Advances in Colloid and Interface Science 318 (2023) 102970

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Advances in Colloid and Interface Science

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Historical Perspective

Nanocellulose-stabilized Pickering emulsions: Fabrication, stabilization,

and food applications
Chuye Ji, Yixiang Wang *
Department of Food Science and Agricultural Chemistry, McGill University, Ste Anne de Bellevue, Quebec H9X 3V9, Canada

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

Keywords: Pickering emulsions have been widely studied due to their good stability and potential applications. Nano­
Pickering emulsion cellulose including cellulose nanocrystals (CNCs), cellulose nanofibrils (CNFs), and bacterial cellulose nanofibrils
Nanocellulose (BCNFs) has emerged as sustainable stabilizers/emulsifiers in food-related Pickering emulsions due to their
Stabilization
favorable properties such as renewability, low toxicity, amphiphilicity, biocompatibility, and high aspect ratio.
Food application
Nanocellulose can be widely obtained from different sources and extraction methods and can effectively stabilize
Pickering emulsions via the irreversible adsorption onto oil-water interface. The synergistic effects of nano­
cellulose and other substances can further enhance the interfacial networks. The nanocellulose-based Pickering
emulsions have potential food-related applications in delivery systems, food packaging materials, and fat sub­
stitutes. Nanocellulose-based Pickering emulsions as 3D printing inks exhibit good injectable and gelling prop­
erties and are promising to print spatial architectures. In the future, the utilization of biomass waste and the
development of “green” and facile extraction methods for nanocellulose production deserve more attention. The
stability of nanocellulose-based Pickering emulsions in multi-component food systems and at various conditions
is an utmost challenge. Moreover, the case-by-case studies on the potential safety issues of nanocellulose-based
Pickering emulsions need to be carried out with the standardized assessment procedures. In this review, we
highlight key fundamental work and recent reports on nanocellulose-based Pickering emulsion systems. The
sources and extraction of nanocellulose and the fabrication of nanocellulose-based Pickering emulsions are
briefly summarized. Furthermore, the synergistic stability and food-related applications of nanocellulose-
stabilized Pickering emulsions are spotlighted.

1. Introduction systems, protein and polysaccharide particles can function as stabilizers

Pickering emulsions are solid colloidal particle-stabilized emulsions
where solid particles are partly wetted by oil and water and adsorbed
onto the oil-water interface [1]. Compared to the conventional emul­
sions, the irreversible adsorption of the nano-sized particles at the oil/
water interface makes the Pickering emulsions preserve the droplets
against coalescence and Ostwald ripening. In particular, there has been
an increase in the replacement of common synthetic surfactants in food
products with natural and sustainable substitutes such as plant-derived
materials [2]. The demand for food-grade colloidal particles to formu­
late Pickering emulsions is growing because of the significant role of
Pickering emulsions in food industries [3]. The basic prerequisite for
food-grade colloidal particles to function as Pickering emulsion stabi­
lizers is to remain insoluble and intact in both phases throughout the
lifetime of the emulsion system. In multi-phase food Pickering emulsion
in the form of nano− /micro-particles [4,5]. Protein nanoparticles can
generate a viscoelastic interface layer for preventing the aggregation
and coalescence of droplets via electrostatic and/or steric repulsive
forces [6–8]. However, many protein particles may cause structural
instability and aggregation due to their environmental sensitivities
[9,10]. Unlike the fast interfacial adsorption kinetics and high surface
activity of protein, polysaccharide particles usually exhibit poor emul­
sifying performance and surface activity [11].
The extraction, characterization, and application of nanocellulose
(including CNCs, CNFs, and BCNFs) have been widely studied. Nano­
cellulose has potential applications in food stabilizers [12], food thick­
eners [13], food packaging [14], and dietary fibers to reduce the caloric
value of good [15]. According to the search results of Web of Science,
6648 research papers on nanocellulose have been published from 2017
to 2022, of which 175 papers (around 3%) are about Pickering

* Corresponding author.
E-mail address: yixiang.wang@mcgill.ca (Y. Wang).

https://doi.org/10.1016/j.cis.2023.102970
Received in revised form 13 June 2023;
Available online 26 July 2023
0001-8686/© 2023 Elsevier B.V. All rights reserved.
C. Ji and Y. Wang
Fig. 1. Sources and main extraction methods of nanocellulose employed for
preparing Pickering emulsions.
emulsions. Nanocellulose possesses the unique properties of high aspect
ratio, wettability, and good mechanical properties, and can form the
strongly entangled and disordered interfacial structure to effectively
prevent emulsion droplets from coalescence, which make it an excellent
stabilizer of Pickering emulsions [16,17]. The surface wettability of the
particles as Pickering emulsifiers is considered as one of the most
important factors because it directly influences the stabilizing capacity.
For the emulsions stabilized by a single layer of solid particles, oil-in-
water (o/w) emulsions are stable with a contact angle of 15◦ < θ <
90◦ , while water-in-oil (w/o) emulsions require a contact angle of 90◦ <
θ < 165◦ [18]. The contact angle of nanocellulose is in the range of 15◦
< θ < 90◦ [19–26], which is promising to stabilize oil-in-water Pickering
emulsions [18]. Its emulsifying capacity can be further improved by
physical/chemical modifications and/or the incorporation of other
synergistic stabilizers, and the resultant nanocellulose-based Pickering
emulsions have potential food-related applications such as the
controlled release of bioactive compounds (e.g., encapsulation of
β-carotene [27] and curcumin [28]), construction of active food pack­
aging materials (e.g., paper coated by nanocellulose-stabilized Pickering
emulsions [29]), and substitution of fat in food (e.g., nanocellulose-
stabilized Pickering emulsions as fat alternatives in low-calorie chewy
biscuit products [30]). However, their further commercial applications
are limited by: (i) the potential safety issues of nanocellulose-based
Pickering emulsions and the lack of the standardized assessment pro­
cedures to evaluate the immunotoxicity, genotoxicity, nutrient uptake,
etc.; (ii) the lack of the standardized nanocellulose products because the
source and extraction methods can largely affect the size and properties
of nanocellulose; and (iii) the very few nanocellulose manufacturers and
the high production cost.
There is a lack of comprehensive overview of recent development in
nanocellulose-based Pickering emulsions for food-related applications
[31,32]. Therefore, this review focuses on the recently reported
nanocellulose-stabilized Pickering emulsions, and summarizes their raw
materials, traditional/emerging technologies, and food-related appli­
cations. It aims to provide perspectives on future study to promote the
full potential of nanocellulose-based Pickering emulsions.
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Advances in Colloid and Interface Science 318 (2023) 102970
2. Source and extraction of nanocellulose
Nanocellulose has been obtained from various sources, including
woody and non-woody plants [33] (asparagus [34], bamboo pulp [35],
cotton [35], eucalyptus [36], Miscanthus floridulus straw [37], and wood
pulp [35]), marine organisms (tunicate [38] and algae [39]), and bac­
teria [40–43] (Fig. 1). The extraction of CNCs and CNFs is usually based
on top-down chemical and/or mechanical strategies, while BCNFs are
directly synthesized by bacteria. Herein, the different sources and
extraction methods of recently reported nanocellulose for the prepara­
tion of food-related Pickering emulsions are summarized in Table 1.
Compared with woody biomass, agricultural resources contain a lower
content of lignin, and thus require less chemicals and energy con­
sumption during the extraction process [33]. Especially, agricultural
wastes are excellent cellulosic feedstock because of their low-cost and
sustainability, and much attention has been attracted to the extraction of
nanocellulose from agricultural wastes, such as banana peels [44],
defatted rice bran [45], ginkgo seed shells [46], lemon seeds [47,48], oil
palm fruit bunch [49], pistachio shells [50], and sweet potato residue
[51]. Meanwhile, BCNFs can be more easily obtained from certain
bacterial species (i.e., Komagataeibacter xylinus [40], Gluconacetobacter
xylinum ATCC23767 [41], Komagataeibacter hansenii CGMCC 3917 [42],
and Acetobacter xylinum CGMCC5173 [43]), because of the high purity
of bacterial cellulose [52]. The extraction of CNCs usually consists of
pretreatments (for removal of dirt and soluble substances), purifications
(for removal of hemicellulose and lignin), and fragmentation (for gen­
eration of nanostructure via hydrolysis or mechanical destruction)
[31,53]. The use of recyclable chemicals, such as organic acids (e.g.,
formic acid [54], acetic acid [55], oxalic acid [56], maleic acid [57],
citric acid [58]) and inorganic acids (e.g., phosphotungstic acid [59] and
recycled sulfuric acid [60]), shows eco-friendly advantages and has been
applied to extract nanocellulose. However, strong acid hydrolysis causes
the harsh equipment corrosion and environmental pollution and is less
favorable. Generally, CNCs extracted by acid hydrolysis exhibit the
higher crystallinity and shorter length than those by mechanical
methods, and the properties of CNCs can be controlled and modulated
by appropriate adjustments of hydrolysis or mechanical conditions [31].
Different from CNCs, CNFs can be prepared facilely through high-
pressure homogenization, microfluidization [61], or ultrasonication.
Particularly, CNFs prepared by aqueous counter collision (ACC-CNFs)
showed enhanced exposure of inherent hydrophobic surface planes,
which contributed to the better emulsifying capacity [62,63]. Both the
source and extraction methods determine the characteristics of nano­
cellulose, such as size, crystallinity, and functional groups. The main
differences between CNCs and CNFs lie in their size and the proportion
of amorphous phase. Compared with CNCs (hundreds of nanometers in
length), CNFs have higher aspect ratios and greater length of several or
tens of micrometers [64]. At the same time, CNFs are mainly produced
by mechanical disintegration and have lower crystallinity, while CNCs
are typically produced after the removal of amorphous domains [65].
Owing to the higher aspect ratio, CNFs can form entangled nanofibril
networks at low concentrations during emulsification to limit the oil
droplet transport and collision, but the emulsion droplets are usually
larger than those stabilized by CNCs [66,67]. CNCs with lower aspect
ratio can pack densely on the oil/water interface [68]. Moreover,
nanocellulose can be further modified, for example by charged surfac­
tants such as octenyl succinic anhydride (OSA) [24,69], polymeric dia­
ldehyde [70], cinnamoyl chloride, and butyryl chloride [71], to adjust
its hydrophobicity [72].
3. Fabrication of nanocellulose-stabilized Pickering emulsions
Rotor-stator homogenization, high-pressure homogenization, and
sonication are the most widely used techniques to prepare Pickering
emulsions. Almost half of the Pickering emulsions presented in this re­
view were prepared by the rotor-stator homogenizer (Tables 1 and 2),
C. Ji and Y. Wang Advances in Colloid and Interface Science 318 (2023) 102970

Table 1
Summary of recent Pickering emulsions stabilized by nanocellulose for food-related applications. (N/R: not reported).
Cellulose source Extraction of Size of Contact Oil phase [ratio of Droplet diameter Emulsification Refs.
nanocellulose nanocellulose angle oil to water] method
Lignocellulosic Asparagus Sulfuric acid hydrolysis Length of N/R Palm oil [30:70] 3 μm Sonication [34]
sources (Woody 178.2–261.8 nm (v/v)
and non-woody
plants)
Bamboo pulp Phosphoric acid Length of tens of N/R Caprylic/capric 10–20 μm Sonication [35]
Cotton fabric hydrolysis, high- nanometers triglyceride 15–47 μm
Wood pulp pressure [10:90–50:50] 4–12 μm
homogenization (v/w)
Eucalyptus fibers High-pressure Diameter of 236 N/R Soybean oil 2.6–3.3 μm Sonication [36]
homogenization, high- nm, length higher [10:90] (v/v)
intensity ultrasound than 1 μm
Miscanthus High-pressure Width of N/R Dodecane 10.62 μm Homogenization [37]
floridulus straw homogenization 33.27–49.79 nm, [10:90] (v/v)
height of
3.68–7.55 nm
Banana peels High-pressure Diameters of 23.6 ± Sunflower oil 2 μm High-pressure [44]
homogenization, high- 3.3–3.5 nm, 2.0◦ [10:90] (w/w) homogenization or
intensity ultrasound length of 1.5–3.4 sonication
μm
Defatted rice bran Sulfuric acid (55 wt%) Diameter of N/R Rice bran oil 4.45–18.03 μm Homogenization or [45]
hydrolysis under 5.7–12.7 nm, [50:50] (v/v) sonication
sonication process length of
152–327 nm
Ginkgo seed shells Sulfuric acid (62 wt%) Length of 41.5◦ - Corn oil 10–32 μm High-pressure [46]
hydrolysis, high- 406–1500 nm 65.6◦ [10:90–70:30] homogenization
pressure (v/v)
homogenization
Lemon seeds Sulfuric acid (64 wt%) Diameter of 61.82◦ Sunflower oil 70–80 μm Homogenization [47]
hydrolysis 14.43 nm, length [50:50] (v/v)
of 155 nm
Ball milling using ionic Diameter of 62.17◦
liquids [BMIM]Cl 42.84 nm, length
of 1782 nm
Lemon seeds Sulfuric acid hydrolysis Diameter of N/R Sunflower oil N/R Homogenization [48]
(64 wt%), APS (1 M) 26–42 nm, length [50:50] (w/w)
and TEMPO oxidation of 340–380 nm
Oil palm fruit Soda pulping, Width of 4 nm, 57.5◦ Dodecane 10–18 μm Sonication [49]
bunch bleaching, TEMPO length of few [20:80] (w/w)
oxidation, and high- microns
pressure
homogenization
Pistachio shells Hydrolysis with 3 M Average diameter N/R Corn oil [10:90] 2.5–17 μm Sonication [50]
HCl, Modification with of 68.8 nm (v/v)
OSA
Sweet potato Dealing with deionized N/R 35.9 ± MCT [10:90] (v/ 8–30 μm Homogenization [51]
residue water, methanol, and 2.49◦ v)
DMAC
Algae Macroalgae Phosphoric acid Length within the N/R Paraffin oil 1.6–4.0 μm Homogenization [39]
(Eucheuma cottonii) hydrolysis sub-micrometer [10:90] (v/v)
range, diameters
of 17–157 nm
Bacteria Komagataeibacter Disintegration using a Length of 100 64.46 ± Peanut oil 15 ± 0.82 nm High-pressure [40]
xylinus blender at 15,000 rpm nm-several μm, 3.56◦ [15:85] (v/v) homogenization
for 5 min, hydrolysis diameters of
with 2.5 M HCl at 70 ◦ C 30–80 nm
for 1 h
Gluconacetobacter Activation of bacterial Diameters of 70.8 ± Olive oil [20:80] 2.6–4.1 μm Sonication [41]
xylinum ATCC cellulose at 35 ◦ C for 7 56.2–74.1 nm 1.5◦ (v/v)
23767 days
Komagataeibacter Disintegration using a Length of several N/R Sunflower seed 947.70–1409.47 Homogenization [42]
hansenii CGMCC blender at 15,000 rpm μm, diameter of oil [75:25] (v/v) nm
3917 for 5 min, hydrolysis 35–80 nm
with 2.5 M HCl at 70 ◦ C
for 1 h
Acetobacter xylinum Sulfuric acid hydrolysis Diameters of N/R CH2Cl2 [10:90] 8.8–18.5 μm Sonication [43]
CGMCC 5173 259.6 nm (v/v)

which consists of a rotor with blades and a stator with openings [73].
Rotor-stator homogenization has many advantages such as low oper­
ating cost, ease of setting-up, rapid process, and small amount of liquid
required, but also drawbacks of the lack of uniformity, limited energy
input, and risks of temperature increase and high shear rate [73–75].
High-pressure homogenizer has been frequently used in the industry and
consists of a standard homogenizing nozzle and a high-pressure pump
with pressure ranging from tens to hundreds of MPa (Fig. 2). The pri­
mary coarse emulsion prepared at the pre-emulsification step is injected
in the homogenizing nozzle to reduce the emulsion droplet size even
down to the nanometer range. Compared with rotor-stator homogeni­
zation, high-pressure homogenization can prepare smaller droplets,

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C. Ji and Y. Wang Advances in Colloid and Interface Science 318 (2023) 102970

Table 2
Summary of recent Pickering emulsions stabilized synergistically by nanocellulose and co-stabilizers. (N/R: not reported).
Co-stabilizers Nanocellulose (size) Oil phase [ratio of oil to Droplet diameter Emulsification Refs.
water] methods

Protein Soy protein isolate BCNFs (length: several μm; Sunflower seed oil [75:25] 947.70–1409.47 nm Homogenization [42]
diameter: 35–80 nm) (v/v)
Soy protein isolate BCNFs (length: several μm; Canola oil [50:50] (v/v) 10–40 μm High-pressure [87]
diameter: 50–100 nm) homogenization
Soy protein isolate BCNFs (length: <10 μm; Dodecane [20:80–50:50] 10–15 μm Homogenization [88]
diameter: 38.8 ± 3.26 nm) (v/v)
Soybean protein CNCs (length: 80–100 nm; Soy oil [20:80–50:50] (v/v) <0.1 μm Homogenization [171]
diameter: 6–12 nm)
Soybean protein isolate BCNFs (length: <10 μm; width: Dodecane [50:50–74:26] 10–15 μm at pH 3.0; High-pressure [172]
112 nm; height: 11.8 nm) (v/v) 15–40 μm at pH 7.0 homogenization
Soy protein isolate Bacterial cellulose (N/R) Sunflower seed oil [74:26] 20.71 ± 2.03 μm Homogenization [135]
(v/v)
Bovine serum albumin CNCs (length: 200 nm; Soy oil [80:20] (v/v) 52–84 μm Homogenization [89]
diameter: 10 nm)
Peanut protein isolate CNCs (length: 145.7 ± 2.6 nm) Rapeseed oil [30:70] (v/v) 5–7 μm High-pressure [173]
homogenization
Polysaccharide Chitosan CNCs (length: 156 ± 4 nm) Oleic acid [3:97] (v/v) 888 ± 127 nm Homogenization [130]
GTMAC-Chitosan Phosphorylated CNCs (length: Olive oil [30:70] (v/v) 3–5 μm Sonication [90]
around 200 nm)
Chitin CNFs (length: 18.6 ± 1.2 μm; Corn oil [10:90] (v/v) <45 μm Sonication [92]
width: 20 ± 8 nm)
Chitin nanofibril CNFs (length: 18.6 ± 1.2 μm; Corn oil [10:90] (v/v) 3.1–7.6 μm Sonication [91]
width: 20 ± 8 nm)
Sodium alginate BCNCs (width: 56.2–74.1 nm) Olive oil [20:80] (v/v) 2.6–4.1 μm Sonication [41]
Inorganic Fe3O4 CNCs (length: 50–450 nm; Flaxseed oil [10:90] (v/v) 1.14–2.68 μm Homogenization [102]
particle width: 6–26 nm)
Fe3O4 CNCs (length: 165 ± 35 nm; Paraffin oil [30:70] (v/v) 2.9–3.3 μm Sonication [104]
width: 8 ± 2 nm)
Fe3O4 CNCs (length: 150–200 nm; Palm oil [30:70] (v/v) 11.90–109.00 μm Sonication [103]
diameter: 5 nm)
Hectorite nanoplatelets BCNFs (length: 3 μm; width: 20 Silicone oil [10:90–90:10] 2– 5 μm High-pressure [99]
nm) (v/v) homogenization
Polyphenol Tea polyphenols BCNFs (N/R) Dodecane (or camellia seed <30 μm Homogenization [95]
oil) [10:90] (v/v)
Tannic acid CNCs (length: 128 nm; width: 7 Corn oil [20:80] (v/v) <5 μm Sonication [96]
nm)
(− )-Epigallocatechin-3- CNCs (N/R) Corn oil [40:60] (v/v) 7.9–9.45 μm Sonication [116]
gallate (EGCG)
Lignin CNFs (length: 700 nm; Dodecane [20:80] (v/v) ~16 μm Sonication [61]
width: 8 nm)
CNCs (N/R) Dichloromethane <30 μm Homogenization [93]
[16.7:83.3] (v/v)
Surfactant Lauric arginate (food CNCs (length: 368 ± 131 nm; Soybean oil [5:95] (v/v) ~1 μm Homogenization [97]
grade) diameter: 7.4 ± 1.5 nm)
Triterpene saponins (green) CNCs (length: 99.97 ± 3.86 Soapberry oil [30:70] (v/v) 221.2–581.3 nm Sonication [98]
nm; width: 3.68 ± 0.22 nm)

process large volume samples in a continuous manner, and control
droplet size by adjusting the pressure or the number of homogenizing
cycles [73,74]. However, the high-pressure homogenization has several
disadvantages such as high energy consumption (leading to high oper­
ating cost), difficulty of cleaning (inducing the cross-contamination),
and increase of temperature (inducing the destabilization of particles
and/or emulsions) [75,76]. High intensity ultrasound with frequencies
>20 kHz, as a green technology, has been utilized for emulsification
[77]. The titanium probe transmits the ultrasonic energy to surrounding
samples, inducing emulsification by cavitation and ultrasonic force
(Fig. 2). The ultrasonic emulsification possesses high energy efficiency,
good emulsion stability, and ease of operating and cleaning [73,78].
However, it suffers from the risks of the disruption of fragile particles or
particle aggregates and the deposition of trace amount of titanium into
the sample during ultrasonic emulsification. Besides these three tech­
niques, membrane emulsification and microfluidization have also been
applied to prepare Pickering emulsions (Fig. 2), but the industrial scale
production is still a challenge [79].
Pickering emulsions are stabilized by solid particles at the oil-water
interface, showing higher resistance to droplet coalescence and better
elastic responses than surfactant-stabilized emulsions [80]. As the
amphiphilic property plays an important role in conventional emulsions,
the wettability of solid particles is the key factor governing the forma­
tion and stabilization of Pickering emulsions [17]. Contact angle (θ) can
be used to determine the location of the solid particle with respect to oil-
water interface and thus the emulsion type (o/w and w/o). These solid
particles (stabilizers/emulsifiers) have partial wettability in both
immiscible phases, and the energy required to remove a particle from
the interface is orders of magnitude greater than the thermal energy
[81], manifesting their irreversible adsorption processes. This is essen­
tially different from the dynamic equilibrium process between the
interface and the bulk phase of conventional emulsifier-stabilized
emulsions [17]. In addition to the wettability of solid particles, their
size and shape are two other important parameters of their emulsifying
and stabilizing capacities [1]. The particle size should be substantially
smaller (at least one order of magnitude) than the targeted emulsion
droplet size. The non-spherical particles such as nanocellulose theoret­
ically have stronger ability to generate a densely packed network at the
interface because they have high energy of attachment even at a low
concentration [82–84]. It has been proved that the longer particles with
higher-aspect-ratio presented the larger emulsion volume and better
stability [68]. Nanocellulose has been widely applied to stabilizing
Pickering emulsions. CNCs (hundreds of nanometers in length) can
provide a steric barrier at the interface to prevent emulsion droplets

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C. Ji and Y. Wang
Fig. 2. Schematic diagram of fabrication methods of nanocellulose-stabilized Pickering emulsions.
from coalescence, while CNFs and BCNFs (several or tens of micrometers
in length) generate the network at the interface as well as in the
continuous phase, resulting in the hindrance for coalescence and the
increase of emulsion viscosity.
3.1. Synergistic stability
Nanocellulose is usually modified to improve the emulsifying ca­
pacity [85]. Compared to chemical modifications, the generation of
cellulose-based complexes by hydrogen bonding and/or electrostatic
interaction is an facile, cost-effective, and eco-friendly way to adjust the
hydrophobicity and interfacial tension of nanocellulose [86]. The
recently reported Pickering emulsions stabilized synergistically by
nanocellulose and co-stabilizers are summarized in Table 2. Liu et al.
[42] reported the preparation of BCNF-soy protein isolate (SPI) com­
posite colloidal particles via antisolvent approach. The addition of
BCNFs exposed the hidden hydrophobic regions of SPI through
hydrogen bonding and resulted in the improved viscoelasticity (gel-like
behavior), crystallinity, thermal stability, and interfacial wettability.
The high internal phase Pickering emulsions (HIPEs) stabilized by
BCNF-SPI (7:25, w/w) composite particles exhibited high stability dur­
ing 2-month storage. Similarly, the SPI-BCNF complexes were prepared
by triggering the electrostatic interaction [86,87]. The addition of SPI
increased the surface hydrophobicity and emulsifying capacity of
TEMPO-oxidized BCNFs, and allowed the formation of uniform and
highly stable Pickering emulsions [87]. Moreover, the Pickering emul­
sions showed good oxidative stability because of the steric barrier built
by SPI-BCNF complexes, and the increased viscosity of the emulsions
inhibited the movement of oxidative free radicals. The SPI-TEMPO-
oxidized BCNF complexes were also applied to construct edible foams
with excellent mechanical properties, porous structure, and good energy
absorption capacity [88]. Besides, CNC-bovine serum albumin (BSA)
complexes were formed via strong electrostatic interaction at pH 3.0,
which showed the increased stiffness, low surface charge, and improved
amphiphilic properties, and were suitable for the formation of HIPEs
[89]. The prepared HIPEs exhibited gel-like properties and controllable
shapes due to the 3D network bedded with close-packed emulsion
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Advances in Colloid and Interface Science 318 (2023) 102970
droplets.
Polysaccharides such as chitosan, chitin, and sodium alginate have
also been employed as co-stabilizers of Pickering emulsions. Baek et al.
[90] modified chitosan (GCh) with glycidyl trimethylammonium chlo­
ride (GTMAC) and formed the complexes with phosphorylated-CNC (P-
CNC) by ionic gelation under mild sonication. The GCh-P-CNC nano­
complexes showed a hard sphere morphology and an average diameter
of 300–500 nm, and the obtained Pickering emulsions were highly stable
without creaming, coalescence, or phase separation during 3-month
storage. A similar thick particulate layer formed by CNF-chitin com­
plexes could also prevent the emulsion droplets (3.1–7.6 μm in diam­
eter) from coalescence and creaming [91], while the complexes of high
molecular weight sodium alginate (SA) and BCNCs had better amphi­
pathicity compared to the original BCNCs [41]. Interestingly, besides
the design of nanocellulose-based complexes, Zhou et al. [92] prepared
the Pickering emulsions by blending anionic CNF-stabilized droplets
with cationic nanochitin-stabilized droplets. The adsorption of CNFs
increased the interfacial elasticity, and the formed network with inter­
linked CNFs improved the resistance of emulsion droplets to coalescence
and flocculation. At high levels of nanochitin-coated droplets (≥50%),
the cationic droplets formed a shell layer around the anionic droplets,
resulting in the reduced viscosity and increased stability.
Lignin has functional properties such as anti-oxidation and ultravi­
olet adsorption, and has been applied to modulate the hydrophilicity of
nanocellulose [93,94]. Recently, Guo et al. [61] reported the prepara­
tion of CNFs containing residual lignin. The content of lignin on CNFs
could be modulated during extraction process, and the emulsions with
more residual lignin showed the higher emulsion ratio and stability.
Polyphenols with promising bioactivities (antioxidative, antibacterial,
anti-inflammatory, etc.) can bind to nanocellulose through hydrogen
bonds and hydrophobic and cation-π interactions [95]. For example, the
complexation of tea polyphenols increased the interfacial diffusion rate
constant and surface activity of BCNFs, and the Pickering emulsions
exhibited great free-radical scavenging activity [95], while the addition
of tannic acid (TA) as a co-stabilizing agent condensed the interfacial
layer and formed the insoluble shell with CNCs and methyl cellulose to
realize the dispersibility of freeze-dried emulsions [96].
C. Ji and Y. Wang
Fig. 3. (A) Schematic illustration of the fabrication of β-carotene loaded Pickering emulsion co-stabilized by ZCPs and CNCs; (B) Cryo-SEM microstructures of
different Pickering emulsions; (C) Retention rate of β-carotene entrapped in different Pickering emulsions; (D) Digestion time dependence of FFA release (%) from
different Pickering emulsions. ZCPs: zein colloidal particles; CNCs: cellulose nanocrystals; MCT: medium-chain triglyceride; 1Z4C, 1Z1C, and 4Z1C: m(ZCPs):m
(CNCs) = 1:4, 1:1, and 4:1, respectively; 1C4Z, 1C1Z, and 4C1Z: m(CNCs):m(ZCPs) = 1:4, 1:1, and 4:1, respectively. Modified from Wei et al. [111] with permission.
Besides them, food-grade surfactants can also promote the interfacial
adsorption of nanocellulose [97]. Food-grade lauric arginate (LAE) was
combined with CNCs at a mass ratio of 2:1 (LAE: CNCs), and the Pick­
ering emulsions stabilized by positively charged LAE-CNCs complexes
showed small droplet size (~1 μm) and good stability. Pelegrini et al.
[98] created a novel green strategy to improve the hydrophobicity of
CNCs by using triterpene saponins (SAP). The high energy of adsorption
prevented the release pf particles from the fluid-fluid interface and thus
endowed the Pickering emulsions with high steric hindrance and me­
chanical resistance to coalescence and deformation.
Inorganic particles such as positively charged hectorite nanoparticles
(AHNPs) could be combined with negatively charged BCNFs to effec­
tively prevent the coalescence of emulsion droplets [99]. Meanwhile,
the incorporation of Fe3O4 magnetic nanoparticles could not only
improve the stability of Pickering emulsions [100], but also endow them
with a dual responsive behavior towards magnetic field and pH changes
[101]. Magnetic cellulose nanocrystals (MCNCs) were prepared by
ultrasound-assisted in-situ coprecipitation and exhibited the increased
wettability and higher surface charge compared to original CNCs
[102,103]. Mikhaylov et al. [104] found that the oil-in-water Pickering
emulsions stabilized by Fe3O4-CNCs complexes possessed high dynamic
viscosity and thixotropic properties. The surface charge of the com­
plexes was reduced and controlled by varying CNC-Fe3O4 ratio, so the
electrostatic mutual repulsive forces between emulsion droplets became
weaker to allow the formation of aggregates and the increase of dynamic
viscosity and thixotropic behavior. Moreover, the obtained palm olein-
in-water Pickering emulsions stabilized by Fe3O4-CNCs complexes
were stable under pH 3–6 and maintained the morphology and size of
emulsion droplets at pH 6 for at least 14 days [103]. The dense MCNC
layer formed at oil/water interface acted as a physical steric barrier to
preventing droplets from aggregation and coalescence, as well as
showing a dual responsive behavior towards magnetic field and pH
change.
3.2. Double pickering emulsions
Double emulsions can be applied to improve the encapsulation and
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Advances in Colloid and Interface Science 318 (2023) 102970
protection of multiple bioactive compounds with different solubilities
[105]. They also provide the possibility of fabricating reduced-fat
emulsion products by the replacement of conventional oil-in-water (o/
w) emulsions with the equivalent water-in-oil-in-water (w/o/w) emul­
sions that have lower actual oil contents but similar in-mouth perceived
texture [106]. Cunha et al. [107] designed novel surfactant-free oil-in-
water-in-oil (o/w/o) Pickering emulsions stabilized by the combination
of natural and partially esterified nanocellulose from softwood sulfite
pulp fibers. The double emulsions were prepared by adding the modified
nanocellulose suspensions in hexadecane to the primary o/w emulsions,
exhibiting remarkable stability without coalescence during one-month
storage [107].
4. Food-related applications
Due to their sustainability, biocompatibility, cost-efficiency, low
toxicity, and high stability, nanocellulose-stabilized Pickering emulsions
have many potential applications such as the delivery vehicles for bio-
medicine design, pest control, and optoelectronic devices. Especially,
more and more attention has been paid to their food-related applica­
tions, and recent research work has mainly focused on the encapsulation
of bioactive compounds, construction of functional food packaging
materials, and preparation of fat substitutes.
4.1. Delivery systems of bioactive compounds
The delivery systems with encapsulated bioactive compounds have
promising applications in food and pharmaceutical fields, ranging from
improving stability to controlling the release for enhanced functionality
[108]. Nanocellulose can generate a solid layer as the physical barrier at
the interface of emulsified droplets. This layer inhibits the interaction of
lipase with lipids in the gastric environment to protect the encapsulated
bioactive compounds against degradation, while in the small intestine,
the release of fatty acids during lipid digestion and the further formation
of mixed micelles with the incorporation of bioactive compounds
facilitate the delivery at target sites [109].
Qi et al. [27] prepared oleogel-in-water Pickering emulsions
C. Ji and Y. Wang
Fig. 4. Schematic illustration of (A) the fabrication of beeswax-in-water Pickering emulsion and the casting process of oleofilm with enhanced hydrophobicity
(reprinted from Li, et al. [129] with permission); (B) the preparation of CNCs-stabilized soybean oil-in-water Pickering emulsions and the emulsion coating process on
paper (reprinted from Liu, et al. [29] with permission); (C) the preparation of antimicrobial and antioxidant CNF films containing microcapsules (reprinted from Wu
et al. [136] with permission).
stabilized by CNCs to encapsulate β-carotene, which exhibited the
remarkable stability over a wide range of pH (4–8) and salt concentra­
tions (0.05–0.60 M). Patel et al. [110] developed Pickering emulsions
from lauric acid modified CNCs for the encapsulation of β-carotene at
different pH levels, dilution factors, and storage periods. The modified
CNCs stabilized the morphology of lipid droplets and effectively pre­
vented the creaming of lipid phase during storage. Wei et al. [111]
designed the complexes of zein colloidal particles (ZCPs) and CNCs (the
mass ratio of ZCPs and CNCs was 1:4) as the stabilizers for Pickering
emulsions to encapsulate β-carotene (Fig. 3). A high retention rate
(60.23%) was observed over 28-day storage period, and the multilayer
structure could delay the lipolysis of emulsions during digestion. The
inner CNC layer and the outer layer formed of ZCPs synergistically
restricted the penetration of lipases and bile salts due to the high
desorption energy while maintaining the high bioaccessibility of
β-carotene (20%).
Curcumin has various biological and pharmacological activities
(including antimicrobial, antioxidant, anti-inflammatory, and anti­
cancer properties), but its therapeutic applications are limited because
of its low stability and bioavailability. Recently, numerous research
works reported that nanocellulose-stabilized Pickering emulsions could
be used as delivery systems of curcumin [28,112–115]. Pickering
emulsions stabilized by aminated CNCs for coumarin and curcumin
showed high encapsulation efficiency (>90%), increased bioavail­
ability, and low toxicity [28]. Some other Pickering emulsions stabilized
by nanocellulose with encapsulated curcumin also exhibited good sta­
bility during long-term storage [114]. The stability of curcumin could be
further improved with the incorporation of (− )-epigallocatechin-3-
gallate (EGCG), which was added in the pineapple peel CNC-stabilized
Pickering emulsions and interacted with CNCs through hydrogen
bonding [116]. The interaction between EGCG and CNCs led to a denser
barrier that inhibited the isolation of serum layer, and the antioxidant
and antiultraviolet properties of EGCG prevented free radicals from
attacking on the encapsulated curcumin during storage. In the CNC/
EGCG emulsions, curcumin was highly stable with 70% retention over a
30-day storage period and was not affected under thermal and ultravi­
olet light treatments. The release of encapsulated curcumin could be
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Advances in Colloid and Interface Science 318 (2023) 102970
controlled by the incorporation of Fe3O4 [113]. Upon exposure to
external magnetic field (EMF), the adsorbed CNCs at oil/water interface
turned magnetized and got attracted and concentrated towards the field
direction, leading to the dilatational deformation of the fluid interface
and then the escape of CNCs from oil/water interface to the aqueous
phase. The de-stabilization of emulsion droplets under EMFs resulted in
the accelerated release rate of curcumin, and thus an inhibition of the
growth of human colon cancer cells (down to 18%) was observed.
Vitamin D3 is a required nutrient to control numerous important
biochemical pathways but has poor water-solubility and low bio­
accessibility. It was encapsulated in CNF stabilized Pickering emulsions
to improve the stability and bioaccessibility [117,118]. The effect was
related to the CNF concentration, where a significant decrease in the
bioaccessibility and stability of vitamin D3 was observed at the high
level of CNFs (7:10 CNF-to-oil) [117]. The high level of CNFs partially
inhibited the lipid digestion, so some vitamin D3 molecules remained in
the indigested oil phase and less mixed micelles were generated to sol­
ubilize the vitamin. In the Pickering emulsions stabilized by CNC-
pseudoboehmite heterocoagulates, 83% of initial vitamin D3 was
encapsulated and the target release in the small intestine was achieved
[119].
Some other bioactive compounds were also employed as the models.
For example, TEMPO-oxidized CNFs (TOCNFs) extracted form Undaria
pinnatifida, one of brown algae, were used to stabilize astaxanthin
(AXT)-loaded Pickering emulsions and enabled better storage stability
and thermal stability (retention rate of AXT was 57.23% at 75 ◦ C)
compared with the Pickering emulsions stabilized by whey protein
isolate and xanthan gum (retention rate of AXT was about 30% at 70 ◦ C)
[120]. BCNFs stabilized Pickering emulsions were applied to load and
sustained release alfacalcidol (a hydrophobic drug) [43]. Lu et al. [121]
used milled-cellulose-particles stabilized Pickering emulsions to deliver
aged citrus peel extract, which exhibited a range of biological activities
and improved bioaccessibility.
4.2. Food packaging films
Biopolymer-based edible films have gained much interest, but the
C. Ji and Y. Wang
hydrophilic nature of polysaccharides and proteins displays high mois­
ture adsorption [122]. Pickering emulsions stabilized by nanocellulose
have been applied as reinforcements or coatings to improve the func­
tional properties of edible films, such as moisture barrier property,
mechanical properties to resist the external destructive forces [123],
antioxidant property to protect foods from oxidation [124], and anti­
microbial property to prevent food spoilage [125–128]. Li et al. [129]
prepared edible oleofilms by casting beeswax-in-water Pickering emul­
sions stabilized by hybrid particles of BCNFs and carboxymethyl chito­
san (Fig. 4A). Beeswax as a natural wax is a remarkable barrier to
moisture, while the addition of carboxymethyl chitosan enhanced the
interactions within BCNFs network. With the increase of hybrid particle
contents, the oleofilms showed the reduced water vapor permeability
(<1.1 × 10− 7 g⋅m− 1⋅h− 1⋅Pa− 1), enhanced surface hydrophobicity (the
contact angle increased from 113.1◦ to 139.1◦ ), and improved re-
dispersibility.
Deng et al. [130] prepared an edible coating from CNC-stabilized
Pickering emulsions and chitosan (2 wt%). CNC Pickering emulsions
enhanced the water vapor barrier property and stability of chitosan
coatings, and an optimized oleic acid concentration of 20 g/kg retained
peel chlorophyll, decreased senescent core browning, and delayed
ethylene production and respiration peaks of postharvest pears [131].
Liu et al. [29] fabricated thermally curable Pickering emulsions stabi­
lized by CNCs (3.0 wt%) (Fig. 4B). CNCs as a coating agent uniformly
formed a film after drying and filled the pores of paper to realize a low
permeability against air, contributing to enhanced tortuosity of water
molecules through the film and raised water vapor barrier properties of
the Pickering emulsion coating.
Moreover, nanocellulose was used as the reinforcement to provide
structural support for nanocomposite polymer materials via the forma­
tion of nanocellulose-stabilized Pickering emulsions. Reinforcing pearl
millet starch-based films with Kudzu CNCs increased tensile strength
(from 3.9 to 16.7 MPa) and Young’s modulus (from 90 to 376 MPa) with
the lower water vapor permeability (from 9.60 to 7.25 × 10− 10
g⋅m− 1⋅s− 1⋅Pa− 1) [132]. The improvement in mechanical properties was
likely due to the strong interaction and increased number of bonds be­
tween CNCs and starch molecules in the film matrix, while the lower
water vapor permeability was because of the relatively higher hydro­
phobicity and more rigid crystalline structure of CNCs compared to
starch molecules that increased the tortuosity of the diffusion path. Xu
et al. [133] prepared dialdehyde-oxidized CNCs (DCNC) as stabilizers of
Pickering emulsions. The DCNC-stabilized Pickering emulsions loaded
with dihydromyricetin were incorporated into gelatin matrix to prepare
gelatin-based edible films, resulting in strong UV-light barrier ability
with high transparency, low water vapor permeability (~3.62 × 10− 10
g⋅m− 1⋅s− 1⋅Pa− 1), and enhanced mechanical property. Interestingly,
DCNC was also used to crosslink gelatin by Schiff’s base reaction to
generate a compact and dense network structure, which improved the
stiffness of active films and provoked certain tortuosity to reduce the
diffusion of water molecules. Moreover, transparent films were obtained
from gelatin and agar with the incorporation of clove essential oil
Pickering emulsions stabilized by TEMPO-CNFs, and showed strong
antioxidant activity and good UV blocking property without sacrificing
the transparency (T280 decreased from 26.9% to 1.7%) [127]. The
polystyrene/CNF composite film prepared by polymerizing the encap­
sulated styrene monomer inside the CNF-stabilized Pickering emulsions
showed good thermal dimensional stability and improved mechanical
properties due to the reinforcement by homogenously distributed CNFs
[134]. During the polymerization process, the polystyrene nanoparticles
precipitated in the aqueous medium, and each individual particle was
surrounded by CNFs. After filtration and subsequent drying, a unique
film structure was formed where the polystyrene nanoparticles were
confined by the CNF framework. Liu et al. [135] developed high-barrier
konjac glucomannan (KGM) based emulsion films by incorporating
HIPEs, where HIPEs were stabilized by the complexes of BCNFs and soy
protein isolation. With the increase of HIPEs concentration in KGM
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Advances in Colloid and Interface Science 318 (2023) 102970
films, the HIPEs/KGM films exhibited the improved thermal stability,
and the reduced moisture content, water solubility, water vapor
permeability, and oxygen permeability.
Nanocellulose-stabilized Pickering emulsions have been considered
as promising carriers of oil-soluble bioactive compounds to prepare
edible films for keeping food quality and safety. Recently, researchers
have successfully prepared novel antimicrobial packaging films by using
nanocellulose-based solid particles, which contributed to the mechani­
cal strength and stability of Pickering emulsions. Plant essential oils are
natural bioactive compounds with excellent antimicrobial properties.
The combination of cellulose particles and essential oils in Pickering
emulsions provided a simple way to improve water vapor barrier
property and antimicrobial activity. Wu et al. [136] fabricated CNF-
based active films with remarkable antimicrobial and antioxidant ac­
tivities by adding oregano essential oil Pickering emulsions stabilized by
functionalized solid particles (Fig. 4C). Notably, the particles were
complexes of polyethyleneimine (PEI) and TEMPO-oxidized CNCs
(TOCNCs). The combination of PEI-TOCNCs (contact antimicrobial) and
oregano essential oil (volatile antimicrobial) brought the enhanced
antimicrobial properties (the inhibition rates: 97.28 ± 0.20% of L.
monocytogenes, 97.23 ± 0.53% of E.coli) into CNF films. Souza et al.
[125] fabricated starch-based thermoplastic films containing CNF-
stabilized Pickering emulsions of ho wood essential oils, showing the
improved mechanical resistance and less crystallinity (~2%) than the
pristine starch films. The hydrogen bonding interactions between
polyphenols of essential oils and amylopectin chains contributed to the
stiffness of the active films, the integrity of internal network, the
improvement of cohesiveness, and low melting temperatures (50 ◦ C).
These interactions also helped retain essential oil in the active films to
prolong the release, showing a promising application in active food
packaging to inhibit food spoilage and extend the shelf lives of foods.
4.3. Fat substitutes
Fat substitutes can mimic the taste, functions, and sensory properties
of fats and reduce the use of saturated and trans-fat in various foods.
They can also inhibit or slow down lipid digestion to effectively promote
obesity reduction and decrease calorie intake. Recently, the application
of Pickering emulsions as fat substitutes has attracted considerable in­
terest. Sensory evaluation and physicochemical characterization have
shown that Pickering emulsions could reduce the calories of food
products and extend their shelf life without changing color and texture
[137,138]. Nanocellulose-stabilized soybean oil emulsions were used in
meat sausages to replace pork back-fat [139]. The dense network formed
by nanocellulose contributed to the enhanced texture and rheological
behavior of emulsion-type sausages, which presented the improved
water holding capacity and increased hardness, springiness, and chew­
iness. The addition of CNF-stabilized Pickering emulsions as a fat sub­
stitute in biscuits increased the springiness and cohesiveness of dough
due to the promoted complexity of CNF/gluten network, and inhibited
the release of free fatty acid during digestion as the strong irreversible
adsorption of CNFs to the oil droplet surface restricted the lipases to
access lipids [140].
Pickering emulsions as fat substitutes are more advantaged in
modulating oil digestion process and reducing lipid oxidation, which
have been applied in the manufacture of various food products, such as
in ice cream to replace cream and in cakes to replace butter [138,141].
Nanocellulose has been considered as the suitable stabilizers to modu­
late the lipid digestion and the suitable structuring agent for liquid oil
during emulsification to maintain the texture of food products. The
physical barrier generated by the irreversible adsorption of nano­
cellulose at the oil/water interface can induce the resistance to enzymes
(pepsin and gastric lipase) and bile salt displacement [142–144].
Nanocellulose can better prevent the diffusion of lipase to the droplet
surface than the proteinaceous Pickering emulsion stabilizers, resulting
in slow initial lipid digestion [142]. The stability of nanocellulose-
C. Ji and Y. Wang Advances in Colloid and Interface Science 318 (2023) 102970

Fig. 5. (A) (a) Droplet size distribution of oil-in-water emulsions stabilized by PPM (PPM-E) at pH 7.0 and pH 3.0, and PPM + SGF mixture at pH 3.0 without the
addition of pepsin; (b) Cryo-SEM micrograph of PPM-E at pH 7.0. Scale bar: 10 μm; (c) Schematic illustration of the gastric digestion of Pickering emulsions
(reprinted from Zhang, et al. [149] with permission). (B) Schematic illustration of the in vitro gastrointestinal digestion of nanocellulose-stabilized Pickering
emulsions by altering cellulose lengths (reprinted from Ni et al. [154] with permission). (C) (a) The color and appearance of biscuits prepared with different fat
replacement rates; (b) The hardness and springiness of biscuit doughs prepared with different fat replacement rates. Different letters indicate statistically significant
differences between groups (p < 0.05) (reprinted from Xie et al. [30] with permission).

stabilized Pickering emulsions at the preceding stage of digestion
(gastric phase) is important for the subsequent restriction of interfacial
digestive dynamics [145]. At the intestinal stage, the lipolysis of emul­
sions should be delayed and/or hindered by restricting the coupling of
bile salts to reduce calorie adsorption. Since the lipolysis is governed by
interfacial property, the packed interfacial layer formed by nano­
cellulose can provide resistance to the displacement by bile salts in the
digestive environment and play an important role in the design of fat
substitutes [146–148]. It was worth noting that the lipid type influenced
the digestion profiles of emulsions significantly. The rate and degree of
lipolysis of Pickering emulsions with medium chain triglycerides (MCT)
were greater than those with long chain triglycerides (such as soybean
oil and canola oil) [114], and the restricted lipolysis could increase the
bioavailability of lipid-soluble components because of the effective de­
livery to specific sites [146].
Recently, Le et al. [142] reported the Pickering emulsions stabilized
by hydrophobically modified CNCs that decreased the release of short-
chain fatty acids (SCFA) and showed the stability as a delivery vehicle
for SCFAs to targeted regions of the digestive tract. CNCs are not
responsive to human proteolytic enzymes, such as pepsin in the gastric
phase. In addition, the resistance to bile salt displacement due to the
electrostatic repulsion of CNCs to bile salts and the high desorption
energy of irreversibly absorbed CNCs slowed the rate of initial lipid
digestion. Zhang et al. [149] also confirmed that CNCs added to pea
protein-particle-stabilized Pickering emulsions acted as a barrier and
restricted the access of pepsin to the interfacial available substrate sites
in the Pickering emulsions (Fig. 5A). Moreover, the composite layers of
Pickering emulsions could modulate the intestinal digestion of oil
droplets. Mackie et al. [150] found that CNCs were entrapped in the
intestine mucosal layer (a barrier to nanoparticle uptake) and failed to
reach the underlying epithelium, resulting in the reduction of lipid ab­
sorption. The elastic interface layer formed by BCNFs-SPI complexes
also enhanced the anti-digestion of Pickering emulsions [87]. The
adsorption of BCNFs generated physical hindrance to bile salts and
lipase, resulting in reducing the possibility of lipolysis and delaying the
lipid digestion. Liu et al. [151] found that TEMPO-oxidized CNFs-sta­
bilized Pickering emulsions exhibited stable colloidal properties in the
simulated intestinal environment and stronger inhibitory behavior to
lipid digestion compared with Tween 80 and cholate. Meanwhile, the
larger oil droplets reduced the surface area of triacylglycerol exposed to
the lipase enzymes, which could contribute to the inhibition of lipid
digestion [152].
Similarly, the coalescence and flocculation of droplets and the
accumulation of free fatty acids at the droplet surfaces also resulted in
the inhibition of lipolysis [152,153]. Besides the above-mentioned fac­
tors, the length of cellulose nanoparticles could affect the in vitro
gastrointestinal digestion of Pickering emulsions (Fig. 5B), where the
Pickering emulsions stabilized by longer cellulose nanoparticles showed
the lower release of free fatty acids due to the gel structure formed at the
gastric stage [154]. A few potential applications have been evaluated.
For example, Xie et al. [30] designed oil-in-water Pickering emulsions
stabilized by bacterial cellulose and water-insoluble dietary fibers as fat
alternatives to meet the demands of low-calorie baked foods (Fig. 5C).
The replacement of fat with emulsion could induce more interparticle
interactions to improve the development of the gluten network. The
reduction of oil content and the introduction of dietary fiber led to the
decreased hardness and increased springiness of the dough and the
higher hardness of the resultant biscuit products. Similarly, Wang et al.

9
C. Ji and Y. Wang
[155] used CNF-stabilized Pickering emulsions as fat alternatives in
emulsified sausage. When 30% fat was replaced, the low-fat sausage
exhibited brighter color and enhanced texture properties, such as higher
hardness, springiness, and chewiness, because of the rigid network
formed by CNFs with high strength and stiffness. Compared with the
full-fat sausage, the overall acceptability of the low-fat sample had no
significant difference.
5. Nanocellulose-stabilized Pickering emulsions for 3D printing
With the development of technology and materials, three-
dimensional (3D) printing has revolutionized the conventional
manufacturing. Pickering emulsions stabilized by nanocellulose have
gained an increasing demand as suitable 3D printing inks due to their
outstanding internal phase capacity and remarkable rheological be­
haviors (flowability as well as solidity against collapse) in terms of
injectable and gelling properties [32,156]. Nanocellulose stabilized
HIPEs inks benefit from high storage modulus and shear-thinning
property and are promising to print spatial architectures with high
shape fidelity, high structural integrity, and low shrinkage [157,158].
Ma et al. [159] prepared 0.5 wt% unmodified CNC-stabilized HIPEs
with 80% internal phase fraction, which exhibited high printing reso­
lution and fidelity at pH 7. The addition of NaCl could partially screen
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Advances in Colloid and Interface Science 318 (2023) 102970
Fig. 6. (A) (a) Schematic illustration of the fabrica­
tion of all-in-one, two-phase emulgels; (b) Confocal
image of PLA/CHCl3-in-water Pickering emulsion
stabilized by nanofibrils isolated from cellulose (CNF)
and chitin (ChNF); (c) Confocal and bright field im­
ages of two-phase emulgels before (upper) and after
(bottom) alkali-induced gelation. The dashed line
highlights a region depleted of droplets after gelation.
(B) Top view of cubic grids printed with ESR4 ink
using a needle size of (a) 0.25, (b) 0.41, (c) 0.63, and
(d) 0.84 mm. SEM images of the grids printed by
using (e) 0.41 and (f) 0.84 mm needles. Top view of
the 3D-printed model of (g) a human dental and (h)
the letters Aalto. Reprinted from Huan et al. [163]
with permission.
the surface charge of CNCs to promote their interfacial packing, and thus
contribute to the stability of HIPEs [160]. The continuous phase of CNC-
stabilized Pickering emulsions constructed by host-guest hydrogels
enabled the high storage modules (reached up to ~113 kPa) and
maintained the shear-thinning behavior, resulting in high-quality 3D
printed structures with high shape fidelity, structural integrity, and
excellent dimensional stability (volume shrinkage of 7% ± 2% after
freeze-drying) [158]. Hydrocarbon chain grafted CNCs (HCNCs) stabi­
lized HIPEs could be used for 3D printing to construct ultra-lightweight
porous scaffolds with tunable structures by adjusting the oil phase
fraction and aqueous conditions (such as pH value, ionic strength, and
HCNCs content). Stimuli-responsive materials could be further incor­
porated in the printed hydrogels to control the release rate of internal oil
phase in response to surrounding environment [161].
TEMPO-oxidized bacterial cellulose (TOBC) reinforced poly(lactic-
acid) (PLA) was used to produce fully biodegradable nanocomposites
through 3D printing [162]. The uniform dispersion of 1.5 wt% TOBC in
PLA matrix enabled the strong network to transmit stress and hinder the
diffusion of cracks when the composites were subjected to the external
force, and thus they showed the improved mechanical properties (tensile
strength and elongation at break increased by 9.2% and 202%) and
crystallization rate (increased by 116%) of PLA. Huan et al. [163]
designed the two-phase emulgel systems for 3D printing that consisted
C. Ji and Y. Wang Advances in Colloid and Interface Science 318 (2023) 102970

Table 3 generated by the irreversible adsorption of nanocellulose at oil/water

Summary of recent toxicological studies on nanocellulose. interface could slow down the digestion of lipid to promote obesity
Nanocellulose Test methods Results Refs. reduction. Nanocellulose also provided the excellent rheological prop­
erties of Pickering emulsions for 3D printing. Nanocellulose-stabilized
CNC, CNF Embryonic zebrafish Low toxicity (mortality rate [174]
assay < 10% at concentration HIPEs inks had high storage modulus and shear-thinning property,
below 1000 mg/L CNC/CNF) which enabled the printed architectures with high shape fidelity and
CNC, CNF Pierce LDH assay No toxicity (insignificant [175] structural integrity.
increase in LDH release at In future studies, the utilization of agricultural waste/by-products
1.5% (w/w) of CNC/CNF)
CNC MTT assay (HEK293 No cytotoxicity (cell [176]
and the development of “green” and facile extraction methods for
cells) viability > 85% with 5 μg nanocellulose production deserve more attention. The current utmost
CNC in the nanocomplex) challenge is to maintain the stability of nanocellulose-based Pickering
CNC Cytokine/chemokine Inflammatory effects (level [177] emulsions in multi-component food systems and at various conditions
level value increased at 200 μg of
(different pH values, temperatures, ionic strengths, etc.). Considering
CNC)
CNC MTT and LDH assays No cytotoxicity (no cytotoxic [178] the controllable digestion of the loaded lipid, it is promising to develop
(HBMEC, bEnd.3, RAW effects at 50 μg/mL of CNC) composite interface layer based on rational design to further delay lipid
264.7, MCF-10A, MDA- digestion and/or site-dependently release lipidic bioactive molecules. It
MB-231, MDA-MB-468, has been reported that soluble dietary fibers can promote metabolic
KB, PC-3 and C6 cells)
CNC Cell culture and MTT Low cytotoxicity (cell [179]
benefits on body weight and glucose control [164], and higher intakes
assay (L929 cells) viability > 80% at 0.2% (w/ (25– 29 g per day) of dietary fiber are suggested to protect against
w) of CNC) cardiovascular diseases, colorectal and breast cancer [165]. Despite the
CNF MTT and LDH assays Cytocompatibility (MTT [180] great promise, more studies on translating the knowledge of
(L929 cells) activity > 80% at 500 μg/mL
nanocellulose-based Pickering emulsions into new and useful applica­
of CNC, no LDH release at 1
mg/mL of CNC) tions are still expected. Additionally, it is worth noting that CNCs and
CNF Animal experiments Few effects on the fecal [181] BCNFs have not been listed as the “generally regarded as safe” sub­
(male Wistar Han rats) microbiome and stances by the food and drug administration (FDA) [166]. The safety and
metabolome in the presence biological effects of nanocellulose depend on various factors (size,
of 1% (w/w) CNF
shape, surface chemistry, charge, etc.) [67,167,168]. Table 3 summa­
CNF Animal experiments No harmful effects in the [182]
(male Rattus norvegicus animal metabolism when the rizes the recent results of toxicological studies on nanocellulose. Multi­
albinus mice) diets for mice contained 21% ple cell lines and animal models have suggested that nanocellulose
CNF exhibited limited cytotoxicity and genotoxicity [169,170]. However,
CNF Human and mice No cytotoxicity and non- [170]
considering the complexity of emulsion and food systems, the case-by-
macrophages (RAW inflammatory (cell viability
264.7 cells), cytokine/ > 90% and no expression of case studies on the potential safety issues of nanocellulose-based Pick­
chemokine level inflammatory cytokine at ering emulsions need to be carried out.
300 μg/mL of CNF)
BCNF CellTiter-Glo™ and LDH No cytotoxicity (cell [183] Declaration of Competing Interest
assays (L929 cells) viability > 70% and LDH
release < 7.5% at 1 mg/mL
of BCNF) The authors declare that they have no conflicts of interest to this
BCNF MTT assay (NIH/3 T3 Low cytotoxicity and good [184] work.
cells) biocompatibility (relative
growth rate > 80% on the
Data availability
BCNF films)
BCNF MTT assay (HUVECs) No immunogenicity and no [185]
and animal experiments toxicity (cell viability > 85% No data was used for the research described in the article.
(male C57/Bl6 mice) on the BCNF films)
Acknowledgments

of PLA/chitin/CNF Pickering emulsions and CNF/silica particles

We would like to acknowledge financial support from the Natural
hydrogels (Fig. 6). The hierarchical layers generated by precise and
Sciences and Engineering Research Council of Canada (NSERC RGPIN-
simultaneous phase separation of immiscible but metastable emulsion/
2019-04498), and Natural Sciences and Engineering Research Council of
hydrogel phases allowed the regulation of diffusion and permeability of
Canada Discovery Launch Supplement (NSERC DGECR-2019-00472). C.
cargos, thereby controlling the mass transport and channeling to outer
J. would like to thank the China Scholarship Council (CSC NO.
environment.
202106790028) for financial support of her Ph.D. program.

6. Conclusions and perspective

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stabilization. Nanocellulose-stabilized Pickering emulsions have offered
promising advantages in food applications. Especially, they could
improve the stability of encapsulated bioactive compounds and control
the release for enhanced bioavailability. Functional food packaging
films formed with nanocellulose-stabilized Pickering emulsions had
good moisture barrier property, mechanical strength, antioxidant ac­
tivities, and antimicrobial properties. Moreover, the physical barrier
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