise the testing to the
instrument such that the forces and displacements are
appropriately scaled in terms of both range and
resolution.60,61 At large length scales, a Universal Test-
frame can be adapted for execution of an indentation
test. More recently, ‘nanoindentation’, or indentation
testing at small length scales, has emerged as a leading
technique for the mechanical investigation of a wide
range of materials.62,63 For this small scale indentation,
there are different approaches to take with respect to the
testing instrumentation. There are a number of com-
mercial stand alone nanoindentation systems that have
been developed for small scale quantitative testing and
that have been used for hydrogel characterisa-
tion.19,22,58,59 For historical reasons, these instruments
are largely optimised for testing stiff engineering
materials, and adaptations of typical test techniques
(such as the use of larger indenter and contact radii) are
employed for testing compliant polymers and hydrogels.
An alternative approach is to utilise atomic force
microscopes (AFMs) for gel indentation testing.12,13,27,41
Oyen Mechanical characterisation of hydrogel materials
The primary difference between commercial nano
indentation systems and AFMs for indentation testing
lies in the different physical principle governing trans-
ducer operation: commercial nanoindenters utilise
speaker coils or capacitance gages to directly actuate
the indenter probe into the sample, while AFMs actuate
the tip indirectly via a calibrated cantilever. The
cantilever based systems are thus easy to optimise for
testing different classes of materials with different
stiffness simply by switching the cantilever stiffness.
However, the quantitative nature of the test results is
extremely sensitive to cantilever calibration. In both
cases, feedback control is required for quantifying time
dependent deformation. In practice, neither nanoinden-
tation nor AFM indentation utilising existing commer-
cial equipment is ideally suited for hydrogel testing but
both are likely to grow in popularity in coming years as
the need for gel property quantification at small length
scales continues to increase.
Frequency based testing
Frequency based sinusoidal testing is commonly carried
out using ‘black-box’ testing systems (Fig. 5e and f),
using rheometry in shear20,43,64,65 or in a range of DMA
test modes including shear, tension or bending.51,66 The
sample, often in a geometry specified for the instrument
and test type, is loaded into the instrument and different
types of ‘sweep’ measurements can be performed. Most
common is a frequency sweep (Fig. 6c), where the
sinusoidal oscillation is at a fixed (and usually quite
small) strain but the oscillation frequency is varied. The
test output is the storage (G9) and loss (G0) components
of the complex modulus G* as a function of frequency,
where
G  ~G’ziG’’ (8)
The temperature can also be varied to investigate
transition temperatures such as the glass transition,
and also to take advantage of time temperature super-
position to extend the range of the experimental data far
beyond that which is experimentally practicable in terms
of very slow and very fast oscillation rates. These
frequency based methods have their user base largely in
polymer chemistry and polymer science programmes;
rheometry grew to be the technique for studying physics
of polymer melts, while DMA was used for interroga-
tion of solid polymers. Because rheometry can be used
International Materials Reviews 2014 VOL 59 NO 1 49
Oyen Mechanical characterisation of hydrogel materials

on viscoelastic liquids as well as viscoelastic solids, it has

been employed for studying the gelation process.64
New developments have expanded the use of frequency
based techniques beyond traditional rheometry or DMA.
Sinusoidal indentation testing utilising frequency sweeps
has gained some recent traction for polymer and hydrogel
characterisation41,59,67 and dynamic confined compres-
sion measurements have been used,51 for comparison
with traditional confined compression creep measure-
ments. For small scale dynamic measurements, a novel
technique termed ‘microrheology’ has emerged, in which
tracer particles are embedded within a gel and their
motion monitored.12 This allows for local probing of
viscoelastic behaviour throughout a gel’s thickness.
Non-contact
The techniques described above have all involved
physically extending or compressing a sample. There
are some additional mechanisms for measurement of
physical properties without substantially deforming the
sample. Permeability can be measured by flowing fluid
through a gel at known rate and measuring pressure
with a pressure transducer20 or by varying the pressure
and measuring the flowrate.68 This type of measurement
is sometimes denoted the ‘direct permeability’ since no
material model is required to interpret the measurement.
Other measures rely on indirect mechanical estimates of
mechanical properties based on the pioneering models of
Flory and Rehner in the 1940s.47,48 The actual measure-
ments can come from Gel Permeation Chromatography
(GPC)69 or from light scattering.70–73
Models for interpretation of mechanical
results
Once experimental data have been collected, the load–
displacement–time (F–x–t, or P–h–t for indentation) or
stress–strain–time (s–e–t) data must be interpreted
within some mechanical framework in order to obtain
mechanical property values. The simplest case, for
linearly elastic and isotropic materials, is when the slope
of the linear, small strain, stress–strain (s–e) curve from
a uniaxial (tensile or compressive) test is taken as the
elastic modulus E. For hydrogels, the fundamental
constitutive response is more complicated, and as such
more complicated analyses must be considered, includ-
ing elasticity under large strains and the presence of time
dependent deformation. Both analytical and numerical
computational approaches will be considered here.
Elastic and hyperelastic
The polymer network in hydrogels is often treated using
concepts originally developed for the study of rubber
elasticity. This is a particularly good approximation for
highly cross-linked chemical gels (Fig. 2a) in which the
polymer network is in effect a single molecule due to
the high connectivity. For such a rubber-like network,
entropic elasticity is dominant, and the elastic modulus G
(note this is the terminology utilised in the rubber elasticity
literature, and is not necessarily meant to indicate shear
modulus) in the unswollen state can be simply expressed
using the theory of rubber elasticity15,47,48,74,75
G~Np kT
where k is the Boltzmann constant, T is temperature, Np is
the number of polymer chains per unit volume, and where
7 Stress–stretch relationships for rubber elasticity, illustrat-
ing the tensile relationship for both true stress and engi-
neering stress (equations (10) and (11) respectively)
a chain is defined as a segment of polymer between cross-
link points. This can also be rewritten as
rRT
G~ (10)
M
-c
where r is the density and R is the ideal gas constant. Both
of these expressions for the elastic modulus are based on
the assumption that the network deforms in an affine
manner, that is, that the deformations at the macroscopic
level translate directly to the molecular level. Thus, the
modulus is effectively a direct function of the cross-linking
density75 and can be related directly to M - c , which is then
related to Q and j via equations equations (3) and (4)
respectively. Whether the assumption of affine deforma-
tion holds for a hydrogel is an open question, and this
assumption has been directly challenged for polyacryla-
mide gels.76
For the swollen state, characteristic of a hydrogel with
volume swelling ratio Q, the solvent swollen modulus is
the product of the unswollen (pure polymer) modulus
and the linear swelling ratio
Gswollen ~GQ{1=3 (11)
The assumption in all of these cases is that the network
is perfect (Fig. 2a), and the expected modulus for a real
network would be decreased proportionally to the
number of network defects, such as loops and free chain
ends (Fig. 2b).
For simple uniaxial extension or compression, the
relationship between true stress t, and stretch,
l~lcurrent =loriginal is simply
 
t~G l2 {l{1 (12)
The use of stretch, instead of the more classical
mechanical parameter strain (e), and the use of true
(9)
stress are both useful in treating rubbery polymers with
very large deformations to failure. The engineering
stress-stretch relationship

50 International Materials Reviews 2014 VOL 59 NO 1


a lumped parameter models for viscoelasticity, with series and parallel combinations of springs and dashpots, here
representing generalised Kelvin and Maxwell models; b schematic illustration of water motion and pore pressure during
poroelastic deformation
8 Time dependent deformation mechanisms
 
s~G l{l{2 (13)
is quite different from the true stress–stretch expression
(Fig. 7), and is clearly Hooke’s law (using G instead of
E) in the limit of small stretches. A key difference
between rubber elasticity and elasticity theory is that
there is only one constant here, G, whereas in generalised
isotropic elasticity there are two (often E and v).
The elastic modulus is an equilibrium parameter, and
to this point we have considered only elastic deforma-
tion. An approximate equilibrium modulus is measured
with the use of relatively slow strain rates in tensile or
compressive testing in many studies of hydrogel
mechanical behaviour. However, there is intrinsic time
dependent deformation present in hydrogels due both to
the viscoelasticity of the polymer network and due to the
motion of fluid within the gel, and there is thus an
opportunity to gain additional structural information
about a gel via the examination of this dissipative
deformation. Thus, we will next consider hydrogel non-
equilibrium behaviour.
Time dependent
Time dependent mechanical behaviour is fundamental
to the physics of polymers, including hydrogels. The
treatment of this time dependence ranges from relatively
simple empirical measurements through to complex
Oyen Mechanical characterisation of hydrogel materials
models treating the solid and fluid phases and their
interactions explicitly using the coupled equations of
poroelasticity. Here, viscoelasticity will be considered
first, followed by poroelasticity.
Viscoelastic
The basic premise of viscoelasticity is that it parallels the
framework of elasticity but with time dependent
functions replacing the elastic constants.77 It is quite
typical to assume that that volumetric deformation is
time independent, and there is a solitary time dependent
shear function G(t). For very simple cases, physical
lumped parameter models (Fig. 8a) are used and explicit
spring and dashpot coefficients are reported. However,
more generally, polymer viscoelasticity in the experi-
mental time domain is treated with the assignment of an
empirical Prony series (summed exponential function) to
describe the material’s time dependent constitutive
response, for example
sðtÞ X
G ðtÞ~ ~D0 z Dk expð{t=tk Þ (14)
e0
This expression describes the time dependent change in
modulus during the relaxation of stress under fixed
strain e0. The complementary function, for describing
extensional creep under fixed stress s0 is written in a
similar form
International Materials Reviews 2014 VOL 59 NO 1 51
Oyen Mechanical characterisation of hydrogel materials
X characterises the multiphase nature of hydrated materi-
eðtÞ
J ðtÞ~ ~C0 { Ck expð{t=tk Þ
s0
The functions are not explicitly inverses in the time
domain but are in the Laplace domain [G(s)J(s)51],
although the limiting values do invert in the time
domain [G(0)51/J(0) and G(‘)51/J(‘)]. Given a known
testing geometry, it is relatively straightforward to fit
these functions to experimental data, even for compli-
cated testing modalities such as indentation.61 Fre-
quently reported parameters would include the instan-
taneous and equilibrium modulus values, G(0) and G(‘)
or E(0) and E(‘), and the time constants tk. The ratio of
modulus values f5G(‘)/G(0)5E(0)/E(‘) can also be
reported as a measure of elasticity, where f51 signifies a
perfectly elastic solid and f50 a perfectly viscous liquid.
For complicated load–time or displacement–time his-
tories, Boltzmann hereditary integrals can be used to
find full F–x–t or P–h–t time histories.77
Further, viscoelastic experiments can be conducted in
the frequency domain, as in the case of DMA66 and
rheometry.64 Spring and dashpot models can be used to
interpret the data, or more sophisticated approaches such
as fractional derivative models can be employed.66
Viscoelastic textbooks77 show how data can be converted
between the frequency domain and the time domain, as
there are not different physical functions describing
viscoelastic behaviour in the frequency versus time domain
but a single physical viscoelastic response for the material.
However, in practice, it is relatively infrequent to find the
conversion done routinely. DMA and rheometry data are
frequently reported as a plot of storage and loss moduli (G9
and G0) versus frequency (v) without further analysis. To
date, there has been relatively little success in trying to
assign any of the viscoelastic coefficients to explicit
physical processes or to derive expected values from first
principles based on polymer structure.
Poroelastic
The time dependence demonstrated by hydrated materi-
als has been described using multiphasic poroelastic
models. Poroelastic78 (also called biphasic79) constitutive
behaviour is when time dependence arises due to the
flow of a fluid through an elastic (or viscoelastic), porous
solid (Fig. 8b). The material properties of interest in
poroelastic material characterisation include the elastic
properties of the porous skeleton: the (shear) elastic
modulus G and the (drained) Poisson’s ratio v, the
undrained Poisson’s ratio vu and a parameter a which
ranges from 0 to 1 and describes fluid-solid interactions.
The fifth constitutive parameter is the Darcy perme-
ability k which is a function of the fluid viscosity g and
the intrinsic permeability k5kg. The permeability can be
thought of as analogous to the diffusion coefficient, but
under the motive of a mechanical, rather than a
chemical, gradient. The intrinsic permeability is also
indicative of the effective pore size (j) of the porous
skeleton k5Cj2 where C is a constant. A frequently
reported parameter for poroelastic data analysis is the
aggregate modulus HA
E ð1{nÞ
HA ~
ð1znÞð1{2nÞ
An advantage of such an approach compared with linear
viscoelasticity is twofold. First the model directly
52 International Materials Reviews 2014 VOL 59 NO
(15) als such as gels. Second, the material parameters are
physical constants related to the material microstruc-
ture, and not empirical fitting parameters as in typical
linear viscoelastic analysis (equations (14) and (15)). In
particular, the intrinsic permeability k can also be
calculated from first principles related to the total solid
fraction (v2,s5w in the poroelastic literature) and a
characteristic length scale a, typically taken as a fibre
diameter, such that k5a2f(w) where the function f(w)
depends on the arrangement of the fibrous network. For
a random, isotropic network, which is a sensible
assumption for a hydrogel material, an expression was
derived by Jackson and James80 based on extrapolation
of two-dimensional fibre arrays
3
f(w)~ ð{lnw{0:931Þ (17)
20w
Recently, numerical simulation has been undertaken to
perform detailed three dimensional computations for
permeability of three-dimensional networks,81,82 which
agree well with the Jackson and James calculation.80
There are also classic solutions for flow of fluid parallel
and perpendicular to oriented fibre arrays, and as
expected, the isotropic network prediction falls between
these two extremes.
Poroelastic data analysis is complicated by the fact
that the governing equations are coupled, and as such
there are relatively few types of mechanical tests for
which simple, closed form solutions exist and can be fit
directly to data. The exception is the case of one-
dimensional consolidation, or confined compression
(Fig. 5c). For both the assumption of a step load or a
constant rate ramp followed by a hold period at
constant load (stress), relatively simple solutions exist
and are in the form of summed exponentials.83 However,
for more complicated loading schemes, such as indenta-
tion testing, numerical or computational models are
often required. Several recent papers have considered
the measurement of poroelastic properties, particularly
the elastic modulus and hydraulic or intrinsic perme-
ability, of hydrogels with good success using indentation
techniques.19,44,83–87 Permeability has been measured
directly by flowing fluid through a gel, and there was
good agreement between direct permeability measure-
ments and values obtained via indentation testing.19,88
However, compared with elastic modulus values, the
permeability is less frequently measured and reported
for hydrogel materials.
Compiled mechanical results for
common hydrogels
Next, material property results from a number of studies
will be compared as a function of gel polymer concen-
tration for two common hydrogel systems, one physical
gel and one chemical gel. Where tabular data were
unavailable, data were digitised using open source
software.89
Agar
(16) Agar gels are largely made of the linear polysaccharide
molecules that form from the disaccharide agarose. They
are physical gels that form on cooling, with a gel
temperature near 35 C.39
1
 Oyen Mechanical characterisation of hydrogel materials

10 Plot of compiled agar gel permeability values as a

function of gel concentration. Data compiled from
Refs. 51, 56, 57, 92 and 93

size is also decreasing. However, the data do not seem

to indicate a simple change in pore size at fixed fibre size,
9 Plot of compiled agar gel elastic modulus values as a
as might be expected. The trend is steeper than would
function of gel concentration. Scaling laws for modulus
be predicted from a 23=2 power law in polymer
with polymer (gel) concentration according to cellular
concentration.73 The data are also not consistent with
solids (power law 290) and polymer physics (power law a single prediction of permeability based on molecule
2?2591) are shown for reference. Data compiled from size and solid fraction:80 using a value of a51?9 nm80
Refs. 39–41, 51 and 56–59. only the data at large polymer concentrations agree well
with the prediction from equation (17). The data at
smaller polymer concentrations are consistent with a
larger effective fibre size, on the order of 3a, as shown in
Fig. 11. It is useful to note that the simple 23=2 scaling
Elastic modulus values for agar gels have been
compiled from a number of sources39–41,51,56–59 and are
plotted as a function of gel concentration in Fig. 9.
Owing to the large range in values, and the uncertainties
associated with assuming Poisson’s ratio values, the data
are plotted as reported, without attempt to convert any
reported shear (G), storage (E9 or G9), reduced (ER) or
aggregate modulus (HA) to a Young’s modulus (E)
value. As can be seen from the plot, for any single value
of the gel concentration, the reported modulus values
vary by up to three orders of magnitude. There are
factors that are likely confounding the simple plot, such
as agarose chain molecular weight.39 However, within
any single set of data, the scaling with gel concentration
is consistent with either of two predicted concentration
dependencies proposed in the literature, either quadratic90
or in concentration to the 9=4 power.91 The difference
between different studies is a vertical translation of entire
data sets to larger modulus at smaller gel concentration.
There is no clear trend with test method when different
studies were compared, as there is agreement between
indentation and confined compression41,51 at small
modulus and between indentation, tension and compres-
sion at large modulus.39,58 11 Plot of agar gel permeability data from Fig. 10, com-
Data for hydraulic permeability of agar gels as a pared with analytical permeability scaling laws for a
function of gel concentration are much more consistent fibre size of a51?9 nm and 3a55?7 nm (equa-
than modulus data across studies,51,56,57,92,93 as shown in tion (15)80) and with a 23=2 power law.73 The perme-
Fig. 10. As the polymer concentration increases, the ability data do not appear to follow any one of these
permeability decreases, implying that the effective pore predicted trends with gel concentration

International Materials Reviews 2014 VOL 59 NO 1 53

Oyen Mechanical characterisation of hydrogel materials
12 Plot of compiled acrylamide gel elastic modulus
values as a function of the total polymer concentra-
tion (%T). Scaling laws for modulus with polymer (gel)
concentration according to cellular solids (power law
290) and polymer physics (power law 2?2591) are
shown for reference. Data compiled from Refs. 19–23,
26 and 27
power law73 closely approximates the trend of much of
the more complicated Jackson and James expression.80
To summarise the agar data (Figs. 9–11), the modulus
increases and the intrinsic permeability decreases with
increased polymer content, or conversely the modulus
decreases and intrinsic permeability increases with
increased water content. There is substantial scatter in
the modulus data, and less scatter in the permeability
data. The modulus data seems to agree well with
predicted behaviour from scaling arguments, but perme-
ability does not. This may be because agar is more
appropriately treated as poroviscoelastic rather than
poroelastic, which was only considered in one of the
cited studies.56 Recent work has confirmed that agar is
poroviscoelastic, and that with multiple experiments the
contributions from different time dependent deforma-
tion mechanisms can be decoupled.94 To establish the
generalisability of the agar results here, a second gel is
next examined using the same methodology, allowing
for consideration of bond type in contrasting the
physical agar gel with a chemically cross-linked gel.
Polyacrylamide
Polyacrylamide (PAAm) hydrogels could be considered
to be the best characterised of all gels, being ubiquitous
across biomedical research, including being used in
biology for gel electrophoresis to separate proteins by
size and charge.95 As with many polymer systems, the
acrylamide monomer is extremely toxic in its unpoly-
merised form but benign once polymerised. For this
reason, much effort is usually undertaken to allow for
outward diffusion of unpolymerised monomer prior to
using the gels for a wide range of biomedical applica-
tions. The monomer is polymerised in aqueous solution
in the presence of small amounts of bifunctional cross-
linker, frequently N,N9-methylenebisacrylamide (BIS or
bis-acrylamide), resulting in an acrylamide network with
54 International Materials Reviews 2014 VOL 59 NO
13 Acrylamide gel elastic modulus values from Fig. 12,
replotted as a function of cross-linker concentration
(%C) for data grouped according to the total polymer
concentration (%T)
BIS links. This reaction occurs via a free-radical mechan-
ism, where ammonium persulfate (APS, N2H8S2O8) is
used as an initiator, and a small volume of TEMED
(N,N,N,N-tetramethylethylenediamine) is used to stabi-
lise the reaction. This is typically held at fixed concentra-
tion, such that there are two key parameters determining
the overall nature of the hydrogel. These have a reason-
ably consistent nomenclature used across the literature as
follows: T represents the total polymer concentration (w/
v) where this is the sum of the monomer and BIS; C is the
percentage (w/w) of the polymer that is cross-linker. Thus,
%T5100%[BIS (gm)zmonomer (gm)]/100 mL solution
and %C5100%[BIS/(BISzmonomer)] (gm/gm). Because
the cross-linker is typically present in much smaller
amounts than the monomer, it is common for the polymer
concentration %T to be calculated based on the monomer
only, and for the cross-linker concentration %C to be
calculated from the ratio of BIS to monomer.
The relationship between pore size and both total
polymer concentration and cross-linker concentration is
complex. When %T is fixed, the pore size decreases with
increasing %C up to a point, but then larger %C causes a
more macroporous structure due to polymer chains
agglomerating.69 When %T is varied at fixed cross-linker
concentration, the pore size decreases with increasing
%T. Thus, a full picture for the mechanical behaviour of
PAAm hydrogels requires a large data set with a range
of both polymer and cross-linker concentration.
Polyacrylamide elastic modulus values (E,G or G)
have been compiled from a range of sources19–23,26,27
and are plotted in Fig. 12 as a function of total polymer
concentration (%T). As was described above for the agar
gels, different moduli were plotted on the same axes
without conversion, as the differences between these for
a single material are small compared with the scale
extending over seven orders of magnitude in modulus.
Two scaling lines, one corresponding to modulus going
as the square of the concentration90 and the second
using the 9=4 scaling law,91 are again shown to agree with
the basic trend in the data for sharply increasing
1

14 Plot of compiled acrylamide gel permeability values
as a function of total polymer concentration (%T). In
contrast with the agar data (Fig. 11), the data taken as
a group do appear to follow a -3/2 scaling law for per-
meability with gel concentration.73 Data compiled from
Refs. 19,20, 68, 69, 73, 96 and 97
modulus with increased %T. As with the agar data, there
is considerable variation in the modulus values reported
at a single polymer concentration, by as much as several
orders of magnitude. As some of this is likely
attributable to different cross-linker concentration, the
modulus data from the same authors and studies was
binned by %T and plotted as a function of the cross-
linker concentration (%C) in Fig. 13. The effect of %C is
relatively minor compared with that of %T in determin-
ing elastic modulus values when looking at the data
across such a wide range of %T.
Intrinsic permeability values for PAAm gels, assum-
ing a linearly poroelastic response, have been compiled
from a number of sources where measurements were
made19,20,68,73,96,97 and one study for which permeability
is here calculated as the square of the estimated pore size
from GPC measurements,69 and are plotted in Fig. 14
(note that it was assumed that the y-axis scale for Fig. 8
in Tokita and Tanaka’s work73 is mislabeled, based on
comparisons with their Figs. 6 and 7, and the data
have been shifted accordingly). There is good agree-
ment between the computed values69 and the measured
values for some of the studies,19,68,73,96 but the data
from two other studies20,97 are substantially different,
namely, larger by several orders of magnitude. The
data that all agree are also consistent with a roughly
23=2 scaling law with total polymer concentration
(%T)73 as shown on the plot. The scatter in perme-
ability values for the same total polymer concentration,
at about an order of magnitude across a range of
studies, is less than the scatter in reported modulus
values shown in Fig. 14. Further, there is even less
Oyen Mechanical characterisation of hydrogel materials
15 Acrylamide gel permeability values from Fig. 14,
replotted as a function of cross-linker concentration
(%C) for data grouped according to the total polymer
concentration (%T)
evidence of any trend for permeability values with
cross-linker concentration (Fig. 15), which is interest-
ing and slightly perplexing, since the %C is thought to
have such important and not straightforward influ-
ences on the hydrogel pore size.69
Conclusion and outlook
In this review, two complementary aspects of the
mechanical properties of hydrogels have been consid-
ered. For many applications, the hydrogel properties are
intrinsically important, and in many structural applica-
tions, they present a major limitation for utilisation in
real world applications. Mechanical measurements are
further important for inferring difficult to measure
hydrogel structural characteristics such as mesh size.
Both of these aspects are important moving towards the
introduction of hydrogels into clinical applications, such
as in tissue engineering applications. Avascular con-
nective tissues, such as the spinal disc57 and cornea98 are
obvious targets for tissue engineering due to their
relatively simple microstructures, but in both cases, the
tissues are subject to mechanical loads in vivo. If
hydrogels are to succeed as tissue engineering scaffolds,
their mechanical properties must be sufficient and robust
and repeatable methods for measuring these properties
must be established.
In compiling sets of data from the literature,
variability in reported elastic property values (E, ER,
G, G9, HA) for the same hydrogel compositions has been
shown to be quite large at any given gel concentration.
For example, reported modulus values for a 2% agar gel
range from just over 1–300 kPa (Fig. 9). For a 3%
acrylamide gel values ranged from 10 Pa to 100 kPa,
although the 100 kPa value is perhaps an outlier
(Fig. 12). With that value excluded, a two order of
magnitude range remains, from 10 Pa to 1 kPa. These
comparisons in Figs. 9–15 show significant but not
systematic variation between different studies, in which
different testing modalities were utilised. This raises the
question as to whether there is consistency when
multiple test modalities have been compared within the
same study. Good quantitative agreement between
International Materials Reviews 2014 VOL 59 NO 1 55
Oyen Mechanical characterisation of hydrogel materials
frequency based rheometry and DMA results has been
demonstrated for dextran methacrylate hydrogels.99 In
one study where compression and indentation testing
were used for the same 3% agar gel, the indentation
measurements of modulus were 52% larger than for
compression testing.40 Indentation data, both nanoin-
dentation and microindentation, also gave larger mod-
ulus values than either compression or DMA testing for
20% acrylamide gels, and there was only limited
agreement between time and frequency based test
methods.19 A third study also compared compression
and AFM indentation of hyaluronic acid gels and found
no systematic variation between the test modalities with
gel concentration; at small gel concentrations the
indentation values were smaller, whereas at large gel
concentrations the indentation values were larger.13
These results taken together highlight potential chal-
lenges for the field. Uncertainties for the measurement of
elastic modulus values in structural engineering materi-
als are typically on the order of a few percent, not a few
orders of magnitude. Significant questions remain as to
both the accuracy and precision of the modulus
measurements for these compliant hydrogels. Taken as
a whole, the data illustrate that more studies are needed
in which mechanical testing modalities are system-
atically compared for the same gels, in order to establish
best practices for both experimental protocol and data
analysis methodology.
The gels considered here, particularly those simple
gels such as agar and polyacrylamide, were single
component, in bulk form and subject to bulk mechanical
characterisation. The modulus and permeability both
scaled with gel concentration in a manner largely
consistent with theoretical predictions.73,80,90,91 Modulus
increased with increasing polymer fraction (and decreased
proportion of water) and permeability decreased with
increasing polymer fraction (and decreased water frac-
tion). There is thus limited scope for dramatically
improving hydrogel properties in these simple systems.
Recent attempts have aimed at designing unique micro-
structures in which modulus and permeability are
decoupled.100,101 Composite gels have been made, includ-
ing gel–gel systems, such as agar–gelatine,57,102–105 agar–
collagen106–108 or gelatine–PAAm.109 Gels have also been
reinforced with a non-gel phase, such as particles of
hydroxyapatite,110 clay111 or electrospun polymer
fibres.112,113 Multilayer gels have been formed in three
dimension, including onion-like structures with multiple
layers and internal interfaces.114,115 Slide ring gels with
mobile cross-links have been envisaged.111 Superporous
hydrogels that can quickly imbibe large quantities of
water have been developed for drug delivery applications
in the gut.116 Functional gels, such as ‘responsive’ gels7,
gels that are able to undergo a change in response to a
change in local chemical or physical stimulus such as a pH
change5,117 or a change in temperature.118 Such gels can be
used as sensors to detect local environmental changes.9
Novel properties of gels, such as electroconductivity,119
may allow for the formation of artificial muscles9 with
good biocompatibility.
Although this review has largely focussed on the
limitations of hydrogel stiffness, for demanding applica-
tions such as structural tissue engineering, the failure
characteristics, strength and toughness, are also critical.
There has been significant recent interest in mechanically
56 International Materials Reviews 2014 VOL 59 NO
tough double network gels such as PAAm with poly(2-
acrylamido-2-methylpropanesulfonic acid) (PAMPS),
for which the toughness of the composite is orders of
magnitude greater than the toughness of single compo-
nent PAAm or PAMPS.120–122 The mechanisms for
extreme toughening have been investigated using mole-
cular modelling.123
The double network gels in particular show promise
for mechanical applications, but as has been demon-
strated here, there is a need for a frank discussion within
the research community on measurement accuracy and
precision when measuring gel mechanical properties. In
examination of the papers reviewed here, hardly any
mention was found of calibration, or of verification of
the testing technique using an independent standard with
known physical properties. In general, there is a need for
broad consensus to be developed across the field on best
practice mechanical characterisation techniques, includ-
ing both execution of experiments and analysis of data.
This is perhaps surprising given the ubiquity of commer-
cially available hydrogels with obvious needs for mechan-
ical stiffness and good transport properties, such as
contact lenses. Going forward, in order to develop
advanced hydrogel systems for demanding structural
tissue engineering applications, and in which the perme-
ability is critical for cell viability, greater confidence in,
and even possibly standardisation of, hydrogel mechan-
ical characterisation will be required.
Acknowledgements
The author acknowledges the efforts of DGT Strange
and JM Shapiro in reading versions of this manuscript
and making helpful corrections and suggestions, and
DGTS for drawing Fig. 8b based on an original design
of M. Galli, EPFL. The author also acknowledges K
Main for hydrogel SEM work in the course of her final
year project work (Fig. 4). Finally, Ed Berger, of the
University of Virginia, is gratefully acknowledged for
first showing the author a plot illustrating the variability
of agar modulus with concentration, which inspired this
review.
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