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Janus nanoparticles in cancer diagnosis, therapy

Cite this: Biomater. Sci., 2019, 7,
and theranostics
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1262
Yifan Zhang, Kai Huang, Jing Lin and Peng Huang *

Received 23rd November 2018,
Accepted 16th January 2019
DOI: 10.1039/c8bm01523f
rsc.li/biomaterials-science
Anisotropic Janus nanoparticles (JNPs), due to their several distinct merits, have been widely investigated
for cancer theranostics. Firstly, the asymmetric modification of JNPs allows the independent release of
multiple drugs. Secondly, the spatial separation of imaging materials effectively diminishes the signal
interference and attenuation that usually take place in core–shell or alloy nanoparticles. Thirdly, their
asymmetric composition contributes to the photothermal or potential gradient that could drive the
motion of JNPs for dynamic drug delivery. This review focuses on the state-of-the-art progress of JNPs
for cancer theranostics, including their categorization, fabrication strategies, and biomedical applications
in cancer theranostics. The performances of JNPs and conventional core–shell nanoparticles are com-
pared. Last but not least, the challenges and opportunities of JNPs in cancer theranostics are also
discussed.

1. Introduction In the biomedical field, JNPs have also attracted ever-

increasing research interest, because their anisotropic struc-
Janus, an ancient Roman god of beginnings and gates, has two tures can accommodate more versatile and relatively indepen-
faces in opposite directions. In 1991, Pierre-Gilles de Gennes dent diagnostic/therapeutic functions compared with conven-
first proposed the concept of “Janus particles” to the scientific tional hybrid nanoparticles (e.g. core–shell nanoparticles and
community in his Nobel Laureate Lecture entitled Soft Matter.1 alloy nanoparticles). By the construction of JNPs using
Janus particles are described as particles with two sides of imaging materials (e.g. MnO2 and Fe3O4 for magnetic reso-
opposite polarity. The amphiphilic Janus particles were specu- nance imaging (MRI), gold for photoacoustic imaging (PAI)
lated to form a monolayer in an air–water interface such as and silver for surface-enhanced Raman scattering (SERS)) or
surfactant molecules, and this particulate film would allow incorporation of imaging agents into the JNPs (e.g. fluorescent
small molecules to transport. Over the past three decades, the dyes and quantum dots), various imaging modalities can be
concept of Janus nanoparticles (JNPs) has been extended to
the nanoparticles that have two or more districts that differ in
chemical compositions or surface properties. Generally, JNPs
can be divided into three main categories: polymeric, in-
organic, and polymeric–inorganic JNPs. They have not only
symmetric shapes such as sphere, disk, rod, etc., but also
diverse hybrid shapes such as dumbbell, snowman, mush-
room and raspberry (Fig. 1).2 The compositions of JNPs range
from organic materials (e.g. polymers) to inorganic materials
(e.g. gold, silica and silver).3–5 The unique anisotropic features
of JNPs in compositions and functions endow them with intri-
guing surface, optical, electronic, magnetic, and catalytic pro-
perties, allowing for applications in a wide range of fields,
such as particulate stabilizers, optical sensors, catalysis, and
nanomotors.6–9

Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging,

Carson International Cancer Center, Laboratory of Evolutionary Theranostics, School
of Biomedical Engineering, Health Science Center, Shenzhen University, Shenzhen
518060, China. E-mail: peng.huang@szu.edu.cn Fig. 1 Schematic illustration of typical Janus nanostructures.

1262 | Biomater. Sci., 2019, 7, 1262–1275 This journal is © The Royal Society of Chemistry 2019

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achieved for diagnosis and monitoring. Compared to core–
shell nanoparticles, the interference between different
imaging materials is significantly diminished in JNPs. On the
other hand, a combination of JNPs with therapeutic agents
can realize various therapeutic approaches such as chemo-
therapy, phototherapy, magnetolytic therapy, radiotherapy,
gene therapy, immunotherapy, and so on. JNPs provide mul-
tiple independent districts for drug loading, which allows the
spatiotemporally independent release of multiple drugs.10
Furthermore, JNPs can act as nanomotors to deliver drugs or
physically destroy tumor cells owing to the propulsion gener-
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ated from the physical–chemical reactions towards external
fields.11 Therefore, JNPs have great potential to become next
generation nanomedicines for cancer diagnosis, treatment and
theranostics.
In this review, recent progress of JNPs in cancer theranos-
tics will be summarized. First, we will introduce some main
fabrication strategies of theranostic JNPs. Then, the appli-
cations of JNPs in cancer therapy and theranostics will be illus-
trated in detail. Finally, the challenges and prospects of JNPs
for biomedical applications will be discussed.
2. Synthesis of Janus nanoparticles
for cancer theranostics
One primary focus of the research on JNPs has been the optim-
ization of fabrication strategies. In recent years, various syn-
thesis approaches have been developed for Janus structures,
such as masking, assembly, emulsion polymerization, phase
separation, Pickering emulsion interfacial synthesis, surface
nucleation and growth, etc.4,12–14 Previous review articles have
introduced preparation methods and procedures of Janus
particles.2,10,15–20 In this section, we will emphasize the syn-
thetic strategies of JNPs relevant to cancer theranostic appli-
cations, based on the compositions of polymeric, inorganic
and polymeric/inorganic materials (Table 1).
2.1 Polymeric Janus nanoparticles
A number of studies have employed polymeric JNPs for drug
delivery, including Janus dendrimers and spherical polymeric
nanoparticles. Dendrimers are repetitively branched macro-
molecules with roughly spherical symmetry around the core,
and the repetitive unit is defined as a dendron. Dendrimers
are potential drug carriers due to their monodispersity, hydro-
philicity, encapsulation ability, and abundant functionalizable
groups.21 Janus dendrimers are composed of two or more
chemically distinct dendrons, thus forming an asymmetric
branched architecture. Due to the composition heterogeneity,
Janus dendrimers could exert more theranostic functions than
symmetrical dendrimers. As an example, Acton et al. syn-
thesized polyethylene glycol (PEG)-based Janus dendrimers for
multi-drug loading by conjugating two model drugs, benzyl
alcohol (BA) and 3-phenylpropionic acid (PPA), to the den-
drons.22 Meanwhile, BA and PPA could be released at different
speeds owing to the different linkages between the dendrons
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and drugs. When simultaneously conjugating with hydro-
phobic and hydrophilic dendrons, Janus dendrimers can func-
tion as surfactants to stabilize phase interfaces. In an appro-
priate ratio between dendrons of opposite hydrophilicity,
amphiphilic Janus dendrimers could form self-supra-assem-
blies named dendrimersomes or supramolecular hydro-
gels.23,24 Different from Janus dendrimers, the poly-lactic-co-
glycolic acid (PLGA) JNPs are synthesized via the microfluidic
nanoprecipitation method (Fig. 2a).25 The two kinds of PLGA
polymers with different PLA/PGA ratios were injected separ-
ately through two side-by-side channels into the dispersing
stream containing 1% poly(vinyl alcohol) (PVA) in water. The
two PLGA polymers precipitated separately and formed a Janus
droplet at the exit of the inlet streams.
2.2 Inorganic Janus nanoparticles
JNPs consisting of inorganic materials (e.g. gold, silica, and
Fe3O4) have been widely investigated for cancer theranostics
owing to their intrinsic advantages such as magnetism,
surface plasmonic resonance (SPR), photothermal conversion
ability, easy functionalization, and so on.26 The fabrication of
these inorganic JNPs could be roughly divided into two cat-
egories: masking and surface nucleation/seeded growth. The
representative synthesis methods for inorganic JNPs mainly
include flat interfacial synthesis, Pickering emulsion inter-
facial synthesis, controlled nucleation and seeded growth, and
hard-templating methods.
Masking is an essential technique to produce JNPs, and it
mainly involves the protection of one portion of a nanoparticle
until the unprotected portion is modified by a chemical reac-
tion, a polymerization, or a functionalization step. This
straightforward method mainly includes three steps: (1) the
symmetric nanoparticle is temporarily immobilized at a
surface in order to expose one portion for further modifi-
cation; (2) the surface of the exposed region is modified; (3)
the nanoparticle is released from the surface. An optional
fourth step, modifying the previously masked region, can be
performed at the end. The immobilization of inorganic nano-
particles on flat substrates has been reported. For instance,
Lee et al. pressed a monolayer of azide-functionalized silica
nanoparticles on a polydimethylsiloxane (PDMS) elastomer
covered with biotinylated bovine serum albumin (BSA), so that
the exposed surface of the nanoparticles was conjugated with
alkyne-tagged anti-CD28 (Fig. 2b).27 After detaching from the
PDMS elastomer, biotin-BSA remained on the masked surface,
leaving it available for the conjugation of anti-CD3 antibodies.
The bull-eye shape JNPs were then obtained for further T cell
stimulation.
Besides the straightforward surface modification of in-
organic nanoparticles immobilized on flat surfaces, employing
nanoparticle aggregates at flexible liquid–liquid or gas–liquid
interfaces is a common approach for inorganic JNP fabrication
in cancer theranostics. For example, Cao et al. synthesized
dumbbell-like CMR–MS/Au–6MP JNPs by Pickering emulsion
interfacial synthesis (DOX: doxorubicin, CMR: 7-hydroxy-cou-
marin-3-carboxylate, MS: mesoporous silica, Au: gold, and
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Table 1 Summary of composition–morphology–fabrication–application of Janus nanoparticles in cancer theranostics

Type Composition Morphology Fabrication method Applications Ref.

Polymeric Four PEG-based dendrons Bow-tie dendrimer Covalent conjugation Chemotherapy 22

PLGA nanoparticles-DOX-PTX Sphere Microfluidic synthesis Chemotherapy 25
Inorganic Silica-antibodies Sphere Surface modification Immunotherapy 27
FA–Au–mesoporous silica–DOX Spindle Surface nucleation and seeded growth CT, chemotherapy, 28
radiotherapy
GNRs–mSiO2 Rod Surface nucleation and seeded growth FLI, PTT 29
GNRs@mSiO2–DOX Lollipop Surface nucleation and seeded growth Chemotherapy, PTT 30
Au–silica Sphere Surface nucleation and seeded growth PAI 31
Au@MnO@SiO2 Snowman Surface nucleation and seeded growth Two-photon FLI 32
Dox–CMR–MS/Au–6MP Dumbbell Pickering emulsion interfacial synthesis FLI, SERS, 33
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chemotherapy
Au–IONP/Dye@MS Snowman Pickering emulsion interfacial synthesis FLI, CT, MRI 34
Fe3O4–MSNs–P@GCV@pTK Rod Surface nucleation and seeded growth MRI, magnetic 35
hyperthermia, gene
therapy
Fe3O4–SiO2 Bullet Surface nucleation and seeded growth Magnet field-enhanced 36
chemotherapy
Fe3O4–TiO2 Dumbbell Surface nucleation and seeded growth MRI, PDT 37
Mn3O4–TiO2/ZnO/Fe3O4 Dumbbell Surface nucleation and seeded growth MRI, PDT 38
Fe3O4–Au Octahedron-sphere Surface nucleation and seeded growth FLI, MRI, 39
chemotherapy
Fe3O4–Au Star Surface nucleation and seeded growth CT, MRI, PAI, SERS 40
Branched PEG–Au–Fe3O4 Dumbbell Surface nucleation and seeded growth CT, MRI, PAI, PTT 41
FA–Au/Fe3O4@C Dumbbell Surface nucleation and seeded growth MRI, CT, PTT, 42
chemotherapy
Au–Fe2C–Z (ZHER2:342) Dumbbell Surface nucleation and seeded growth CT, MRI, PAI, PTT 43
PEG–CuS–Au–MnO2 Dumbbell Surface nucleation and seeded growth CT, MRI, 44
chemotherapy, NIR-II
PTT
MnFe2O4–NaYF4 Dumbbell Surface nucleation and seeded growth Upconversion 45
luminescence, PTT
Ag–MSN–DOX Bullet Surface nucleation and seeded growth Chemotherapy 46
FA–Ag–MSN–ICG–DOX Bullet Surface nucleation and seeded growth PTT, chemotherapy 47
Ag–Fe@Fe3O4 Snowman Surface nucleation and seeded growth Two-photon FI, MRI 48
Esterase/GOx–Au–mesoporous silica Dumbbell Pickering emulsion interfacial synthesis Chemotherapy 49
(MS)–DOX/CD
Acetylcholinesterase–Au– Dumbbell Pickering emulsion interfacial synthesis Chemotherapy 50
mesoporous silica (MS)–DOX/CD
Polymeric– Fe3O4–PS16-b-PAA10 Sphere Phase separation FLI, magnetolytic 51
inorganic therapy
Fe3O4–PS16-PAA10 Sphere Phase separation Chemotherapy 52
FA–polystyrene/Fe3O4@SiO2–DOX Sphere Surface nucleation and seeded growth Chemotherapy 53
FA–Au–PAA/mCaP Dumbbell Surface nucleation and seeded growth CT, chemotherapy 54
PCL–AuNC/Fe(OH)3–PAA Sphere Surface nucleation and seeded growth CT, MRI, PTT, 55
chemotherapy
PEG–ICG–PDA/mCaP Hollow sphere Surface nucleation and seeded growth PAI, PTT, 56
chemotherapy
Au–bis-pyrene@SiO2–PEG Snowman Surface nucleation and seeded growth Two-photon FLI, PTT 57
PEG–Au–PAA/mSiO2–lactobionic Octopus Surface nucleation and seeded growth PTT, chemotherapy 58
acid
DOX/HCPT loaded CD–PdNS/ZIF-8 UFO Surface nucleation and seeded growth NIR-II PTT, 59
chemotherapy
UCNP@SiO2@mSiO2&PMO Cube + sphere Surface nucleation and seeded growth Chemotherapy 60
Hollow Janus MON Stone, cashew/ Hard-templating USI, chemotherapy 61
shrunken vesicle,
cube
Au–polydivinylbenzene–curcumin Sphere Surface nucleation and seeded growth Chemotherapy, PTT 62
GNP–BCP–MNP Hemisphere Coassembly PAI, MRI 63
Fe3O4@PFODBT–COOH Sphere Nanoprecipitation FLI, MPI 64

6MP: 6-mercaptopurine).33 The CMR–MS nanoparticles were
fixed at the interface of a Pickering emulsion, wherein paraffin
was the oil phase. The exposed region of the nanoparticle
surface was modified with (3-mercaptopropyl) trimethoxysi-
lane before the attachment by Au–6MP nanoparticles.
Another important method for inorganic JNP fabrication is
surface nucleation and seeded growth. This method involves
controlled growth or attachment of one inorganic nanoparticle
on the surface of another nanoparticle via heterogeneous
nucleation. The precise control of epitaxial growth can be

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Fig. 2 Fabrication flow diagrams of Janus nanoparticles (JNPs). (a) Schematic illustration of the synthesis process of PLGA JNPs via the nanoprecipi-
tation method. Reprinted with permission from ref. 25. Copyright 2012 American Chemical Society. (b) Schematic illustration of the preparation of
silica JNPs conjugated with anti-CD3/anti-CD28 via the masking method. Reprinted with permission from ref. 27. Copyright 2017 Royal Society of
Chemistry. (c) Schematic illustration of the fabrication of the DOX-loaded Ag–MSN JNPs via the surface nucleation and seeded growth method.
Reprinted with permission from ref. 46. Copyright 2016 American Chemical Society. (d) Schematic illustration of the construction of Janus vesicles
consisting of GNP, free BCP and MNPs via the coassembly method. Reprinted with permission from ref. 63. Copyright 2016 Wiley-VCH.

achieved by using appropriate surfactants. For instance, Shao
et al. synthesized bullet-like Janus silver–mesoporous silica
nanoparticles (Ag–MSNs) in which an MS stick was formed on
the silver nanosphere by using the surfactant hexadecyltri-
methylammonium bromide solution (CTAB) as the template
and tetraethylorthosilicate (TEOS) as the silica source
(Fig. 2c).46 After introducing a carboxylate group on the pore
surface of the MS stick and loading DOX into the pores, the
pH-responsive release of DOX could be achieved. In addition
to surfactants, ligands are also common regulators for the
nucleation and growth on a specific region.
As the most widely used strategies for the fabrication of in-
organic Janus composites in cancer theranostics, both
masking and surface nucleation/seeded growth methods have
several technical problems to be resolved. For instance, the
surface modification techniques require the nanoparticles to
be aggregated in monolayers, which limited the production
quantities of JNPs. The elaborate control of optimal reaction
conditions, the thickness regulation of the deposited layers
and the process complexity of fabrication are obstacles for
large-scale production using surface nucleation/seeded growth
methods.
2.3 Polymeric–inorganic Janus nanoparticles
In analogy with inorganic JNPs, surface nucleation/seeded
growth is the primary strategy of polymeric–inorganic JNP fab-
rication. For instance, Wang et al. constructed dumbbell-like
Au–PAA/mesoporous calcium phosphate (mCaP) JNPs for pH-
responsive drug delivery.54 In the typical synthesis, PAA–Ca
was firstly modified on the side of Au nanoparticles to obtain
Au–PAA/Ca nanoparticles, and was then used as the template
for the further growth of mCaP to obtain the final Au–PAA/
mCaP JNPs. The exposed Au domain allowed the conjugation
of folic acid–PEG–thiol (FA–PEG–SH) as a cancer cell targeting
unit, while DOX was loaded into the mesoporous structure of
mCaP. The in vitro release experiments showed that while only
16% of DOX was released in 9 h in the neutral buffer, the drug
release percentage reached 90% in the acid buffer ( pH 5.3).
Besides the above method, phase separation, coassembly
and nanoprecipitation methods have also been used for the
fabrication of polymeric/inorganic JNPs. Phase separation
requires two incompatible substances to be dissolved as a
mixture solution by two or more solvents, and then the sub-
stances are separated into independent domains, which
remain as one part of an intact nanoparticle. Hu et al. used
solvent evaporation to separate the mixture of Fe3O4 nano-
particles and poly(styrene-block-allyl alcohol) (PS16-PAA10). As
the evaporation of chloroform occurs, Fe3O4 nanoparticles
with low solubility precipitate out first and form clusters, fol-
lowed by solidification of the PS16-PAA10 matrix.52 Phase separ-
ation enables large-scale production, but a delicately designed
separation trigger is required to obtain JNPs with complex
shapes. Due to the incorporation of amphiphilic polymers, the
coassembly of several compositions into one single JNP is also

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proven to be feasible. To be specific, Liu et al. fabricated

magneto-plasmonic Janus-like vesicles by coassembling a
mixture of hydrophobic Fe3O4 nanoparticles, amphiphilic
polystyrene-b-poly(acrylic acid) (PS-b-PAA), and GNPs grafted
with polystyrene-b-poly(ethylene oxide) (GNPs-PS-b-PEO)
(Fig. 2d).63 By adjusting the size and mass fraction of Fe3O4/
GNPs, hemispherical Janus vesicles could be synthesized for
PAI and MRI. The nanoprecipitation method is applied for
polymeric–inorganic JNP fabrication. Nanoprecipitation, also
known as the solvent displacement method, is based on the
interfacial deposition because of the displacement of a solvent
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with a non-solvent, which results in the formation of nano-

particles in the interface between the two immiscible sol-
vents.65 For example, Song et al. employed the nanoprecipita-
tion method to prepare spherical Fe3O4–PFODBT ( poly[2,7-
(9,9-dioctylfluorene)-alt-4,7-bis(thiophen-2-yl)benzo-2,1,3-thia-
diazole]) JNPs for fluorescence imaging (FLI) and magnetic
particle imaging (MPI).64 In this case, PFODBT in tetrahydro-
furan (THF), Fe3O4 nanoparticles in oleic acid and PSMA were
added into H2O under sonication. The Fe3O4–PFODBT JNPs
were instantaneously formed and dissolved in water after the
evaporation of THF. Next, Fe3O4–PFODBT–COOH labeled HeLa
cells were implanted into living mice for whole-body FLI and
MPI. After depilation, about 30 000 labeled cells subcu-
taneously injected onto the abdomen area could be distin-
guished only by FLI at the front view, while the same mice
were visualized by MPI with a signal/noise ratio of 1.3. This
method is facile, time-saving and allows large-scale pro-
duction, but its application in JNPs is still scarce.
3. Janus nanoparticles in cancer
diagnosis
JNPs composed of multiple imaging materials (e.g. gold,
Fe3O4, MnO2, etc.) have been widely used as contrast agents for
cancer diagnosis, such as FLI, PAI, CT, MRI, surface enhanced
Raman scattering (SERS) and magnetic particle imaging
(MPI).29,31,32,40,48,63,64 Multimodal imaging provides more com-
prehensive and accurate information on tumor regions and
drug distribution than any single imaging modalities by inte-
grating information from various imaging modalities.66 For
instance, Reguera et al. prepared Janus magnetic nanostars
(JMNSs) via the nucleation and seeded growth method,
wherein Fe3O4–Au nanodumbbells were first synthesized as
seeds for the directional growth of asymmetric gold nanostars
(Fig. 3a).40 The obtained JMNs could perform MRI/CT/PAI/
SERS quadri-modal imaging. The 16 nm Fe3O4 nanoparticles
in JMNSs induced higher relaxivity values than commercially
available Fe3O4 nanoparticles such as Ferumoxytol (r2 of 89
s−1 mM−1) and Ferumoxtran-10 (r2 of 65 s−1 mM−1) (Fig. 3b).
By adjusting to a high Au to Fe ratio, both strong CT and MRI
signals are achieved (Fig. 3c and d). The gold nanostar tips
induced a red shift of the absorption window of the original
Fe3O4–Au nanodumbbell; thus the JMNSs exhibited strong PA
signals with a maximum around 700 nm (Fig. 3e and f ). In
Fig. 3 Janus magnetic nanostars (JMNSs) for MRI/CT/PAI/SERS quadri-
modal imaging. (a) TEM image of JMNSs comprising a 16 nm Fe3O4
domain and 28 nm gold nanostars. (b) Quantification of relaxivities for
JMNSs with a Fe3O4 core of 16 nm (in red) or 20 nm (in blue), plotted as
a function of the gold-to-iron ratio. (c) MRI images of phantoms of
JMNSs.16.28 aqueous solutions at different concentrations. (d) CT
images of JMNS.16.28 solutions at different concentrations and X-ray
energies. (e) UV-Vis-NIR spectra of JMNSs with a 16 nm Fe3O4 domain
and different gold nanostar sizes. (f ) B-ultrasound and PA images of
JMNS aqueous solutions at different gold concentrations. (g) SERS
image of A549 cells after incubation with 4-MBA-JMNS.16.28. (h) SERS
spectrum of the pixel inside the cell indicated by the white cross in (g).
Reprinted with permission from ref. 40. Copyright 2017 Royal Society of
Chemistry.
addition, the JMNSs were functionalized with the Raman
reporter 4-mercaptobenzoic acid (4-MBA). The as-prepared
4-MBA-JMNSs achieved efficient SERS imaging inside the A549
cells at 1078 cm−1, corresponding to the ring stretch vibration
mode of 4-MBA (Fig. 3g and h). In the conventional core–shell
nanostructure, the original signals of the imaging cores were
diminished or even hidden by the shell part, which could be
prevented by the spatial separation of materials in JNPs.59 As
an example, Park et al. modified the surface of gold nano-
particles (GNPs) with two ligands, polyacrylic acid (PAA) and
4-MBA, via different bonds, but only 4-MBA-coated GNPs facili-
tated silicification.31 By adjusting an appropriate ratio between
PAA and 4-MBA and adding the silica source TEOS, GNPs with
anisotropic silica shells as gold–silica JNPs (GSJNPs) were suc-
cessfully prepared. Notably, the anisotropic silica shells in

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GSJNPs caused a red-shift in the SPR of GNPs for highly
efficient PAI. The heterogeneous structure of JNPs also enables
complicated imaging functions such as FRET (Förster reso-
nance energy transfer). In this case, Schick et al. synthesized
Au@MnO@SiO2 JNPs for two-photon FLI, where only the MnO
domain was covered with a silica layer while the Au domain
was exposed.32 The gold rod exhibited strong green FLI signals
within HeLa cells (Ex: 970 nm). By incorporating a green FL
dye into the silica shell, the FRET between the dye and the
gold rod could produce both green and red signals (Ex:
970 nm).
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4. Janus nanoparticles in cancer
treatment
JNPs have been widely investigated for cancer therapeutic
applications including chemotherapy, magnetolytic therapy,
phototherapy and a combination of multiple treatments.
Compared with conventional core–shell or alloy nanoparticles,
JNPs have some distinct advantages including heterogeneous
structures, little interference among the elements, and distinct
surfaces for different functionalizations and modifi-
cations.40,48 Therefore, JNPs are deemed as potential “second-
generation nanoparticles” for the development of multifunc-
tional nanomedicines in cancer treatment.
4.1 Chemotherapy
Nanoparticles are ideal carriers of chemotherapeutic drugs
because of their passive/active tumor targeting, water solubility
and smart drug release/penetration profiles.67 Conventional
core–shell nanoparticles have homogeneous inner cavities that
could only load drug molecules with similar properties (e.g.
polarity and charge). In contrast, JNPs, due to their intrinsic
heterogeneity and separation, are suitable for delivering drugs
with different physicochemical properties and independent
release kinetics.61 In 2012, Xie et al. synthesized PLGA JNPs
with diameters of ∼300 nm via a nanoprecipitation method for
the co-delivery of hydrophobic paclitaxel (PTX) and hydrophilic
DOX. The encapsulation efficiencies of PTX and DOX were
80% and 19%, respectively.25 Similarly, Li et al. prepared
bicompartmental UCNP@SiO2@mSiO2&PMO JNPs (UCNP:
upconversion nanoparticle = NaGdF4:Yb,Tm@NaGdF4, mSiO2:
mesoporous silica shell, PMO: periodic mesoporous organo-
silica) for co-delivery of PTX and DOX as model drugs (Fig. 4a
and b).60 The hydrophilic DOX was encapsulated in the meso-
pore channels of the UCNP@SiO2@mSiO2 domain and
blocked by a photosensitive azobenzene (Azo) switch, and then
the hydrophobic PTX was loaded in the PMO part. Due to the
existence of heat sensitive phase change materials (1-tetradeca-
nol) surrounding the UCNP@SiO2@mSiO2&PMO JNPs, PTX
could be released from the PMO part upon heating (∼39 °C).
Under NIR laser irradiation, the UCNP cores could emit
photons in the Ultraviolet–visible (UV-Vis) region. The short
wavelength light induced the continuous rotation-inversion
movement of azo molecules, which could transform from the
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Fig. 4 UCNP@SiO2@mSiO2&PMO JNPs for heat/NIR light dually trig-
gered release of DOX and PTX. (a) Schematic of the dual-stimuli respon-
sive drug release process (guest-1: PTX; guest-2: DOX). (b) TEM images
of the obtained UCNP@SiO2@mSiO2&PMO JNPs. (c) Cell viabilities of
PTX and DOX coloaded UCNP@SiO2@mSiO2&PMO JNPs upon exposure
to heat (H) and NIR light (L) (S means sample). (d) Confocal laser scan-
ning microscopy (CLSM) observations of the HeLa cells after incubation
with the Rh123 (green) and DAPI (blue) coloaded JNPs with or without
heat (H) and NIR light (L) stimuli. Reprinted with permission from ref. 60.
Copyright 2014 American Chemical Society.
trans-isomer to the cis-isomer under UV irradiation (∼360 nm),
and vice versa under irradiation of visible light (∼450 nm).
This wagging motion of the azo molecules provided the
impetus for the release of DOX inside the mesopore channels
of UCNP@SiO2@mSiO2. This heat and NIR light dually trig-
gered drug release was verified in HeLa cells using Rh123 (rho-
damine 123) and DAPI (4′,6-diamidino-2-phenylindole) to
replace PTX and DOX, respectively (Fig. 4c). The combination
of PTX and DOX caused significant death of HeLa cells (>50%)
(Fig. 4d). These dual-compartment Janus mesoporous silica
nanocomposites allow co-loading of hydrophobic/hydrophilic
drugs and independent release of each drug from separated
domains. This work provides an intriguing choice in the
design of nanocarriers for multidrug delivery.
To achieve a stimulus-responsive drug release, a control
unit is necessary to transform the input stimuli into new
chemical signals via an effector to trigger the drug release
from the gating system. The homogeneous surfaces of nano-
particles can hardly integrate the effectors and gate moieties
into one entity, which somehow limits the efficiency of signal
transmission and response. The multiple surfaces of JNPs with
distinct composition and surface chemistry are promising to
realize highly efficient intelligent drug release.53 Díez et al.
designed a series of Janus Au–MS nanoparticles in which the
gate moieties in the MS surface were integrated with effectors
on the gold side.49,50 In one case, a pH-responsive
β-cyclodextrin (β-CD)-based supramolecular nanovalve was
used as the gating system, while glucose oxidase and esterase
were the two effectors.50 In the presence of enzyme substrates
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(i.e. glucose or ethyl butyrate), gluconic acid ( pKa = 3.6) or
butyric acid ( pKa = 4.82) was generated under the catalysis of
enzymes. The acids resulted in a decrease of pH and the sub-
sequent opening of the β-CD-based nanovalve. As the model
drug, the burst release of Ru(bpy)32+ was observed in the pres-
ence of either glucose or ethyl butyrate, or a combination of
both substrates.
4.2 Phototherapy
Phototherapy such as photothermal therapy (PTT) and photo-
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dynamic therapy (PDT) employs photothermal conversion
agents (PTCAs)/photosensitizers (PSs) to generate heat/singlet
oxygen (SO) to kill tumor cells upon laser irradiation, respect-
ively. Compared to chemotherapeutic drugs, the photothera-
peutic agents usually show little toxicity in the dark, endowing
phototherapy with high selectivity and low systemic toxicity.68
JNPs composed of inorganic materials (e.g. gold, TiO2, CuS,
and Pb) have been investigated for phototherapy. Some of
these studies have indicated a better therapeutic performance
of JNPs than that of conventional core–shell nanoparticles. For
instance, Yang et al. attached gold stars (Au) to bis-pyrene (BP)
nanoaggregates and then coated the Au–BP with a PEGlated
silica shell (SP) to obtain Au–BP@SP JNPs for PTT.57 Notably,
Au–BP@SP had higher photothermal conversion efficiency
(PCE) (29.4%) than Au@SP (25.5%). This phenomenon was
explained by the higher kinetic energy of Janus Au–BP@SP
than that of Au@SP. The kinetic energy is generated by the
motion of GNPs, which is then transformed into heat through
collision with a surrounding medium. In this case, the active
motion of Au–BP@SP JNPs driven by a temperature gradient
between the hot Au and cold BP domain upon NIR exposure
exhibited higher kinetic energy than the Brownian motion of
Au@SP, leading to enhanced heat generation. Zeng et al. fabri-
cated dumbbell-like Fe3O4–TiO2 JNPs with a size distribution
of 30–40 nm for PDT.37 Under UV light exposure, TiO2 pro-
duced free radicals and SO to kill the majority of MCF-7 cells.
The limited tissue penetration depth (<1 cm) of light
employed for phototherapy severely limits its clinical appli-
cations.69 Recently, phototherapy in the second near-infrared
NIR (NIR-II) window has aroused tremendous research interest
because of the deep penetration depth (>5 cm) of NIR-II light.
In 2018, Li et al. synthesized dumbbell-like PEG–CuS–Au–
MnO2 ternary JNPs with an average diameter of ∼135 nm for
NIR-II PTT (Fig. 5a and b).44 The PCE (η) of PEG–CuS–Au–
MnO2 JNPs was determined as 28.0%. Under NIR-II laser
irradiation (1064 nm, 1 W cm−2, 5 min), PEG–CuS–Au–MnO2
showed surprisingly less attenuation of photothermal conver-
sion capability with the increase of pork muscle thickness
(0–10 mm) compared to the NIR-I laser (Fig. 5c). The PEG–
CuS–Au–MnO2 plus NIR-II laser could efficiently kill >50% of
HepG-2 cells and inhibit the subcutaneous tumor growth
(Fig. 5d). Compared to plain GNPs, the Janus nanocomposite
of Au, CuS and MnO2 domains, Au cores exhibited an obvious
red shift due to the high refractive index of MnO2 and CuS sur-
rounding the Au core.70
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Fig. 5 PEG–CuS–Au–MnO2 ternary JNPs for NIR-II PTT. (a) Schematic
of the ternary JNP. (b) TEM images of the obtained PEG–CuS–Au–
MnO2 ternary JNPs. (c) Cell viabilities of PTX and DOX coloaded
UCNP@SiO2@mSiO2&PMO JNPs upon exposure to heat (H) and NIR
light (L) (S means sample). (d) The cytotoxicity of PEG–CuS–Au–MnO2
JNPs and CST-loaded PEG–CuS–Au–MnO2 JNPs against HepG-2 cells
with or without NIR irradiation. Reprinted with permission from ref. 44.
Copyright 2018 American Chemical Society.
4.3 Magnetolytic therapy
In 2010, Hu et al. proposed the concept of magnetolytic
therapy, which uses magnetic nanoparticles to mechanically
destroy cancer cell membranes under a magnetic field.51,71 The
application of an external magnetic field in cancer treatment
has some distinct advantages such as deep-penetration, low
toxicity, facile manipulation, etc. For magnetolytic therapy,
magnetic Fe3O4–PS16-b-PAA10 JNPs were fabricated and they
acted as a nanomotor to attach to the cell membrane under
the exposure of a magnetic field.51 Once placed in a spinning
magnetic field (50 rpm) for 15 min, the Trypan blue staining
images showed that most of the tumor cells were killed.
Afterward, the magnetic field was used to enhance the thera-
peutic efficacy of chemotherapy. For example, the same group
used a high frequency magnetic field (HFMF, 50 kHz) to
realize an immediate burst release of the drug from Fe3O4–
PS16–PAA10 JNPs.52 Shao et al. showed that the cellular uptake
of Fe3O4–SiO2–DOX JNPs was significantly enhanced upon
exposure to an NdFeB permanent magnetic field (0.2 T) for
about 0.5 h (Fig. 6a and b).36 After intravenous injection to
orthotopic H-22-tumor-bearing mice, the JNP plus external
magnetic field group exhibited significant tumor growth inhi-
bition and extended the survival rate of the Janus group from
50% to 62.5% (Fig. 6c and d).
4.4 Combination therapy
JNPs consisting of materials with inherent photothermal con-
version properties (e.g. Au and Pb) have been widely used as
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Fig. 6 Fe3O4–SiO2–DOX JNPs for magnetolytic therapy. (a) Schematic
illustration of the fabrication strategy of Fe3O4–SiO2–DOX JNPs. (b)
CLSM images of HepG-2 cells after 3, 12, and 24 h of exposure (0.5 h
with or without exposure to a magnetic field followed by 2.5, 11.5 and
23.5 h of incubation) to Fe3O4–SiO2–DOX. (c) Tumor weight and (d) sur-
vival rate of the orthotopic H-22-tumor-bearing mice with different
treatments. Reprinted with permission from ref. 36. Copyright 2016
Elsevier Ltd.
drug carriers for chemo-photothermal therapy.30,47,59,62
Compared with traditional chemo-photothermal therapy,
nanoparticle-induced chemo-photothermal therapy allows site-
specific hyperthermia and light-controllable drug release.72
For instance, Zhang et al. synthesized octopus-type PEG–Au–
Fig. 7 Octopus-type PEG–Au–PAA/mSiO2–lactobionic acid (PEG–OJNP–LA) JNPs for chemo-photothermal therapy. (a) Schematic illustration of
the synthesis of PEG–OJNP–LA JNPs via the surface nucleation and seeded growth method. (b) TEM imaging of PEG–OJNP–LA JNPs; the inset
shows the magnified SEM image of the mSiO2 surface. (c) MTT cell viability assay of HepG-2 cancer cells with different treatments. (d) Tumor
volume of the H22-tumor-bearing Kunming mice with different treatments. Reprinted with permission from ref. 58. Copyright 2016 Wiley-VCH.
This journal is © The Royal Society of Chemistry 2019
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PAA/mSiO2–lactobionic acid (PEG–OJNP–LA) JNPs to integrate
NIR laser/pH dual-stimuli responsive drug release and PTT
(Fig. 7a and b).58 Without loading of drugs or light exposure,
PEG–OJNP–LA JNPs showed little cytotoxicity to HepG-2 cancer
cells. The PCE (η) of PEG–OJNP–LA JNPs was 49.5% for
808 nm, and the loading efficiency of DOX into PEG–OJNP–LA
was about 90%. The deprotonation of polymer PAA in an
acidic environment and remote heating caused by Au together
could significantly enhance the release rate of DOX from 9%
to 95%, and induce nearly 94% of cell death in vitro (Fig. 7c).
After intravenous injection to H-22 tumor-bearing mice, the
specific targeting of LA to hepatoma cells ensured a high
tumor accumulation of JNPs. After the chemo-photothermal
treatment, the tumor suppression rate of JNPs was around
98%, while those of single PTT and chemotherapy treatment
were 91% and 81%, indicating the effective synergistic efficacy
in vivo (Fig. 7d). Recently, to combine the NIR-II PTT of a
much deeper tissue penetration depth with chemotherapy, the
same group prepared unidentified flying object (UFO)-like CD–
palladium nanosheet/zeolitic imidazolate framework-8 (CD–
PdNS/ZIF-8) JNPs.59 To prepare CD–PdNS/ZIF-8 JNPs, the PAA
coating on one of the flat surfaces of 2D PdNS was followed by
further formation of ZIF-8 on a PAA template, and CDs were
subsequently conjugated onto the exposed flat surface of
PdNS. The CD–PdNS and PAA-ZIF-8 domains are designed to
load hydrophobic 10-hydroxycamptothecin (HCPT) and hydro-
philic DOX for synergistic chemotherapy (combination index:
0.65). Meanwhile, the PbNS exhibited excellent photothermal
performance in the NIR-II window, and the PCE (η) was calcu-
lated to be 20% for 1064 nm. Due to the deprotonation of the
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pH-sensitive polymer PAA and the dissolution of ZIF-8 in an

acidic environment, the enhanced release profiles of both
HCPT and DOX at pH = 5.3 were observed. Upon NIR-II laser
exposure, the temperature increase induced by CD–PdNS/
ZIF-8 JNPs broke the bond between thiol-CDs and PdNS, and
also changed the physicochemical properties of drugs includ-
ing their solubility, fluidity, and viscosity.73 The dual-stimuli
of low pH and NIR-II light could elevate the release rate of
HCPT and DOX to about 74% and 83%, respectively. The
chemo-photothermal therapy mediated by CD–PdNS/
ZIF-8 JNPs induced highly efficient tumor suppression in the
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H22-tumor-bearing mice model.

5. Janus nanoparticles in cancer

theranostics
Cancer theranostics integrate diagnostic and therapeutic func-
tions in a single nanoparticle and thus realize imaging-guided
cancer treatment, allowing real-time monitoring of drug distri-
bution and therapeutic outcomes.74 JNPs have some inherent
advantages for cancer theranostics owing to their asymmetric
structure and distinct surfaces modified with various function-
alities and thus permit bioimaging, pH/NIR-responsive drug
release, phototherapy, radiation therapy, and magnetolytic
therapy.
5.1 FLI-guided therapy
FLI uses visible to NIR wavelengths of light to activate fluoro-
phores and emit strong fluorescence signals. Recently, JNPs
that simultaneously load fluorescent dyes and therapeutic
agents have been applied for FLI-guided cancer therapy.29,57 As
an example, rod-like gold nanorod–mesoporous SiO2 (GNR–
mSiO2) JNPs were fabricated and Rhodamine B (RdB) was
incorporated into the mSiO2 part. Compared to core–shell
nanoparticles, the SPR absorption peak of GNRs underwent a
much weaker red-shift in the GNR–mSiO2 JNPs.29 As potential
theranostic agents, the photothermal effect of GNRs could
effectively kill HepG-2 cells, and the red fluorescence of RdB
could effectively image the cytoplasm of HepG-2 cells in vitro.
Except for fluorescent dyes, JNPs consisting of fluorescent in-
organic materials (e.g. GNRs, upconversion nanomaterials) are
also potential candidates for FLI-guided cancer therapy.32,45
For instance, Wu et al. synthesized dumbbell-like MnFe2O4–
NaYF4 JNPs with diameters of ∼13 nm for upconversion
luminescence (UCL)-guided PTT (Fig. 8a and b).45 The PCE (η)
of the MnFe2O4–NaYF4 JNPs was estimated to be 16.7% for
808 nm (Fig. 8c). At the same time, the green UCL of JNPs was
observed inside Eca-109 cells after 0.5 h of incubation
(Fig. 8d). The PTT effect of MnFe2O4 caused over 99% of cell
death in vitro, as well as thorough eradication of the sub-
cutaneous tumors in vivo (Fig. 8e). According to these results,
the authors claimed that the interference between the photo-
thermal effect of MnFe2O4 and the UCL of NaYF4 in the core–
shell nanoparticles was minimized in a Janus nanostructure.
Fig. 8 Dumbbell-like MnFe2O4–NaYF4 JNPs for UCL-guided PTT. (a)
Schematic illustration of MnFe2O4–NaYF4–FA JNPs. (b) TEM imaging of
MnFe2O4–NaYF4–FA JNPs. (c) Temperature curves of distilled water and
MnFe2O4–NaYF4 JNP aqueous solutions at different concentrations. (d)
CLSM imaging of Eca-109 cells after incubation for 0.5 and 2 h with
MnFe2O4–NaYF4–FA JNPs. (e) The cytotoxicity of MnFe2O4–NaYF4 JNPs
against Eca-109 cells with and without 808 nm laser irradiation at
different concentrations. (f ) Tumor volume of the Eca-109 tumor-
bearing mice with different treatments. Reprinted with permission from
ref. 45. Copyright 2017 Wiley-VCH.
5.2 MRI-guided therapy
JNPs consisting of inorganic magnetic materials (e.g. MnO2,
Fe3O4, and Mn3O4) could realize MRI-guided cancer
therapy.35,37,38 For example, Iqbal et al. prepared dumbbell-like
Mn3O4–TiO2 JNPs for MRI-guided PDT (Fig. 9a).38 The dia-
meters of Mn3O4 and TiO2 particles were about 5 and 10 nm.
The r1 relaxivity of Mn3O4–TiO2 JNPs at 0.47 T was as high as
2.95 s−1 mM−1 owing to the ultrasmall nanoparticle size, and
the r2/r1 ratio was 4.3, indicating an excellent T1-weighted MRI
performance. After intravenous injection, profound contrast
enhancement in normal tissues such as heart, liver, and
kidney was observed (150 μL of Mn: 554 μg mL−1 and Ti:
443 μg mL−1). A similar contrast enhancement was also exhibi-
ted in tumors after intratumoral injection of Mn3O4–TiO2 JNPs
(Fig. 9b). Furthermore, the PDT effect mediated by TiO2 under
UV light irradiation for 30 min was shown to effectively kill
∼50% MCF-7 cells and effectively inhibited the tumor growth
in vivo (Fig. 9c and d).
5.3 PAI-guided therapy
PAI is used to image the acoustic waves at MHz frequencies
that were generated from substrates due to thermal expansion
after absorption of short laser pulses.75 The photon-to-heat

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Fig. 9 Dumbbell-like Mn3O4–TiO2 JNPs for MRI-guided PDT. (a)
Schematic illustration of Mn3O4 for MRI and TiO2 for PDT. (b) T1 MRI of
MCF-7 tumor-bearing mice before injection and after intratumoral
injection. (c) The cytotoxicity of Mn3O4–TiO2 JNPs against MCF-7 cells
with and without UV light at different concentrations for 24 h. (d) Tumor
volume of the MCF-7 tumor-bearing mice with different treatments.
Reprinted with permission from ref. 38. Copyright 2017 Tsinghua
University Press and Springer-Verlag Berlin Heidelberg.
conversion process is the premise of PAI. Therefore, most of
the photothermal materials (e.g. Au, CuS, and NIR dyes)
exhibit a strong PAI contrast. Recently, JNPs composed of
these materials have been investigated for PAI-guided cancer
therapy.31,56 Among these photothermal materials, organic
NIR dyes have the best biocompatibility and degradability, and
some dyes such as indocyanine green (ICG) have already been
approved for clinical use.76 Hence, Zhang et al. employed
spherical polydopamine/mesoporous calcium phosphate
hollow JNPs (PDA/mCaP H-JNPs) for the surface modification
of indocyanine green (ICG) and PEG on the PDA domains for a
superior PAI capability and to improve the stability, thus
obtaining PEG–ICG–PDA/mCaP H-JNPs.56 The PCE (η) of
H-JNPs was calculated to be 45.3% for 808 nm. DOX was then
loaded in the hollow cavity of mCaP with a high loading
efficiency of 92%. The pH/NIR dually responsive DOX release
was observed due to the dissolution of mCaP in the acidic
environment and the thermal effect of H-JNPs. After intrave-
nous injection, the PA signals of H-JNPs in the tumor tissues
were enhanced with time, indicating an excellent PAI contrast.
The synergistic therapeutic effect of pH/NIR responsive
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chemo-photothermal therapy was verified on HepG-2 cells and
HepG-2 tumor-bearing BALB/c nude mice.
5.4 Computed tomography (CT)-guided therapy
JNPs constituted of Au were utilized in CT-guided cancer
therapy since GNPs have strong X-ray attenuation pro-
perties.28,34,54 For instance, Wang et al. developed spindle-like
FA–Au–mesoporous silica nanoparticles (GSJNs) for CT-guided
chemotherapy and radiotherapy.28 The loading efficiency of
DOX in the carboxylate-functionalized JNPs was around 61.7%,
much higher than that of the core–shell-type mesoporous
silica nanoparticles (FA–GSCNs) (about 49.5%) owing to the
large surface area of the Janus structure. The pH-responsive
DOX release from FA–GSJNs at pH 5.5 (>40%) was attributed to
the protonation and dissociation of the amine groups in acidic
environments. As evaluated in SMMC-7721 cells, the IC50
values of chemotherapy and radiotherapy were 3.83 μg mL−1
and 1.53 Gy, respectively. The FA–GSJNs in the tumor region of
the mice exhibited distinguished CT signals 24 h after intrave-
nous injection based on transverse section images. The in vivo
therapeutic efficacy was also evaluated in SMMC-7721 xeno-
grafts. The nude mice were intravenously injected with drugs
or/and received irradiation at 5 Gy every 3 days for a total of six
administrations. Both the FA–GSJNs–DOX plus radiotherapy
group and free DOX group achieved prominent tumor inhi-
bition in 23 days, but the biotoxicity of FA–GSJNs was almost
negligible compared to the severe acute toxicity of free DOX.
5.5 Multimodal imaging-guided therapy
Multimodal imaging provides comprehensive imaging infor-
mation on the therapeutic process via integrating multiple
imaging modalities.77,78 JNPs constituted of Au and magnetic
materials (e.g. Fe(OH)3, Fe3O4, and MnO2) have been widely
used for CT/MRI/PAI multimodal imaging-guided cancer
therapy.39,41–44,55 For example, Ju et al. synthesized dumbbell-
like Au–Fe2C JNPs with a size of 12 nm for CT/MRI/PAI-guided
PTT (Fig. 10a and b). To promote the tumor accumulation of
Au–Fe2C JNPs to HER2+ tumors, the affibody ZHER2:342 was con-
jugated to the surface of JNPs. The real-time T2-weighted MR
images exhibited a signal enhancement for 12 and 24 h after
intravenous injection of Au–Fe2C–ZHER2:342, respectively
(Fig. 10c). Likewise, PAI signals of Au–Fe2C–ZHER2:342 JNPs
appeared at ∼4 h after injection and gradually increased until
24 h postinjection. The CT contrast in the tumor site was also
enhanced after intratumoral injection of Au–Fe2C according to
the 3D CT images of the mouse. After 24 h of incubation with
NIH3T3 and MDA-MB-231 cells, Au–Fe2C–ZHER2:342 showed a
remarkable PTT effect and caused more than 50% of cell death
(Fig. 10d). Under 8 min laser exposure (808 nm, 1 W cm−2),
Au–Fe2C–ZHER2:342 induced a profound temperature increase
(27.21 °C) in vivo, and eliminated the tumors of two-thirds of
the group (Fig. 10e). Furthermore, some chemotherapy drugs
per se exhibit imaging functions. JNPs that deliver these drugs
could also realize multimodal imaging-guided cancer therapy.
As an example, Cao et al. employed the dumbbell-like DOX–
CMR–MS/Au–6MP JNPs (DOX: doxorubicin, CMR: 7-hydroxy-
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Fig. 10 Au–Fe2C JNPs for CT/MRI/PAI guided photothermal therapy (PTT). (a) Schematic illustration of the fabrication of Au–Fe2C JNPs via the
surface nucleation and seeded growth method. (b) High resolution TEM (HRTEM) imaging of Au–Fe2C JNPs. (c) In vivo MRI/CT/PAI/thermal imaging
of mice after injection of Au–Fe2C JNPs. (d) The cytotoxicity of Au–Fe2C–PEG and Au–Fe2C–ZHER2:342 JNPs against MDA-MB-231 cells with or
without laser irradiation (808 nm, 1 W cm−2) at different concentrations. (e) Relative tumor volume of the MDA-MB-231 tumor-bearing mice with
different treatments. Reprinted with permission from ref. 43. Copyright 2017 American Chemical Society.

coumarin-3-carboxylate, MS: mesoporous silica, Au: gold, 6MP:
6-mercaptopurine) to monitor dual drug release in vitro.33
While 6MP was conjugated to the Au surface via the Au–S
bond, DOX was attached to MS via a pH-responsive linker
hydrazone. The FRET between CMR and DOX effectively
quenched the fluorescence of CMR, and the fluorescence
gradually recovered during the release of DOX. Therefore, the
release of DOX was monitored in real-time by the fluorescence
recovery. On the other hand, the SERS intensity of 6MP was
enhanced by gold containing JNPs; therefore the decline of
SERS signals was an efficient indicator for the release of 6MP.
The pH/redox-responsive dual drug chemotherapy had a much
lower IC50 (DOX: 1.15 µg mL−1, 6MP: 1.27 µg mL−1) for HeLa
cells compared with free drugs.
6. Challenges and opportunities
Although the anisotropic JNPs have shown their superiority
over core–shell nanoparticles and drawn ever-increasing inter-
est on cancer theranostics, several challenges are yet to be
tackled for broader applications and future clinical translation.
The first issue lies in the complicated synthesis process of
JNPs due to their heterogeneous structure in nanosize. For
most of the existing fabrication techniques, the precise control
of size and morphology as well as large-scale production are
still challenging. To be specific, the surface modification could
only produce a limited amount of JNPs because the spatially
selective modifications require immobilization of nano-
particles in monolayers. The surface nucleation and seeded
growth method allows the second material to grow on the
surface of one material, but the thickness and shape of epitax-
ial growth are difficult to be precisely controlled. The second
challenge arises from the biosafety of theranostic JNPs. Most
of the above studies on JNPs for cancer theranostics focused
on the fabrication strategies to obtain a high drug loading
amount or a specific composition to enhance their theranostic
efficiency. Nevertheless, the biosafety of nanoparticles is the
crucial premise for further clinical applications, which lacks
systematic investigation in these studies. Therefore, more
aspects of biosafety including the half lethal dose (HLD), phar-
macokinetics, biodistributions, elimination mechanisms, and
long-term bio-toxicity, should be comprehensively evaluated
on several mammal models to evaluate the feasibility of JNPs
for clinical cancer diagnosis and treatment. Thirdly, most of
these JNPs are composed of inorganic materials, while few
polymeric JNPs have been utilized for cancer theranostics.
Among the materials introduced above, PLGA, PEG, PAA, PS
and Fe3O4 have been approved by the Food and Drug
Administration (FDA). Nevertheless, most of these FDA
approved materials are polymers, but inorganic materials, the
most widely used materials (e.g. gold, silica and Ag), have not
been approved for clinical use mainly due to the biosafety con-
cerns. To accelerate the clinical translation, the development
of polymeric or biomacromolecular JNPs (e.g. protein and
DNA) for cancer imaging and therapy should not be neg-
lected.79 Fourthly, the exploration of JNPs in cancer theranos-
tics is still in its infancy, and most of the studies are proof-of-
concept studies. A number of studies only demonstrated
in vitro tests, while some others only focused on imaging or
therapy. In addition to the above obstacles, the tremendous
potential of JNPs for synergistic cancer therapy has not been
fully recognized. As a super-additive combination of several
treatments, synergistic therapy may overcome the inherent
limitations of the single treatment modality and achieve pro-
minent therapeutic effects.80 A myriad of synergistic treatment

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paradigms has been developed and achieved impressive thera-
peutic efficacy in preclinical tests and clinical trials, such as
immuno/radiotherapy, immuno/chemotherapy, immuno/
phototherapy, chemo/phototherapy, etc. The anisotropic struc-
ture and various surfaces may enable JNPs to integrate mul-
tiple therapeutic modalities in a sophisticated or mutually
independent manner. Therefore, the application of JNPs to
enhance the synergy effect among cancer treatment modalities
is an intriguing research field.
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7. Conclusions
Janus nanoparticles (JNPs) have anisotropic structures and
multiple surfaces with distinct physicochemical properties.
Currently, most theranostic JNPs are fabricated by surface
nucleation and seeded growth, Pickering emulsion interfacial
synthesis, surface modification, phase separation, and coas-
sembly methods. JNPs have some distinct superior properties
over core–shell nanoparticles for cancer theranostics, such as
an anisotropic architecture for independent drug release,
minimum signal interruptions among imaging materials, and
active drug delivery as nanomotors. Nevertheless, the mass
production, precise control of the size and morphology, biosaf-
ety and sophisticated synergy of multiple therapeutic modal-
ities could be the focus of future research for broader appli-
cations and clinical translation.
Conflicts of interest
There are no conflicts to declare.
Acknowledgements
This work is financially supported by China Postdoctoral
Science Foundation (2018M633139 and 2018M630987), the
National Natural Science Foundation of China
(31771036, 51703132, and 51573096), the Basic Research
Program of Shenzhen (JCYJ20170412111100742 and
JCYJ20160422091238319), the Guangdong Province Natural
Science Foundation of Major Basic Research and Cultivation
Project (2018B030308003), and the Fok Ying-Tong Education
Foundation for Young Teachers in the Higher Education
Institutions of China (161032).
Notes and references
1 P. G. de Gennes, Science, 1992, 256, 495–497.
2 A. Walther and A. H. Muller, Chem. Rev., 2013, 113, 5194–
5261.
3 L. Cheng, G. L. Hou, J. J. Miao, D. Y. Chen, M. Jiang and
L. Zhu, Macromolecules, 2008, 41, 8159–8166.
4 R. H. Deng, F. X. Liang, X. Z. Qu, Q. Wang, J. T. Zhu and
Z. Z. Yang, Macromolecules, 2015, 48, 750–755.
This journal is © The Royal Society of Chemistry 2019
View Article Online
Review
5 X. Wang, X. Feng, G. Ma, L. Yao and M. Ge, Adv. Mater.,
2016, 28, 3131–3137.
6 X. Ma, A. Jannasch, U. R. Albrecht, K. Hahn, A. Miguel-
Lopez, E. Schaffer and S. Sanchez, Nano Lett., 2015, 15,
7043–7050.
7 G. Rucinskaite, S. A. Thompson, S. Paterson and R. de la
Rica, Nanoscale, 2017, 9, 5404–5407.
8 T. Yang, L. Wei, L. Jing, J. Liang, X. Zhang, M. Tang,
M. J. Monteiro, Y. I. Chen, Y. Wang, S. Gu, D. Zhao,
H. Yang, J. Liu and G. Q. M. Lu, Angew. Chem., Int. Ed.,
2017, 56, 8459–8463.
9 M. N. Popescu, W. E. Uspal, C. Bechinger and P. Fischer,
Nano Lett., 2018, 18, 5345–5349.
10 L.-T.-C. Tran, S. Lesieur and V. Faivre, Expert Opin. Drug
Delivery, 2014, 11, 1061–1074.
11 M. Luo, Y. Feng, T. Wang and J. Guan, Adv. Funct. Mater.,
2018, 28, 1706100.
12 R. H. Deng, H. Li, J. T. Zhu, B. H. Li, F. X. Liang, F. Jia,
X. Z. Qu and Z. Z. Yang, Macromolecules, 2016, 49, 1362–
1368.
13 J. B. Fan, Y. Song, H. Liu, Z. Lu, F. Zhang, H. Liu, J. Meng,
L. Gu, S. Wang and L. Jiang, Sci. Adv., 2017, 3, e1603203.
14 Y. Wang, S. Gong, D. Gomez, Y. Ling, L. W. Yap,
G. P. Simon and W. Cheng, ACS Nano, 2018, 12, 8717–8722.
15 M. Lattuada and T. A. Hatton, Nano Today, 2011, 6, 286–
308.
16 J. Hu, S. Zhou, Y. Sun, X. Fang and L. Wu, Chem. Soc. Rev.,
2012, 41, 4356–4378.
17 F. Liang, C. Zhang and Z. Yang, Adv. Mater., 2014, 26,
6944–6949.
18 X. Ma, K. Hahn and S. Sanchez, J. Am. Chem. Soc., 2015,
137, 4976–4979.
19 X. Fan, J. Yang, X. J. Loh and Z. Li, Macromol. Rapid
Commun., 2018, 1800203.
20 Z. Y. Wu, L. Li, T. Liao, X. Q. Chen, W. Jiang, W. Luo,
J. P. Yang and Z. Q. Sun, Nano Today, 2018, 22, 62–82.
21 E. R. Gillies and J. M. Frechet, Drug Discovery Today, 2005,
10, 35–43.
22 A. L. Acton, C. Fante, B. Flatley, S. Burattini, I. W. Hamley,
Z. W. Wang, F. Greco and W. Hayes, Biomacromolecules,
2013, 14, 564–574.
23 V. Percec, D. A. Wilson, P. Leowanawat, C. J. Wilson,
A. D. Hughes, M. S. Kaucher, D. A. Hammer, D. H. Levine,
A. J. Kim, F. S. Bates, K. P. Davis, T. P. Lodge, M. L. Klein,
R. H. DeVane, E. Aqad, B. M. Rosen, A. O. Argintaru,
M. J. Sienkowska, K. Rissanen, S. Nummelin and
J. Ropponen, Science, 2010, 328, 1009–1014.
24 S. Nummelin, V. Liljestrom, E. Saarikoski, J. Ropponen,
A. Nykanen, V. Linko, J. Seppala, J. Hirvonen, O. Ikkala,
L. M. Bimbo and M. A. Kostiainen, Chem. – Eur. J., 2015,
21, 14433–14439.
25 H. Xie, Z. G. She, S. Wang, G. Sharma and J. W. Smith,
Langmuir, 2012, 28, 4459–4463.
26 I. Schick, S. Lorenz, D. Gehrig, S. Tenzer, W. Storck,
K. Fischer, D. Strand, F. Laquai and W. Tremel, Beilstein J.
Nanotechnol., 2014, 5, 2346–2362.
Biomater. Sci., 2019, 7, 1262–1275 | 1273

Review
27 K. Lee and Y. Yu, J. Mater. Chem. B, 2017, 5, 4410–4415.
28 Z. Wang, D. Shao, Z. Chang, M. Lu, Y. Wang, J. Yue,
D. Yang, M. Li, Q. Xu and W. F. Dong, ACS Nano, 2017, 11,
12732–12741.
29 Y. S. Wang, D. Shao, L. Zhang, X. L. Zhang, J. Li, J. Feng,
H. Xia, Q. S. Huo, W. F. Dong and H. B. Sun, Appl. Phys.
Lett., 2015, 106, 173705.
30 L. Fang, W. Q. Wang, Y. Liu, Z. G. Xie and L. Che, J. Mater.
Chem. B, 2017, 5, 8833–8838.
31 J. H. Park, D. S. Dumani, A. Arsiwala, S. Emelianov and
R. S. Kane, Nanoscale, 2018, 10, 15365–15370.
Published on 16 January 2019. Downloaded by IOCB Prague on 1/29/2020 11:48:59 PM.
32 I. Schick, S. Lorenz, D. Gehrig, A. M. Schilmann, H. Bauer,
M. Panthofer, K. Fischer, D. Strand, F. Laquai and
W. Tremel, J. Am. Chem. Soc., 2014, 136, 2473–2483.
33 H. Cao, Y. Yang, X. Chen and Z. Shao, Nanoscale, 2016, 8,
6754–6760.
34 A. Sanchez, K. O. Paredes, J. Ruiz-Cabello, P. Martinez-
Ruiz, J. M. Pingarron, R. Villalonga and M. Filice, ACS Appl.
Mater. Interfaces, 2018, 10, 31032–31043.
35 Z. Wang, Z. Chang, M. Lu, D. Shao, J. Yue, D. Yang,
X. Zheng, M. Li, K. He, M. Zhang, L. Chen and W. F. Dong,
Biomaterials, 2018, 154, 147–157.
36 D. Shao, J. Li, X. Zheng, Y. Pan, Z. Wang, M. Zhang,
Q. X. Chen, W. F. Dong and L. Chen, Biomaterials, 2016,
100, 118–133.
37 L. Zeng, W. Ren, L. Xiang, J. Zheng, B. Chen and A. Wu,
Nanoscale, 2013, 5, 2107–2113.
38 M. Z. Iqbal, W. Ren, M. Saeed, T. Chen, X. Ma, X. Yu,
J. Zhang, L. Zhang, A. Li and A. Wu, Nano Res., 2018, 10,
5735–5750.
39 M. V. Efremova, V. A. Naumenko, M. Spasova,
A. S. Garanina, M. A. Abakumov, A. D. Blokhina,
P. A. Melnikov, A. O. Prelovskaya, M. Heidelmann, Z. A. Li,
Z. Ma, I. V. Shchetinin, Y. I. Golovin, I. I. Kireev,
A. G. Savchenko, V. P. Chekhonin, N. L. Klyachko, M. Farle,
A. G. Majouga and U. Wiedwald, Sci. Rep., 2018, 8, 11295.
40 J. Reguera, D. Jiménez de Aberasturi, M. Henriksen-Lacey,
J. Langer, A. Espinosa, B. Szczupak, C. Wilhelm and
L. M. Liz-Marzán, Nanoscale, 2017, 9, 9467–9480.
41 X. Chen, G. Li, Q. Han, X. Li, L. Li, T. Wang and C. Wang,
Chem. – Eur. J., 2017, 23, 17204–17208.
42 Q. Zhang, L. Zhang, S. Li, X. Chen, M. Zhang, T. Wang,
L. Li and C. Wang, Chem. – Eur. J., 2017, 23, 17242–17248.
43 Y. Ju, H. Zhang, J. Yu, S. Tong, N. Tian, Z. Wang, X. Wang,
X. Su, X. Chu, J. Lin, Y. Ding, G. Li, F. Sheng and Y. Hou,
ACS Nano, 2017, 11, 9239–9248.
44 S. Li, L. Zhang, X. Chen, T. Wang, Y. Zhao, L. Li and
C. Wang, ACS Appl. Mater. Interfaces, 2018, 10, 24137–24148.
45 Q. Wu, Y. Lin, F. Wo, Y. Yuan, Q. Ouyang, J. Song, J. Qu and
K. T. Yong, Small, 2017, 13, 1701129.
46 D. Shao, X. Zhang, W. Liu, F. Zhang, X. Zheng, P. Qiao,
J. Li, W. F. Dong and L. Chen, ACS Appl. Mater. Interfaces,
2016, 8, 4303–4308.
47 Z. Wang, Z. Chang, M. Lu, D. Shao, J. Yue, D. Yang, M. Li
and W. F. Dong, ACS Appl. Mater. Interfaces, 2017, 9, 30306–
30317.
1274 | Biomater. Sci., 2019, 7, 1262–1275
View Article Online
Biomaterials Science
48 H. Zhang, Z. Yang, Y. Ju, X. Chu, Y. Ding, X. Huang,
K. Zhu, T. Tang, X. Su and Y. Hou, Adv. Sci., 2018, 5,
1800271.
49 P. Díez, A. Sánchez, M. Gamella, P. Martínez-Ruíz,
E. Aznar, C. de la Torre, J. R. Murguía, R. Martínez-Máñez,
R. Villalonga and J. M. Pingarrón, J. Am. Chem. Soc., 2014,
136, 9116–9123.
50 A. Llopis-Lorente, P. Díez, C. de la Torre, A. Sánchez,
F. Sancenón, E. Aznar, M. D. Marcos, P. Martínez-Ruíz,
R. Martínez-Máñez and R. Villalonga, Chem. – Eur. J., 2017,
23, 4276–4281.
51 S. H. Hu and X. Gao, J. Am. Chem. Soc., 2010, 132, 7234–
7237.
52 S. H. Hu, S. Y. Chen and X. Gao, ACS Nano, 2012, 6, 2558–
2565.
53 F. Wang, G. M. Pauletti, J. Wang, J. Zhang, R. C. Ewing,
Y. Wang and D. Shi, Adv. Mater., 2013, 25, 3485–3489.
54 H. Wang, S. Li, L. Zhang, X. Chen, T. Wang, M. Zhang, L. Li
and C. Wang, Nanoscale, 2017, 9, 14322–14326.
55 L. Zhang, M. Zhang, L. Zhou, Q. Han, X. Chen, S. Li, L. Li,
Z. Su and C. Wang, Biomaterials, 2018, 181, 113–125.
56 M. Zhang, L. Zhang, Y. Chen, L. Li, Z. Su and C. Wang,
Chem. Sci., 2017, 8, 8067–8077.
57 P. P. Yang, Y. G. Zhai, G. B. Qi, Y. X. Lin, Q. Luo, Y. Yang,
A. P. Xu, C. Yang, Y. S. Li, L. Wang and H. Wang, Small,
2016, 12, 5423–5430.
58 L. Zhang, Y. Chen, Z. Li, L. Li, P. Saint-Cricq, C. Li, J. Lin,
C. Wang, Z. Su and J. I. Zink, Angew. Chem., Int. Ed., 2016,
55, 2118–2121.
59 L. Zhang, S. Li, X. Chen, T. Wang, L. Li, Z. Su and C. Wang,
Adv. Funct. Mater., 2018, 1803815.
60 X. Li, L. Zhou, Y. Wei, A. M. El-Toni, F. Zhang and D. Zhao,
J. Am. Chem. Soc., 2014, 136, 15086–15092.
61 G. J. Tao, Z. Y. Bai, Y. Chen, H. L. Yao, M. Y. Wu, P. Huang,
L. D. Yu, J. M. Zhang, C. Dai and L. Zhang, Nano Res., 2017,
10, 3790–3810.
62 Y. M. Wang, X. Ji, P. Pang, Y. F. Shi, J. Dai, J. K. Xu,
J. P. Wu, T. B. Kirk and W. Xue, J. Mater. Chem. B, 2018, 6,
2481–2488.
63 Y. Liu, X. Yang, Z. Huang, P. Huang, Y. Zhang, L. Deng,
Z. Wang, Z. Zhou, Y. Liu, H. Kalish, N. M. Khachab,
X. Chen and Z. Nie, Angew. Chem., Int. Ed., 2016, 55,
15297–15300.
64 G. Song, M. Chen, Y. Zhang, L. Cui, H. Qu, X. Zheng,
M. Wintermark, Z. Liu and J. Rao, Nano Lett., 2018, 18,
182–189.
65 S. Hornig, T. Heinze, C. R. Becer and U. S. Schubert,
J. Mater. Chem., 2009, 19, 3838–3840.
66 T. H. Shin, Y. Choi, S. Kim and J. Cheon, Chem. Soc. Rev.,
2015, 44, 4501–4516.
67 Q. Sun, Z. Zhou, N. Qiu and Y. Shen, Adv. Mater., 2017, 29,
1706100.
68 L. Cheng, C. Wang, L. Feng, K. Yang and Z. Liu, Chem. Rev.,
2014, 114, 10869–10939.
69 K. Huang, Y. Zhang, J. Lin and P. Huang, Biomater. Sci.,
2018, DOI: 10.1039/c8bm00642c.
This journal is © The Royal Society of Chemistry 2019

Biomaterials Science
70 X. Ding, C. H. Liow, M. Zhang, R. Huang, C. Li, H. Shen,
M. Liu, Y. Zou, N. Gao, Z. Zhang, Y. Li, Q. Wang, S. Li and
J. Jiang, J. Am. Chem. Soc., 2014, 136, 15684–15693.
71 D. Liu, L. Wang, Z. Wang and A. Cuschieri, Nano Lett.,
2012, 12, 5117–5121.
72 Z. Zhang, J. Wang and C. Chen, Adv. Mater., 2013, 25,
3869–3880.
73 M. Borzenkov, G. Chirico, L. D’Alfonso, L. Sironi,
M. Collini, E. Cabrini, G. Dacarro, C. Milanese,
P. Pallavicini, A. Taglietti, C. Bernhard and F. Denat,
Langmuir, 2015, 31, 8081–8091.
Published on 16 January 2019. Downloaded by IOCB Prague on 1/29/2020 11:48:59 PM.
74 H. Chen, W. Zhang, G. Zhu, J. Xie and X. Chen, Nat. Rev.
Mater., 2017, 2, 17024.
This journal is © The Royal Society of Chemistry 2019
View Article Online
Review
75 S. Wang, J. Lin, T. Wang, X. Chen and P. Huang,
Theranostics, 2016, 6, 2394–2413.
76 C. Chi, Y. Du, J. Ye, D. Kou, J. Qiu, J. Wang, J. Tian and
X. Chen, Theranostics, 2014, 4, 1072–1084.
77 J. Lin, M. Wang, H. Hu, X. Yang, B. Wen, Z. Wang,
O. Jacobson, J. Song, G. Zhang, G. Niu, P. Huang and
X. Chen, Adv. Mater., 2016, 28, 3273–3279.
78 J. Kim, Y. Piao and T. Hyeon, Chem. Soc. Rev., 2009, 38,
372–390.
79 O. G. Hayes, J. R. McMillan, B. Lee and C. A. Mirkin, J. Am.
Chem. Soc., 2018, 140, 9269–9274.
80 W. Fan, B. Yung, P. Huang and X. Chen, Chem. Rev., 2017,
117, 13566–13638.
Biomater. Sci., 2019, 7, 1262–1275 | 1275