Food Research International 122 (2019) 209–221

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Food Research International

journal homepage: www.elsevier.com/locate/foodres

Considerations for readdressing theoretical descriptions of particle- T

reinforced composite food gels
Andrew J. Gravelle, Reed A. Nicholson, Shai Barbut, Alejandro G. Marangoni
⁎

Department of Food Science, University of Guelph, Guelph, ON N1G 2W1, Canada

ARTICLE INFO ABSTRACT

Keywords: The aim of this work was to address the ability of established theoretical models to describe the small de-
Particle reinforcement formation mechanical properties of particle-filled food protein gels. To this end, the effect of incorporating glass
Particle filled gel microspheres on the elastic modulus of heat-set whey protein isolate/xanthan gum gels is reported. Filler size
Composite gel and polydispersity strongly influenced the observed reinforcement with increasing filler content; however, these
Whey protein isolate
effects were also strongly correlated to the ionic strength of the gelator phase (0–200 mM NaCl). Fillers with
van der Poel
General power law
greater polydispersity provided less reinforcement at high filler content, which was associated with improved
packing efficiency. Increasing ionic strength reduced the extent of filler/matrix binding, drastically reducing the
impact of the smaller glass microspheres (4 μm, 7–10 μm). Larger particles increased the elastic modulus at high
salt content due to interfacial stress concentration and particle-particle contacts. Theoretical fits could not sa-
tisfactorily describe the general trend in reinforcement observed with increasing filler content, despite em-
ploying various methods to account for the effects of filler self-crowding. Using an empirical approach, we
propose an alternative functional form which provides improved fits over the entire range of filler content
investigated. This general power law (GPL) model provided physically reasonable values for the maximum
packing fraction through an empirically-derived expression for the scaling exponent. A weighted average ap-
proach was also proposed to incorporate effects of imperfect filler/matrix adhesion. This method incorporates
contributions of both bound and unbound fillers, providing a means to model the effect of increasing ionic
strength.

1. Introduction
Incorporating particulate fillers in a polymer matrix is a commonly
employed strategy to manipulate or tailor the mechanical properties of
the resulting composite material. Particle-filled composites are used in
a broad range of applications in numerous industries, including phar-
maceutical, cosmetics, medical, food, and plastics, among others (Pal,
2002a; Rueda et al., 2017). Various desirable properties of engineered
materials are obtained through the addition of particulate fillers. The
mechanical strength and toughness of elastomeric materials can be
greatly enhanced though the addition of rigid particles such as carbon
black (Sohn, Kim, Hong, & Kim, 2003). The addition of either soft fillers
(e.g. rubbers) or rigid particulates is a commonly used method for
modulating the properties of plastics (Liang, 2002). Restorative mate-
rials used in dental applications often consist of particle-filled resins
containing a high filler phase volume (Braem, Van Doren, Lambrechts,
& Vanherle, 1987). Incorporating hollow, rigid fillers to a solid matrix
can provide enhanced mechanical properties, while reducing weight
and density of the composite (Porfiri & Gupta, 2009; Shutov, 1991).
This latter class of materials are commonly referred to as “syntactic
foams”, and are used in numerous marine and aerospace applications.
Various food products can also be regarded as particle-filled com-
posite materials. In particular, fat-containing foods such as dairy pro-
ducts (cheese, processed cheese products, yoghurt, etc.) and emulsified
meats (e.g. bologna and frankfurters) consist of a dispersed fat phase
embedded in a protein matrix. In such systems, the fat droplets often
play major functional roles, influencing the structural, rheological, and
sensory properties of the food product (Scholten, 2017). However, a
continual shift in dietary guidelines and evolving consumer perception
regarding dietary fats has led to an increase in demand for low satu-
rated fat and fat-reduced foods. Simply decreasing the total fat content
or replacing solid fat with liquid oil can have deleterious effects on the

Abbreviations: CLSM, confocal laser scanning microscopy; GPL, general power law; vdP, van der Poel; WPI, whey protein isolate; WPI-X, whey protein isolate/
xanthan gum
⁎
Corresponding author at: Marangoni, 50 Stone Road, E. Food Science Building, Guelph, ON N1G 2W1, Canada.
E-mail address: amarango@uoguelph.ca (A.G. Marangoni).

https://doi.org/10.1016/j.foodres.2019.03.070
Received 7 February 2019; Received in revised form 28 March 2019; Accepted 30 March 2019
Available online 01 April 2019
0963-9969/ © 2019 Elsevier Ltd. All rights reserved.
A.J. Gravelle, et al.
functional and sensory properties of food products, and can negatively
impact consumer acceptability. Therefore, developing new methods for
tailoring the macroscopic properties of fat-filled protein-based food
matrices would be of particular relevance to the food sector.
One prospective strategy which has recently garnered interest in the
literature has been to optimize the contribution of the dispersed phase
in a predictive manner using well established particle reinforcement
theories (Chen & Dickinson, 1998; Kim, Renkema, & van Vliet, 2001;
Manski, Kretzers, van Brenk, van der Goot, & Boom, 2007; Oliver,
Berndsen, van Aken, & Scholten, 2015; Rosa, Sala, van Vliet, & Van De
Velde, 2006; Thionnet, Havea, Gillies, Lad, & Golding, 2017; van Vliet,
1988). This method could be used to alter the properties of the com-
posite gel based solely on the relative properties of the matrix and filler,
and the total filler content (Lewis & Nielsen, 1970; Pal, 2005; Smith,
1975). Additionally, it has been recognized that both the spatial dis-
tribution of the filler particles (Oliver, Berndsen, et al., 2015; van Aken,
Oliver, & Scholten, 2015) and extent of matrix/filler interactions (Chen
& Dickinson, 1999; Gravelle, Barbut, & Marangoni, 2015) also sig-
nificantly impact the rheological behavior of the resulting composite.
Such factors could be used as additional parameters for optimizing the
macroscopic properties of fat-filled food systems. However, there are
several theories which are commonly adapted in the literature to de-
scribe particle-filled food gels, and each model predicts similar, but
distinct trends in the rheological properties with increasing filler con-
tent. This can lead to the arbitrary selection of a particular model,
which will influence the physical interpretation of experimental data.
Furthermore, established theories are presently unable to account for
particle clustering, polydispersity, and imperfect adhesion to the ma-
trix. Each of these effects can significantly influence the observed re-
inforcement in the modulus of the composite. Such behavior has been
particularly noted for filler clustering in emulsion-filled gels, in which
the magnitude of reinforcement induced by the addition of fat droplets
far exceeds that predicted by theory (Chen & Dickinson, 1998; Kim
et al., 2001; Oliver, Berndsen, et al., 2015; van Vliet, 1988).
The goal of this investigation was to compare the predictive cap-
abilities of various established particle reinforcement theories when
modulating properties of the matrix and filler in a model particle-filled
food gel. To this end, rigid spherical inclusions were incorporated in a
biopolymer gel matrix to accentuate the reinforcing effect of the dis-
persed phase. This strategy served to enhance any differences in the
trends predicted by various theoretical models. We note that such dif-
ferences are commonly masked due to a marginal reinforcement when
using fillers with a comparable modulus to that of the continuous phase
(such as in emulsion-filled gels). Experimental data was thus collected
on heat-set whey protein isolate/xanthan gum (WPI-X) mixed biopo-
lymer gels filled with glass microspheres. The impact of the filler phase
on the elastic modulus of the resulting composite was modulated by
varying both particle size and ionic strength. The latter parameter
provided a means of maintaining a consistent gelator, while altering the
microstructural organization and rheological properties of the protein
matrix. In the following section, a brief overview of the relevant par-
ticle reinforcement theories is presented, and further justification for
the selected model system is provided.
2. Present theoretical approaches
As noted by Pal, exact analytical solutions describing the mechan-
ical properties of particle-filled composites exist only for the limit of
infinite dilution (Pal, 2005). Existing models used to derive exact so-
lutions for concentrated systems are based on approximations which
extend such models beyond the dilute regime (Kerner, 1956; Krieger &
Dougherty, 1959; Mooney, 1951; Pal, 2005; Palierne, 1990; Smith,
1975). However, these equations can be far less cumbersome to work
with, and have thus seen continued use in the literature, as well as
further empirical modifications to improve agreement with experi-
mental data (Lewis & Nielsen, 1970; van Aken et al., 2015). Therefore,
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Food Research International 122 (2019) 209–221
the purpose of this section is to provide an overview of established
particle reinforcement theories, and subsequent adaptations which
have been proposed to address the influence of filler self-crowding. The
latter refers to methods which incorporate effects of filler-filler con-
tacts, as well as introducing an upper limit on the filler content, arising
due to geometrical packing constraints.
Numerous theories describing the mechanical properties of parti-
culate composites have been adapted from equations describing the
viscosity of a dilute suspension of monodisperse, rigid spherical parti-
cles. The earliest such theory describing the relative change in the shear
or elastic (Young's) modulus as a function of volume fraction filler
particles (ϕf) follows from Einstein's equation (Einstein, 1911):
Ec c
Er = = 1 + 2.5 f
Em (1)
m
Here, and throughout, the subscripts m, f, and c respectively denote
the properties of the matrix, filler, and composite, while Er is defined as
the relative elastic modulus of the composite. The proportionality is
valid provided the fillers are rigid, spherical, monodisperse, and per-
fectly bound to the surrounding matrix (Guth, 1945; Smallwood, 1944).
This equation is limited to highly dilute systems, as no consideration is
made for perturbations resulting from the presence of neighboring
particles. Due to this constraint, various methods have been proposed to
extend the Einstein equation beyond the dilute regime (Eilers, 1941;
Guth, 1945; Krieger & Dougherty, 1959; Mooney, 1951). One such
method is the so-called ‘differential effective medium’ approach, which
addresses the effects of filler self-crowding resulting from the incre-
mental increase in filler particles by incorporating the maximum
packing fraction (ϕmax) into the expression for Er. The role of this
function is to correct for the actual available space remaining to in-
corporate additional particles, and the value of ϕmax is thus dependent
on the spatial distribution of the dispersed particles (Pal, 2005). This
approach was employed in a purely empirical extension of Eq. 1 to
develop the well-known Mooney equation, which can be written as
(Mooney, 1951):
2.5 f
Er = exp
f
1
max (2)
For monodisperse particles, values of ϕmax vary from 0.74 for a
hexagonal lattice packing to ~0.64 for a random close packing ar-
rangement (Scott & Kilgour, 1969). Experimental evidence has also
been reported suggesting a random loose packing arrangement results
in ϕmax ≈ 0.56 (Onoda & Liniger, 1990; Pal, 2008). However, due to
the requirement that the particulates be rigid, all models derived from
Eq. 1 are unable to incorporate contributions of viscoelastic fillers,
particularly limiting their applicability in food systems such as emul-
sion-filled gels.
Alternative methods have been used to develop models describing
the rheological behavior of composites containing particles with a finite
modulus. Perhaps the most notable is that developed by van der Poel
(Smith, 1974, 1975; van der Poel, 1958), who adapted the theoretical
construct developed by Fröhlich and Sack to describe the macroscopic
mechanical properties of a sufficiently dilute composite (Fröhlich &
Sack, 1946). This model incorporates the relative modulus of the filler
and matrix, as well as the compressibility of the two components (i.e.,
the Poisson ratio). The van der Poel model considers the effects of
neighboring particles on the rheological behavior of the composite;
however, the impact of particle crowding was not considered, and this
model is thus valid only up to intermediate ϕf. The full form of van der
Poel's model can be expressed as a second order polynomial:
(Er 1)2 + (Er 1) + =0 (3)
The coefficients are dependent on ϕf, the relative modulus of the
filler and matrix (M = Ef/Em), and the Poisson ratio of these two
A.J. Gravelle, et al.
components (νm, νf). For the interested reader, the full form is provided
in the Appendix A (Supplementary material). Generally, the matrix is
assumed to be incompressible, and νm is thus taken to be 0.5, and ex-
perimental evidence has suggested this is a reasonable approximation
for food gel systems (van Aken et al., 2015). Further, it was proposed by
van Vliet (1988) that for liquid or semi-solid fillers, such as in emulsion-
filled gels, Ef could be sufficiently approximated using the Laplace
pressure of the droplets; ΔP = 2σ/r. Here, σ and r respectively denote
the interfacial tension and average radius of the embedded droplets.
A second order simplification of van der Poel's model was also
proposed, which provides reasonable agreement when the mechanical
properties of the filler and matrix are comparable (Smith, 1975):
15(1 m )(M 1) f max
Er = 1 +
(8 10 m)M + 7 5 (8 10 m )(M 1)
m f
An equivalent expression to this simplification was first derived
through an alternative approach by Kerner (Kerner, 1956), and is
therefore generally referred to as the Kerner model. The relative sim-
plicity of Eq. 4 has led to its continued use for interpreting the re-
inforcement of composite food gels (Kim et al., 2001; Manski et al.,
2007; Oliver, Berndsen, et al., 2015; Rosa et al., 2006; Sala, van Aken,
Stuart, & van de Velde, 2007; Yang, Rogers, Berry, & Foegeding, 2011).
However, this equation is generally accepted as being a lower bound of
the van der Poel model (Hashin & Shtrikman, 1963; Smith, 1975), and
the discrepancy between Eqs. 3 and 4 increases with increasing values
of M. A comparison of these two expressions is presented in Fig. 1a,
showing the extreme case, where M→∞. Furthermore, the Kerner
model has also generally been found to under-predict experimental data
(Kim et al., 2001; Thionnet et al., 2017; van Aken et al., 2015; van Vliet,
1988). Efforts to improve the accuracy of this model have been made by
employing an effective volume fraction approach. Here, a ‘self-
crowding’ function ψ(ϕf) is used to describe the effective volume occu-
pied by filler particles, which has an explicit dependence on ϕfmax to
account for geometrical packing constraints (Lewis & Nielsen, 1970).
This method is analogous to the differential effective medium approach
noted above, in which a term describing the incremental change in
volume available for the continued addition of filler particles is ex-
plicitly introduced in the derivation. In contrast, the effective volume
Food Research International 122 (2019) 209–221
approach addresses the effects of filler crowding by simply replacing all
instances of ϕf with ψ(ϕf) in the final equation form (van Aken et al.,
2015).
Two self-crowding functions satisfying necessary boundary condi-
tions were first reported as a correction to the Kerner equation, and take
the following forms (Lewis & Nielsen, 1970):
f
1( f) =1 exp
f
1
max (5)
1 max
2( f) = 1+ 2 f f
(6)
(4) These functions effectively act as scaling factors, enhancing the
contribution of the filler, as would be expected when particle crowding
occurs. This can be seen in Fig. 1a, which compares the Kerner model
adapted to incorporate filler crowding using ψ2(ϕf) with the original
forms of both the Kerner and exact van der Poel models. In addition to
these self-crowding functions, it was also previously noted that the
disparity between the Kerner model (Eq. 4) and experimental data on
emulsion-filled gels could be reasonably overcome by introducing a
scaling constant (Kim et al., 2001; van Vliet, 1988). Therefore, in a
rather simplistic manner, but analogous to that reported by Mooney,
the effective volume fraction occupied by the filler (ϕeff) could alter-
natively be expressed as:
f
eff =
max (7)
A comparison of these three forms for the self-crowding factor are
presented in Fig. 1b, which depicts their impact on the effective volume
occupied by dispersed phase with increasing filler content. Further,
Fig. 1b illustrates the relative increase in the elastic modulus resulting
from the inclusion of rigid particles (Ef ≫ Em), as predicted by the exact
solution to the van der Poel model, the Kerner model, and the effect of
implementing ψ2(ϕf) into the latter, with ϕmax=0.64 (i.e., a random
close packing arrangement). The asymptotic increase in the modified
Kerner equation is due to the rigid nature of the particles (M = 104).
Fig. 1. Comparison of established theore-
tical models describing the relative elastic
modulus (Er) of particle-filled composites as
a function of filler volume fraction (ϕf; a, c,
d), and various forms for self-crowding
functions assuming a maximum packing
fraction of 0.64 (b). For panels a, c, and d,
legends indicate the model (van der Poel,
Kerner, or Pal) and expression employed for
filler self-crowding. vdP refers to the van
der Poel equation, and vdP-exact indicates
no self-crowding function. Pal-Exp and Pal-
Power indicate the exponential and power-
law forms of the models proposed by Pal.
For all fits, Ef/Em=104.
211
A.J. Gravelle, et al.
Although this condition results in the greatest disparity between these
two approaches, it is relevant to the experimental portion of this in-
vestigation. Furthermore, it also serves to highlight the fact that al-
though the Kerner model is more approachable, it also represents a
simplified form of the van der Poel model (Smith, 1975), and is known
to under-predict experimental data. As such, employing the Kerner
equation with subsequent modification to improve fits can influence the
physical interpretation of the data, and caution should thus be ex-
ercised.
Fig. 1c demonstrates the effect on Er when implementing the three
variations on the self-crowding function (Eqs. 5–7) in exact van der Poel
model (Eq. 3), again assuming Ef ≫ Em. Both forms of ψ(ϕf) have rela-
tively little effect at low filler content (ϕf < 0.2), while the predicted
reinforcement deviates quite substantially as the concentration of the
rigid inclusions increases. As also noted in Fig. 1b, the scaling factor
approach produces a greater increase in Er at low filler content, fol-
lowed by a somewhat more gradual enhancement with continued filler
addition. Selection of the most appropriate form for the crowding factor
has generally not been discussed, and is likely selected based on
achieving agreement with experimental data. This method may be
justified based on the empirical nature of these modifications.
All of the approaches thus far outlined use the bulk properties of the
filler and matrix, and consequently neglect any contribution of the in-
terface. This deficiency was first addressed by Palierne, who developed
a model to describe particulate composites with incompressible, vis-
coelastic inclusions (Palierne, 1990). This model is particularly relevant
to emulsion-filled gels, as it incorporates the interfacial properties oc-
curring at the filler surface separate from the modulus of the interior
(van Aken et al., 2015). Although Palierne's approach incorporated fi-
nite filler concentrations, it did not consider the effects of filler
crowding. This formalism was expanded by Pal to incorporate ϕmax
using a so-called “differential effective medium” approach which pre-
dicts the macroscopically averaged modulus composite upon incre-
mental addition of filler particles (Pal, 2002b, 2005). Assuming the
space available for further particle addition decreases as 1 − ϕf/ϕmax,
Pal developed two alternative expressions for Er(ϕf). Adapting the ex-
pression proposed by Mooney to describe the incremental increase in
the effective ϕf (Mooney, 1951) led to the following equation:
Er M 2.5 2.5 f fully solubilized (minimum 2 h). This stock was split into smaller por-
(Er ) = exp
1 M 1
f
max (8)
Using the same approach, but considering a slightly different form
for the function describing the incremental increase in filler particles
(Krieger & Dougherty, 1959), Pal also derived an expression having a
power law-dependence on the maximum packing fraction:
Er M 2.5
f
2.5 max
(Er ) = 1
1 M max (9)
Fig. 1d shows a comparison of the exact van der Poel solution with
the two equations developed by Pal, again assuming ϕmax=0.64. Al-
though the magnitude of the effect differs substantially between the two
approaches, the general trend in reinforcement is quite similar for both
equations. That is, a relatively minor reinforcement in the mechanical
properties of the composite is observed up to intermediate ϕf, followed
by a rapid increase at higher filler content. This behavior is analogous
to the van der Poel model, when adapting the effective volume fraction
approach (Fig. 1c).
In the remainder of this work, we will present new experimental
data on a model composite food protein gel consisting of heat-set WPI-X
mixed biopolymer gels filled with glass microspheres of varying size
ranges. This particular class of filler was selected for its well defined
physical characteristics, such as the size, modulus, and Poisson ratio.
Further, the use of rigid fillers minimizes the number of variables which
need to be considered (e.g., interfacial tension, Laplace pressure, M,
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Food Research International 122 (2019) 209–221
etc.), allowing for a direct comparison of the various theoretical
models. In this work, two main parameters were explored; the impact of
filler size, and altering the physical interactions between the filler and
matrix by modulating electrostatic interactions. Presently, particle size
and imperfect adhesion are not considered in any of the models out-
lined above.
It is noteworthy that the various functional forms and proposed
adaptations to address filler crowding show similar, yet distinct trends
in reinforcement (Fig. 1c, d). However, there has been no conclusive
evidence to validate the use of one particular model over the others. As
a result, the arbitrary selection of one such model will introduce con-
siderable ambiguity in the physical interpretation of experimental data.
In this work, our intent was to address this issue by comparing the
various forms of particle reinforcement models to determine if any
particular equation accurately described the experimental data when
varying either filler size or ionic strength.
3. Materials and methods
3.1. Materials
Whey protein isolate (WPI; BiPro™, 91% protein) was obtained from
Davisco Foods International (Le Sueur, MN, USA). Xanthan gum was
purchased from TIC Gums Inc. (White Marsh, MD, USA), and used as a
viscosity enhancer to avoid particle sedimentation prior to gelation.
Sodium chloride was obtained from Fisher Scientific (Ottawa, ON,
Canada). Glass microspheres were obtained in four size ranges; ~4 μm
soda lime microspheres were purchased from Cospheric LLC (Santa
Barbara, CA, USA), and 7–10 μm, 45–90 μm, and 150–210 μm glass
beads were obtained from Potters Industries (Ballotini glass micro-
spheres; Potters Industries LLC, Valley Forge, PA, USA). All glass mi-
crospheres were solid soda lime glass, ρ = 2.5, and were used as re-
ceived.
3.2. Sample preparation
WPI (10 wt%) was added to distilled water and mixed with a
magnetic stir bar for a minimum of 30 min to ensure full dissolution of
the protein. Xanthan gum (0.8 wt%) was then added and mixed until
tions, to which NaCl was added (0, 50, 100, and 200 mM), and mixed
for several min with a magnetic stir bar. Glass beads were incorporated
by hand at volume fractions (ϕf) of 0, 0.025, 0.05, 0.10, 0.15, 0.20,
0.33, 0.43, and 0.50. Due to a small amount of alkaline residue present
on the glass beads, the final pH of particle-filled gels was ~7.5. The
protein stock used to prepare control samples was thus pH corrected to
7.5 using 0.5 N NaOH. Each sample was portioned in two 10 ml dis-
posable beakers (Fisher Scientific), covered with plastic food wrap, and
heated for 30 min in a water bath (Haake W-26, Haake; Berlin,
Germany) maintained at 80 °C. Upon removal from the water bath,
samples were placed in a refrigerator (4 °C) for 2 h prior to testing.
3.3. Confocal laser scanning microscopy (CLSM)
Prior to imaging, WPI-X gels were stained by incorporating
Rhodamine B into the thickened protein stock solution prior to gelation.
A total stain concentration of 15 μl/ml was added to the biopolymer
stock solution. Samples were then prepared as outlined in Section 3.2.
For all samples prepared with filler particles, ϕf= 0.10. CLSM imaging
was conducted using a Leica TCS SP5 Confocal laser scanning micro-
scope (Leica Microsystems CMS GmHB, Mannheim, Germany) equipped
with a DM6000 B upright microscope using a Green Helium Neon laser
(GreHeNe). Images were acquired with a 63× glycerol objective using
a laser excitation wavelength of 543 nm, and the signal was collected
over an emission range of 600–680 nm. Both fluorescence and trans-
mitted light images were acquired simultaneously to improve visibility
A.J. Gravelle, et al.
of the glass microspheres.
3.4. Mechanical testing
A uniaxial compression test was performed immediately after re-
moving each sample from the refrigerator. Samples were removed from
the disposable beakers and cut to a height of 10 mm. Due to the slight
conical shape of the molds, the average diameter of the pucks was
22 mm. Each gel was subjected to a compression strain of 60% at a fixed
rate of 0.83 mm/s through two successive compressions between par-
allel plates, using a texture analyzer (Model TA.XT2, Stable Micro
Systems, Texture Technologies Corp., Scarsdale, NY, USA) outfitted
with a 30 kg load cell. Data was collected and analyzed using the
Exponent Lite Express software (Stable Micro Systems). The elastic
constant was taken as the slope of the linear region at the onset of
compression; i.e., strain ≤10% (Oliver, Berndsen, et al., 2015), and the
elastic (Young's) modulus was determined based on the geometry of the
sample.
3.5. Model fitting and data analysis
In both the van der Poel and Kerner models, the protein network
was assumed to behave as an incompressible, elastic material, and the
Poisson ratio of the matrix (νm) was therefore taken to be 0.5. For the
van der Poel model, the Poisson ratio of the filler was taken to be 0.25.
Experimental data was fit to theoretical models using GraphPad Prism
5.0 (GraphPad Software, Inc., San Diego, CA, USA). Fitted parameters,
standard errors, and corresponding goodness of fits were obtained using
non-linear regression analysis. Error bars in all figures indicate the
standard error of the mean (SEM).
4. Results and discussion
4.1. Microstructural analysis
CLSM images of the heat-set WPI-X gels prepared at varying ionic
strength are presented in Fig. 2. Although xanthan gum was used as a
thickening agent to ensure the filler remained suspended during gela-
tion, interactions between whey proteins and xanthan gum have been
well documented (Bertrand & Turgeon, 2007; Bryant & McClements,
2000). The pH conditions used in this work (pH 7.5) were significantly
above the isoelectric point of whey proteins (~4.6). This produces a
higher net negative charge on the protein surface, which is known to
induce segregative micro-phase separation (Bertrand & Turgeon, 2007).
Altering the ionic strength is a well-known method of influencing the
microstructural organization and resulting mechanical properties of
WPI gels (Barbut, 1995; Foegeding, Davis, Doucet, & McGuffey, 2002),
but will also impact the segregative behavior.
Gels prepared with 0, 100, and 200 mM NaCl (without filler) are
depicted in Figs. 2a-c, respectively, and display distinct microstructural
differences. In the absence of salt (Fig. 2a), a relatively homogeneous
gel structure was observed, indicative of a dense, fine-stranded protein
network. This was further supported by the translucent nature of the
gels (not shown). The presence of small pores throughout the gels in-
dicates only a marginal extent of phase micro-separation occurred.
These pores are expected to be protein-excluded, carbohydrate rich
regions (Bertrand & Turgeon, 2007). The phase micro-separation was
particularly pronounced in the mixed biopolymer gels prepared at in-
termediate ionic strength (100 mM NaCl); however, these gels were
considerably firmer than those prepared without salt, and were opaque
in nature. The increase in mechanical strength is likely due to a com-
bination of two factors. First, the greater prevalence of xanthan-rich
pores serves to concentrate the proteins in the remainder of the con-
tinuous phase. Second, the presence of monovalent ions induces charge
screening in the protein phase, which facilitates more extensive protein-
protein interactions (Barbut, 1995; Kuhn & Foegeding, 1991).
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Food Research International 122 (2019) 209–221
The gels prepared with 200 mM NaCl displayed a more aggregated,
particulate-like structure, and were visually opaque and white at the
macroscopic level. The high concentration of salt ions causes repulsive
interactions, promoting the formation of particulate aggregates. This
microstructure has previously been associated with decreased gel
strength and water binding properties (Barbut, 1995). Due to the
drastic salt-induced changes in microstructure, it is difficult to observe
any distinct voids indicative of the xanthan-rich regions noted in the
gels prepared with 0 mM and 100 mM NaCl. However, based on pre-
vious investigations it is expected that segregative phase separation still
contributes to the microstructural organization of these particulate
protein/carbohydrate mixed gels (Bertrand & Turgeon, 2007).
The microstructure of WPI-X gels containing glass microspheres
(ϕf=0.10) are shown in Fig. 2d-j. The transmitted CLSM images are
presented in Fig. 2d-g (upper half of each panel), while the lower half
these panels show the same images overlayed with the reflected light
channel to highlight the location and appearance of filler particles.
Composite WPI-X gels containing 4 μm glass microspheres are pre-
sented in Figs. 2d-f at 0, 100, and 200 mM ionic strength, respectively.
An additional sample was prepared at 200 mM with the 45-90 μm mi-
crospheres, and is shown in Fig. 2g. In all cases, the presence of the
filler particles appeared to have little effect on the microstructure of the
mixed biopolymer gel network. The 4 μm particles appeared to be in-
corporated into the gel matrix, and were relatively homogeneously
distributed throughout the network. The larger glass beads also showed
evidence of a protein layer at the filler/matrix interface, which can be
seen to extend into the continuous phase. Therefore, these images in-
dicate the protein had some affinity for the model filler particles used in
this study.
4.2. Mechanical properties of composite gels: effect of ionic strength
The influence of glass beads on the elastic modulus of the particle-
filled WPI-X composites was investigated as a function of both ionic
strength and filler size. To separate the contribution of these two
parameters, the effect of filler size was contrasted at each of the salt
concentrations used to produce the microstructurally distinct networks
shown in Fig. 2. The relative elastic modulus of composites (Er = Ec/Em)
prepared with 0, 100, and 200 mM NaCl are depicted in Fig. 3a-c, re-
spectively (*note the differences in scales). Irrespective of either filler
size or ionic strength, the rigid microspheres provide little reinforce-
ment below ϕf= 0.2. This is consistent with previous studies on other
particle-filled protein gels (Manski et al., 2007; Oliver, Scholten, & van
Aken, 2015; Yang et al., 2011), as well as with established theoretical
models.
In the absence of added salt (0 mM; Fig, 3a), all filler sizes produce a
similar trend in Er, where a rapid increase was observed at high ϕf. This
behavior is somewhat intuitive, as it would be expected that at ϕf=0.5,
the fillers approach the maximum packing threshold, as discussed
above. Further, although the extent of reinforcement was negatively
correlated with increasing filer size, this can be rationalized by noting
that the size distribution (i.e., polydispersity) also increased with in-
creasing diameter. This would have a direct impact on the packing ef-
ficiency, leading to a higher ϕmax, thus reducing the effect of filler
crowding. The consistent trend in reinforcement for all filler sizes,
combined with the apparent dependence on polydispersity (ϕmax) sug-
gests that the glass beads behaved as an ideal bound filler in the 0 mM
WPI-X gels.
The WPI-X gels prepared with 100 mM NaCl exhibited substantially
less reinforcing behavior with increasing filler content (Fig. 3b). It is
noteworthy that in the absence of filler, these gels were considerably
firmer than those prepared without added salt. Therefore, the decrease
in reinforcement with increasing ϕf can be (partly) attributed to the
higher elastic modulus of the matrix (Em). However, particle re-
inforcement models such as those proposed by van der Poel, Kerner,
and Pal all suggest that for rigid inclusions, Er should no longer depend
A.J. Gravelle, et al. Food Research International 122 (2019) 209–221

Fig. 2. Confocal laser scanning microscope (CLSM) images of the heat-set whey protein isolate/xanthan gum gels prepared at varying ionic strength (0, 100, or
200 mM), in the absence of filler (a-c) or with the addition of glass microspheres (ϕf = 0.10; d-g). The lower half of panels (d-g) show overlays of the transmitted
light, highlighting the location of glass microspheres.

Fig. 3. Relative elastic modulus (Er) as a

function of volume fraction filler (ϕf) for
whey protein isolate/xanthan gum gels
containing glass microspheres and prepared
at varying ionic; 0 mM NaCl (a), 100 mM
NaCl (b), and 200 mM NaCl (c, d). Filler size
ranges are indicated in the legend, and da-
shed lines are intended as a visual guide.

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A.J. Gravelle, et al.
on the physical properties of the matrix (i.e., when Ef ≫ Em). This point
is addressed further in Section 4.5.
When considering the general trend in reinforcement, only the
100 mM gels filled with 4 μm and 7–10 μm particles displayed a similar
behavior to that of the 0 mM composites. In contrast, the larger mi-
crospheres produced a greater reinforcement at intermediate ϕf, which
appeared to reach a plateau at ϕf > 0.33. Furthermore, it can be seen
that the 4 μm particles caused a small, but notable depression in the
modulus at intermediate ϕf (i.e. Er < 1). As previously suggested by van
Vliet, fillers which are not bound to the surrounding matrix are unable
to provide mechanical support to the continuous phase (van Vliet,
1988). These so-called “passive” fillers interrupt the continuity of the
gel matrix, and can also contribute to interfacial stress concentration,
ultimately producing a decrease in Er.
At intermediate ϕf, the macroscopic dimensions of the 45–90 μm
and 150–210 μm glass beads may cause them to deviate from the as-
sumption of homogeneous, non-interacting particles, resulting in a
greater likelihood of filler-filler contacts. The deformation of matrix
material surrounding rigid filler particles also leads to stress con-
centration at the interface, which can propagate outward from the in-
clusions (Gao & Lelievre, 1994; van Vliet & Walstra, 1995). As larger
particles represent more expansive discontinuities on the continuous
phase, the 45–90 μm and 150–210 μm fillers enhance the interfacial
stress concentration experienced under loading. These effects manifest
in an artificial increase in Er at lower ϕf than would be theoretically
expected, and is reflected in the experimental data (Fig. 3b). The ob-
served trends in Er for the various filler sizes were thus be attributed to
a combined effect of the stiffer matrix, decreased filler-matrix interac-
tions, and effects of particle-particle contacts associated with filler
crowding.
The impact of glass microspheres on Er of WPI-X gels prepared with
200 mM NaCl is depicted in Fig. 3c. The same data is also presented in
Fig. 3d with a rescaled y-axis, for direct comparison with the 100 mM
gels. Overall, these figures suggest the trend in reinforcement was si-
milar for gels prepared at intermediate and high salt content, despite
the drastic difference in the magnitude of reinforcement produced by
the larger particles. Fig. 3c demonstrates that both the 45–90 μm and
150–210 μm fillers resulted in a substantial increase in Er, roughly an
order of magnitude larger than that observed in the 100 mM gels. In
contrast, comparison of Fig. 3b and d shows that Er(ϕf) produced by the
4 μm and 7–10 μm microspheres was nearly identical for WPI-X gels
prepared with 100 mM and 200 mM NaCl. This result indicates smaller
particles behave more as ideal fillers which are homogeneously dis-
tributed throughout the matrix, and the observed impact on Er is thus
closer to that expected by theory, due to particle crowding.
The discrepancy in Er(ϕf) seen in the 100 mM and 200 mM NaCl gels
observed with the larger glass beads can be attributed to two factors; i)
the differences in microstructural organization of the gel network, and
ii) the ability to dissipate mechanical energy within this biopolymer
matrix. At the gelation conditions used in this study (pH 7.5), no sy-
nergistic interactions are expected between WPI and xanthan gum.
Therefore, the mechanical properties of the mixed biopolymer gels are
predominantly attributed to the structure of the WPI network and the
extent to which these proteins are concentrated by dissociative phase
micro-separation (Bertrand & Turgeon, 2007). The gels prepared at
intermediate ionic strength consisted of a highly phase separated
system in which the WPI-rich phase was made up of relatively thick
protein strands with lower porosity. In contrast, at high ionic strength
there was less distinction between the two biopolymer phases, and the
protein-rich phase consisted of a more porous, aggregated particulate
network (Barbut, 1995; Bertrand & Turgeon, 2007).
Energy dissipation within the protein matrix is highly dependent on
the structure, organization, and associative interactions of the mole-
cular building blocks, which will directly impact the large deformation
mechanical attributes of the resulting composite (van Vliet & Walstra,
1995). Electrostatic interactions have previously been reported to
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Food Research International 122 (2019) 209–221
directly influence the fracture properties and elastic modulus of heat-set
WPI gels (McGuffey & Foegeding, 2001). A comparison of fine-stranded
and particulate heat-set WPI gels was also reported to impact the strain
at fracture (Gwartney, Larick, & Foegeding, 2004). Emulsion-filled gels
were prepared with 25 mM NaCl, and the particulate structure was
induced via the addition of 10 mM CaCl2. In the absence of filler, the
fracture stress of stranded and particulate gels was nearly equivalent,
while the former produced a 2.5-fold increase in the fracture strain.
These results suggest the fine-stranded network is more effective at
dissipating mechanical energy within the gel matrix, allowing it to
adapt to the presence of the larger microspheres which effectively act as
discontinuities in the continuous phase. In contrast, the particulate gels
are more susceptible to interfacial stress concentration, resulting in
stress translation between neighboring filler particles during loading.
Furthermore, the larger particles would have a higher propensity to
incur filler-filler contacts at lower ϕf. Due to the rigid nature of the
fillers used in this investigation, this effect would result in the accu-
mulation of mechanical stress in the particulate gel network, thus
producing a more rapid increase in Er with increasing filler content.
4.3. Modelling Er of bound fillers in WPI-X gels
As noted above, both the clear trend in reinforcement, and the ne-
gative correlation between Er and filler size distribution observed in the
0 mM NaCl gels suggest that in the absence of salt, the glass micro-
spheres behave as ideal bound fillers in heat-set WPI-X gels. The in-
teraction between the matrix and filler are expected to arise due to the
presence of terminal functional groups on the surface of the glass
(predominantly silanol groups). The pH of the biopolymer solution used
in this investigation was 7.5, which is above the isoelectric point of
whey proteins. Consequently, these proteins had a greater prevalence of
negatively charged functional groups available to interact with the
positively charged species present on the glass surfaces, resulting in a
strong filler/matrix adhesion. At higher salt concentrations, charge
screening interferes with these interactions, while promoting the for-
mation of particulate aggregates. As a result, the fillers may be im-
perfectly bound to the continuous phase, thus complicating the theo-
retical analysis (discussed in more detail in Section 4.6). Therefore, a
comparison of the experimental data with established theoretical
models was initially restricted to the gels prepared with no added salt.
As there has been no conclusive agreement regarding which theoretical
model provides the most accurate description of experimental data (in
food gels, or otherwise), an exploratory fitting procedure was con-
ducted using those theories outlined in Section 2. It is again noted that
the use of rigid fillers was intended to enhance the contribution of the
dispersed phase in the model gel system. This is an important distin-
guishing feature, as it served to accentuate any differences in the ex-
perimentally observed reinforcement and that predicted by the various
established models. In contrast, discrepancies between experiment and
theory are generally marginalized in model food systems such as
emulsion-filled gels, as Ef is often comparable to Em.
The trends in reinforcement predicted by several of the established
models outlined in Section 2 are shown in Fig. 4a. Theoretical curves
were generated assuming M = 104 to satisfy the condition Ef ≫ Em, and
ϕmax= 0.64 (i.e., random close packing). These curves are overlaid with
the experimental data corresponding to the 4 μm glass beads, which
displayed the lowest polydispersity and greatest reinforcement. The
theoretical models presented include the exact solution of the van der
Poel model (vdP-exact; Eq. 3), as well as modifications using the ef-
fective volume fraction approach to replace ϕf with either ψ1(ϕf) or ϕeff
(Eqs. 5 and 7, respectively). Finally, the exponential form for Er pro-
posed by Pal is also presented (Eq. 8), as this equation deviates more
significantly from the exact van der Poel solution than the corre-
sponding power-law form (i.e., Eq. 9; see Fig. 1d).
Consistent with findings reported in previous studies on various
food gels (Kim et al., 2001; Oliver, Berndsen, et al., 2015; van Vliet,
A.J. Gravelle, et al.
Fig. 4. Experimental data and theoretical fits for whey protein isolate/xanthan
gum composite gels prepared at 0 mM NaCl. (a) Relative elastic modulus (Er) of
composites containing 4 μm glass microspheres, compared to various estab-
lished particle reinforcement theories, as indicated in the legend (See Fig. 1 for
description of abbreviations). For all fits, Ef/Em=104. (b) Ln-ln transformation
of Er as a function of volume fraction filler (presented as 1-ϕf) for gels con-
taining glass microspheres of varying size (ranges indicated in legend). Solid
lines in (b) correspond to linear fits of the transformed data.
1988), vdP-exact drastically under-predicted the impact of the rigid,
bound fillers. Both vdP-ψ1 and Pal-Exp produced similar trends,
showing little deviation from vdP-Exact at low ϕf, and progressing to a
near-asymptotic increase as the filler content approached ϕmax. The
more rapid increase of vdP-ψ1 is due to the form of the function de-
scribing the effective filler volume (see Fig. 1b). In either case, these
models do not follow the same general trend in reinforcement observed
in the experimental data; both theories under-predict the data at low- to
intermediate ϕf, while the contribution of filler packing appears to
produce a much greater increase in Er than is physically realized. The
effective volume approach proposed in this work (vdP-ϕeff) was ex-
pected to dampen these effects, due to the greater increase in ϕeff at low
filler content. Although this modification provided a minor increase in
the theoretical curves in this regime, the same general trend in re-
inforcement was observed, resulting in too great an increase as ϕf →
ϕmax.
From this comparison, it is apparent that established particulate
reinforcement theories which adapt a maximum packing term do not
accurately predict the general trend in reinforcement displayed by the
model composite gels. However, it was noted that Er closely followed a
simple power-law relation for all particle sizes, as indicated by the
linear trend observed in the ln-ln transformation presented in Fig. 4b
(data plotted as a function of ϕm = 1 − ϕf). It is noted that a power-law
relation was also previously derived by Pal to describe the elastic
modulus of composites containing fillers with finite modulus, as dis-
cussed above (see Eq. 9). This approach was also generalized by van
Aken and coworkers (van Aken et al., 2015). However, these equations
were derived using the approach of Krieger and Dougherty, who at-
tested the incremental increase in the dispersed phase due to the ad-
dition of filler particles as dϕf/(1 − ϕf/ϕmax), thus introducing a de-
pendence on the maximum packing fraction in ϕf. In the present work,
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Food Research International 122 (2019) 209–221
from a purely empirical perspective, we have found that an improved fit
was achieved when ϕmax was neglected in this term. The general form of
the best fit equation was
Er = (1 f) (10)
which will subsequently be referred to as the general power law (GPL)
model. The prefactor is ideally assumed to be unity, as Er describes the
relative modulus, and the scaling factor γ thus fully describes the trend
in reinforcement. This assumption is expected to be valid in the limit of
rigid inclusions investigated here (Pal, 2005).
4.4. Proposed functional form for the GPL model
As an extension of the empirical approach adapted here, an ex-
pression for γ was derived in the following manner: i) It was ac-
knowledged that the influence of filler crowding must still be con-
sidered, and should thus be incorporated in some form. ii) Analogous to
the approach taken by Mooney, it was further assumed that the con-
stant 2.5 be explicitly expressed in the scaling factor. This term was
selected to maintain consistency with the functional form proposed by
Pal (Eq. 9), which was first introduced in the Einstein equation for rigid,
spherical inclusions (Mooney, 1951; Pal, 2005). Finally, the value of
ϕmax was used as the sole fitting parameter, as filler polydispersity was
expected to directly influence the packing efficiency.
From these assumptions, quite reasonable values of ϕmax were ob-
tained when the scaling factor was expressed as γ = 2.5/ϕmax2. The
modified version of the GPL equation can thus be proposed based on the
experimental data presented here:
2
2.5/ max
Er = (1 f) (11)
The best fit values for γ (and associated values for ϕmax) are pre-
sented in Table 1, and demonstrate excellent agreement with the ex-
perimental data for all filler sizes employed (R2 ≥ 0.98). The theore-
tical fits generated by this empirical model are overlaid with the
experimental data in Fig. 5. In each panel, the associated value of ϕmax
obtained from Eq. 11 is reported. Further, the established particle re-
inforcement models presented in Fig. 4a are again included for com-
parison. The latter curves were generated using the associated value of
ϕmax derived from Eq. 11 for each filler size. Although these theoretical
curves look quite similar in the various panels, only vdP-exact is un-
changed, and all other fits exhibit a gradual reduction in reinforcement
as ϕmax increased.
From Eq. 11, a maximum packing fraction of 0.585 was obtained for
the 4 μm particles, which approaches the experimentally reported value
for a random loose-packed filler arrangement (~0.56-) (Onoda &
Liniger, 1990; Pal, 2008). The best fit values for ϕmax increased with
filler size, reaching 0.642 for the gels filled with 150–210 μm micro-
spheres. This value notably coincides with that of a random close
packing arrangement of uniform spheres (Pal, 2002b). The positive
correlation between ϕmax and filler size is again attributed to the as-
sociated increase in polydispersity (and thus packing efficiency).
Therefore, the values obtained for ϕmax using the empirical power-law
relation proposed here appear to justify the functional form in the limit
Table 1
Linear regression analysis of ln-ln transformation of Er(ϕf) for the 0 mM whey
protein isolate gels containing glass microspheres of varying size.
Corresponding goodness of fits are reported as R2. *Note that ϕmax was calcu-
lated using the empirical expression γ = 2.5/ϕmax2.
Filler size γ ( ± SEM) R2 *ϕmax
4 μm 7.31 ± 0.14 0.98 0.585
7–10 μm 7.06 ± 0.14 0.98 0.595
45–90 μm 6.68 ± 0.11 0.99 0.612
150–210 μm 6.07 ± 0.12 0.98 0.642
A.J. Gravelle, et al. Food Research International 122 (2019) 209–221

Fig. 5. Relative elastic modulus (Er) as a function of volume fraction filler (ϕf) for whey protein isolate/xanthan gum composite gels prepared at 0 mM NaCl and
containing glass microspheres of varying size. Theoretical fits are presented using the general power law (GPL) method with the empirical expression γ = 2.5/ϕmax2.
The maximum packing fraction obtained from each fit is displayed in the bottom right corner of the corresponding panel. A comparison of select particle re-
inforcement theories is also provided, as indicated in the legend (See Fig. 1 for description of abbreviations). For all fits, Ef/Em=104.

of bound, rigid inclusions. Further work will need to be carried out to
confirm this functional form can be obtained using a strict mathema-
tical derivation.
As a practical consideration, it is noted that the GPL does not pro-
duce an asymptotic increase in Er, suggesting the equation continues to
predict a finite reinforcement when ϕf > ϕmax. However, both the
various modified van der Poel equations and the models derived by Pal
all break down beyond this limit, indicating that none of these models
can be applied beyond the geometrical constraint. The requirement that
Er → Ef at the filler packing limit is likely unrealistic when the matrix is
significantly softer than the filler particles. This expectation is likely
drawn from parallels to models describing the viscosity of particulate
suspensions, in which fillers can be in direct contact with one another.
In contrast, models describing the mechanical properties of particulate
composites are built on the assumption that each filler particle is sur-
rounded by a shell of the matrix material. Although this condition may
not be completely satisfied at higher ϕf, it can be assumed the matrix
forms a continuous, 3-dimensional network.
As the filler content approaches the geometrical packing limit, the
protein may not be able to maintain sufficient continuity and structural
integrity to support the embedded particles under loading. This phy-
sical constraint suggests that the modulus of a soft particle-filled com-
posite containing rigid inclusions will never equal that of the filler; i.e.,
Er(ϕf → ϕmax) < Ef. From this argument, the trend in reinforcement
predicted by the empirical GPL may be quite reasonable. Therefore, the
comparison in Fig. 5 suggests that the GPL model predicts the general
trends in Er observed for heat-set WPI-X gels containing bound glass
microspheres more accurately than previously proposed models of in-
corporating filler self-crowding. Finally, the expectation that the filler
particles are surrounded by a shell of matrix material suggests that a
random close packing arrangement may represent an overestimate of
ϕmax. This rationale is in line with the theoretical ϕmax values obtained
from the GPL fit reported in Table 1.
4.5. Extension of the GPL approach to non-ideal systems
To determine if the GPL approach might provide an accurate de-
scription of biopolymer gels other than the model system used in this
study, we have considered two previously reported particle-reinforced
food gel systems. Our group previously reported on the use of estab-
lished particle reinforcement theories in finely comminuted poultry
meat gels, which are predominantly structured by salt-extracted myo-
fibrillar proteins (Gravelle et al., 2015). In this work, high-melting
point wax particles were used to model fat particles which could be
separated into distinct size ranges. These fillers were also selected for
their hydrophobic nature, allowing them to interact strongly with the
protein gel matrix. In this work, we reported that a GPL provided an
improved fit over both the exact van der Poel equation (Eq. 3) and the
modified Kerner equation which implements the effective filler volume
approach (Eqs. 4, 5). The transformed data set is reported in Fig. S1, as
well as corresponding fits to the empirical GPL approach presented in
this work (Eq. 10). For all three filler size ranges, the GPL model pro-
duced excellent fits (R2 ≥ 0.98); however, values for ϕmax are not re-
ported due to the irregular filler shapes, which significantly deviate
from the assumption of spherical inclusions. The scaling exponents
obtained for the small, medium, and large wax particles were 4.35,
3.88, and 3.18, respectively, and are notably lower than those reported
for heat-set WPI-X gels (see Table 1). This may be due to the variable
packing arrangement and irregular interfaces formed by the wax par-
ticles. It is noted that these values would predict a maximum packing
fraction in the range of 0.75–0.89 based on the form of Eq. 12, which
represents a physically reasonable result. Therefore, although the
physical interpretation is limited, the GPL approach appears to be ap-
plicable to myofibrillar-based protein gels containing rigid bound fil-
lers.
To gauge the applicability of extending the GPL approach to de-
scribe more practical food protein gel systems, approximate data was

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extrapolated from that previously published by Oliver et al. (Oliver,
Berndsen, et al., 2015). The data used for fitting was obtained from
emulsion-filled gelatin gels containing beef fat, which were allowed to
set under conditions which encouraged filler clustering. The protein
content of the matrix was adjusted to observe the impact of Em on the
reinforcing behavior of these composites. The theoretical fits and as-
sociated scaling factors obtained using the GPL model (Eq. 10) are
presented in Fig. S2 for each protein content reported. The gels pre-
pared at 4, 6, and 8% protein all fit extremely well to this model (Figs.
S1a-c), and the scaling factors obtained from these fits were 3.72, 3.50,
and 3.42, respectively. The goodness of fit is not reported, as the ex-
perimental values were manually extracted from a previously published
figure. Although the GPL model could not accurately predict the trend
in reinforcement observed for the 10% protein gels (Fig. S2d), a similar
issue was noted when using the filler clustering approach proposed by
these authors (Oliver, Berndsen, et al., 2015).
The values obtained for the scaling factors of the emulsion-filled
gelatin gels are considerably lower than that observed for the glass-
filled WPI-X gels in the present study. This result suggests that addi-
tional factors such as a finite filler modulus and contributions of filler
clustering would need to be adapted into the GPL approach to provide a
more complete description of food composite gels. The power-law re-
lation derived by Pal (Eq. 9) demonstrates that Em and Ef should in fact
contribute to the scaling factor when the moduli of these two respective
phases are comparable (Pal, 2005). Based on the empirical approach
proposed here, we anticipate the functional form of the GPL should thus
be expanded to incorporate an additional factor which incorporates this
dependence:
2
Er 2.5/ max
(Em , Ef ) (1 f) (12)
Presently, the form of δ is unknown, but as with the model proposed
by Pal (Eq. 9), we anticipate that δ(Ef, Em) approaches unity in the case
of rigid inclusions (Ef ≫ Em). Although the fits presented in Fig. S2 serve
only as a rough approximation to assess the suitability of the GPL model
in a practical food gel system, the success of this approach warrants
further investigation using a more stringent theoretical derivation.
Fig. 6. Relative (Er; a) and absolute (Ec; b) elastic modulus as a function of
volume fraction filler (ϕf) for whey protein isolate/xanthan gum gels containing
4 μm glass microspheres and prepared at varying ionic strength (0–200 mM
NaCl; see legend). Inset in (a) displays the reinforcement at high ϕf for com-
posites containing salt, while the inset in (b) highlights the depression in Ec with
increasing ionic strength. Dashed lines are intended as a visual guide.
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Food Research International 122 (2019) 209–221
4.6. Imperfect filler adhesion: the interaction parameter
The effect of ionic strength on Er(ϕf) of WPI-X gels was discussed in
Section 4.2. The observed differences in reinforcement with varying salt
content were predominantly attributed to changes in electrostatic in-
teractions between the protein building blocks within the mixed bio-
polymer gels. It was further noted that the presence of salt ions will
induce a charge screening effect at the glass/protein interface, thus
diminishes the filler/matrix interactions. Therefore, the addition of
NaCl will induce a concentration dependent decrease in the adhesion
between the glass microspheres and protein gel network. To further
investigate the effect of imperfect adhesion in this model composite
system, Fig. 6 compares the gels containing 4 μm particles prepared at
varying ionic strength. This filler size was selected because it most
accurately satisfies the theoretical expectation of monodispersed,
homogeneously distributed filers. As such, these composites displayed
the expected trend in reinforcement over the entire range of ionic
strength evaluated; i.e., a gradual increase in Er at low- to intermediate
ϕf, followed by a more pronounced reinforcement as the filler content
approaches ϕmax.
An additional data set was collected at 50 mM NaCl to fill in the
relatively broad gap in reinforcement observed between the 0 mM and
100 mM gels at high filler content. Fig. 6a demonstrates the effect of
salt content on Er(ϕf), with the inset showing a rescaled y-axis to
highlight the trends of the 50–200 mM gels. There is a distinct negative
correlation between ionic strength and the extent of reinforcement
observed as ϕf → ϕmax. This trend is consistent with the hypothesis that
filler-matrix interactions weaken with increasing ionic strength in glass-
filled WPI-X gels. A slight depression in Er was also observed in the gels
containing salt, and may further suggest the glass behaves (at least in
part) as a passive filler.
To highlight the decrease in Er at low ϕf, Fig. 6b shows the effect of
increasing salt content on the magnitude of Ec, as opposed to the re-
lative values. The depression in Ec is particularly notable in the inset of
Fig. 6b, which becomes more pronounced in the composites prepared at
higher ionic strength. In contrast, when ϕf≥ 0.2, no appreciable dif-
ferences were observed between the gels, irrespective of salt content.
Rigid inclusions such as those used in the present study should produce
equivalent reinforcement, irrespective of Em, provided the particles are
sufficiently bound to avoid slipping during the initial stages of de-
formation. These findings therefore strongly suggest that increasing
ionic strength reduces filler/matrix adhesion, while at high filler con-
tent, particle-particle contacts dominate the observed reinforcement.
The observed trends in Ec presented in Fig. 6a and b therefore indicate
the glass microspheres are either bound (0 mM NaCl), or partially
bound (50–200 mM NaCl) to the gel matrix.
The particle reinforcement theories discussed above universally
assume perfect adhesion between the dispersed and continuous phases
(Yang et al., 2011). It has been proposed that incompatible fillers pre-
sent in an unbound state could be modelled by assuming Ef = 0, ef-
fectively treating the inclusions as voids in the continuous matrix (Ring
& Stainsby, 1982; van Vliet, 1988). Using this approach, fillers can be
classified as either active or passive (i.e., bound or unbound; see Fig. 7a,
b), and it has been experimentally demonstrated that the latter will
cause a decrease in Er (van Vliet, 1988). However, the presence of filler
particles in these “pseudo-voids” will lead to stress concentration at the
interface as the matrix undergoes deformation around the filler (Gao &
Lelievre, 1994), particularly when Ef > Em (Fig. 7c). This would cause
significant deviation from the assumption that unbound fillers can be
effectively neglected from the theoretical description, particularly at
high filler content. Furthermore, the active vs. passive approach cannot
account for imperfect (or partial) adhesion between the matrix and
filler, which should produce an intermediate effect (Fig. 7d). Although
several micromechanistic models have recently been proposed to ad-
dress imperfect interfacial adhesion (Wang, Zhu, Chen, Jin, & Zhang,
2014; Yanase & Ju, 2012; Zhu et al., 2015), these approaches are quite
A.J. Gravelle, et al. Food Research International 122 (2019) 209–221

Fig. 7. Schematic representation of particle-filled gels containing rigid inclusions, depicting various types of particle-matrix interactions; (a) strongly bound (active)
fillers, (b) unbound (passive) fillers modelled as voids, (c) passive rigid fillers present in “pseudo-voids”, and (d) fillers which exhibit imperfect filler/matrix adhesion.
Note that the white void space around the fillers in (c) and (d) is exaggerated for visual clarity.

mathematically involved and computationally intensive. In contrast,
the possibility of incorporating partial or imperfect adhesion between
the dispersed and continuous phases into existing closed-form particle
reinforcement theories with has not been addressed.
The data presented in Fig. 6b suggests that with the addition of
50–200 mM NaCl, the mechanical properties of the WPI-X gels filled
with 4 μm glass beads exhibit two distinct regimes. At low filler content,
a passive component causes a depression in Ec, which becomes more
pronounced at higher ionic strength. Above ϕf = 0.2, filler crowding
dominates, and Ec becomes solely dependent on the filler content. Both
the observed differences in negative reinforcement in the passive re-
gime and the equivalent reinforcement in the filler crowding regime are
indicative of imperfect adhesion, modulated by the ionic strength.
As a first attempt at describing the effect of imperfect adhesion, the
two distinct regimes were modelled using a linear weighted average
approach. We have therefore expanded the GPL model to be expressed
as
Er = (1 f) + (1 )(1 f ) (13)
The two terms correspond to the bound and unbound contributions,
respectively, and λ denotes the fractional weighting, or the “interaction
parameter”. The scaling factor γ’ thus represents the negative con-
tribution resulting from the unbound component. In the case of per-
fectly bound inclusions, this equation simplifies to the GPL discussed
above (Eq. 10). As the composites all displayed equivalent reinforce-
ment at high ϕf (Fig. 6b), the contribution of the glass microspheres was
expected to be identical when filler crowding dominates, irrespective of

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A.J. Gravelle, et al.
Fig. 8. Relative elastic modulus (Er) as a function of volume fraction filler (ϕf)
for whey protein isolate/xanthan gum gels containing 4 μm glass microspheres
and prepared at varying ionic strength (0–200 mM NaCl; see legend). Solid lines
represent theoretical fits using the interaction parameter approach to model the
effect of imperfect filler/matrix adhesion (Eq. 12). Panel (b) is rescaled to
highlight the trends in reinforcement at low ϕf.
Table 2
Fitting parameters obtained for the whey protein isolate gels prepared with
varying ionic strength (50–200 mM) and filled with 4 μm glass microspheres.
Fits were using the weighted average expression of the generalized power law
model (Eq. 12). For all fits, it was assumed γ = 7.3.
Ionic strength λ ( ± SEM) γ’ ( ± SEM) R2
50 mM 0.871 ± 0.007 14.66 ± 34.07 0.83
100 mM 0.934 ± 0.002 7.74 ± 4.66 0.92
200 mM 0.955 ± 0.001 6.59 ± 1.33 0.98
the extent of filler-matrix adhesion. Theoretical fits using the interac-
tion parameter approach were thus carried out using γ = 7.3; i.e., the
scaling factor obtained for the 0 mM gels, which were assumed to be
perfectly bound.
The fits obtained using the interaction parameter approach are
presented in Fig. 8a, and Fig. 8b highlights the depression in Er at low-
to intermediate ϕf. The corresponding fitting parameters λ and γ’ are
presented in Table 2. For each of the 50, 100, and 200 mM gel systems,
the fits described the experimental data relatively well across the entire
range of filler content employed. In particular, Fig. 8b demonstrates
how these fits were able to mimic the mechanical properties of the
composites as they transitioned from a passive-dominated to an active-
dominated regime. To the best of our knowledge, the proposed inter-
action parameter represents the first attempt at considering the impact
of imperfect filler/matrix adhesion in food composites. Furthermore,
the fits presented in Fig. 8 demonstrate that the GPL approach is cap-
able of being adapted to incorporate such a physically realistic scenario.
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Food Research International 122 (2019) 209–221
5. Conclusion
Glass microspheres of varying sizes were used as model fillers to
characterize the influence of well-defined particulate inclusions in heat-
set WPI-X gels prepared at varying ionic strength. The observed trends
in the elastic modulus of these composites could be rationalized based
on the extent of filler-matrix interactions, filler size, and the corre-
sponding maximum packing fraction. In the absence of salt, the glass
microspheres were strongly bound to the gel matrix, thus allowing for
direct comparison with established particle reinforcement theories.
However, these widely used theories did not follow the same general
trend in reinforcement as that observed in the experimental data, de-
spite employing adaptations to account for filler self-crowding.
For bound fillers, an improved fit for Er(ϕf) could be achieved for all
filler sizes by using the GPL model. This functional form also followed
the observed trend in reinforcement of emulsion-filled gelatin gels
which exhibited filler clustering, previously reported elsewhere (van
Aken et al., 2015). Through a purely empirical approach, the scaling
factor describing the elastic modulus of WPI-X gels with rigid inclusions
could be expressed as a function of ϕmax. We therefore anticipate that
using an approach similar to Pal (Pal, 2002b, 2005) will allow for the
expansion of this form to incorporate fillers of finite modulus, which
would reflect the observed changes in the scaling exponent. Finally, the
GPL expression was adapted to incorporate imperfect matrix/filler ad-
hesion using a weighted average approach. This strategy allowed for
both active and passive contributions from the fillers, each expressed as
an independent power law. Experimental data indicated that increasing
ionic strength decreased the matrix/filler adhesion in glass-filled WPI-X
gels, and the trends in Er could be sufficiently described using the
weighted average approach. This work demonstrated that the GPL
model provides a robust approach to describing the impact of fillers in
particulate-reinforced composite gels. This functional form was able to
follow the general trend in reinforcement with increasing filler content
more accurately than other commonly adapted particle reinforcement
theories. Further work will be required to determine if the GPL model
can be derived using a more rigorous theoretical approach, which may
provide further insight into the factors which influence the scaling
exponent.
Acknowledgements
The authors would like to acknowledge the financial support of the
Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA).
AJG also acknowledges financial support for this work through the
National Science and Engineering Research Council of Canada (NSERC
Postgraduate Scholarship-Doctoral) and the Canadian Dairy
Commission (Doctoral Scholarship). We would also like to acknowledge
the assistance of Dr. Michaela Strüder-Kypke (Molecular and Cellular
Imaging Facility, Advanced Analysis Centre, University of Guelph) for
her assistance with conducting confocal imaging.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.foodres.2019.03.070.
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