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Cite This: ACS Cent. Sci. 2020, 6, 100−116

Size-Tunable Strategies for a Tumor Targeted Drug Delivery System

Wenqi Yu,# Rui Liu,# Yang Zhou, and Huile Gao*
Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry and Sichuan Province, Sichuan Engineering
Laboratory for Plant-Sourced Drug and Sichuan Research Center for Drug Precision Industrial Technology, West China School of
Pharmacy, Sichuan University, Chengdu 610041, China

ABSTRACT: Nanoparticles have been widely used in tumor

targeted drug delivery, while the antitumor effects are not
always satisfactory due to the limited penetration and
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retention. As we all know, there is a paradox that nanoparticles

with large sizes tend to distribute around tumor blood vessels
rather than penetrate into tumor parenchyma, while smaller
sizes can penetrate deeply but with poor tumor retention. In
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recent days, an intelligent, size-tunable strategy provided a

solution to determine the size problem of nanoparticles and
exhibited good application prospects. In this review, we
summarize series of stimuli-induced aggregation and shrinkage
strategies for tumor targeted drug delivery, which can
significantly increase the retention and penetration of
nanodrugs in tumor sites at the same time, thus promoting treatment efficacy. Internal (enzymes, pH, and redox) and
external (light and temperature) stimuli are introduced to change the morphology of the original nanodrugs through
protonation, hydrophobization, hydrogen bond, π−π stacking and enzymolysis-resulted click reactions or dissociation, etc.
Apart from applications in oncotherapy, size-tunable strategies also have a great prospect in the diagnosis and real time
bioimaging fields, which are also introduced in this review. Finally, the potential challenges for application and future directions
are thoroughly discussed, providing guidance for further clinical transformation.

1. INTRODUCTION consideration, which is a very tricky problem to keep in

After years of research and development, nanoparticles have
been widely used in antitumor research because of their high
specific surface area, easy modification, and strong targeting
properties.1,2 To passively deliver nanoparticles to tumor sites,
the enhanced permeability and penetration (EPR) effect is the
strategy that is mostly used, which is specific only in tumors
due to the rapid proliferation of tumor cells and the abnormal
tumor vasculature system.3−5 However, more and more studies
found that only delivering nanodrugs to target sites is far from
enough, and accumulation and penetration problems still
influence the intratumoral delivery efficacy to a great extent.6
Therefore, scientists have tried to design nanodrugs with both
good accumulation and penetration capacity in tumor tissues
to achieve in situ therapeutic concentrations and good
treatment efficacy.
Among all the strategies, designing nanoparticles with
tunable sizes is the most intuitive and controllable approach.
Many studies have found that there is a close correlation
between the antitumor effect and the size of nanodrugs.7,8
Usually, the diameter of nanodrugs is designed according to
the pore size of a leaky tumor vasculature.9 Though differences
may occur owing to the variety of tumor models, subcutaneous
tumors always exhibit a characteristic pore cutoff size ranging
from 200 nm to 1.2 μm, and the size is further reduced in
tumors that grow in the cranium such as glioma.10 Then, size-
related accumulation and penetration abilities are taken into
balance. Because of the special structure and environment of
tumor tissues, there is a contradictory effect of a nanoparticle’s
size on drug delivery. That is, nanoparticles with large sizes
tend to be more capable of retention in tumor tissue than
those with smaller sizes.11−13 As for the permeability, things
become reversed, smaller sizes have a better penetration ability
in tumor tissues.14 To fully utilize the existing paradox,
researchers have designed a series of nanoparticles with
intelligent tunable sizes, including intelligent size aggregation,
size shrinkage, and reversible size-changing strategies, which
are systematically discussed in this review.
Among all the strategies, design-
ing nanoparticles with tunable
sizes is the most intuitive and
controllable approach.
In this review, we will summarize intelligent size-tunable
strategies including size aggregation, size shrinkage, as well as
reversible size changes. Each section is divided through
different stimuli such as enzyme, pH, redox, light, temperature,
etc. In addition to the enhanced retention and penetration, we
Received: November 6, 2019
Published: January 21, 2020

© 2020 American Chemical Society 100 DOI: 10.1021/acscentsci.9b01139

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Scheme 1. Brief Illustration of Stimuli-Induced Size-Tunable Strategies with Their Potential Applications
also focus on other potential applications in different ways.
Aggregation strategies can be used in enhanced cellular uptake,
antimetastasis, and tumor diagnosis (photoacoustic imaging
(PA), positron emission computed tomography (PET),
surface-enhanced Raman scattering, and enhanced magnetic
resonance imaging (MRI)), while shrinkage strategies exhibit
advantages in nuclear delivery, drug release (Scheme 1), etc. In
the end, we conclude with the future application of size-
tunable nanoparticles and existing problems that need to be
solved for better treatment.
2. SIZE IMPACT ON DELIVERY EFFICACY
As one of the most important characteristics of nanoparticles,
size greatly influences the efficiency of tumor targeted drug
delivery in many ways, including circulation, biodistribution,
tumor accumulation and penetration, as well as cellular uptake
and subcellular distribution. A thorough understanding of size
will be introduced first to help better elucidate the importance
of size-tunable strategies.
After entering into the body, the circulation time of
nanodrugs basically determines the efficacy of tumor targeting
as the clearance by mononuclear phagocytic system (MPS) or
filtration by the liver and spleen happens very quickly and
sequesters the majority of nanodrugs. There is a correlation
between the circulation and particle size. The MPS clearance
exhibits a size-dependent behavior such that nanoparticles with
small sizes are less likely to be taken up by macrophages than
large ones.15,16 The biodistribution is also greatly influenced by
the size of nanodrugs because of the different cutoff size of
organs. The renal filtration cutoff size is 5.5 nm,17 and the
vascular fenestrations in liver are 50−100 nm. Particles lower
than 5.5 nm are much more easily excreted through urine, and
sizes smaller than 50 nm could easily penetrate the endothelial
and get trapped in the liver.18,19
To effectively accumulate in tumor sites, there is one more
barrier to cross: the leaky tumor vasculature. As for the rapid
growth of tumor cells, the pore cutoff size of the tumor vessel
ranges from 200 nm to 1.2 μm, depending on the type of
tumor.20,21 Diameters of nanoparticles below the cutoff size are
101
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required for effective passive targeting. Generally, after entering
the tumor region, nanoparticles with a large size are capable of
being well retained in the tumor surroundings but it is hard for
them to penetrate deeply in the dense matrix,22,23 while the
small-sized ones remain sufficiently penetrated in the tumor,
whereas they can be easily pumped back into the bloodstream
by the high interstitial fluid pressure of the tumor.24−26
Although there are investigations exploring strategies to
modulate the tumor microenvironment, consequently improv-
ing nanoparticles’ intratumoral distribution,27,28 optimizing the
nanoparticles’ size is deemed as a critical solution. Also,
regarding the principle of the size-dependent tumor distribu-
tion, it is defined that the unique and ideal size remains exactly
the equilibrium point between penetration and retention. To
optimize the situation, size-tunable nanoparticles show their
extraordinary talent. Compared with nanoparticles that are
unique in size, size-tunable nanoparticles can reach both better
penetration and retention, so long as they change their sizes at
the proper time.
Nanoparticles with large sizes
tend to be more capable of
retention in tumor tissue than
those with smaller sizes. As for
the permeability, things become
reversed, and smaller sizes have
better penetration ability in
tumor tissues.
After nanoparticles enter into tumor sites, their size also
influences the following cellular uptake process in which
nanoparticles with varied sizes exhibit different internalization
rates.29−31 Sizes were found to be the determining factor for
the pathway of cellular internalization. Most of the nano-
particles were internalized into cells through two pathways:
phagocytosis and pinocytosis. The phagocytosis pathway was
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only in some special cells such as macrophages, neutrophils, or
dendritic cells, or particles larger than 500 nm in diameter.
Also, the pinocytosis pathway can be further divided into four
different types based on the mediated proteins: caveolae-
mediated endocytosis, clathrin-mediated endocytosis, non-
caveolin nonclathrin mediated endocytosis, and macropinocy-
tosis.32 Nanoparticles between 20 and 100 nm are internalized
through caveolae-mediated endocytosis,33 120 to 150 nm are
mediated by clathrin,34 and macropinocytosis occurs in the
internalization process of nanoparticles larger than 1 μm.35 As
a reverse mechanism of endocytosis, exocytosis also plays an
important role in enhanced cellular accumulation of nano-
particles. Decreased exocytosis avoids nanoparticles secretion
to the extracellular space and therefore improves the
therapeutic efficacy.36 Quantities of research studies found
that there was a negative correlation between the size of
nanoparticles and the exocytosis process, and nanoparticles
with relatively large sizes exhibit decreased exocytosis and
enhanced intracellular retention.37,38
As with a brain for a human, the nucleus in the cell plays the
most important role among all the organelles and is responsible
for controlling genetics and metabolism.39−41 Various therapies
aim at the nucleus to achieve an anticancer effect, such as
anthraquinone derivatives and short hairpin RNA, which
restrictively exhibit activities inside the nucleus. However, the
major obstacle to nuclear delivery is the nuclear membrane, in
which nuclear pores are generally ∼9 nm in size.42,43 Even
though there are other strategies to promote the nuclear
delivery of nanoparticles with relatively larger sizes, such as
shape modulation,44 ligand modification for active targeting,45
or artificially opening the nuclear envelope by singlet
oxygen,46,47 the simplest and most effective way yet is to
control the nanoparticles’ size below 9 nm. However, the
ultrasmall size ensuring the passage across nuclear pores is
comparable with the renal filtration cutoff, risking nano-
particles’ circulation and distribution at the organ level. To
address the contradiction, size-tunable nanoparticles arise at
the appropriate time, gathering both good blood circulation
and nuclear distribution.
3. AGGREGATION STRATEGIES
As a contradictory behavior of varied sizes described above,
small-sized nanoparticles tend to have good penetration but
are cleared away rapidly, while nanoparticles with large sizes
have enhanced retention time but cannot penetrate deeply into
tumors. Researchers proposed an aggregation strategy to
enhance the accumulation and penetration of nanoparticles in
the meantime. The initial small-sized nanoparticles were first
used for deep penetration, and form large agglomerates for
enhanced retention once deep into the tumor after specific
stimulations. The stimulations can be divided into several types
due to the tumor heterogeneity, such as hypoxia, slightly acidic
microenvironment, and specifically upregulated enzymes. In
addition, some external stimuli such as light and temperature
are also utilized to design the intelligent, size-aggregatable
nanoparticles (Table 1). Under external or internal stimuli,
initial nanoparticles with relatively small sizes interact with
each other through click reaction, self-assembly, electrostatic
interaction, or phase transition to form aggregations.
3.1. Enzyme-Induced Aggregations. The enzyme-
induced strategy has great potential because of its highly
specific substrate selectivity and varied expression of enzyme in
different organs. There are many specific upregulated enzymes
102
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Table 1. Mainly Used Aggregation Strategies
stimuli mechanism ref
enzyme click reaction (cyano and 1,2-thiolamino, azide 51−56
and alkyne, amine and acyl)
self-assembly (amphiphilic block copolymers) 60−63
pH electrostatic interactions (zwitterionic 65−71
compounds, hemoglobin)
light C−C, C−H, O−H, X−H insertions (azobenzene, 73−75
diazirine, triphenylmethane, cinnamenyl)
temperature phase transition (PPCs, PNIPAm, PDEAm, PEO, 81−92
PPO, polyphosphoesters)
redox click reaction and host−guest interaction 99, 101
(disulfide bond, Fc+)
in tumor tissues such as matrix metalloproteinase (MMP),
legumain, hyaluronidase (HAase), gelatinase,48 furin,49 caspase
3/7,50 etc. Click reactions are commonly used in this strategy
because of their easy and quick procedure, which is perfectly
suitable for in vivo synthesis. Many functional groups can be
utilized such as azide and terminal alkynes,51 sulfhydryl and
maleimide,52 1,2-thiolamino and cyano groups,53 etc. By
decorating the above corresponding chemical groups on the
surface of nanoparticles, aggregation will happen very quickly
once the chemical groups are exposed and interact with each
other. Rao et al. first used the click reaction between the cyano
group of 2-cyano-benzothiazole (CBT) and the 1,2-aminothiol
group of cysteine (Cys), which was the last step of the
synthesis of D-luciferin and displayed biological friendliness.54
Our group utilized the substrate of legumain (Ala-Ala-Asn↓
Cys-Lys) and codelivered 2-cyano-6-amino-benzothiazole
(CABT) to tumor sites where legumain was found overex-
pressed,55 which generated aggregations of GNPs from 35.6 to
309.6 nm in 12 h (Figure 1A) and endowed GNPs not only
the prolonged tumor accumulation but also bioimaging of
multispectral optoacoustic tomography (MSOT). In addition
to enzymes that are upregulated in tumor sites, there are
circumstances when the triggered enzymes of click reactions
are not or are lowly expressed in tumors. In that case, enzymes
can be delivered together with nanocarriers. Gu et al.
codelivered transglutaminase (TG) in a hyaluronic acid
(HA)-made nanogel shell on the surface of a carrier (Figure
1B).56 The protected nanogel shell collapsed after entering
into the tumor and released sufficient TG to catalyze the
formation of an isopeptide bond between free amine group
from lysine and the acyl group from glutamine, which led to
aggregation from 10 to 120 nm and prolonged tumor retention
time to more than 72 h.
Unlike click reactions which need to modify other functional
chemical groups to make aggregation happen, the enzyme-
induced self-assembly makes full use of the character of the
nanocarrier itself. Such nanocarriers are usually composed of
amphiphilic block copolymers, which can lead to formation,
destruction, or morphological transformation after changes in
the chemical of physical nature.57−59 Gianneschi et al. designed
a polymer−peptide block copolymer amphiphiles system
containing substrates for four different cancer-associated
enzymes: protein kinase A (PKA), protein phosphatases-1
(PP1), MMP-2, and MMP-9 (Figure 1C).60 Phosphorylation
by PKA caused a 50-fold increase in hydrodynamic diameter
together with the appearance change of amorphous structure,
and the aggregation could be reversed after being successively
treated with PP1 for dephosphorylation, providing feasibility
for enzymatically size-switchable strategy. Alkaline phospha-
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Figure 1. (A) Diagram depicting the legumain-triggered aggregation and composition of GNPs-DOX-A&C. Copyright 2016, American Chemical
Society. (B) Schematic of tumor microenvironment-mediated construction of combination drug-delivery depots for sustained drug release using
CS-NG. Copyright 2016 American Chemical Society. (C) A variety of morphologies of polymeric amphiphile aggregates depending on the design
of the peptide substrate and enzymes added. Copyright 2011, American Chemical Society.

Figure 2. (A) Schematic illustration of pH-responsive “smart” gold nanoparticles (SANs). Copyright 2013, Royal Society of Chemistry. (B) TEM
images of the Hb-IR780 complex at pH 7.4 (top, the red arrows highlight the Hb-IR780 nanoparticles) and at pH 6.5 (bottom). (C) Increase in
hydrodynamic size of the Hb-IR780 complex from normal tissue pH 7.4 to tumor acidic pH 6.5. Copyright 2018 American Chemical Society.

tases (ALPs) is another enzyme widely used to instruct the
self-assembly of nanofibers in vivo,61 which was found to be
highly expressed on the cell membrane or secreted out of some
cancer cells with the ability to responsively cleave the
phosphate groups. Xue et al. and Liang et al. both utilized
ALPs to induce nanofibers through π−π stacking.62,63
3.2. pH-Triggered Aggregations. Different parts of the
human body exhibit different pH levels as we all know. Because
of the rapid proliferation of tumor cells, the microenvironment
of tumors is slightly acidic at around pH 6.5, and after being
internalized into cells, some cytoplasmic organoids such as
endosomes and lysosomes exhibit an even lower pH degree at
103
around pH 5.0−5.5, while blood and normal tissues are
maintained at 7.4.64 Different from enzyme-induced aggrega-
tions, pH-triggered reactions possess the advantages of quick
response and ultrasensitivity. Upon the electrostatic inter-
actions of decorated pH-sensitive surface molecules65,66 or
natural pH-sensitive proteins,67 the nanocarriers will aggregate
and prolong retention in tumors due to the increased
sedimentation-driven uptake and decreased extracellular efflux.
pH-responsive molecules usually are zwitterionic com-
pounds, such as hydrolysis-susceptible citraconic amide,68,69
11-mercaptoundecanoic acid, and (10-mercaptodecyl)-
trimethylammonium bromide).14 Citraconic amides are rapidly
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Figure 3. (A) Schematic illustration of light-triggered assembles of dGNPs. (B) TEM images of dGNPs before and after illuminated with 405 nm
laser in different periods of time. Copyright 2016, John Wiley and Sons.
Figure 4. (A) LCST profiles of F1−F3 before and after treatment with GSH, caspase-3, and Atg4B. (B) Specific responsive nanoaggregation in
cells. Confocal images and bio-TEM images of MCF7 cells treated with PPCs and modulated agents. Copyright 2017, American Chemical Society.
converted to positively charged primary amines by hydrolysis
at a mildly acidic pH, which would in turn react with the
negatively charged carboxyl groups through electrostatic
attraction and lead to aggregation. However, it should be
noted that mixed charged ligands with the same strong
electrolytes would not aggregate and remain stable over the
entire pH range.70 Therefore, disrupting the balanced electro-
lytes was a feasible way to form aggregation (Figure 2A),71 and
the commonly used combination is a weak acid and strong
base such as a weak electrolytic 11-mercaptoundecanoic acid
and strong electrolytic (10-mercaptodecyl)trimethyl-
ammonium bromide.
Though the commonly used pH-sensitive ligands are
synthesized artificially, natural proteins with acidic or basic
amino acid residues are deemed to be pH-sensitive nanoplat-
forms. It is generally known that proteins remain stable at
different pH values, while aggregating around the isoelectric
point (pI).72 If the protein is properly designed with pI around
the physiological pH of tumor, the natural protein could be
chosen as the carrier for enhanced tumor accumulation. Li et
al. employed hemoglobin as a smart pH-sensitive nanocarrier
for near-infrared dye IR780 (Hb-IR780).67 Dynamic light
scattering (DLS) results showed that the Hb-IR780 was well-
dispersed as a singular protein at pH 7.4 while aggregated
severely after incubation at pH 6.5 (Figure 2B,C).
3.3. Light-Induced Aggregations. The light-induced
strategy has shown great advantages in noninvasiveness,
remote manipulation, and easy operation. The photothermal
therapy even shows an extraordinary antitumor effect and is
being used widely. In the aggregation strategies, light-
104
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responsive polymers are supplied with photoactive groups
such as azobenzene, spirobenzopyran,73 triphenylmethane,74 or
cinnamenyl that can undergo reversible structural changes
under UV−Vis light (Figure 3A,B).75 However, the application
of this strategy is only limited in experiments on the cellular
level due to the weak penetration ability of UV−vis and blue
light. Upconversion nanoparticles (UCNPs) are novel
materials consisting of rare-earth elements, which can convert
a near-infrared (NIR) light to UV−vis radiation via two
photons or a multiphoton mechanism.76 The application of
UCNP has been well developed so far and has aroused great
interest of researchers as the extraordinary tissue penetration of
long-wavelength NIR light exactly provides a solution to solve
the long-standing problem of limited tissue penetration of
visible light.77 Liu et al. observed an enhanced photodynamic
treatment efficacy when loaded with a photosensitizer in
UCNP.78 Zhao et al. reported that UCNP can trigger the
photoisomerization of azobenzene,79 which is a commonly
used chemical group for light-induced aggregation as
mentioned above. Therefore, the further application of light-
induced aggregation in vivo can be achieved with great
prospects by utilizing UCNP as the carrier.
3.4. Temperature-Triggered Aggregations. To con-
struct temperature-responsive nanoparticles, the balance
between segment−segment interactions and segment-solvent
intermolecular interactions can be reversed by temperature
changes. Some thermosensitive polymers can be designed to
typically undergo the coil−helix transition upon decreasing
temperature below the upper critical solution temperature
(UCST), while others specifically respond to the increasing
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Figure 5. (A) Schematic drawing of the GSH-responsive antiphagocytosis 99mTc-labeled Fe3O4 nanoparticles for forming particle aggregates
through the interparticle cross-linking reaction. (B) Temporal hydrodynamic size profiles of nonresponsive probe and (C) responsive probe in
reaction with the GSH treatment. Copyright 2017, John Wiley and Sons. (D) Illustration of intracellular host−guest assembly of gold nanoparticles
triggered by GSH. Copyright The Royal Society of Chemistry 2016.
temperature above the lower critical solution temperature
(LCST). Mostly used functional pairs are polymer−peptide
conjugates (PPCs) (Figure 4A,B),80 proteins, polylactic acid
(PLA), and polysaccharides.81−83 Polymers with the merit of
thermoresponsive behaviors such as poly(N-isopropylacryla-
mide) (PNIPAm),84 polyphosphoesters,85 poly(N-diethylacry-
lamide) (PDEAm),86 copolymers of poly(ethylene oxide)
(PEO) and poly(propyleneoxide) (PPO),87 and poly(N-
vinylcaprolactam)88 have been widely used. However, this
strategy always needs the combination with other stimuli such
as enzymes or pH to destroy the inherent interactions and
expose the thermosensitive parts so as to respond to different
temperatures.89−91 In other cases, an additional local temper-
ature increase was given through introducing photothermal
molecules for specific responsiveness.92
3.5. Redox-Induced Aggregation. Attributed to the high
proliferation nature of invasive tumors, tumor tissues and cells
show elevated oxidative stress, resulting in a defined high level
of reactive oxygen species (ROS).93 As a consequence of
handling the oxidative stress, the redox environment of tumor
tissues and cells increases in a way. For example, the
intracellular GSH level is elevated in tumors for increased
antioxidant capacity and resistance against oxidative stress, as
well as regulating cell differentiation, proliferation, and
apoptosis.94 According to research data, the concentration of
GSH in tumor regions, especially the intracellular part, reaches
2−10 mM, whereas that in normal tissue remains 2−20
μM.95−97 The disulfide bond is mostly used GSH-specific
chemical groups that can be reduced to sulfhydryl, and
therefore is widely used in redox environment responsive drug
delivery.98 Gao et al. utilized the dysregulation of GSH for
aggregation strategies and tumor imaging.99 After entering into
tumor sites, the self-peptide was cleaved by GSH, and the
exposed sulfhydryl interacted with maleimide on the end of
PEG of the adjacent nanoparticles according to the click
reaction (Figure 5A). The reaction rate between sulfhydryl and
maleimide happened extremely rapidly so that the nano-
particles aggregated from 7.5 to 295 nm in 5 h (Figure 5B,C).
105
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As an important antioxidant and free radical scavenger in the
body,100 GSH could not only reduce disulfide but also other
species at an oxidative state. Therefore, Wang et al. used GSH
as a reducing agent to specifically respond to reductive
aggregation.101 They modified GNPs with β-CD and incubated
them with Fc+-PEG-Fc+, and aggregation was seen in HepG2
cells after 12 h (Figure 5D). The GSH triggered aggregation
caused significant apoptosis of cancer cells, and because of the
specific express site of GSH, this strategy was also promising in
reducing unexpected side effects induced by traditional
chemotherapy. However, the aggregation efficacy was detected
only at the cellular level as it remained difficult to codeliver β-
CD-GNPs and Fc+-PEG-Fc+ to tumor sites.
3.6. Salt-Induced Aggregation. As is known to all, the
fluid in vivo can be seen as a buffer salt system, and a great
majority of nanoparticles are unstable and easy to aggregate
irreversibly in a high concentration of aqueous salts due to the
disruption of the shielding layer on the surface. Researchers
tried every possible way to prolong the circulation time of
nanocarriers in vivo such as PEGylation and biomimetic cell
membrane coating,102,103 etc. However, Sun et al. first took this
defect as a strategy to design salt-induced GNPs aggrega-
tion.104 After intratumoral injection, GNPs formed irregular
aggregation, while PEG-GNPs did not aggregate at all. The
salt-induced aggregation got rid of complicated surface
modifications and was completed instantaneously. Yet this
strategy was only applied in superficial solid tumors treatment
and needed precise operation because of the specific
requirement of intratumoral injection.
4. SIZE-SHRINKAGE STRATEGIES
With regard to the relationship between nanoparticles’ size and
intratumoral behaviors mentioned before, the high fluid
pressure and dense matrix inside solid tumors always impede
the deep penetration and consequently homogeneous dis-
tribution of nanoparticles inside the tumor;105 therefore, it is
essential for nanoparticles to shrink their size and enhance
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penetration for homogeneous delivery.106 Besides, the size
(reported to be ∼10 nm and up to 39 nm when amplified)107
of nuclear pores limits the nuclear-targeting nanoparticles to be
in an engaged size. The postshrinkage nanoparticles not only
retain a small size for enhanced penetration, but also
contribute to many other properties, such as drug release,108
rapid renal clearance,109 secondary distribution,110 etc. By now,
plenty of nanoparticles have been investigated based on the
endogenous acidic pH, overexpressed enzymes, redox con-
ditions, and exogenous physicochemical stimuli, which are
summarized in Table 2 and discussed in the following.
Table 2. Mainly Used Shrinkage Strategies
stimuli responsive chemical groups refs
pH amino polymers, DMA, Schiff base 114, 115,
119−121
enzyme MMPs, HAase, amylase, thrombin 126−130,
136−141
redox disulfide bond 142, 143
ROS thioketal, thioester, polypropylene sulfide, 148−151
phenylboronic ester
4.1. Size Shrinkage Triggered by Acidic pH. Amino
polymers are a kind of polymer functionalized by amino
groups, which are generally nonprotonated and exhibit
hydrophobicity.111−113 However, the amino groups in amino
polymers change into hydrophilic when protonated at acidic
pH, resulting in the disassembly of the hydrophobic core. The
dramatic protonation of histamine during pH decreasing
provides poly(histamine) with the same property.66 Yuan et al.
reported a kind of nanoassembly composed of nanomicelles
and nanogel,114 which were both self-assembled by amphi-
philic copolymers (PDPA30-b-PAMA15 and P(EGMA-GMA-
PDSEMA), respectively, in aqueous solution (Figure 6A). The
micelle retained a hydrophobic core made up of PDPA, which
was able to respond to intratumoral acidic pH and
consequently transferred to a hydrophilic moiety (Figure 6B)
with the size apparently shrinking from 35 nm to about 10 nm
(Figure 6C). Another similar work recently reported by Ray et
al. is much more suitable to address the penetration problem,
since the size shrinkage is from 100 to 150 nm to 2−5 nm.115
Figure 6. (A) The polymerization of PDPA30-b-PAMA15 and P(EGMA-GMA-PDSEMA), and consequent construction of micelle and nanogel. (B)
Illustration of nanosystem fabrication and responsive shrinkage. (C) TEM images of nanosystem before and after shrinkage, scale bars: 100 nm.
Copyright The Royal Society of Chemistry 2014.
106
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2,3-Dimethylmaleicanhydride (DMA) reacts with many
amines to form acid amine, which can further respond to
slightly acidic condition and break into amine and 2,3-
dimethylmaleic acid.116−118 Wang’s group further constructed
a PCL-CDM-PAMAM/Pt system with a size shrinking from
100 to 5 nm under intratumoral acidic pH (Figure 7A).119 In
addition to directly decomposing contents to reduce size,
DMA also showed another property, charge reversal, for the
design of size-shrinkable nanoparticles (Figure 7B,C).120 The
dramatic size shrinkage and charge reversal were essential for
the penetration and cellular internalization inside the tumor.
Schiff base is another widely used pH-sensitive chemical group,
characterized by a double bond formed between carbon and
nitrogen atoms (-CN-),which is unstable under acidic pH
and easily broken by hydrolysis.121 Although the critical acidic
condition and pervasive proton make a pH-triggered strategy
sensitive and specific, the acidic and hypoxic regions are
commonly far from blood vessels,122 which becomes a major
concern for pH-triggered shrinkage.
4.2. Size Shrinkage Triggered by Overexpressed
Enzyme. MMPs and HAase are the most common tumor-
specific enzymes used as recognizing markers in responsive
drug delivery. MMPs include a large family of proteinases, in
which the MMP-2 and MMP-9 are the most known, playing an
important role in tissue remodeling associated with various
physiological or pathological processes.123−125 The MMP-2
and MMP-9 are generally overexpressed and secreted by
tumors for matrix digestion, recognizing plenty of substrates
involving gelatin and collagens, which retain MMPs’
responsiveness when designed as a component of a drug
carrier. The very first size-shrinkable nanovehicle based on
MMPs was designed by Wong and co-workers.126 They
covered the large gelatin nanoparticles (GelNPs) with small
quantum dots (QDs) to form QDGelNPs, which rapidly
shrank from 100 nm to 9.7 nm when responding to
intratumoral MMP-2 and MMP-9, as a consequence of
GelNPs dissociation (Figure 8A). And taking advantage of
the gelatin nanoparticles, our group functionalized small-sized
dendrigraft polylysine (DGL) on the surface of gelatin
nanoparticles (GelNP), 127−129 with DOX loaded and
angiopep-2 decorated on DGL, 130 resulting in deep
penetration and homogeneous tumor therapy (74.1% growth
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Figure 7. (A) Illustration of construction and pH-responsive size shrinkage of DMA-based nanomicelle. Copyright 2016 National Academy of
Sciences. (B) Schematic illustration of DMA-based PNV, with charge reversal for dissociation. (C) The dissociation of PNV and release of
polymer-Dox inside the tumor. Copyright 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Figure 8. (A) Schematic of 100 nm QDGelNPs shrinking size to 10 nm QDs by cleaving away the gelatin scaffold with MMP-2. Copyright 2011
National Academy of Sciences. (B) Schematic illustration of the design and synergistic effects for deep tumor penetration and therapy effects of
IDDHN. Copyright 2018 Elsevier Ltd.

inhibition). Unlike MMPs with a range of substrates, HAase
exclusively hydrolyzes HA to short chains or modified glucose
units.131 The natural negative charge on HA makes itself easy
to conjugate positively charged materials, and the specific
combination between HA and CD44 also provides HA with
active-targeting ability.132−135 For application of superior HA,
our group cross-linked HA with positively charged DGL to
form size-shrinkable HAase-responsive nanoparticles (Figure
8B) that exhibited extraordinary penetration ability with the
further assistance of an NO donor on an HA shell.136,137
As instanced, the enzyme-triggered size shrinkage is
commonly achieved by shell dissociation of core−shell
nanoparticles and detachment of small-sized decorations
from large nanoparticles.138 The rule is always applicable,
and a recent-report nanosystem is also representative, which
can release small-sized cargo nanoparticles when lactate
oxidase digests its shell.139 Other enzymes-dependent nano-
particles also obeyed this designing principle, such as α-
107
amylase-digested hydroxyethyl starch140 and thrombin-induced
depegylation.141 The enzyme responsiveness is of super-
selectivity owing to exclusive enzyme−substrate recognition,
which is essential for improving the therapeutic efficacy and
reducing the adverse effects of cancer therapy. However,
uneven levels of enzyme expression in different tumors restrict
the application scope of certain responsive nanoparticles.
4.3. Size Shrinkage Triggered by Redox Condition.
Since the GSH level is critical in tumor tissue and cytoplasm,
its responsiveness is also evident in size shrinkage strategies.
Guo et al. constructed a nanomicelle composed of pegylated
polylactide (PEG−PLA) and DMA-modified polythylenimine
(PEI−DMA),142 which was linked by a disulfide bond, forming
PEG−PLA−S−S-PEI−DMA (PELEss-DA, see structures in
Figure 9A). The high level of intracellular GSH reacted with
the disulfide bond and deshielded PEI shell. Also, the dramatic
size shrinkage allowed the remaining nanoparticles’ entry into
nucleus (Figure 9B) and consequent DOX release for DNA
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Figure 9. (A) Schematic design of the nucleus entry of size-shrinkable polymer micelles (PELEss-DA) to overcome MDR. (B) Cellular uptake and
intracellular distribution analysis. Copyright 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. (C) Schematic illustration of the
cooperative dimensional strategy for anticancer drug delivery mediated by hybrid micelle PSPD/P123-Dex. (D) Evaluation of HeLa cells
internalizing different micelles, the number represents: 1. Free DOX; 2. PPD/DOX; 3. PSPD/DOX; 4. PPD/P123-Dex/DOX; 5. PSPD/P123-Dex/
DOX. Copyright 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

interruption. The nucleus delivery ensured the drug’s activity
toward the target, avoiding drug resistant transporters’
function, which is essential for nucleus-targeting chemo-
therapeutic agents. A similar investigation was found in
Wang’s work, in which a PSPD/P123-Dex nanomicelle was
designed (see structures in Figure 9C).143 The two materials
formed 120 nm PSPD/P123-Dex and dramatically shrank into
approximately 30 nm P123-Dex under an intracellular GSH-
abundant condition. Regarding the suitable small size and
assistance of dexamethasone, P123-Dex pithily delivered
encapsulated DOX into the nucleus for pronounced cytotox-
icity (Figure 9D).
4.4. Size Shrinkage Triggered by ROS. Although it is
widely known that there is a relatively high level of ROS in
108
cancer cells (∼10 μM) compared with normal tissues (∼10
nM),144 it is still not sufficient enough to trigger hydrolysis of
ROS-responsive chemical groups instantly. NIR light retains
better tissue penetration compared with ultraviolet and visible
light.145 Importantly, plenty of photosensitizers are able to
convert NIR light into ROS, which has been widely explored as
photodynamic therapy for surficial cancer treatment.146,147 A
thioketal linker is easy to synthesize and respond to ROS,
resulting in two broken parts. Cao et al. designed a PCL−PEG
nanomicelle with poly(thioketal phosphoester) (TK-PPE)
mixed core.148 The chlorin e6 (Ce6) loaded in the micelle
could transfer NIR irradiation into ROS, further cleaving the
TK-PPE and dissociating the core of micelle (Figure 10A). As
a consequence, the nanomicelle shrank its size from 154 to 72
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Figure 10. (A) Preparation and function of the light-activated shrinkable nanoparticle TK-PPE@NPCe6/DOX. (B) TEM images of TK-PPE@
NPCe6 after 660 nm laser irradiation at different times. (C) Plasma DOX concentration versus time after intravenous injection of free DOX,
TKPPE@NPCe6/DOX, and TK-PPE@NPCe6/TK-PPE@NPDOX. (D) Quantification of DOX content in tumors by HPLC. Tumors were
excised 12 and 24 h after intravenous injection. (E) MDA-MB-231 tumor growth curves of various groups after intravenous administration, *p <
0.05; **p < 0.01. Copyright 2017 American Chemical Society.
nm (Figure 10B), with DOX released. By comparison to free
drug, the preshrinkage large micelle ensured its prolonged
circulation (Figure 10C), while the postshrinkage small one
promoted its distribution and deep penetration inside the
tumor (Figure 10D). Together with the transferred ROS and
released DOX facilitated by NIR irradiation, the shrinkable
nanomicelle performed chemo- photodynamic therapy in deep
tumor region, resulting in remarkable antitumor effect (Figure
10E). Other ROS-responsive structure or materials, such as
polypropylene sulfide,149 phenylboronic ester150 and thioest-
er151 are also of talent in designing NIR/ROS-triggered
shrinkage. Unlike other endogenous stimuli, ROS triggered by
external NIR irradiation is desirable and controllable, as well as
tumor-specificity. Meanwhile, no activity remains in the
nonirradiation or nondrug region, ensuring the safety and
avoiding adverse-effect aroused by cancer therapy. Besides,
dual stimuli seem like one case with two locks, ensuring the
specificity of unfolding.
5. MAJOR CHALLENGES OF SIZE-TUNABLE
STRATEGIES FOR APPLICATION
Though great progress has been made in size-tunable
nanoparticles, there are still many challenges that hinder the
efficacy, which need to be further optimized and solved.
5.1. Protein Corona May Shield the Responsive
Ligand. Protein corona acts as the main issue to affect the
targeting efficiency. The corona is quickly formed once
nanoparticles are introduced into biological fluid. As we have
discussed previously, the composition, size, shape, and surface
chemistry of nanoparticles, and even the different fluid
environment in vivo, influence the formation of protein
corona.152 In addition to acting as opsonin to mark a nanodrug
and alter its biodistribution, the adsorbed protein corona on
the surface may also shield the responsive ligand from stimuli
and affect the sensitivity of size changing. Among which, the
mechanism that need direct contact such as enzymatic
109
Outlook
reaction, pH-induced protonation and GSH-dependent redox
may suffer from the protein corona a lot, while the external
stimuli such as light and temperature would not be bothered.
Though PEGylation has been confirmed to facilitate nano-
particles with the “stealth” property, which can reduce the
protein adsorption and decrease the effect of corona, there
comes a new question whether the enzymatic reaction would
also be affected by PEGylation, as the nature of enzyme is
protein as well. The response sensitivity may be affected in the
enzyme-induced size change after PEGylation.
5.2. Accuracy of Measurement. As we are doing research
studies about size changing, the detection of nanoparticles’ size
undoubtably requires very high accuracy. However, the size
measured by DLS always displays a relatively wide peak.153
Nanoparticles with an average DLS size of 100 nm probably
have a peak ranging from 50 to 400 nm. It is not that
convincing to confirm the success of size shrinkage (from 100
to 50 nm) or size aggregation (from 100 to 400 nm) as there is
an overlap of size during the size change process. The
corresponding distribution intervals can be compared sepa-
rately instead of just comparing the average number, which is
meaningless. On the other hand, nanoparticles with a good
polydispersity index are highly recommended in the study of
size change, such as silica nanoparticles, gold nanoparticles,
and polystyrene nanoparticles, as these nanoparticles have
already been well industrialized and have quite uniform size,
and the narrow size range would partly solve the problem in a
way.
5.3. Off-Target Effect. The location accuracy of the
responsive reaction needs to be precisely designed. Nano-
particles are designed to specifically target tumor sites and
enhance the accumulation. However, the off-target effects
always attenuate the antitumor efficacy and cause damage to
normal organs. For example, P-aminophenyl-α-D-mannopyr-
anoside (MAN) is a widely used glucose analogue modified on
nanoparticles to help cross the blood−brain barrier and target
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brain tumor cells,154 while its receptor, the mannose receptor,
is also expressed on Kupffer cells of liver.155 The majority of
nanoparticles are trapped in the liver, limiting the effective
delivery to tumor sites. On the other hand, since the pH and
enzyme environments are somehow different on both sides of
the biological barrier, it remains to be explored whether the
responsiveness still exists after crossing barriers, or the
responsiveness is crippled by prereacting with enzyme on the
barriers. Furthermore, the most commonly used targeting
strategy is ligand−receptor binding; ligands are modified on
nanoparticles to help actively target the tumor sites. However,
the chemical modification would destroy the innate structure
of ligands and attenuate the binding efficiency to some extent.
The recently rising bio-orthogonal reactions improve the
targeting specificity very well.156,157 By first giving a primary
exogenous tag to label target cells, the secondary tag could
specifically recognize the labeled cells and avoid the off-target
effect.158−160 However, the existing primary tag delivery still
depends on the interaction of ligand and receptor, and relevant
studies are only at the cellular level; further work needs to be
done to make it better applied in vivo.
5.4. Complicated Tumor Microenvironment. Because
of the heterogeneous nature of tumor sites, the tumor
microenvironment should be taken into consideration. The
dense ECM and high interstitial pressure of the tumor tissue
attenuate nanoparticle delivery efficiency. However, the
intelligent size change of nanoparticles together with
remodeling of the tumor microenvironment could synergisti-
cally improve the accumulation and penetration of nano-
particles at the tumor sites, such as degradation of ECM and
normalization of tumor vasculature system, which could
enhance the penetration ability of nanoparticles and signifi-
cantly reduce the reflux of nanoparticles leading to promoted
accumulation at tumor sites. However, there also exists
unpredicted adverse effects. Disruption of ECM may also
lead to tumor migration and metastasis, and untimely
normalization of blood vessels may make it difficult for
nanoparticles to passively target tumor tissues through an EPR
effect. The extent and timing of remodeling of microenviron-
ment need to be carefully balanced.
5.5. Sensitivity of Transformation. Because of the high
interstitial pressure of tumor sites, nanoparticles that enter
tumor sites are easily pumped back to vessels, and the
retention time of the original nanoparticles will not be too
long. Therefore, the sensitivity and response time with stimuli
are a key factor in designing size-tunable nanoparticles. As
demonstrated above, pH-triggered reactions possess the
advantages of quick response and ultrasensitivity, while the
reaction time between enzyme and substate is not that fast.
The enzyme-induced aggregations are usually observed at the
12 or 24 h time point, and a majority of the original
nanoparticles may be eliminated before aggregations happen.
5.6. Possibility of Clinical Translation. In the last few
decades, an EPR effect was considered to be the basic principle
for passive drug delivery to tumor sites.161 However, there are
more and more controversies and doubts about the EPR effect
because of the unsatisfactory therapeutic efficacy in clinical
trials. Also, it has been reported that only 0.7% nanoparticles
can be finally delivered to tumor sites.162 One of the most
important reasons is the difference between human and model
animals,163 and there is no single animal model that can fully
reproduce human cancers. In addition, the EPR effect only
exhibits good results in animal models, which is because of the
110
Outlook
requirement of tumor volume. In tumor-bearing mice, it is
until the tumor grows to 200 mm3 that it exhibit good EPR
effect, which is nearly impossible for a human to have a such
big tumor. Recently, scientists are more prone to intervene in
tumors at an early stage after inoculation in mice or choose
active-targeted strategies to treat cancers. Moreover, there are
still other problems that limit the clinical translation such as
the biocompatibility of nanodrugs and scalable manufacturing
for industry, which is a long way to go.
6. OUTLOOK AND FUTURE DIRECTIONS
In addition to the above strategies, there are many other novel
approaches to make nanocarriers aggregate or shrink in tumor
tissues, such as using enzyme-instructed self-assembly to
generate intracellular supramolecular nanofibers to achieve
the aggregation goal,164−167 or using the change of shape to
achieve the purpose of aggregation in a round-about way.168,169
Intracellular aggregation will decrease the efflux of nanodrugs
in a way, which has potential in overcoming multidrug
resistance.49 Except prolonging the retention of nanocarriers
after aggregation, properties of aggregated nanodrugs tend to
be changed, and most of them exhibit a good photothermal,
photoacoustic effect and enhanced contract of MRI, which
provide a possibility for real time imaging and photothermal
therapy leading to better antitumor treatment prospects.170
Aggregation-induced emission (AIE) fluorophores are the
application of aggregation strategies to solve the aggregation-
caused quenching (ACQ) problems and give a new long-term
tracer for in vitro or in vivo imaging.171,172 Also, because of the
good results achieved in antitumor therapy, aggregation
strategies are increasingly being used in other areas, such as
brain tumor surgery guidance173 and myocardial infarction.174
With more and more applications
of size aggregation and size-
shrinkage strategies, there comes
a question of which strategy is
better.
Moreover, the shrinkage strategy also shows capabilities
more than penetration promotion and nucleus delivery. The
shrinking procedure always accompanies shell dissociation or
volume squeezing of nanoparticles, which have potential for
specific drug release. Besides, nanoparticles’ size influences
their distribution not only in tumor tissue, but also in normal
organs. Small-sized nanoparticles tend to accumulate in organs
such as kidney, spleen, and lung, which generate different
potential functions and strategies for nanoparticle-based
therapy, such as secondary distribution in spleen for immune
activation, rapid renal clearance for elimination of toxic
materials, and lung-targeting function for alveolus remodeling.
With more and more applications of size aggregation and
size-shrinkage strategies, there comes a question of which
strategy is better. In fact, though the two strategies are totally
opposite from the methodology, the ultimate goal and
therapeutic efficacy are the same, which are to prolong the
penetration and retention time in tumor sites at the same time.
Besides, the emergence of the size-reversible strategies further
optimizes the strategies of size change.175,176 And recently,
nanoparticles were produced that could simultaneously change
the tumor microenvironment along with the size change,177,178
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such as hypoxia relief,179 normalization of vasculature,180 and
extracellular matrix modification,181 endowing a better
therapeutic effect.
In short, it is trendy to carefully design responsive, size-
changeable nanoparticles based on a certain tumor, even a
certain part of the tumor tissue. We look forward the time
when multiresponsive nanoparticles with multistage size
changes are developed and optimize current treatment,
which seems hard to achieve today but will be of great value.
■ AUTHOR INFORMATION
Corresponding Author
*E-mail: gaohuile@scu.edu.cn; gaohuilescu@163.com.
ORCID
Huile Gao: 0000-0002-5355-7238
Author Contributions
#
W.Y. and R.L. contributed equally to this work.
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
This work was supported by National Natural Science
Foundation of China (31571016), Research Foundation of
Sichuan Science and Technology Department (19YYJC2250),
the Young Elite Scientists Sponsorship Program by CAST
(2017QNR001), the Fundamental Research Funds for the
Central Universities, and 111 Project (B18035).
■
REFERENCES
(1) Pan, L.; He, Q.; Liu, J.; Chen, Y.; Ma, M.; Zhang, L.; Shi, J.
Nuclear-targeted drug delivery of TAT peptide-conjugated mono-
disperse mesoporous silica nanoparticles. J. Am. Chem. Soc. 2012, 134
(13), 5722−5725.
(2) Pan, L.; Liu, J.; He, Q.; Shi, J. MSN-mediated sequential
vascular-to-cell nuclear-targeted drug delivery for efficient tumor
regression. Adv. Mater. 2014, 26 (39), 6742−6748.
(3) Gao, H. Shaping Tumor Microenvironment for Improving
Nanoparticle Delivery. Curr. Drug Metab. 2016, 17 (8), 731−736.
(4) Miao, L.; Lin, C. M.; Huang, L. Stromal barriers and strategies
for the delivery of nanomedicine to desmoplastic tumors. J. Controlled
Release 2015, 219, 192−204.
(5) Setyawati, M. I.; Tay, C. Y.; Bay, B. H.; Leong, D. T. Gold
Nanoparticles Induced Endothelial Leakiness Depends on Particle
Size and Endothelial Cell Origin. ACS Nano 2017, 11 (5), 5020−
5030.
(6) Li, C.; Wang, J.; Wang, Y.; Gao, H.; Wei, G.; Huang, Y.; Yu, H.;
Gan, Y.; Wang, Y.; Mei, L.; Chen, H.; Hu, H.; Zhang, Z.; Jin, Y.
Recent progress in drug delivery. Acta Pharm. Sin. B 2019, 9 (6),
1145−1162.
(7) Carnovale, C.; Bryant, G.; Shukla, R.; Bansal, V. Size, shape and
surface chemistry of nano-gold dictate its cellular interactions, uptake
and toxicity. Prog. Mater. Sci. 2016, 83, 152−190.
(8) Talamini, L.; Violatto, M. B.; Cai, Q.; Monopoli, M. P.; Kantner,
K.; Krpetic, Z.; Perez-Potti, A.; Cookman, J.; Garry, D.; Silveira, C. P.;
Boselli, L.; Pelaz, B.; Serchi, T.; Cambier, S.; Gutleb, A. C.; Feliu, N.;
Yan, Y.; Salmona, M.; Parak, W. J.; Dawson, K. A.; Bigini, P. Influence
of Size and Shape on the Anatomical Distribution of Endotoxin-Free
Gold Nanoparticles. ACS Nano 2017, 11 (6), 5519−5529.
(9) Hashizume, H.; Baluk, P.; Morikawa, S.; McLean, J. W.;
Thurston, G.; Roberge, S.; Jain, R. K.; McDonald, D. M. Openings
between Defective Endothelial Cells Explain Tumor Vessel Leakiness.
Am. J. Pathol. 2000, 156 (4), 1363−1380.
(10) Moghimi, S. M.; Hunter, A. C.; Murray, J. C. Nanomedicine:
current status and future prospects. FASEB J. 2005, 19 (3), 311−330.
111
Outlook
(11) Perrault, S. D.; Walkey, C.; Jennings, T.; Fischer, H. C.; Chan,
W. C. Mediating tumor targeting efficiency of nanoparticles through
design. Nano Lett. 2009, 9 (5), 1909−1915.
(12) Sykes, E. A.; Chen, J.; Zheng, G.; Chan, W. C. W. Investigating
the Impact of Nanoparticle Size on Active and Passive Tumor
Targeting Efficiency. ACS Nano 2014, 8 (6), 5696−5706.
(13) She, W.; Luo, K.; Zhang, C.; Wang, G.; Geng, Y.; Li, L.; He, B.;
Gu, Z. The potential of self-assembled, pH-responsive nanoparticles
of mPEGylated peptide dendron-doxorubicin conjugates for cancer
therapy. Biomaterials 2013, 34 (5), 1613−1623.
(14) Liu, X.; Chen, Y.; Li, H.; Huang, N.; Jin, Q.; Ren, K.; Ji, J.
Enhanced retention and cellular uptake of nanoparticles in tumors by
controlling their aggregation behavior. ACS Nano 2013, 7 (7), 6244−
6257.
(15) Champion, J. A.; Walker, A.; Mitragotri, S. Role of particle size
in phagocytosis of polymeric microspheres. Pharm. Res. 2008, 25 (8),
1815−1821.
(16) He, C.; Hu, Y.; Yin, L.; Tang, C.; Yin, C. Effects of particle size
and surface charge on cellular uptake and biodistribution of polymeric
nanoparticles. Biomaterials 2010, 31 (13), 3657−3666.
(17) Choi, H. S.; Liu, W.; Misra, P.; Tanaka, E.; Zimmer, J. P.; Itty
Ipe, B.; Bawendi, M. G.; Frangioni, J. V. Renal clearance of quantum
dots. Nat. Biotechnol. 2007, 25 (10), 1165−1170.
(18) Liu, D.; Mori, A.; Huang, L. Role of liposome size and RES
blockade in controlling biodistribution and tumor uptake of GM1-
containing liposomes. Biochim. Biophys. Acta, Biomembr. 1992, 1104
(1), 95−101.
(19) Chen, L.-T.; Weiss, L. The Role of the Sinus Wall in the
Passage of Erythrocytes Through the Spleen. Blood 1973, 41, 529−
537.
(20) Hobbs, S. K.; Monsky, W. L.; Yuan, F.; Roberts, W. G.; Griffith,
L.; Torchilin, V. P.; Jain, R. K. Regulation of transport pathways in
tumor vessels: role of tumor type and microenvironment. Proc. Natl.
Acad. Sci. U. S. A. 1998, 95 (8), 4607−4612.
(21) Yuan, F.; Dellian, M.; Fukumura, D.; Leunig, M.; Berk, D. A.;
Torchilin, V. P.; Jain, R. K. Vascular permeability in a human tumor
xenograft: molecular size dependence and cutoff size. Cancer Res.
1995, 55 (17), 3752−3756.
(22) Poh, S.; Chelvam, V.; Low, P. S. Comparison of nanoparticle
penetration into solid tumors and sites of inflammation: studies using
targeted and nontargeted liposomes. Nanomedicine 2015, 10 (9),
1439−1449.
(23) Wang, R.; Shen, Q.; Li, X.; Xie, C.; Lu, W.; Wang, S.; Wang, J.;
Wang, D.; Liu, M. Efficacy of inverso isomer of CendR peptide on
tumor tissue penetration. Acta Pharm. Sin. B 2018, 8 (5), 825−832.
(24) Tang, L.; Yang, X.; Yin, Q.; Cai, K.; Wang, H.; Chaudhury, I.;
Yao, C.; Zhou, Q.; Kwon, M.; Hartman, J. A.; Dobrucki, I. T.;
Dobrucki, L. W.; Borst, L. B.; Lezmi, S.; Helferich, W. G.; Ferguson,
A. L.; Fan, T. M.; Cheng, J. Investigating the optimal size of
anticancer nanomedicine. Proc. Natl. Acad. Sci. U. S. A. 2014, 111
(43), 15344−15349.
(25) Maman, S.; Witz, I. P. A history of exploring cancer in context.
Nat. Rev. Cancer 2018, 18 (6), 359−376.
(26) Danhier, F. To exploit the tumor microenvironment: Since the
EPR effect fails in the clinic, what is the future of nanomedicine? J.
Controlled Release 2016, 244, 108−121.
(27) Carpenter, A. W.; Schoenfisch, M. H. Nitric oxide release: part
II. Therapeutic applications. Chem. Soc. Rev. 2012, 41 (10), 3742−
3752.
(28) Papageorgis, P.; Stylianopoulos, T. Role of TGFβ in regulation
of the tumor microenvironment and drug delivery (review). Int. J.
Oncol. 2015, 46 (3), 933−943.
(29) Gratton, S. E.; Ropp, P. A.; Pohlhaus, P. D.; Luft, J. C.;
Madden, V. J.; Napier, M. E.; DeSimone, J. M. The effect of particle
design on cellular internalization pathways. Proc. Natl. Acad. Sci. U. S.
A. 2008, 105 (33), 11613−11618.
(30) Jiang, W.; Kim, B. Y.; Rutka, J. T.; Chan, W. C. Nanoparticle-
mediated cellular response is size-dependent. Nat. Nanotechnol. 2008,
3 (3), 145−150.
DOI: 10.1021/acscentsci.9b01139
ACS Cent. Sci. 2020, 6, 100−116
ACS Central Science
(31) Huang, J.; Bu, L.; Xie, J.; Chen, K.; Cheng, Z.; Li, X.; Chen, X.
Effects of nanoparticle size on cellular uptake and liver MRI with
polyvinylpyrrolidone-coated iron oxide nanoparticles. ACS Nano
2010, 4 (12), 7151−7160.
(32) Doherty, G. J.; McMahon, H. T. Mechanisms of endocytosis.
Annu. Rev. Biochem. 2009, 78, 857−902.
(33) Wang, Z., Caveolae-mediated Delivery of Therapeutic Nano-
particles across Blood-endothelial Barrier. Austin J. Anal Pharm. Chem.
2014, 1 (4).
(34) Chithrani, B. D.; Ghazani, A. A.; Chan, W. C. W. Determining
the size and shape dependence of gold nanoparticle uptake into
mammalian cells. Nano Lett. 2006, 6 (4), 662−668.
(35) Swanson, J. A. Shaping cups into phagosomes and macro-
pinosomes. Nat. Rev. Mol. Cell Biol. 2008, 9 (8), 639−649.
(36) Wang, T.; Wang, L.; Li, X.; Hu, X.; Han, Y.; Luo, Y.; Wang, Z.;
Li, Q.; Aldalbahi, A.; Wang, L.; Song, S.; Fan, C.; Zhao, Y.; Wang, M.;
Chen, N. Size-Dependent Regulation of Intracellular Trafficking of
Polystyrene Nanoparticle-Based Drug-Delivery Systems. ACS Appl.
Mater. Interfaces 2017, 9 (22), 18619−18625.
(37) Chithrani, B. D.; Chan, W. C. Elucidating the mechanism of
cellular uptake and removal of protein-coated gold nanoparticles of
different sizes and shapes. Nano Lett. 2007, 7 (6), 1542−1550.
(38) Mei, L.; Rao, J.; Liu, Y.; Li, M.; Zhang, Z.; He, Q. Effective
treatment of the primary tumor and lymph node metastasis by
polymeric micelles with variable particle sizes. J. Controlled Release
2018, 292, 67−77.
(39) Gorlich, D.; Kutay, U. Transport between the cell nucleus and
the cytoplasm. Annu. Rev. Cell Dev. Biol. 1999, 15, 607−660.
(40) Rippe, K. Dynamic organization of the cell nucleus. Curr. Opin.
Genet. Dev. 2007, 17 (5), 373−380.
(41) Penman, S. RNA metabolism in the HeLa cell nucleus. J. Mol.
Biol. 1966, 17 (1), 117−130.
(42) Huang, K.; Ma, H.; Liu, J.; Huo, S.; Kumar, A.; Wei, T.; Zhang,
X.; Jin, S.; Gan, Y.; Wang, P. C.; He, S.; Zhang, X.; Liang, X. J. Size-
dependent localization and penetration of ultrasmall gold nano-
particles in cancer cells, multicellular spheroids, and tumors in vivo.
ACS Nano 2012, 6 (5), 4483−4493.
(43) Huo, S. D.; Jin, S. B.; Ma, X. W.; Xue, X. D.; Yang, K. N.;
Kumar, A.; Wang, P. C.; Zhang, J. C.; Hu, Z. B.; Liang, X. J.
Ultrasmall Gold Nanoparticles as Carriers for Nucleus-Based Gene
Therapy Due to Size-Dependent Nuclear Entry. ACS Nano 2014, 8
(6), 5852−5862.
(44) Hinde, E.; Thammasiraphop, K.; Duong, H. T.; Yeow, J.;
Karagoz, B.; Boyer, C.; Gooding, J. J.; Gaus, K. Pair correlation
microscopy reveals the role of nanoparticle shape in intracellular
transport and site of drug release. Nat. Nanotechnol. 2017, 12 (1),
81−89.
(45) Pan, L. M.; Liu, J. A.; He, Q. J.; Wang, L. J.; Shi, J. L.
Overcoming multidrug resistance of cancer cells by direct intranuclear
drug delivery using TAT-conjugated mesoporous silica nanoparticles.
Biomaterials 2013, 34 (11), 2719−2730.
(46) Zhu, Y. X.; Jia, H. R.; Pan, G. Y.; Ulrich, N. W.; Chen, Z.; Wu,
F. G. Development of a Light-Controlled Nanoplatform for Direct
Nuclear Delivery of Molecular and Nanoscale Materials. J. Am. Chem.
Soc. 2018, 140 (11), 4062−4070.
(47) Pan, L.; Liu, J.; Shi, J. Cancer cell nucleus-targeting
nanocomposites for advanced tumor therapeutics. Chem. Soc. Rev.
2018, 47 (18), 6930−6946.
(48) Zhang, D.; Qi, G. B.; Zhao, Y. X.; Qiao, S. L.; Yang, C.; Wang,
H. In Situ Formation of Nanofibers from Purpurin18-Peptide
Conjugates and the Assembly Induced Retention Effect in Tumor
Sites. Adv. Mater. 2015, 27 (40), 6125−6130.
(49) Yuan, Y.; Wang, L.; Du, W.; Ding, Z.; Zhang, J.; Han, T.; An,
L.; Zhang, H.; Liang, G. Intracellular Self-Assembly of Taxol
Nanoparticles for Overcoming Multidrug Resistance. Angew. Chem.,
Int. Ed. 2015, 54 (33), 9700−9704.
(50) Julien, O.; Wells, J. A. Caspases and their substrates. Cell Death
Differ. 2017, 24 (8), 1380−1389.
112
Outlook
(51) Wang, Z.; An, H. W.; Hou, D.; Wang, M.; Zeng, X.; Zheng, R.;
Wang, L.; Wang, K.; Wang, H.; Xu, W. Addressable Peptide Self-
Assembly on the Cancer Cell Membrane for Sensitizing Chemo-
therapy of Renal Cell Carcinoma. Adv. Mater. 2019, 31 (11),
e1807175.
(52) Liu, Y.; Yu, Y.; Lam, J. W.; Hong, Y.; Faisal, M.; Yuan, W. Z.;
Tang, B. Z. Simple biosensor with high selectivity and sensitivity:
thiol-specific biomolecular probing and intracellular imaging by AIE
fluorogen on a TLC plate through a thiol-ene click mechanism. Chem.
- Eur. J. 2010, 16 (28), 8433−8438.
(53) Ai, X.; Ho, C. J. H.; Aw, J.; Attia, A. B. E.; Mu, J.; Wang, Y.;
Wang, X.; Wang, Y.; Liu, X.; Chen, H.; Gao, M.; Chen, X.; Yeow, E.
K. L.; Liu, G.; Olivo, M.; Xing, B. In vivo covalent cross-linking of
photon-converted rare-earth nanostructures for tumour localization
and theranostics. Nat. Commun. 2016, 7, 10432.
(54) Liang, G.; Ren, H.; Rao, J. A biocompatible condensation
reaction for controlled assembly of nanostructures in living cells. Nat.
Chem. 2010, 2 (1), 54−60.
(55) Ruan, S.; Hu, C.; Tang, X.; Cun, X.; Xiao, W.; Shi, K.; He, Q.;
Gao, H. Increased Gold Nanoparticle Retention in Brain Tumors by
in Situ Enzyme-Induced Aggregation. ACS Nano 2016, 10 (11),
10086−10098.
(56) Hu, Q.; Sun, W.; Lu, Y.; Bomba, H. N.; Ye, Y.; Jiang, T.;
Isaacson, A. J.; Gu, Z. Tumor Microenvironment-Mediated
Construction and Deconstruction of Extracellular Drug-Delivery
Depots. Nano Lett. 2016, 16 (2), 1118−1126.
(57) Chien, M. P.; Rush, A. M.; Thompson, M. P.; Gianneschi, N. C.
Programmable shape-shifting micelles. Angew. Chem., Int. Ed. 2010, 49
(30), 5076−5080.
(58) Jain, S.; Bates, F. S. On the origins of morphological complexity
in block copolymer surfactants. Science 2003, 300 (5618), 460−464.
(59) Sundararaman, A.; Stephan, T.; Grubbs, R. B. Reversible
restructuring of aqueous block copolymer assemblies through
stimulus-induced changes in amphiphilicity. J. Am. Chem. Soc. 2008,
130 (37), 12264−12265.
(60) Ku, T.-H.; Chien, M.-P.; Thompson, M. P.; Sinkovits, R. S.;
Olson, N. H.; Baker, T. S.; Gianneschi, N. C. Controlling and
Switching the Morphology of Micellar Nanoparticles with Enzymes. J.
Am. Chem. Soc. 2011, 133 (22), 8392−8395.
(61) Feng, Z.; Zhang, T.; Wang, H.; Xu, B. Supramolecular catalysis
and dynamic assemblies for medicine. Chem. Soc. Rev. 2017, 46 (21),
6470−6479.
(62) Zhou, J.; Du, X.; Yamagata, N.; Xu, B. Enzyme-Instructed Self-
Assembly of Small D-Peptides as a Multiple-Step Process for
Selectively Killing Cancer Cells. J. Am. Chem. Soc. 2016, 138 (11),
3813−3823.
(63) Zheng, Z.; Chen, P.; Xie, M.; Wu, C.; Luo, Y.; Wang, W.; Jiang,
J.; Liang, G. Cell Environment-Differentiated Self-Assembly of
Nanofibers. J. Am. Chem. Soc. 2016, 138 (35), 11128−11131.
(64) Engin, K.; Leeper, D. B.; Cater, J. R.; Thistlethwaite, A. J.;
Tupchong, L.; Mcfarlane, J. D. Extracellular Ph Distribution in
Human Tumors. Int. J. Hyperthermia 1995, 11 (2), 211−216.
(65) Wang, Y.; Lin, Y. X.; Qiao, Z. Y.; An, H. W.; Qiao, S. L.; Wang,
L.; Rajapaksha, R. P.; Wang, H. Self-assembled autophagy-inducing
polymeric nanoparticles for breast cancer interference in-vivo. Adv.
Mater. 2015, 27 (16), 2627−2634.
(66) Yang, P.; Luo, Q.; Qi, G.; Gao, Y.; Li, B.; Zhang, J.; Wang, L.;
Wang, H. Host Materials Transformable in Tumor Microenvironment
for Homing Theranostics. Adv. Mater. 2017, 29 (15), 1605869.
(67) Li, H.; Chen, Y.; Li, Z.; Li, X.; Jin, Q.; Ji, J. Hemoglobin as a
Smart pH-Sensitive Nanocarrier To Achieve Aggregation Enhanced
Tumor Retention. Biomacromolecules 2018, 19 (6), 2007−2013.
(68) Ju, K. Y.; Kang, J.; Pyo, J.; Lim, J.; Chang, J. H.; Lee, J. K. pH-
Induced aggregated melanin nanoparticles for photoacoustic signal
amplification. Nanoscale 2016, 8 (30), 14448−14456.
(69) Song, J.; Kim, J.; Hwang, S.; Jeon, M.; Jeong, S.; Kim, C.; Kim,
S. ″Smart″ gold nanoparticles for photoacoustic imaging: an imaging
contrast agent responsive to the cancer microenvironment and signal
DOI: 10.1021/acscentsci.9b01139
ACS Cent. Sci. 2020, 6, 100−116
ACS Central Science
amplification via pH-induced aggregation. Chem. Commun. (Cam-
bridge, U. K.) 2016, 52 (53), 8287−8290.
(70) Li, S. D.; Huang, L. Pharmacokinetics and biodistribution of
nanoparticles. Mol. Pharmaceutics 2008, 5 (4), 496−504.
(71) Nam, J.; Ha, Y. S.; Hwang, S.; Lee, W.; Song, J.; Yoo, J.; Kim, S.
pH-responsive gold nanoparticles-in-liposome hybrid nanostructures
for enhanced systemic tumor delivery. Nanoscale 2013, 5 (21),
10175−10178.
(72) Krebs, M. R.; Domike, K. R.; Donald, A. M. Protein
aggregation: more than just fibrils. Biochem. Soc. Trans. 2009, 37
(Pt 4), 682−686.
(73) Kundu, P. K.; Samanta, D.; Leizrowice, R.; Margulis, B.; Zhao,
H.; Borner, M.; Udayabhaskararao, T.; Manna, D.; Klajn, R. Light-
controlled self-assembly of non-photoresponsive nanoparticles. Nat.
Chem. 2015, 7 (8), 646−652.
(74) Pieroni, O.; Fissi, A.; Angelini, N.; Lenci, F. Photoresponsive
polypeptides. Acc. Chem. Res. 2001, 34 (1), 9−17.
(75) Cheng, X.; Sun, R.; Yin, L.; Chai, Z.; Shi, H.; Gao, M. Light-
Triggered Assembly of Gold Nanoparticles for Photothermal Therapy
and Photoacoustic Imaging of Tumors In Vivo. Adv. Mater. 2017, 29
(6), 1604894.
(76) Kuk, S.; Lee, B. I.; Lee, J. S.; Park, C. B. Rattle-Structured
Upconversion Nanoparticles for Near-IR-Induced Suppression of
Alzheimer’s beta-Amyloid Aggregation. Small 2017, 13 (11),
1603139.
(77) Zhao, M.; Wang, R.; Li, B.; Fan, Y.; Wu, Y.; Zhu, X.; Zhang, F.
Precise In Vivo Inflammation Imaging Using In Situ Responsive
Cross-linking of Glutathione-Modified Ultra-Small NIR-II Lanthanide
Nanoparticles. Angew. Chem., Int. Ed. 2019, 58 (7), 2050−2054.
(78) Xu, J.; Xu, L.; Wang, C.; Yang, R.; Zhuang, Q.; Han, X.; Dong,
Z.; Zhu, W.; Peng, R.; Liu, Z. Near-Infrared-Triggered Photodynamic
Therapy with Multitasking Upconversion Nanoparticles in Combina-
tion with Checkpoint Blockade for Immunotherapy of Colorectal
Cancer. ACS Nano 2017, 11 (5), 4463−4474.
(79) Zhao, T.; Wang, P.; Li, Q.; Al-Khalaf, A. A.; Hozzein, W. N.;
Zhang, F.; Li, X.; Zhao, D. Near-Infrared Triggered Decomposition of
Nanocapsules with High Tumor Accumulation and Stimuli
Responsive Fast Elimination. Angew. Chem., Int. Ed. 2018, 57 (10),
2611−2615.
(80) Qiao, S. L.; Ma, Y.; Wang, Y.; Lin, Y. X.; An, H. W.; Li, L. L.;
Wang, H. General Approach of Stimuli-Induced Aggregation for
Monitoring Tumor Therapy. ACS Nano 2017, 11 (7), 7301−7311.
(81) Schmaljohann, D. Thermo- and pH-responsive polymers in
drug delivery. Adv. Drug Delivery Rev. 2006, 58 (15), 1655−1670.
(82) Schmaljohann, D.; Beyerlein, D.; Nitschke, M.; Werner, C.
Thermo-reversible swelling of thin hydrogel films immobilized by
low-pressure plasma. Langmuir 2004, 20 (23), 10107−10114.
(83) Jeong, B.; Kim, S. W.; Bae, Y. H. Thermosensitive sol−gel
reversible hydrogels. Adv. Drug Delivery Rev. 2012, 64, 154−162.
(84) Jones, S. T.; Walsh-Korb, Z.; Barrow, S. J.; Henderson, S. L.;
del Barrio, J.; Scherman, O. A. The Importance of Excess Poly(N-
isopropylacrylamide) for the Aggregation of Poly(N-isopropylacryla-
mide)-Coated Gold Nanoparticles. ACS Nano 2016, 10 (3), 3158−
3165.
(85) Wang, Y. C.; Tang, L. Y.; Li, Y.; Wang, J. Thermoresponsive
block copolymers of poly(ethylene glycol) and polyphosphoester:
thermo-induced self-assembly, biocompatibility, and hydrolytic
degradation. Biomacromolecules 2009, 10 (1), 66−73.
(86) Idziak, I.; Avoce, D.; Lessard, D.; Gravel, D.; Zhu, X. X.
Thermosensitivity of Aqueous Solutions of Poly(N,N-diethylacryla-
mide). Macromolecules 1999, 32 (4), 1260−1263.
(87) Loh, X. J.; Goh, S. H.; Li, J. New biodegradable thermogelling
copolymers having very low gelation concentrations. Biomacromole-
cules 2007, 8 (2), 585−593.
(88) Vihola, H.; Laukkanen, A.; Hirvonen, J.; Tenhu, H. Binding and
release of drugs into and from thermosensitive poly(N-vinyl
caprolactam) nanoparticles. Eur. J. Pharm. Sci. 2002, 16 (1−2), 69−
74.
113
Outlook
(89) Klaikherd, A.; Nagamani, C.; Thayumanavan, S. Multi-stimuli
sensitive amphiphilic block copolymer assemblies. J. Am. Chem. Soc.
2009, 131 (13), 4830−4838.
(90) Roy, D.; Cambre, J. N.; Sumerlin, B. S. Triply-responsive
boronic acid block copolymers: solution self-assembly induced by
changes in temperature, pH, or sugar concentration. Chem. Commun.
(Cambridge, U. K.) 2009, No. 16, 2106−2108.
(91) Agut, W.; Brulet, A.; Schatz, C.; Taton, D.; Lecommandoux, S.
pH and temperature responsive polymeric micelles and polymersomes
by self-assembly of poly[2-(dimethylamino)ethyl methacrylate]-b-
poly(glutamic acid) double hydrophilic block copolymers. Langmuir
2010, 26 (13), 10546−10554.
(92) Liu, F. H.; Cong, Y.; Qi, G. B.; Ji, L.; Qiao, Z. Y.; Wang, H.
Near-Infrared Laser-Driven in Situ Self-Assembly as a General
Strategy for Deep Tumor Therapy. Nano Lett. 2018, 18 (10),
6577−6584.
(93) Wu, W. S. The signaling mechanism of ROS in tumor
progression. Cancer Metastasis Rev. 2007, 25 (4), 695−705.
(94) Lee, F. Y.; Vessey, A.; Rofstad, E.; Siemann, D. W.; Sutherland,
R. M. Heterogeneity of glutathione content in human ovarian cancer.
Cancer Res. 1989, 49 (19), 5244−5248.
(95) Liu, T.; Lai, L.; Song, Z.; Chen, T. A Sequentially Triggered
Nanosystem for Precise Drug Delivery and Simultaneous Inhibition of
Cancer Growth, Migration, and Invasion. Adv. Funct. Mater. 2016, 26
(43), 7775−7790.
(96) Shimura, N.; Musya, A.; Hashimoto, T.; Kojima, S.; Kubodera,
A.; Sasaki, T. Usefulness of 99m Tc-d,l-HMPAO for estimation of
GSH content in tumor tissues. Nucl. Med. Biol. 2000, 27 (6), 577−
580.
(97) Yuan, D.; Ding, L.; Sun, Z.; Li, X. MRI/Fluorescence bimodal
amplification system for cellular GSH detection and tumor cell
imaging based on manganese dioxide nanosheet. Sci. Rep. 2018, 8,
No. 1747, DOI: 10.1038/s41598-018-20110-z.
(98) Bardwell, J. C. A.; Mcgovern, K.; Beckwith, J. Identification of a
protein required for disulfide bond formation in vivo. Cell 1991, 67
(3), 581−589.
(99) Gao, Z.; Hou, Y.; Zeng, J.; Chen, L.; Liu, C.; Yang, W.; Gao, M.
Tumor Microenvironment-Triggered Aggregation of Antiphagocyto-
sis (99m) Tc-Labeled Fe3 O4 Nanoprobes for Enhanced Tumor
Imaging In Vivo. Adv. Mater. 2017, 29 (24), 1701095.
(100) Li, F.; Cui, L.; Yu, D.; Hao, H.; Liu, Y.; Zhao, X.; Pang, Y.;
Zhu, H.; Du, W. Exogenous glutathione improves intracellular
glutathione synthesis via the gamma-glutamyl cycle in bovine zygotes
and cleavage embryos. J. Cell. Physiol. 2019, 234 (5), 7384−7394.
(101) Wang, Y.; Li, H.; Jin, Q.; Ji, J. Intracellular host-guest assembly
of gold nanoparticles triggered by glutathione. Chem. Commun.
(Cambridge, U. K.) 2016, 52 (3), 582−585.
(102) Gao, W.; Hu, C. M.; Fang, R. H.; Luk, B. T.; Su, J.; Zhang, L.
Surface functionalization of gold nanoparticles with red blood cell
membranes. Adv. Mater. 2013, 25 (26), 3549−3553.
(103) Suk, J. S.; Xu, Q.; Kim, N.; Hanes, J.; Ensign, L. M.
PEGylation as a strategy for improving nanoparticle-based drug and
gene delivery. Adv. Drug Delivery Rev. 2016, 99 (Pt A), 28−51.
(104) Sun, M.; Liu, F.; Zhu, Y.; Wang, W.; Hu, J.; Liu, J.; Dai, Z.;
Wang, K.; Wei, Y.; Bai, J.; Gao, W. Salt-induced aggregation of gold
nanoparticles for photoacoustic imaging and photothermal therapy of
cancer. Nanoscale 2016, 8 (8), 4452−4457.
(105) McKee, T. D.; Grandi, P.; Mok, W.; Alexandrakis, G.; Insin,
N.; Zimmer, J. P.; Bawendi, M. G.; Boucher, Y.; Breakefield, X. O.;
Jain, R. K. Degradation of fibrillar collagen in a human melanoma
xenograft improves the efficacy of an oncolytic herpes simplex virus
vector. Cancer Res. 2006, 66 (5), 2509−2513.
(106) Minchinton, A. I.; Tannock, I. F. Drug penetration in solid
tumours. Nat. Rev. Cancer 2006, 6 (8), 583−592.
(107) Panté, N.; Kann, M. Nuclear pore complex is able to transport
macromolecules with diameters of about 39 nm. Mol. Biol. Cell 2002,
13 (2), 425−434.
(108) Chen, J. J.; Ding, J. X.; Wang, Y. C.; Cheng, J. J.; Ji, S. X.;
Zhuang, X. L.; Chen, X. S. Sequentially Responsive Shell-Stacked
DOI: 10.1021/acscentsci.9b01139
ACS Cent. Sci. 2020, 6, 100−116
ACS Central Science
Nanoparticles for Deep Penetration into Solid Tumors. Adv. Mater.
2017, 29 (32), 1701170.
(109) Choi, H. S.; Liu, W.; Misra, P.; Tanaka, E.; Zimmer, J. P.; Ipe,
B. I.; Bawendi, M. G.; Frangioni, J. V. Renal Clearance of
Nanoparticles. Nat. Biotechnol. 2007, 25 (10), 1165.
(110) Chen, K.-H.; Lundy, D. J.; Toh, E. K.-W.; Chen, C.-H.; Shih,
C.; Chen, P.; Chang, H.-C.; Lai, J. J.; Stayton, P. S.; Hoffman, A. S.;
Hsieh, P. C.-H. Nanoparticle distribution during systemic inflamma-
tion is size-dependent and organ-specific. Nanoscale 2015, 7 (38),
15863−15872.
(111) Zhou, K.; Wang, Y.; Huang, X.; Luby-Phelps, K.; Sumer, B.
D.; Gao, J. Tunable, ultrasensitive pH-responsive nanoparticles
targeting specific endocytic organelles in living cells. Angew. Chem.,
Int. Ed. 2011, 50 (27), 6109−6114.
(112) Zhu, Z.; Armes, S. P.; Liu, S. pH-Induced Micellization
Kinetics of ABC Triblock Copolymers Measured by Stopped-Flow
Light Scattering. Macromolecules 2005, 38 (23), 9803−9812.
(113) Ma, X.; Wang, Y.; Zhao, T.; Li, Y.; Su, L. C.; Wang, Z.; Huang,
G.; Sumer, B. D.; Gao, J. Ultra-pH-Sensitive Nanoprobe Library with
Broad pH Tunability and Fluorescence Emissions. J. Am. Chem. Soc.
2014, 136 (31), 11085−11092.
(114) Yuan, C.; Raghupathi, K.; Popere, B. C.; Ventura, J.; Dai, L.;
Thayumanavan, S. Composite supramolecular nanoassemblies with
independent stimulus sensitivities. Chem. Sci. 2014, 5 (1), 229−234.
(115) Ray, P.; Alhalhooly, L.; Ghosh, A.; Choi, Y.; Banerjee, S.;
Mallik, S.; Banerjee, S.; Quadir, M. Size-Transformable, Multifunc-
tional Nanoparticles from Hyperbranched Polymers for Environment-
Specific Therapeutic Delivery. ACS Biomater. Sci. Eng. 2019, 5 (3),
1354−1365.
(116) Kaneda, Y.; Yamamoto, Y.; Kamada, H.; Tsunoda, S.;
Tsutsumi, Y.; Hirano, T.; Mayumi, T. Antitumor activity of tumor
necrosis factor alpha conjugated with divinyl ether and maleic
anhydride copolymer on solid tumors in mice. Cancer Res. 1998, 58
(2), 290−295.
(117) Mehrishi, J. N. Positively charged amino groups on the surface
of normal and cancer cells. Eur. J. Cancer 1970, 6 (2), 127−137.
(118) Kozin, S. V.; Shkarin, P.; Gerweck, L. E. The cell
transmembrane pH gradient in tumors enhances cytotoxicity of
specific weak acid chemotherapeutics. Cancer Res. 2001, 61 (12),
4740−4743.
(119) Li, H.-J.; Du, J.-Z.; Du, X.-J.; Xu, C.-F.; Sun, C.-Y.; Wang, H.-
X.; Cao, Z.-T.; Yang, X.-Z.; Zhu, Y.-H.; Nie, S.; Wang, J. Stimuli-
responsive clustered nanoparticles for improved tumor penetration
and therapeutic efficacy. Proc. Natl. Acad. Sci. U. S. A. 2016, 113 (15),
4164−4169.
(120) Xuan, M.; Shao, J.; Dai, L.; He, Q.; Li, J. Macrophage Cell
Membrane Camouflaged Mesoporous Silica Nanocapsules for In Vivo
Cancer Therapy. Adv. Healthcare Mater. 2015, 4 (11), 1645−1652.
(121) Cordes, E. H.; Jencks, W. P. On the Mechanism of Schiff Base
Formation and Hydrolysis. J. Am. Chem. Soc. 1962, 84 (5), 832−837.
(122) Helmlinger, G.; Yuan, F.; Dellian, M.; Jain, R. K. Interstitial
pH and pO2 gradients in solid tumors in vivo: High-resolution
measurements reveal a lack of correlation. Nat. Med. 1997, 3 (2),
177−182.
(123) Coussens, L. M.; Tinkle, C. L.; Hanahan, D.; Werb, Z. MMP-
9 supplied by bone marrow-derived cells contributes to skin
carcinogenesis. Cell 2000, 103 (3), 481−490.
(124) Vu, T. H.; Shipley, J. M.; Bergers, G.; Berger, J. E.; Helms, J.
A.; Hanahan, D.; Shapiro, S. D.; Senior, R. M.; Werb, Z. MMP-9/
Gelatinase B Is a Key Regulator of Growth Plate Angiogenesis and
Apoptosis of Hypertrophic Chondrocytes. Cell 1998, 93 (3), 411−
422.
(125) Oh, J.; Takahashi, R.; Kondo, S.; Mizoguchi, A.; Adachi, E.;
Sasahara, R. M.; Nishimura, S.; Imamura, Y.; Kitayama, H.; Alexander,
D. B.; Ide, C.; Horan, T. P.; Arakawa, T.; Yoshida, H.; Nishikawa, S.-
i.; Itoh, Y.; Seiki, M.; Itohara, S.; Takahashi, C.; Noda, M. The
membrane-anchored MMP inhibitor RECK is a key regulator of
extracellular matrix integrity and angiogenesis. Cell 2001, 107 (6),
789−800.
114
Outlook
(126) Cliff, W.; Triantafyllos, S.; Jian, C.; John, M.; Chauhan, V. P.;
Wen, J.; Zoran, P.; Jain, R. K.; Bawendi, M. G.; Dai, F. Multistage
nanoparticle delivery system for deep penetration into tumor tissue.
Proc. Natl. Acad. Sci. U. S. A. 2011, 108 (6), 2426−2431.
(127) Ruan, S.; Cao, X.; Cun, X.; Hu, G.; Zhou, Y.; Zhang, Y.; Lu,
L.; He, Q.; Gao, H. Matrix metalloproteinase-sensitive size-shrinkable
nanoparticles for deep tumor penetration and pH triggered
doxorubicin release. Biomaterials 2015, 60, 100−110.
(128) Ruan, S.; He, Q.; Gao, H. Matrix metalloproteinase triggered
size-shrinkable gelatin-gold fabricated nanoparticles for tumor micro-
environment sensitive penetration and diagnosis of glioma. Nanoscale
2015, 7 (21), 9487−9496.
(129) Cun, X.; Chen, J.; Ruan, S.; Zhang, L.; Wan, J.; He, Q.; Gao,
H. A Novel Strategy through Combining iRGD Peptide with Tumor-
Microenvironment-Responsive and Multistage Nanoparticles for
Deep Tumor Penetration. ACS Appl. Mater. Interfaces 2015, 7 (49),
27458−27466.
(130) Hu, G.; Chun, X.; Wang, Y.; He, Q.; Gao, H. Peptide
mediated active targeting and intelligent particle size reduction-
mediated enhanced penetrating of fabricated nanoparticles for triple-
negative breast cancer treatment. Oncotarget 2015, 6 (38), 41258−
41274.
(131) Menzel, E. J.; Farr, C. Hyaluronidase and its substrate
hyaluronan: biochemistry, biological activities and therapeutic uses.
Cancer Lett. 1998, 131 (1), 3−11.
(132) Arpicco, S.; De Rosa, G.; Fattal, E. Lipid-Based Nanovectors
for Targeting of CD44-Overexpressing Tumor Cells. J. Drug Delivery
2013, 2013 (4), 860780.
(133) Liu, R.; Hu, C.; Yang, Y.; Zhang, J.; Gao, H. Theranostic
nanoparticles with tumor-specific enzyme-triggered size reduction and
drug release to perform photothermal therapy for breast cancer
treatment. Acta Pharm. Sin. B 2019, 9 (2), 410−420.
(134) Yu, W.; He, X.; Yang, Z.; Yang, X.; Xiao, W.; Liu, R.; Xie, R.;
Qin, L.; Gao, H. Sequentially responsive biomimetic nanoparticles
with optimal size in combination with checkpoint blockade for
cascade synergetic treatment of breast cancer and lung metastasis.
Biomaterials 2019, 217, 119309.
(135) Luo, Z.; Dai, Y.; Gao, H. Development and application of
hyaluronic acid in tumor targeting drug delivery. Acta Pharm. Sin. B
2019, 9 (6), 1099−1112.
(136) Hu, C.; Cun, X.; Ruan, S.; Liu, R.; Xiao, W.; Yang, X.; Yang,
Y.; Yang, C.; Gao, H. Enzyme-triggered size shrink and laser-
enhanced NO release nanoparticles for deep tumor penetration and
combination therapy. Biomaterials 2018, 168, 64−75.
(137) Hu, C.; Yang, X.; Liu, R.; Ruan, S.; Zhou, Y.; Xiao, W.; Yu,
W.; Yang, C.; Gao, H. Coadministration of iRGD with Multistage
Responsive Nanoparticles Enhanced Tumor Targeting and Pene-
tration Abilities for Breast Cancer Therapy. ACS Appl. Mater.
Interfaces 2018, 10 (26), 22571−22579.
(138) Niu, Y.; Zhu, J.; Li, Y.; Shi, H.; Gong, Y.; Li, R.; Huo, Q.; Ma,
T.; Liu, Y. Size shrinkable drug delivery nanosystems and priming the
tumor microenvironment for deep intratumoral penetration of
nanoparticles. J. Controlled Release 2018, 277, 35−47.
(139) Tseng, S. J.; Kempson, I. M.; Huang, K. Y.; Li, H. J.; Fa, Y. C.;
Ho, Y. C.; Liao, Z. X.; Yang, P. C. Targeting Tumor Microenviron-
ment by Bioreduction-Activated Nanoparticles for Light-Triggered
Virotherapy. ACS Nano 2018, 12 (10), 9894−9902.
(140) Li, Y.; Hu, H.; Zhou, Q.; Ao, Y.; Xiao, C.; Wan, J.; Wan, Y.;
Xu, H.; Li, Z.; Yang, X. alpha-Amylase- and Redox-Responsive
Nanoparticles for Tumor-Targeted Drug Delivery. ACS Appl. Mater.
Interfaces 2017, 9 (22), 19215−19230.
(141) Guo, X.; Deng, G.; Liu, J.; Zou, P.; Du, F.; Liu, F.; Chen, A.
T.; Hu, R.; Li, M.; Zhang, S.; Tang, Z.; Han, L.; Liu, J.; Sheth, K. N.;
Chen, Q.; Gou, X.; Zhou, J. Thrombin-Responsive, Brain-Targeting
Nanoparticles for Improved Stroke Therapy. ACS Nano 2018, 12 (8),
8723−8732.
(142) Guo, X.; Wei, X.; Jing, Y.; Zhou, S. Size Changeable
Nanocarriers with Nuclear Targeting for Effectively Overcoming
DOI: 10.1021/acscentsci.9b01139
ACS Cent. Sci. 2020, 6, 100−116
ACS Central Science
Multidrug Resistance in Cancer Therapy. Adv. Mater. 2015, 27 (41),
6450−6456.
(143) Wang, H.; Li, Y.; Bai, H.; Shen, J.; Chen, X.; Ping, Y.; Tang, G.
A Cooperative Dimensional Strategy for Enhanced Nucleus-Targeted
Delivery of Anticancer Drugs. Adv. Funct. Mater. 2017, 27 (24),
1700339.
(144) Pei, Q.; Hu, X.; Zheng, X.; Liu, S.; Li, Y.; Jing, X.; Xie, Z.
Light-Activatable Red Blood Cell Membrane-Camouflaged Dimeric
Prodrug Nanoparticles for Synergistic Photodynamic/Chemotherapy.
ACS Nano 2018, 12 (2), 1630−1641.
(145) Luo, S.; Zhang, E.; Su, Y.; Cheng, T.; Shi, C. A review of NIR
dyes in cancer targeting and imaging. Biomaterials 2011, 32 (29),
7127−7138.
(146) Patel, N.; Pera, P.; Joshi, P.; Dukh, M.; Tabaczynski, W. A.;
Siters, K. E.; Kryman, M.; Cheruku, R. R.; Durrani, F.; Missert, J. R.;
Watson, R.; Ohulchanskyy, T. Y.; Tracy, E. C.; Baumann, H.; Pandey,
R. K. Highly Effective Dual-Function Near-Infrared (NIR) Photo-
sensitizer for Fluorescence Imaging and Photodynamic Therapy
(PDT) of Cancer. J. Med. Chem. 2016, 59 (21), 9774−9787.
(147) Lu, T.; Shao, P.; Mathew, I.; Sand, A.; Sun, W. Synthesis and
photophysics of benzotexaphyrin: a near-infrared emitter and
photosensitizer. J. Am. Chem. Soc. 2008, 130 (47), 15782−15783.
(148) Cao, Z.; Ma, Y.; Sun, C.; Lu, Z.; Yao, Z.; Wang, J.; Li, D.;
Yuan, Y.; Yang, X. ROS-Sensitive Polymeric Nanocarriers with Red
Light-Activated Size Shrinkage for Remotely Controlled Drug
Release. Chem. Mater. 2018, 30 (2), 517−525.
(149) Thomas, S. N.; van der Vlies, A. J.; O’Neil, C. P.; Reddy, S. T.;
Yu, S. S.; Giorgio, T. D.; Swartz, M. A.; Hubbell, J. A. Engineering
complement activation on polypropylene sulfide vaccine nano-
particles. Biomaterials 2011, 32 (8), 2194−2203.
(150) Zhang, T.; Chen, X.; Xiao, C.; Zhuang, X.; Chen, X. Synthesis
of a phenylboronic ester-linked PEG-lipid conjugate for ROS-
responsive drug delivery. Polym. Chem. 2017, 8 (40), 6209−6216.
(151) Zhao, W.; Qiao, Z.; Duan, Z.; Wang, H. Synthesis and self-
assembly of pH and ROS dual responsive Poly (beta-thioester) s.
Huaxue Xuebao 2016, 74 (3), 234−240.
(152) Xiao, W.; Gao, H. The impact of protein corona on the
behavior and targeting capability of nanoparticle-based delivery
system. Int. J. Pharm. 2018, 552 (1−2), 328−339.
(153) Bhattacharjee, S. DLS and zeta potential - What they are and
what they are not? J. Controlled Release 2016, 235, 337−351.
(154) Ruan, S.; Qin, L.; Xiao, W.; Hu, C.; Zhou, Y.; Wang, R.; Sun,
X.; Yu, W.; He, Q.; Gao, H. Acid-Responsive Transferrin Dissociation
and GLUT Mediated Exocytosis for Increased Blood-Brain Barrier
Transcytosis and Programmed Glioma Targeting Delivery. Adv. Funct.
Mater. 2018, 28 (30), 1802227.
(155) Hirata, K.; Maruyama, T.; Watanabe, H.; Maeda, H.; Nakajou,
K.; Iwao, Y.; Ishima, Y.; Katsumi, H.; Hashida, M.; Otagiri, M.
Genetically engineered mannosylated-human serum albumin as a
versatile carrier for liver-selective therapeutics. J. Controlled Release
2010, 145 (1), 9−16.
(156) Oliveira, B. L.; Guo, Z.; Bernardes, G. J. L. Inverse electron
demand Diels-Alder reactions in chemical biology. Chem. Soc. Rev.
2017, 46 (16), 4895−4950.
(157) Devaraj, N. K. The Future of Bioorthogonal Chemistry. ACS
Cent. Sci. 2018, 4 (8), 952−959.
(158) Taichi, M.; Nomura, S.; Nakase, I.; Imamaki, R.; Kizuka, Y.;
Ota, F.; Dohmae, N.; Kitazume, S.; Taniguchi, N.; Tanaka, K. In Situ
Ligation of High- and Low-Affinity Ligands to Cell Surface Receptors
Enables Highly Selective Recognition. Adv. Sci. (Weinh) 2017, 4 (11),
1700147.
(159) Steen, E. J. L.; Edem, P. E.; Norregaard, K.; Jorgensen, J. T.;
Shalgunov, V.; Kjaer, A.; Herth, M. M. Pretargeting in nuclear
imaging and radionuclide therapy: Improving efficacy of theranostics
and nanomedicines. Biomaterials 2018, 179, 209−245.
(160) Westerlund, K.; Vorobyeva, A.; Mitran, B.; Orlova, A.;
Tolmachev, V.; Karlstrom, A. E.; Altai, M. Site-specific conjugation of
recognition tags to trastuzumab for peptide nucleic acid-mediated
radionuclide HER2 pretargeting. Biomaterials 2019, 203, 73−85.
115
Outlook
(161) Li, H.; Li, J.; He, X.; Zhang, B.; Liu, C.; Li, Q.; Zhu, Y.;
Huang, W.; Zhang, W.; Qian, H.; Ge, L. Histology and antitumor
activity study of PTX-loaded micelle, a fluorescent drug delivery
system prepared by PEG-TPP. Chin. Chem. Lett. 2019, 30 (5), 1083−
1088.
(162) Wilhelm, S.; Tavares, A. J.; Dai, Q.; Ohta, S.; Audet, J.;
Dvorak, H. F.; Chan, W. C. W. Analysis of nanoparticle delivery to
tumours. Nature Reviews Materials 2016, 1 (5), 16014.
(163) Shi, J.; Kantoff, P. W.; Wooster, R.; Farokhzad, O. C. Cancer
nanomedicine: progress, challenges and opportunities. Nat. Rev.
Cancer 2017, 17 (1), 20−37.
(164) Li, J.; Kuang, Y.; Shi, J.; Zhou, J.; Medina, J. E.; Zhou, R.;
Yuan, D.; Yang, C.; Wang, H.; Yang, Z.; Liu, J.; Dinulescu, D. M.; Xu,
B. Enzyme-Instructed Intracellular Molecular Self-Assembly to Boost
Activity of Cisplatin against Drug-Resistant Ovarian Cancer Cells.
Angew. Chem., Int. Ed. 2015, 54 (45), 13307−13311.
(165) Huang, P.; Gao, Y.; Lin, J.; Hu, H.; Liao, H. S.; Yan, X.; Tang,
Y.; Jin, A.; Song, J.; Niu, G.; Zhang, G.; Horkay, F.; Chen, X. Tumor-
Specific Formation of Enzyme-Instructed Supramolecular Self-
Assemblies as Cancer Theranostics. ACS Nano 2015, 9 (10),
9517−9527.
(166) Son, J.; Kalafatovic, D.; Kumar, M.; Yoo, B.; Cornejo, M. A.;
Contel, M.; Ulijn, R. V. Customizing Morphology, Size, and Response
Kinetics of Matrix Metalloproteinase-Responsive Nanostructures by
Systematic Peptide Design. ACS Nano 2019, 13 (2), 1555−1562.
(167) Li, L. L.; Qiao, S. L.; Liu, W. J.; Ma, Y.; Wan, D.; Pan, J.;
Wang, H. Intracellular construction of topology-controlled polypep-
tide nanostructures with diverse biological functions. Nat. Commun.
2017, 8 (1), 1276.
(168) Liu, Y.; He, J.; Yang, K.; Yi, C.; Liu, Y.; Nie, L.; Khashab, N.
M.; Chen, X.; Nie, Z. Folding Up of Gold Nanoparticle Strings into
Plasmonic Vesicles for Enhanced Photoacoustic Imaging. Angew.
Chem., Int. Ed. 2015, 54 (52), 15809−15812.
(169) Liu, R.; Yu, M.; Yang, X.; Umeshappa, C. S.; Hu, C.; Yu, W.;
Qin, L.; Huang, Y.; Gao, H. Linear Chimeric Triblock Molecules Self-
Assembled Micelles with Controllably Transformable Property to
Enhance Tumor Retention for Chemo-Photodynamic Therapy of
Breast Cancer. Adv. Funct. Mater. 2019, 29 (23), 1808462.
(170) Xing, R.; Liu, K.; Jiao, T.; Zhang, N.; Ma, K.; Zhang, R.; Zou,
Q.; Ma, G.; Yan, X. An Injectable Self-Assembling Collagen-Gold
Hybrid Hydrogel for Combinatorial Antitumor Photothermal/Photo-
dynamic Therapy. Adv. Mater. 2016, 28 (19), 3669−3676.
(171) Wang, Z.; Yong, T. Y.; Wan, J.; Li, Z. H.; Zhao, H.; Zhao, Y.;
Gan, L.; Yang, X. L.; Xu, H. B.; Zhang, C. Temperature-sensitive
fluorescent organic nanoparticles with aggregation-induced emission
for long-term cellular tracing. ACS Appl. Mater. Interfaces 2015, 7 (5),
3420−3425.
(172) Yang, L.; Fang, W.; Ye, Y.; Wang, Z.; Hu, Q.; Tang, B. Z.
Redox-responsive fluorescent AIE bioconjugate with aggregation
enhanced retention features for targeted imaging reinforcement and
selective suppression of cancer cells. Materials Chemistry Frontiers
2019, 3 (7), 1335−1340.
(173) Gao, X.; Yue, Q.; Liu, Z.; Ke, M.; Zhou, X.; Li, S.; Zhang, J.;
Zhang, R.; Chen, L.; Mao, Y.; Li, C. Guiding Brain-Tumor Surgery via
Blood-Brain-Barrier-Permeable Gold Nanoprobes with Acid-Trig-
gered MRI/SERRS Signals. Adv. Mater. 2017, 29 (21), 1603917.
(174) Nguyen, M. M.; Carlini, A. S.; Chien, M. P.; Sonnenberg, S.;
Luo, C.; Braden, R. L.; Osborn, K. G.; Li, Y.; Gianneschi, N. C.;
Christman, K. L. Enzyme-Responsive Nanoparticles for Targeted
Accumulation and Prolonged Retention in Heart Tissue after
Myocardial Infarction. Adv. Mater. 2015, 27 (37), 5547−5552.
(175) Liang, H.; Ren, X.; Qian, J.; Zhang, X.; Meng, L.; Wang, X.; Li,
L.; Fang, X.; Sha, X. Size-Shifting Micelle Nanoclusters Based on a
Cross-Linked and pH-Sensitive Framework for Enhanced Tumor
Targeting and Deep Penetration Features. ACS Appl. Mater. Interfaces
2016, 8 (16), 10136−10146.
(176) Su, Y.; Yu, T.; Chiang, W.; Chiu, H.; Chang, C.; Chiang, C.;
Hu, S. Hierarchically Targeted and Penetrated Delivery of Drugs to
DOI: 10.1021/acscentsci.9b01139
ACS Cent. Sci. 2020, 6, 100−116
ACS Central Science Outlook

Tumors by Size-Changeable Graphene Quantum Dot Nanoaircrafts

for Photolytic Therapy. Adv. Funct. Mater. 2017, 27 (23), 1700056.
(177) Dong, X.; Liu, H. J.; Feng, H. Y.; Yang, S. C.; Liu, X. L.; Lai,
X.; Lu, Q.; Lovell, J. F.; Chen, H. Z.; Fang, C. Enhanced Drug
Delivery by Nanoscale Integration of a Nitric Oxide Donor To Induce
Tumor Collagen Depletion. Nano Lett. 2019, 19 (2), 997−1008.
(178) Yang, S.; Gao, H. Nanoparticles for modulating tumor
microenvironment to improve drug delivery and tumor therapy.
Pharmacol. Res. 2017, 126, 97−108.
(179) Chen, Q.; Feng, L. Z.; Liu, J. J.; Zhu, W. W.; Dong, Z. L.; Wu,
Y. F.; Liu, Z. Intelligent Albumin-MnO2 Nanoparticles as pH-/
H2O2-Responsive Dissociable Nanocarriers to Modulate Tumor
Hypoxia for Effective Combination Therapy. Adv. Mater. 2016, 28
(33), 7129−7136.
(180) Xiao, W.; Ruan, S.; Yu, W.; Wang, R.; Hu, C.; Liu, R.; Gao, H.
Normalizing Tumor Vessels To Increase the Enzyme-Induced
Retention and Targeting of Gold Nanoparticle for Breast Cancer
Imaging and Treatment. Mol. Pharmaceutics 2017, 14 (10), 3489−
3498.
(181) Hu, X.; He, P.; Qi, G.; Gao, Y.; Lin, Y.; Yang, C.; Yang, P.;
Hao, H.; Wang, L.; Wang, H. Transformable Nanomaterials as an
Artificial Extracellular Matrix for Inhibiting Tumor Invasion and
Metastasis. ACS Nano 2017, 11 (4), 4086−4096.

116 DOI: 10.1021/acscentsci.9b01139

ACS Cent. Sci. 2020, 6, 100−116