REVIEW
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Recent Progress in the Design and Application of
Supramolecular Peptide Hydrogels in Cancer Therapy
Ying Cai, Chao Zheng, Fengqin Xiong, Wei Ran, Yihui Zhai, Helen H. Zhu, Hao Wang,
Yaping Li,* and Pengcheng Zhang*
Supramolecular peptide hydrogel (SPH) is a class of biomaterials
self-assembled from peptide-based gelators through non-covalent
interactions. Among many of its biomedical applications, the potential of SPH
in cancer therapy has been vastly explored in the past decade, taking
advantage of its good biocompatibility, multifunctionality, and injectability.
SPHs can exert localized cancer therapy and induce systemic anticancer
immunity to prevent tumor recurrence, depending on the design of SPH. This
review first gives a brief introduction to SPH and then outlines the major
types of peptide-based gelators that have been developed so far. The
methodologies to tune the physicochemical properties and biological
activities are summarized. The recent advances of SPH in cancer therapy as
carriers, prodrugs, or drugs are highlighted. Finally, the clinical translation
potential and main challenges in this field are also discussed.
1. Introduction
Extracellular matrix (ECM) is a 3D scaffold for cell adhe-
sion and biomolecule binding, providing cells with structural
Y. Cai, C. Zheng, F. Xiong, Dr. W. Ran, Y. Zhai, Prof. Y. Li, Prof. P. Zhang
State Key Laboratory of Drug Research and Center of Pharmaceutics
Shanghai Institute of Materia Medica
Chinese Academy of Sciences
Shanghai 201203, China
E-mail: ypli@simm.ac.cn; pzhang@simm.ac.cn
Y. Cai, Dr. W. Ran, Y. Zhai, Prof. Y. Li, Prof. P. Zhang
University of Chinese Academy of Sciences
Beijing 100049, China
C. Zheng, F. Xiong, Prof. H. Wang
China State Institute of Pharmaceutical Industry
Shanghai 200040, China
Prof. H. H. Zhu
State Key Laboratory of Oncogenes and Related Genes
Renji-Med-X Stem Cell Research Center
Department of Urology
Ren Ji Hospital
School of Medicine and School of Biomedical Engineering
Shanghai Jiao Tong University
Shanghai 200127, China
Prof. P. Zhang
Yantai Key Laboratory of Nanomedicine and Advanced Preparations
Yantai Institute of Materia Medica
Shandong 264000, China
The ORCID identification number(s) for the author(s) of this article
can be found under https://doi.org/10.1002/adhm.202001239
DOI: 10.1002/adhm.202001239
Adv. Healthcare Mater. 2020, 2001239 2001239 (1 of 23)
support, chemical, and biological cues at
a spatiotemporal precision. The ECM can
modulate cell adhesion, cell-to-cell com-
munication, and differentiation, via chang-
ing its composition and therefore physico-
chemical and biological properties.[1] Given
the vital role of ECM in cell modulation,
ECM-based materials have been widely ex-
plored in tissue engineering and cancer
therapy.[2–5] Despite the functionalities of
natural ECM and the development of tissue
decellularization methodology, the biomed-
ical application of ECM is hampered by its
limited source.[6,7] Besides, the complexity
in the composition of ECM also makes fur-
ther engineering challenging.
As an alternative strategy, synthetic hy-
drogels have been explored as mimicry of
natural ECM.[8–10] Biopolymers, synthetic
polymers, and supramolecular hydrogelators have been uti-
lized to create such functional hydrogels.[11–18] Compared with
hydrogels formed by covalent crosslinking, supramolecular
hydrogels formed by non-covalent and dynamic chemical bonds
are unique for their reversibility and simplicity in molecular
design.[18–21] Non-covalent bonds (1–5 kJ mol−1 ) are several
orders of magnitude weaker compared with covalent bonds
(600–4000 kJ mol−1 ), and thus can be tuned to respond to mild
changes in the temperature, ions, and biological cues.[22,23] For
example, DNA oligomers containing moieties can be easily
co-assembled into the supramolecular hydrogel, which could
thus recognize and respond to endogenous nucleic acids such
as microRNA. Supramolecular peptide hydrogels (SPHs) that
are formed by self-assembly of peptide and its derivatives have
additional advantages and broad applications in biomedical
engineering due to their intrinsic and tunable biofunctionality
and biocompatibility.[24–27] SPHs usually have shape-persistent
nanostructure with a high degree of internal order, which
is widely used in living systems. Importantly, short peptides
(e.g., cyclicRGDD FK) are able to exert significant biological
activities (such as promote cell adhesion) and can be easily
integrated into peptide hydrogelator design to create biofunc-
tional SPHs.[22,28] Moreover, small peptide hydrogelators can
be more easily degraded and excreted through kidneys. So far,
SPHs have been developed to mimic natural collagen,[29]
to enhance wound-healing,[30] to realize biochemicals-
responsive drug release,[31] to create optoelectronic biomaterials
and pressure sensors,[32,33] and to exert drug-free cancer cell
eradication.[34]
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Given the unique features, SPHs are ideal for local ther-
apy. Localized therapy is advantageous for high and prolonged
drug exposure in the lesions with low systemic side effects. Ex-
amples are drug-eluting beads and Gliadel wafer, which have
been approved for the treatment of hepatocellular carcinoma and
brain tumor, respectively. Among many local drug delivery sys-
tems, supramolecular hydrogels including SPHs receive increas-
ing attention because of their tunable mechanical properties,
stimuli-responsive drug release, and injectability.[24,27] While ef-
fective and safe in treating primary tumors,[35–37] conventional
supramolecular hydrogels that deliver drugs acting on cancer
cells usually have limited efficacy against metastatic tumors
in distant organs. Recently, the development of in situ pep-
tide gelation technology in response to kinase/phosphatase en-
ables the formation of hydrogels in metastatic sites.[38,39] In
addition, SPH-based localized treatments have been found to
elicit systemic antitumor activity by priming immune responses
against tumors.[40–43] These discoveries boost interest in develop-
ing SPHs for cancer therapy.
In this progress report, we first summarize peptide hydroge-
lators that have been reported so far and then highlight current
strategies that have been explored to modulate the mechanical,
chemical, and biological properties of the hydrogels. We then fo-
cus on the recent progress in SPH-based cancer therapy. The po-
tential of SPHs in clinical cancer therapy and the remaining chal-
lenges are briefly discussed.
2. Design of Supramolecular Peptide Hydrogel
Peptide is a class of biomaterials with high diversity. Eight-
thousand different tri-peptides can be composed using 20 types
of natural amino acids, but only a few of them can self-assemble
and form SPHs in aqueous solution.[44] For biomedical appli-
cations, longer peptides and further derivatization are required,
which make experimental optimization impossible. Thus, it is
necessary to start with known peptide hydrogelators and then
optimize the molecular design for specific purposes. In this sec-
tion, we summarized well-established peptide hydrogelator sys-
tems and the methodologies for SPH property control.
2.1. Hydrogelator Design and Hydrogel Formation
The formation of SPHs starts with self-assembly of peptide hy-
drogelators and requires balanced attractive and repulsive forces
between the resulting assemblies that are dominantly filaments.
In general, the self-assembly of peptide hydrogelators is driven by
intermolecular attraction forces such as hydrogen bonding, hy-
drophobic interaction, chelating effect and 𝜋–𝜋 stacking, which
are mainly balanced by electrostatic and solvation forces. The en-
tanglement of filaments on the contrary mainly mediated by salt
bridge. These forces can be encoded and finely tuned by adjust-
ing peptide sequence and auxiliary moieties.[45] According to the
chemical structure of the building blocks, peptide-based gelators
can be divided into five categories including poly-/oligo-peptide,
peptide amphiphile, aromatic peptide, peptide-drug amphiphile,
and other peptide derivatives (Figure 1).
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2.1.1. Poly- and Oligo-Peptides
Peptides with specific sequences can naturally form hydrogels.
Two such examples are elastin and collagen, which are abun-
dant in connective tissues of animals. Elastin is a highly elastic
protein characteristic for its repeated VPGVG sequence. Chilkoti
and colleagues created an elastin-like peptide (ELP) with repeated
VPGVG segments, and initially used ELP as a tag to facilitate
protein purification through a process termed as “inverse tran-
sition cycling,” as the ELP tag could aggregate and form hydro-
gel under physiological conditions.[46] Given the unique nature
of ELP, the same group recently designed a thermally respon-
sive hydrophobic elastin-like polypeptide (VPGVG)120 , which was
further fused with a death receptor agonist (a hexameric protein
composed of oligomers of the 3rd fibronectin type III domain
of tenascin that is engineered to bind death receptor 5 [DR5])
to create an injectable in situ hydrogel for cancer therapy.[47]
While ELP is an intrinsically disordered peptide, collagen has
well-defined secondary and tertiary structures.[48,49] Collagen is
the most abundant protein in human ECM, and has characteris-
tic proline-hydroxyproline-glycine (Pro-Hyp-Gly) repeating units.
Although as a natural component of ECM, the application of col-
lagen is hindered by its limited source and difficulty in qual-
ity control. To create a synthetic version of collagen, Hartgerink
and colleagues created a class of poly-peptides composed of Pro-
Hyp-Gly repeating units.[50] The most successful version of these
synthetic collagen mimics is (Pro-Lys-Gly)4 (Pro-Hyp-Gly)4 (Asp-
Hyp-Gly)4 , which could form salt-bridged hydrogen bonds be-
tween lysine and aspartate to stabilize the triple-helix structure
in a sticky-ended assembly.[29] The peptides could replicate the
self-assembly of collagen and assemble into nanofibers with char-
acteristic triple-helical packing and length with a lower bound of
several hundred nanometers, which further entangled with each
other to give a hydrogel. The protein-derived peptide hydrogela-
tors typically have good biocompatibility. However, only a small
portion of natural proteins form hydrogel, and thus hydrogelators
with novel sequence must be explored to expand the hydrogelator
library.
In addition to long and protein-derived peptides, short and ra-
tionally designed peptides have been explored as hydrogelators
because of ease in preparation. Since hydrogen bonding in 𝛽-
sheet is directional and can facilitate the formation of 1D fib-
rils, many 𝛽-sheet forming peptides have been designed. The
ionic self-complementary peptide RADA16-I (COCH3 -(RADA)4 -
CONH2 ) designed by Zhang and colleagues is one excellent
example.[51] According to the sliding diffusion model, the pep-
tide will have two distinctive sides, one hydrophobic side with
an array of alanines and the other with alternating aspartic acids
and arginines. Driven by hydrophobic force and inter-molecular
ionic interactions, these peptides could adopt an anti-parallel 𝛽-
sheet structure and self-assemble into nanofibers, which then
formed hydrogel consisting of >99.5% water. Another such ex-
ample is the MAX1 (V(KV)4 D PL PT(KV)4 -NH2 ) peptide that can
adopt 𝛽-hairpin structures with a lateral and facial association,
which can form hydrogel after the addition of Dulbecco’s Mod-
ified Eagle’s Medium (DMEM) into the peptide solution.[52,53]
Schneider and colleagues further reported that the solution
pH required for hydrogel formation could be finely tuned by
engineering net charge of the peptide and exact position of
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Figure 1. Representative sequence and structure of peptide-based hydrogelators. These hydrogelators can be mainly divided into four categories includ-
ing poly-/oligo-peptide, peptide amphiphile, aromatic peptide, and peptide-drug amphiphile.
substitution within the peptide sequence.[54] A point substitu-
tion of one lysine residue of amphiphilic 𝛽-hairpin peptides
MAX1 with glutamic acid (MAX8, V(KV)4 D PPTKVEV(KV)2 -NH2 )
was sufficient to decrease the critical pH at which the peptide
forming hydrogel. Amino acids with phosphorylated side group
could also be integrated into the sequence, which enabled enzy-
matic dephosphorylation to control SPH formation.[55] In addi-
tion to 𝛽-sheet-forming peptides, those adopted 𝛼-helical struc-
tures are also able to form hydrogel with high water content.
First of this class was reported by Woolfson and colleagues,
and the hSAFAAA p1 ((KIAALKA)2 EIAALEAENAALEA) formed
purely 𝛼-helical fibrils, which entangled to give hydrogel with
>99% water.[56]
The discovery that dipeptide such as diphenylalanine (FF)
could also self-assemble into hydrogels propagates researchers
to find more short peptide hydrogelators.[57] As we mentioned
above, the huge sequence space of peptide is one major hurdle
for experimental screening. To address this limitation, Ulijn and
colleagues built a dynamic combinatorial peptide library based
on unprotected homo- and hetero-dipeptides.[58] These dipep-
tides were subjected to continuous enzymatic condensation, hy-
drolysis, and sequence exchange, and self-assembling candidates
would be amplified as a result of free-energy change associated
with the assembly process. More importantly, it was able to pur-
posely select different peptide sequences and nanoscale mor-
phologies by varying environmental conditions of screening. An-
other strategy to explore large peptide sequence space is com-
putational simulation. Ulijn and coworkers first explored the
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whole sequence space of tripeptide using coarse-grained simu-
lation considering the aggregation propensity and hydrophilic-
ity of different tripeptides.[44] The simulation screened out some
new tripeptide hydrogelators such as KYF, KYY, KFF, and KYW,
the gelation capability of which was further confirmed experi-
mentally. Despite its powerful capability in predicting peptide hy-
drogelators, the coarse-grained simulation requires the param-
eters of each residue to be defined, and is thus limited by cur-
rent understanding of the amino acid residues. Although already
a huge sequence space, the peptides can be further functional-
ized through palmitoylation, glycosylation, and phosphorylation,
which are widely adopted by nature.
2.1.2. Peptide Amphiphile
Palmitoylation is one important post-translational modification
of proteins, which regulates protein trafficking and intracellular
signaling.[59] Inspired by this process, researchers have generated
a huge class of peptide derivatives called peptide amphiphiles
(PAs), which were created by conjugating peptides with fatty
acids like myristic acid (C14 ) and palmitic acid (C16 ). Given their
amphiphilic nature, the PAs could spontaneously arrange into
nanosized structures in water, which process was driven by hy-
drophobic interaction. Stupp and coworkers reported the first
PA consisting of a long alkyl tail (C16 ), four consecutive cys-
teine residues, a flexible linker of three glycine residues, a sin-
gle phosphorylated serine residue and an Arg-Gly-Asp (RGD)
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sequence.[60] The resulting PA could form a nanostructured fi-
brous scaffold-like ECM which could be used for tissue regen-
eration. The directional packing of PA molecules was guided by
𝛽-sheets orienting parallel to the long axis of nanofibers, which
could be promoted by using a less branched peptide. The peptide
sequence was also important, but the first amino acid residue
following the alkyl segment plays a vital role in promoting 𝛽-
sheet formation and axial elongation into fibers.[61,62] These find-
ings suggest that the morphology, surface chemistry, and poten-
tial bioactivity can be engineered more freely by changing the
amino acid sequences. Indeed, the morphological of PA-based
fibrils could be tuned from short twisted ribbon, to long twisted
ribbons and to helical ribbons via engineering three phenylala-
nine residues.[63] The PAs could also be tailored to respond to
pH change and divalent ions,[64] and to display bioactive lig-
ands such as 𝛽-alanine-histidine carnosine dipeptide motif and
YEALRVANEVTLN.[65,66] In addition to the increased versatility
of peptide sequence, PAs also provide an additional hydropho-
bic space for potential drug encapsulation, which makes them
extremely useful in drug delivery.
2.1.3. Aromatic Peptide
The study on FF peptide suggested that aromatic interaction con-
tributes to the free energy of self-assembly and also guides di-
rectional molecular stacking. Given the vital role of 𝜋–𝜋 stack-
ing during self-assembly, many aromatic peptide hydrogelators
have been created. The most widely used aromatic moieties
for peptide modification are 9-fluorenylmethoxycarbonyl (Fmoc)
and naphthalene (Nap). Fmoc-FF is an aromatic peptide that
has been vastly studied. Gazit and colleagues first synthesized
Fmoc-FF and found that this molecule could form a rigid ma-
terial with macroscopic characteristics of a gel at a concentra-
tion of <1 wt%.[67] The observation was confirmed by Ulijn and
colleagues, who first suggested that the hydrogel was formed
through 𝜋–𝜋 interlocked 𝛽-sheets.[68] The Fmoc-FF contains
a carboxyl group, and thus requires an alkaline environment
for self-assembly, which may not be physiologically relevant.
Nilsson et al. thus created a set of C-terminal cation-modified
Fmoc-F derivatives that were positively charged across a broad
range of pH values, which could form sheet-based nanotubes
and hydrogel at physiological pH.[69] In addition to Fmoc-FF,
Ulijn et al. also designed a series of Fmoc-dipeptides and found
that both the sequence and position of dipeptide would affect
self-assembly.[70] For tyrosine-containing peptide hydrogelators
(Fmoc-YX-OH), alkaline phosphatase (ALP) responsive hydro-
gel formation could be realized by design a hydrogelator precur-
sor (Fmoc-pYX-OH).[71] Very recently, a novel aromatic dipeptide
(Fmoc-Lys(Fmoc)-Asp) was reported by Gazit and coworkers.[72]
Due to the strong intramolecular and intermolecular aromatic in-
teraction, the reported molecule had an ultra-low critical gelation
concentration (0.002 wt%, 28.3 × 10−6 m). More importantly, it
showed a novel two-step assembly process, starting from sphere-
based opaque hydrogel to filament-based transparent hydrogel.
In addition to Fmoc, other aromatic molecules such as naph-
thalene have also been used to create aromatic dipeptide and
oligopeptide.[73] When phosphorylated tyrosine was introduced
into the molecule, ALP-responsive hydrogelation could also be
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realized.[74] In the above cases, the aromatic groups only con-
tributed to self-assembly. Ulijn and coworkers conjugated an or-
ganic chromophore diketopyrrolopyrrole (DPP) with two amino
acid (Phe, Tyr, Leu) ethyl esters (DPP-(XOMe)2 ), which could
form hydrogel at the presence of 𝛼-chymotrypsin.[75] The result-
ing hydrogel could produce 1 O2 as a result of DPP-mediated pho-
todynamic reaction. Li’s group set up a machine learning sys-
tem and trained it with a structurally diverse hydrogel library
with over 2000 peptides with characterized chemical features,
including topological and physicochemical properties.[76] They
then utilized this trained system to predict the self-assembly be-
havior of new peptide-like molecules, and successfully obtained
81 peptide-like hydrogelators, expanding the library of aromatic
peptide hydrogelator.
2.1.4. Peptide-Drug Amphiphile
Modifications of peptide backbone with fatty acids and aro-
matic moieties mainly affect their self-assembly. By conjugating
functional molecules such as drugs to peptide could introduce
additional bioactivities while modulating self-assembly. Camp-
tothecin (CPT) is one drug that has been widely investigated
for peptide-drug amphiphile development because of its potent
antitumor activity and planar structure facilitating 1D molecu-
lar packing. Cui and coworkers first tested this concept by con-
jugating CPT to a 𝛽-sheet-forming peptide sequence (VQIVY)
derived from the Tau protein through a reducible disulfylbu-
tyrate (buSS) linker.[77] Since the successful proof-of-concept
study, the group dedicated to proving the versatility and tun-
ability of this particular platform. They demonstrated that dif-
ferent numbers of CPT molecules could be conjugated with one
peptide to modulate drug loading and morphology without af-
fecting 1D molecular packing.[77] Since different cancer cells
may express distinct receptors, the formation of hydrogel should
be ideally independent of peptide sequence. Indeed, peptide of
various sequences such as hydrophilic 𝛽-sheet-forming peptide
GV2 Q2 HKD-OH and hydrophilic and cyclic peptide iRGD could
be used instead of hydrophobic 𝛽-sheet-forming peptide VQIVY
to realize pH-responsive hydrogel formation and enhanced tis-
sue penetration of both peptide-drug amphiphiles and encapsu-
lated drugs.[78,79] In addition, linkers that could respond to dif-
ferent stimuli could also be easily integrated into the molecule
design to enable controlled drug release at the presence of glu-
tathione (disulfylbutyrate or disulfanyl-ethyl carbonate linker) or
matrix metalloproteinase-2 (MMP-2 cleavable PLGLAG).[78–80] In
these examples, the hydrophobic interaction and 𝜋–𝜋 stacking
among CPT molecules are critical for peptide-CPT amphiphile
self-assembly into nanosized filaments such as fibers and nan-
otubes, and the salt bridge and depleted electrostatic repulsion
mediated by multivalent ions in the solution are key for hydrogel
formation.
In addition to CPT, other drug molecules have also been ex-
plored. Parquette and coworkers created a 5-fluorouracil (5-Fu)
based peptide-drug amphiphile by conjugating 5-Fu to the 𝜖-
amine of the C-terminal lysine residue of Fmoc-KK via succinate
or acetamide linkage.[81] Driven by the hydrophobic 𝜋–𝜋 associ-
ation between uracil and Fmoc groups, the molecules could self-
assemble into nanofibers or nanotubes, which further entangled
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to form self-supporting hydrogels. The self-assembly of a chlo-
rambucil (CRB)-based peptide-drug amphiphile has been inves-
tigated by Yang and colleagues. By conjugating CRB to a phos-
phorylated peptide GD FD FpD Y, the resulting conjugate, CRB-
GD FD FpD Y, only self-assembled into stable nanofibers and hy-
drogels after the removal of phosphate group by ALP due to
weakened electrostatic repulsion between the molecules.[82] Non-
steroidal anti-inflammatory drugs have also been used to cre-
ate peptide-drug amphiphile-based hydrogels. Xu and colleagues
conjugated naproxen to the N-terminus or 𝜖-amine of lysines
of a set of peptides (D FD F, D FD FD Y, D FD FD K, or D FD FD KD Y).[83]
All the peptide-naproxen amphiphiles self-assembled into fila-
ments and formed self-standing hydrogels due to the strong 𝜋–𝜋
stacking between the drug molecules and side chains of pheny-
lalanines. Zhong and coworkers conjugated naproxen to short
peptide dendrons (E3 FID and E3 FNP), and the resulting conju-
gates could self-assemble and form hydrogel at the assistance
of zinc ions.[84] Similarly, Chen et al. successfully created an
(S)-ketoprofen (Ket) based hydrogel by conjugating Ket to either
L- or D-VEVE peptide.[85] In these cases, all the drugs were of
small molecular weight and planar structure, which would fa-
cilitate molecular packing through 𝜋–𝜋 stacking. Cui and col-
leagues recently conjugated a bulky anticancer drug paclitaxel
to the GGRGDRPLGVRVVV peptide to obtain a peptide-drug
amphiphile, which could also form a self-supporting reverse
bolaamphiphile (RBA) hydrogel at the presence of MMP-2 in
PBS.[86] This result demonstrated that drug of different chemical
properties can all potentially be engineered to become a hydroge-
lator through proper peptide conjugation.
2.1.5. Other Peptide Derivatives
In the above cases, amide bond of peptide contributes to the
self-assembly of the hydrogelators. For instance, Hartgerink
and coworkers showed that hydrogen bond among the first
amino acid residues following hydrocarbon tail is critical for
self-assembly of peptide amphiphiles.[87] Peptidomimetic hydro-
gelators with various backbone amide modifications have been
investigated, giving rise to hydrogels of diverse properties and
increased stability.[88] Given the important role of interpeptide hy-
drogen bonding, Gazit and colleagues replaced the amide bond
in Fmoc-FF peptide with the urea moiety, which increased hy-
drogen bonding capabilities between the molecules. The urei-
dopeptide, Fmoc-F-NHCONH-F-OH (Fmoc-FuF), formed hydro-
gels with improved mechanical properties compared with Fmoc-
FF, with additional benefits such as anti-fouling and controlled
release of urea stemmed from the urea moiety.[88] Another cate-
gory of peptide-mimetic materials is peptoid, the side chain of
which is appended to the nitrogen atom of the peptide back-
bone instead of the 𝛼-carbons. Peptoid is more flexible than pep-
tides since intramolecular CO–HN hydrogen bonds are removed.
After specific engineering, tripeptoids could also form hydrogel
with the assistance of organic solvents.[89] In addition to the nat-
ural peptides, ureidopeptides and peptoids greatly expanded the
territory of peptide-derived materials, providing enormous pos-
sibilities to obtain functional hydrogels for specific biomedical
applications.
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2.2. Hydrogel Characterization and Property Control
Hydrogels formed by different hydrogelators may have very dis-
tinct properties. Besides, different functional molecules could be
encapsulated either covalently or non-covalently. Given that the
ECMs at different organs are distinct from each other in prop-
erties such as mechanical strength, stimuli-responsive decom-
position, and controlled release of active molecules, hydrogels
that can be finely tuned in these properties are highly required
(Figure 2).
2.2.1. Morphology
The morphology of a nanoparticle (e.g., size and shape) has
great influences on its interaction with biomacromolecules and
cells.[90,91] Though most hydrogels are formed by the entangle-
ment of 1D filaments, they can be in the form of nanofiber, nan-
otape, or nanotube, which are distinct in their surface property
and molecular organization. In addition, the filaments are un-
der dynamic changes, which is characteristic of supramolecule
structures. An effective strategy to control the morphology of the
filaments is varying the chemical structure of peptide hydrogela-
tor. For instance, C16 -VEVE self-assembled into nanobelts, while
C16 -VVEE, C16 -EVEV, and C16 -EEVV tended to form singular
nanofibers, ribbons, and interwound nanofibers, respectively.[92]
The addition of GRGD sequence at the C-terminus of the pep-
tide amphiphile did not affect the morphology of the filaments
and further hydrogel formation. Non-peptidic segments could
also influence the morphology of the filaments. Cui and col-
leagues conjugated one, two, or four CPT molecules to one
VQIVY peptide, and the drug amphiphile with four CPT seg-
ments self-assembled into nanotubes, while the rest two formed
nanofibers.[77] Aside from tuning the molecular structure of pep-
tide hydrogelators, drug loading was also able to affect the mor-
phology of the self-assemblies. Cui and colleagues reported that
peptide-paclitaxel conjugate (PTX-buSS-CGN2 Q2 (KOEG )4 , Spax)
could form filaments only at the presence of free PTX.[93] Li and
colleagues showed that the morphology of the filament could also
be modulated by the kinetic control of self-assembly pathway.[94]
The Fmoc-GF2 Q2 NYKD-OH self-assembled into nanofibers and
hydrogel with low mechanical strength at room temperature, but
formed twisted ribbons and self-supporting hydrogels after being
heated for 2 h at 70 °C. The capability of modulating morphology
of the filaments will provide a feasible way to adjust the molecular
display of ligand and anchor points on their surfaces.
2.2.2. Mechanical Property
Hydrogels are distinct from solutions in their mechanical prop-
erty that is important for their biomedical applications. As natural
hydrogels, ECMs in different parts of human body show different
mechanical properties. The storage modulus of the soft tissues
range from 103 Pa (in the brain, lung, and kidney) to 105 Pa (in the
bladder).[95] The stiffness of the ECMs is mainly determined by
the concentrations of ECM fibers including elastic fibers and col-
lagen fibers and is under dynamic change over time in response
to physiological and pathological stimulus.[96] These properties
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Figure 2. Characterization and major properties of SPH. Some methods to modulate the properties are mentioned. The hydrogelators can self-assemble
into different filamentous structures, and form SPH of different and controllable mechanical, chemical, and biological properties.
are associated with cell differentiation and functions in particu-
lar tissues.[97,98] Therefore, it is critical to adjust the mechanical
properties for the purposed biomedical applications. To quantify
the mechanical strength of hydrogels, storage modulus G′ and
loss modulus G″ are the most widely used parameters. G′ reflects
the rigidity of hydrogels, and is closely related to its injectabil-
ity, tissue retention capacity, and cell interaction. It is sensitive to
changes in micro- or nano-structure of viscoelastic systems and
thus can be modulated at both scales. G″ describes the energy
transformed into heat when a material is deforming and reflects
the viscosity. The mechanical property of a hydrogel can be mod-
ulated via changing hydrogelator structure and combination, as
well as components in the solution environment (Figure 3).
Engineering chemical structure of hydrogelator is one strat-
egy to modulate the mechanical property of a hydrogel. Since the
hydrogels were formed by 1D filaments, Stupp and colleagues
explored the effect of strength of 𝛽-sheet on the stiffness of the
resulting hydrogel.[99] They created a set of peptide amphiphiles
with various number of valines and alanines at different positions
relative to the hydrocarbon tails, and found that valine positioned
at proximity to the hydrocarbon tail could increase the stiffness
and mechanical property (e.g., two alanine replaced with valines
in the hydrocarbon tail increased the G′ by tenfold). Similarly, Xu
and colleagues substituted the valines of MAX1 sequence with
isoleucines that had a higher propensity for 𝛽-sheet hydrogen
bonding to obtain MAX1I8 ((IK)4 VD PPT(KI)4 -NH2 ).[100] MAX1I8
showed a faster gelation process than MAX1 (30 s vs 75 min to
reach a G′ of 100 Pa), and formed hydrogel with a G′ as high as
1320 Pa after 2 h. However, Muraoka and coworkers showed that
substitution of alanine with glycine (A8G, RADARADA(RADA)2 )
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in the middle of the RAD16 peptide ((RADA)4 ) increased the stiff-
ness of the hydrogel (G′RADA16 = 265 Pa, G″RADA16 = 42 Pa; G′A8G
= 469 Pa, G″A8G = 56 Pa).[101] The enhanced stiffness of A8G
probably resulted from the formation of twisted 𝛽-sheet struc-
tures and nanofiber bundles due to a transition in the molecu-
lar packing. These results suggested that a balanced rigidity and
flexibility of filament should be reached to achieve high stiff-
ness. In addition to the peptide sequence, Hamley and cowork-
ers also found that terminal modification of a peptide hydrogela-
tor could also significantly affect the G′ of the hydrogel.[102] They
showed that N-acylation and C-termini histidine of peptide H2 N-
VEQLTEEQKNEFKAAFDIFVLGA-OH increased the G′ by three-
fold to a value of ≈13 kPa. Lin et al. explored the effect of iso-
meric hydrocarbons (C7 ) on the stiffness of hydrogel, and found
that the G′ decreased with an increase in the ring size of the tail
through regulating molecular packing at the interface between
the peptide domain and the hydrophobic core.[103] In addition to
the composition of the hydrogelator, Schneider and colleagues
further demonstrated that the chirality of a peptide hydrogelator
could be used as a design tool to control the mechanical rigidity
of peptide-based hydrogels.[104] An enantiomeric peptide mixture
of MAX1 and DMAX1 showed threefold greater rigidity than that
of hydrogel prepared from either of pure enantiomer because of
energetically favorable interactions between enantiomers.
Engineering structure of hydrogelator, while effective in mod-
ulating the stiffness of the hydrogel, may also affect other proper-
ties of the hydrogel. Since the interaction between the filaments
is another important factor that determines the mechanical prop-
erty of the hydrogel, a simple yet effective way to modulate hy-
drogel stiffness is manipulating ionic strength that can affect
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Figure 3. Peptide design for modulation of mechanical property of SPHs.
electrostatic interactions. Indeed, Williams and coworkers found
that G′ of the Fmoc-FRGDF hydrogel increased from 10 to 3200,
7500, and 11 000 Pa in PBS solutions of 0.05, 0.25, 0.5, and 0.75
m, respectively.[105] Given that the concentration of salts must be
limited within a tolerable range for biomedical application, Stupp
and coworkers used oligo-L-lysines of different molecular lengths
to precisely tune anionic gel stiffness and found that G′ increased
by 10.5 Pa for each additional lysine monomer in the oligo-
L-lysine chain.[106] In addition to these small molecular cross-
linkers, macromolecules and even nanoparticles could be used
to modulate the mechanical property of the hydrogels through
non-specific or specific intermolecular interactions. Huang and
coworkers demonstrated that mixing ovalbumin (OVA) protein
with Nap-HHFF peptides can enhance the mechanical property
up to 15-fold.[107] Nilsson and colleagues utilized specific molec-
ular recognition between mannose-modified fibrils with con-
canavalin A (ConA) to modulate hydrogel stiffness, and found
that the G′ of mannose-functionalized Ac-(FKFE)2 -NH2 peptide
hydrogels increased from 148 ± 9 to 466 ± 61 Pa after the ad-
dition of ConA.[108] Ulijn and coworkers further created a spe-
cific and multivalent nano-crosslinker by decorating magnetic
nanoparticles with thermolysin, which could generate hydroge-
lator Fmoc-TF-NH2 in situ.[109] This resulted in up to a tenfold
increase (from 1.9 × 103 to 15 × 103 Pa) in G′ of the hydrogel
compared to a conventional soluble enzyme system. Crosslinkers
could also be introduced into filaments by co-assembly strategy.
For instance, Ulijn and coworkers co-assembled Fmoc-FF with
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surfactant-like Fmoc-S in different concentrations to produce hy-
drogels with tunable stiffness (1, 13, and 32 kPa).[110] Fmoc-R,
another surfactant-like molecule, has also been used to increase
the Fmoc-FF-based hydrogels.[111] The above strategies used non-
covalent crosslinkers, and Guler and colleagues explored the ef-
fect of glutaraldehyde, a covalent crosslinker, on the stiffness of
lauryl-VVAGKK-Am (K2 -PA) hydrogel.[112] The G′ of the hydro-
gel was significantly increased after the addition of glutaralde-
hyde, which could exceed 105 Pa. More importantly, the viscoelas-
tic properties of the hydrogel could further be tuned by changing
ratios of glutaraldehyde or addition other dialdehydes of differ-
ent molecular length. Pyridoxal-5-phosphate (P5P) has also been
investigated by Xu and colleagues to modulate the mechanical
property of the SPH.[113]
2.2.3. Chemical Property
Natural ECMs are under constant chemical modification to ex-
ert desired functions. Therefore, SPHs need to exert stimuli-
responsive chemical property to maximize their activities in
biomedical applications. A feasible strategy to control the
chemical properties of a hydrogel is to introduce stimuli-
responsive moieties into the molecular design (Figure 4). Light-
responsive groups are those under broad investigation, as chem-
ical reactions could be easily triggered by irradiation.[114–116]
Schneider and colleagues reported a self-assembly hydrogel
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Figure 4. Peptide design for modulation of chemical property of SPHs. The letter (X) represents GSH, NO, or H2 S.
composed of 𝛽-sheet-forming peptides, the lysyl sorbamide side
chains of which could polymerize after exposure to ultraviolet
(UV) radiation.[117] Instead of lysyl sorbamide, flavin mononu-
cleotide (FMN) and [3-(3-aminomethylphenylazo)phenyl] acetic
acid (AMPP) have also been used to create peptide hydrogel with
light responsiveness.[118,119]
For biomedical applications, it is ideal to create hydrogels that
can respond to biological cues such as reducing agents, enzymes,
or other biomolecules.[120] Nitric oxide (NO) and hydrogen sul-
fide (H2 S) are molecules that have important biological func-
tions. To create peptide hydrogels that could respond to these
molecules, Yang and coworkers designed a short peptide, GFFY,
with self-immolative aromatic groups to yield supramolecular hy-
drogelators that could form hydrogels. The self-immolative cap-
ping group could be removed in the presence of NO, H2 S, or
GSH, causing gel–sol phase transition.[121] Tumor microenviron-
ment is characteristic for enhanced expression of protease such
as matrix metalloproteinases (MMPs). MMP-responsive hydro-
gels could thus be potentially used in cell behavior control and
tumor-specific drug delivery.[20] Xu and colleagues designed an
MMP-2 responsive peptide hydrogelator (Ac-I3 SLKG-NH2 ) by in-
corporating MMP-2 substrate sequence into molecule design.[122]
Upon exposure to MMP-2 overexpressing HeLa cells, the self-
assembled nanofibers and the entangled hydrogel network can be
significantly disrupted. A similar design was reported by Cui and
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colleagues, and the hydrogel filaments could rapidly break into
fragments prior to reassembling into spherical micelles upon
MMP-2 exposure.[86]
2.2.4. Bioactivity
Hydrogel such as ECM is not merely a scaffold providing
space for cells, but more importantly, provides biological sig-
nals to tune the behavior of cells. Natural ECM fulfills this task
through two strategies. First, ECM incorporates functional lig-
ands such as RGD-containing domain into the filaments. Sec-
ond, ECM recruits cells to functionalize the matrix. Similar
strategies have been employed to create biomimetic hydrogels
(Figure 5). For instance, Stupp and coworkers incorporated a
laminin-mimetic IKVAV sequence into their peptide amphiphile
design (C16 V2 A2 E4 GIKVAV), creating a supramolecular hydro-
gel for neural transdifferentiation.[123] Yang and colleagues mod-
ified their peptide hydrogelator with folic acid (FA) to generate
FA-functionalized hydrogel, which significantly improved the re-
tention and survival of pluripotent stem (iPS) cells.[124] Li and
colleagues reported a new peptide gelator (Nap-FFRGD) to en-
capsulate vascular endothelial growth factor (VEGF), and the re-
sulting hydrogel enhanced blood capillary density in mice.[125]
In addition to a direct chemical modification to the chemical
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Figure 5. Peptide design for modulation of bioactivity of SPHs.
structures,[126] co-assembly strategy was also explored to func-
tionalize SPHs. For instance, Ulijn and coworkers reported the
co-assembly of Fmoc-FF and Fmoc-RGD to construct small-
molecule-based bioactive hydrogels through 𝜋–𝜋 stacking of
Fmoc groups.[127] The same group also co-assembled Fmoc-FF
with aromatic carbohydrate to generate minimalistic proteogly-
can mimics, which could prolong the stability of growth factors
and preserve the viability of cells in the SPH.[128] By the modi-
fication of fatty acids and aromatic rings, the range of gelators
has been extended. Interestingly, Gazit et al. combined Fmoc-FF
peptide with a conductive polymer polyaniline (PAni) to form a
two-component and conductive hydrogel, which could be used
for dynamic range pressure sensing and as a conductive inter-
face for electrogenic cardiac cells.[32]
In contrast to the above strategies, Hartgerink and colleagues
investigated the infiltrated cell-mediated K2 (SL)6 K2 peptide hy-
drogel remodeling.[129] The K2 (SL)6 K2 hydrogel could induce
rapid cell infiltration, and then be enriched with natural collagen-
containing ECM and a wide range of cytokines and growth fac-
tors produced by the infiltrating cells, which could support cells
present in the wound healing process.[130] To study the effect
of ionic charged domains at the N- and C-termini, they further
designed a series of multidomain peptides (MDPs) including
K2 (SL)6 K2 , R2 (SL)6 R2 , E2 (SL)6 E2 , and D2 (SL)6 D2 . All the studied
MDPs showed similar nanostructures and physical properties,
but they trigger markedly different inflammatory responses. The
negatively-charged peptide hydrogels elicited minimal inflam-
mation characterized by tissue-resident macrophage infiltration,
fast remodeling, and no collagen deposition or blood vessel for-
mation within the implants. In contrast, the positively-charged
peptides were highly infiltrated by immune cells, were remod-
eled at a slower rate, would promote angiogenesis, and resulted
in a high degree of collagen deposition.[131] These studies showed
that the biological activity of the peptide-based hydrogels could be
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modulated at different levels from molecular design to hydrogel
composition, which facilitates their biomedical applications.
2.3. Control of Cargo Loading and Release
As we discussed above, peptide hydrogel has intrinsic biological
activities. These activities could be further enhanced by incorpo-
rating additional biologically active components. These compo-
nents can be small-molecule drugs, protein therapeutics, gene
therapeutics, or even cells, and the resulting hydrogels have
been used in cancer therapy, regenerative medicine, and immune
engineering.[62,132,133] Functional components can be introduced
into the hydrogels through either physical entrapment or chemi-
cal conjugation (Figure 6). Effective cargo loading and its control-
lable release are key factors to maximize their efficacy.
2.3.1. Physical Entrapment
SPHs are characteristic of their non-covalent nature, mesh-like
structures, and capability of immobilizing liquid. Therefore, the
cargoes can be easily entrapped within the filament network
through mixing, and their release from the SPHs could be ad-
justed via tuning interactions between the cargoes and filament
network. Schneider and colleagues revealed that the size of the
cargoes and their electrostatic interactions with hydrogel net-
works are two important factors influencing release. They devel-
oped a multi-compartment hydrogel composed of EGFR kinase
inhibitor Erlotinib (ERL) and Doxorubicin (DOX) vesicles, and
found that the ERL was released in early time due to its small
molecular size while the larger DOX vesicles showed a much
slower release. The sequential treatment (first ERL and then
DOX) first inhibited EGFR signaling, which sensitized LN229
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Figure 6. Methods of loading cargos into SPHs. Cargoes including small-molecule drugs, biomacromolecules, nanoparticles, and even cells can be
easily entrapped within the filament network through physical entrapment and chemical conjugation.
glioblastoma cells to the subsequent DOX.[134] Similar result was
also observed when Zhang and colleagues investigated the ef-
fect of protein size on their diffusion out of the SPH using
single-molecule fluorescence correlation spectroscopy (FCS).[135]
To understand the effect of electrostatic interaction on drug re-
lease, the same group created a negatively charged SPH (Ac-
VEVSVSVEVDPLPTEVSVEVEV-NH2 , AcVES3) and a set of en-
hanced green fluorescent protein (EGFP) analogues with dif-
ferent positively charged interaction domains (IDs) at their N-
termini.[136] These IDs exerted weak, medium, or strong affin-
ity for the fibrils, realizing controlled release of EGFP from the
AcVES3 SPH. Wang et al. even showed that a strong electro-
static interaction between the cargoes and dense hydrogel net-
work would even retain cargo enzyme chloroperoxidase (CPO)
in the Fmoc-Y-OH-based SPHs for more than two weeks without
significant release.[137]
In contrast with biomacromolecules and nanoparticles that
are large and multi-valent, small molecular drugs must have a
strong affinity for hydrogelators in order to be retained in and
sustainedly released from SPHs. Nilsson and coworkers cre-
ated an SPH self-assembled from Fmoc-F5 -Phe-DAP (DAP, di-
aminopropane) for localized delivery of diclofenac.[138] Due to
the 𝜋–𝜋 interaction between the altered quadrupole in the aro-
matic benzyl side chain of the gelator and aromatic rings of
the diclofenac, sustained release of the drug from SPH over
two weeks in vivo after a single injection was achieved. Li
and colleagues explored the influence of hydrogelators’ struc-
ture on drug encapsulation efficiency.[94] They created a set
of peptide hydrogelators with different peptide sequences and
hydrophobic tails (C16 -GN2 Q2 NYKD-OH, Fmoc-GN2 Q2 NYKD-
OH, and Fmoc-GF2 Q2 NYKD-OH) for encapsulation of losar-
tan. They showed that the drug loading was the highest in C16 -
GN2 Q2 NYKD-OH, the self-assembly of which contained a large
hydrophobic cavity for losartan. However, Pochan et al. showed
that vincristine, a hydrophobic chemotherapeutics, could also be
encapsulated within MAX8-based SPH with constant drug re-
lease over the course of one month.[139] Since MAX8 did not con-
tain hydrophobic tail, their findings suggested that the side chain
of MAX8 may contribute to shielding vincristine from surround-
ing environment.
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Another strategy to control the release of entrapped car-
goes is to disrupt mesh structure. Given the large size of the
mesh in the SPHs, it is ideal for entrapment and controlled re-
lease of biomacromolecules and nanoparticles. Hamachi’s group
used a H2 O2 -responsive supramolecular hydrogel as the uni-
versal matrix, coupling with enzymatic reactions with the gel
response.[31] The hydrogel could response to input stimuli like
H2 O2 molecules and various disease-related biomarkers, includ-
ing glucose, lactose, and uric acid, which could be converted
into H2 O2 by the hydrogel-enzyme hybrids. They combined two
different gelators BPmoc-F3 and NPmoc-F2 that could respond
to oxidation and reduction, respectively, and built Boolean logic
gates (OR and AND) into materials. The addition of both NADH
and glucose (AND logic-regulated) induced a gel–sol transition
and release about 90% of the initially encapsulated drugs. In com-
parison with hydrogel network limited or hydrophobic interac-
tion controlled drug release mechanism, gel–sol transition me-
diated drug release is more suitable for stimuli-responsive bolus
drug release.
2.3.2. Co-Assembly
Co-assembly is another important strategy to realize supramolec-
ular encapsulation and controlled release. This strategy has
been utilized to combine B and T epitopes by conjugating the
epitopes to Q11 (Ac-QQKFQFQFEQQ-Am) peptide domain re-
spectively, achieving a high-titer antibodies and a modular T-
cell response.[140] Similar strategy has been reported by Yang
and colleagues for delivery of phosphorylated antigens such as
TpSN, CpTW, and KpTL.[141] The hydrogel slowly released the
co-assembled antigens and protect them from early dephos-
phorylation. In addition to peptides, the same group also ex-
plored the co-assembly of proteins with SPH. In one such exam-
ple, they modified antiHER2 affibody with hydrophobic drugs,
which co-assembled with NBD-𝛽A-GD FD FpD Y (NBD: 4-Chloro-
7-nitrobenzo-2-oxa-1,3-diazole) to realize high affibody loading
and prolonged release.[142] In another example, they demon-
strated the co-assembly of flurbiprofen (Fbp)-modified peptide
Fbp-GD FD FD YD K(𝛾E)2 -NH2 with protein antigens such as OVA
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and hepatitis B surface antigen (HBsAg).[143] During the co-
assembly between peptide and protein antigens, OVA or HB-
sAg served as coacervation stimulators to induce hydrogelation
and colocalized onto peptide nanofibers. Compared with pro-
teins that were physically encapsulated in hydrogels, antigen re-
lease from co-assembled nanofibers was much slower. In addi-
tion to biomacromolecules, small molecules could also be loaded
through the co-assembly strategy, in which the co-assembled
drugs also played essential roles in hydrogel formation. For
instance, Zhong and colleagues reported that DOX could co-
assembled with peptide Nap-GFFYGRGDH by electrostatic in-
teraction to form hydrogel which would release the DOX under
weakly acidic conditions.[144] The same group also found that cis-
platin (CDDP) could co-assemble with Nap-FFYERGD peptide
to form a rigid hydrogel.[145] In the above two cases, the two aro-
matic peptides themselves were not able to form rigid hydrogels.
2.3.3. Chemical Conjugation
Physical entrapment, while a feasible strategy, usually shows
a fast drug release. Chemical conjugation provides a precisely
controlled method to achieve high drug loading and prolonged
drug release. In the peptide-drug amphiphile section, the re-
quirement for drug and peptide has been discussed in detail.
In this section, we mainly focus on the design of crosslinker,
which must be reversible and stimuli-responsive, as most drugs
must be released as their parental forms in order to be active.
Hydrolysable ester bond is a widely used bond for drug conju-
gation. 1-naphthaleneacetic acid (NAA) that was conjugated to
peptide G′FFY (NAA-G′FFY) through an ester bond could slowly
release NAA in aqueous solution, which is in sharp contrast with
amide bond.[146] In addition to spontaneous hydrolysis, drugs co-
valently loaded into SPH through an ester bond could also be
specifically released at the presence of carboxylic-ester hydrolase
(CES) in the cells.[147] In these cases, the focus was given to the
release of free drug from the conjugate. Cui and colleagues de-
veloped a set of GSH-responsive peptide-drug amphiphiles, and
found that drug release from SPHs could be divided into two
steps, filament dissociation and free drug release. They first cre-
ated a peptide-paclitaxel conjugate with buSS linker (PTX-buSS-
G2 V3 RGDR), which could form self-supporting hydrogels.[148]
They showed that the release of hydrogelator monomers from
the SPH was controlled by their critical aggregation concentra-
tion (CAC). Since the CAC was constant, a linear release pro-
file could be achieved rather than burst release. The monomers
were converted into free PTX after their internalization into
cells. Given the low CAC of the monomer and limited release
medium, the hydrogel maintained a sustainable drug release
over the period of several months. This observation was further
confirmed on another peptide-drug amphiphile (camptothecin-
buSS-GV2 Q2 HKD, CPT-HKD) based SPH, which also showed
a steady and linear release of CPT.[78] The dis-assembly of fil-
aments could also be engineered to respond to extracellular
biomolecules. The same group further introduced an MMP-2
cleavable sequence into their molecular design.[86] MMP-2 could
accelerate filament fragmentation into spheres and short rods,
which could also be internalized by cells. This design signifi-
cantly enhanced the conversion of peptide-drug amphiphiles into
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PTX. However, the location of the protease-cleavable peptide in
the filament is important. Enzymatic cleavage events could be
inhibited if the peptide substrate was hidden in the assembled
structures.[149] In the above cases, the drug molecules mostly
located at the hydrophobic domain of the filaments. However,
functional peptide and amino acid could be integrated into the
peptidic segment of the hydrogelator. For instance, Hartgerink
and coworkers integrated a hydrophilic drug N6-(1-iminoethyl)-
L-lysine (L -NIL) (an inhibitor of inducible nitric oxide synthase,
iNOS) into K2 (SL)6 K2 MDP to give L -NIL-MDP, which showed
comparable activity in inhibiting inducible nitric oxide syn-
thase (iNOS) in vitro and in vivo.[150] Yang et al. used tuftsin
(TKPR) with Nap-GD FD FD Y to form a hybrid peptide compound
Nap-GD FD FD YTKPR, which formed hydrogel and enhanced the
maturation of DCs and phagocytosis of macrophages.[151] Sim-
ilarly, Ruokolainen et al. conjugated carnosine dipeptide (𝛽-
alanine-histidine, 𝛽AH) onto a KTT sequence from “Matrixyl”
lipopeptides to a lipopeptide C16KTT𝛽AH, which showed dose-
dependent cytotoxicity to MCF-7 cancer cells.[65] It is also possible
to conjugate proteins on the surfaces of filaments, which strategy
could be used to realize stimuli-responsive and sustained drug
release.
3. Supramolecular Peptide Hydrogel in Cancer
Therapy
As we have discussed above, SPHs are advantageous for tun-
able physicochemical properties and biological activity, good bio-
compatibility, and multiple spaces for both hydrophobic and hy-
drophilic molecules. Moreover, they could be further function-
alized to exert stimuli-responsive (dis)-assembly and drug re-
lease, thus making it a perfect candidate for drug delivery.[152]
Given their unique advantages, SPHs have been explored as both
supramolecular drug and biocompatible vector for localized and
systemic cancer therapy.[153]
3.1. Small Molecular Drug Delivery
Small molecular weight drugs are widely used in cancer therapy
due to their well-defined efficacy against a broad spectrum of can-
cers. However, most of them suffer from shortcomings such as
limited anticancer efficacy and serious systematic toxicity, which
are resulted from non-targeted distribution and short half-life of
the drugs in the tumors. Localized small molecular weigh drug
delivery with injectable SPHs could realize prolonged drug expo-
sure in the tumors, which can potentially increase efficacy and
lower systemic toxicity.
3.1.1. Delivery of Chemotherapeutics with SPH
The filament network of the SPHs can hinder the diffusion of
the entrapped molecules out of the depot, the capability of which
can be experimentally quantified by measuring the diffusion con-
stant of a model fluorophore through fluorescence recovery after
the photobleaching (FRAP) method.[154] Aside from the physi-
cal barrier, the rate of drug diffusion is also influenced by non-
covalent interaction between the encapsulated drugs and the net-
work. Nie and colleagues designed a hexapeptide hydrogelator
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(FEF3 K), which could form nanofibers with antiparallel 𝛽-sheet
structure and hydrogel.[155] Due to the strong 𝜋–𝜋 interaction be-
tween the filaments and DOX, a low but effective dose of DOX
could be maintained in the tumor, which significantly reduced
the side-effect of DOX in the treatment of 4T1 breast cancer, and
enhanced the tumor growth inhibition efficacy at the primary tu-
mor site, and prolonged the survival to over 40 days. This design
requires direct intratumoral injection of the SPH. Ulijn et al.
took one step further and created an matrix metalloproteinase-
9 (MMP-9) responsive mixed peptide micelle for DOX delivery
using aromatic moiety-containing PhAc-FFAGLDD and MMP-9
degradable GFFLGLDD and their analogues.[156] These micelles
could convert into fibrous nanostructure after reaching MDA-
MB-231 tumor site with high MMP-9 expression and form DOX
depot for localized therapy of cancer. In addition to DOX, other
chemotherapeutics such as vincristine,[139] tanshinone IIA,[157]
and triptolide[158] have also been physically encapsulated in SPHs
for localized cancer therapy. These hydrogels maintained the con-
centrations of drugs within a suitable range, which greatly en-
hanced the efficacy and safety of chemotherapy.
Physical entrapment provides a feasible way to load
chemotherapeutics into SPH. However, it is difficult to achieve
high drug loading, and the drug release and hydrogel de-
composition are not synchronized. SPH self-assembled from
peptide-drug amphiphile could be one possible strategy to
address this issue. For instance, Cui et al. developed a camp-
tothecin (CPT)-based self-assembling prodrug (SAPD) hydrogel
for the treatment of glioblastoma multiforme (GBM) after max-
imal tumor resection. The covalent linkage yielded a fixed drug
loading as high as 23% drug loading (in weight), and ensured the
synchronous kinetical behaviors of CPT and the HKD hydrogel.
The SAPD hydrogel showed a steady and linear drug release
of more than 30 days in vitro. After intracranial implantation
into the cavity left over by surgical resection, the SAPD hydrogel
penetrated deeply into the brain parenchyma, and inhibited the
recurrence of GBM (Figure 7a).[78] To target inoperable tumors,
Zhong et al. designed a hydrogel precursor (HCPT-SA-FFEssEE)
by conjugating HCPT-SA-NHS with FFEssEE. When exposed
to GSH, HCPT-SA-FFEssEE would be cleaved and converted to
HCPT-SA-FFE, which could form a supramolecular hydrogel in
the tumors. The hydrogel at a concentration of 5.0% w/v HCPT-
peptide could release drugs sustainedly for at least 72 h at the
presence of MD-MBA-231 cells.[159] In addition, SPHs have been
reported to deliver small weight molecules loaded nanoparticles.
Mo and colleagues created PTX-CT by encapsulating PTX into
a cell-penetrating-peptide (CPP) R8 H3 -modified transfersomes.
Then PTX-CTs were entrapped into Fmoc-FFF-Dopa-based
SPH.[160] The hydrogel could be applied as a patch on the skin
for transdermal administration of PTX. Due to its network struc-
ture, the SPH design significantly prolonged PTX-CT retention
and allow a much longer interaction time for PTX-CT to squeeze
through the skin barrier compared with PTX-CT solution.
3.1.2. Delivery of Other Small Molecular Weight Drugs with SPH
In addition to chemotherapeutics, other small molecular weight
drugs have also been explored in cancer therapy, including pho-
tosensitizers, molecule-targeted drugs, and immune modula-
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tors. Photosensitizers such as Chlorin e6 (Ce6) have also been
delivered with (FHFD)2 -based hydrogel for photodynamic ther-
apy of cancer.[161] Liu and colleagues took this work a step fur-
ther by conjugating recombinant elastin-like polypeptide (ELP)
with Ce6 to function as a convection-enhanced delivery (CED)
driven photoradiation-controlled intratumoral depot (PRCITD).
They showed that the penetration of ELP could be enhanced by
CED under a 660 nm laser, and the generated reactive oxygen
species (ROS) catalyzed periodic cysteine residues of ELP and in-
duced their crosslink into a therapeutic stable depot. Compared
with control groups without crosslinking, the PRCITD showed
high stability and significantly developed the retention in tumors,
improving the anti-tumor efficiency in a Ce6 dose-dependent
way.[162] Since the activity of Ce6 must be triggered with localized
irradiation, this strategy could further decrease off-tumor side ef-
fects.
Jeffrey et al. validated the concept of delivering immune
modulators with SPH (Figure 7b).[41] They encapsulated cyclic
dinucleotides dithio-(RP,RP)-[cyclic[A(2′,5′)pA(3′,5′)p]] (CDNs),
a stimulator of interferon genes (STING) agonist, within MDP
to form the “STINGel” through the favorable electrostatic inter-
actions between the CDN’s negative thiophosphate linkages and
the positive lysine residues at the peptide termini. The STINGel
was injected subcutaneously in MOC2-E6E7 murine oral cancer
tumor, realizing localized and sustained release of CDNs, which
was eightfold slower compared with standard collagen hydrogel
group. This local treatment induced tumor rejection and pro-
longed overall survival of model mice.
In the above examples, the small molecular-weight drugs de-
livered by SPHs mainly targeted cancer cells. Li and coworkers
aimed to create an SPH for sustained delivery of losartan, an in-
hibitor of cancer-associated fibroblast (CAF).[94] CAF is one of
the major components of tumor stromal cells and contributes
to ECM remodeling and cancer metastasis. Prolonged losartan
treatment is necessary to block the function of CAF, and thus
a high drug loading and prolonged drug retention are necessary.
To achieve this goal, they designed and screened a set of hydorge-
lators and found that C16 -GN2 Q2 NYKD-OH (C16-N) showed the
highest drug loading because the long hydrocarbon tail formed
a flexible and large hydrophobic cavity for losartan. The losartan-
loaded nanofibers could form hydrogel under physiological con-
ditions, and retained in the orthotopic 4T1 tumor for over nine
days after intratumoral injection, which was in sharp contrast
with losartan-loaded spherical micelles formed by DSPE-PEG.
The SPH significantly inhibited the incidence of CAF and the
synthesis of ECM, and therefore enhanced intratumoral accumu-
lation and penetration of DOX-liposomes, as well as their antitu-
mor efficacy.
3.1.3. Co-Deliver Chemotherapeutics and Other Small Molecular
Drugs with SPH
Combination treatment is widely adopted in clinical cancer ther-
apy to maximize efficacy. SPH is a convenient platform for co-
delivery of multiple drugs through mixing.[163,164] Besides, SPH
could be formed by self-assembling prodrugs, which offers a
new method for combinational therapy. For example, Wang et al.
first created a self-assembling prodrug via conjugating two CPT
© 2020 Wiley-VCH GmbH
www.advancedsciencenews.com www.advhealthmat.de

Figure 7. Schematic illustrations of SPHs loaded with small molecular weight drugs. a) The design and mechanism of action of self-assembling prodrug
(SAPD) hydrogels. Reproduced with permission.[78] Copyright 2020, Elsevier B.V. b) The design of “STINGel” formed by CDN-loaded MDP hydrogel.
Reproduced with permission.[41] Copyright 2018 Elsevier Ltd.

molecules to a tumor homing and penetrating peptide iRGD
(cyclic-CRGDKGPDC)[165] to obtain diCPT-iRGD. This peptide-
drug amphiphile could assemble into SPH, which could entrap
STING agonist cyclic di-AMP (c-di-AMP (CDA)) for co-delivery
of CPT and CDA into the GL-261 brain tumor. The resulting
SPH showed much better tumor penetration throughout the tu-
mor and the tumor periphery compared with the control hydro-
gel composed of diCPT-iRGRD (Figure 8a), which manifested no
signal at the periphery of the tumor. The CDA-diCPT-iRGD also
induced a higher percentage of intratumoral CD8+ T cells and
elicited tumor regression, which was in sharp contrast with CDA-
diC8-iRGD (Figure 8b).[79] Recently, they developed diCPT-iRGD
to serve as a universal dispersing agent for low-molecular-weight
hydrophobic drugs. A second therapeutic agent like DOX or cur-
cumin (Cur) could be co-delivered for combination therapy.[166]
In these examples, the SPH was injected directly into the tumor.

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Figure 8. The design and mechanism of action of SPH loaded with CPT and CDA. a) The chemical structure of diCPT-iRGD. b) The diCPT-iRGD could self-
assemble into nanotubes (NT) with positive surface charge and thus adsorb negatively charged CDA. The resulting CDA-loaded NT solution could form
hydrogel after intratumoral injection, and release CPT and CDA for combinational chemotherapy and immunotherapy. Reproduced with permission.[79]
Copyright 2020, Springer Nature.

In another example, Liu and colleagues developed a novel pro-
drug, namely Taxol-EYSV, which could from SPH in the tumor af-
ter intravenous injection.[167] The Taxol-EYSV molecule was syn-
thesized by conjugating Taxol to a tripeptide tyroservatide (YSV,
histone deacetylase and P-glycoprotein inhibitor) through an es-
ter bond that could be cleaved under hydrolysis. The YSV peptide
then formed hydrogel for Taxol deposition, and inhibited histone
deacetylase and P-glycoprotein. Therefore, the Taxol-EYSV-based
hydrogel overcame Taxol resistance and enhanced its anti-tumor
efficiency. These studies highlight the versatility of SPH in tumor
targeted delivery of small molecules, demonstrating that one or
multiple drugs could be feasibly introduced into SPH either co-
valently or non-covalently to exert synergistic anticancer effects.
3.2. Biomacromolecule Delivery
Cancer therapy using biomacromolecular drugs such as anti-
gens, proteins, antibodies and nucleic acids is becoming pop-
ular. Most of these drugs, while of high potency, suffer from
short half-life due to fast clearance or degradation. Given that
SPH can entrap molecules in a hydrophilic network, many SPHs
have been created and used to protect biomacromolecular drugs
from deactivation and clearance.[107,168,169] For instance, anti-
cancer peptide G(IIKK)3 I-NH2 (G3) would be eliminated from
tumors quickly after systemic administration. To improve its effi-
ciency, Xu and colleagues screened out Ac-I3 SLKG-NH2 through
rational design to form an MMP-2 degradable SPH for G3

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Figure 9. The design and mechanism of action of fibrin-based SPH. A solution containing fibrinogen and aCD47@CaCO3 nanoparticles was applied into
the post-surgery tumor bed with a solution of thrombin, which together formed hydrogel in situ. Under mild acidic tumor microenvironment, CaCO3 was
degraded to elevated pH and release aCD47, which re-polarized TAM and inhibited CD47-mediated immune evasion. Reproduced with permission.[177]
Copyright 2018, Springer Nature.

delivery. The SPH could retain in the tumor for a prolonged pe-
riod, and release G3 in a “cell-demanded” way due to the MMP-2
responsiveness of SPH.[122] In contrast to this study, Mo and col-
leagues used enzyme WQ9-2 to initiate SPH formation.[170] The
enzyme-catalyzed a condensation reaction between two precur-
sors Fmoc-F and FF-Dopa to form hydrogelator Fmoc-FFF-Dopa,
which entrapped hirudin (an antiangiogenic protein), and TRAIL
(an apoptosis-inducing cytokine) in the hydrogel. This hydrogel
showed great protein-loading capacity without bioactivity damage
and provided sustained release in the MDA-MB-231 tumor.
In addition to the anticancer peptides and proteins, SPHs have
also been used to deliver antigens. Premature degradation of
antigen peptides is one challenge that limits immunization ef-
ficiency, which is critical for phosphorylated antigens. Yang et al.
thus created a Ca2+ -induced gelator Nap-GD FD FpD Y to protect
phosphorylated antigens (TpSN, CpTW, and KpTL) from dephos-
phorylation by in vivo phosphatase.[141] While protecting the anti-
gens, the D-tetrapeptide-based hydrogel itself also helped to pro-
duce antibodies and inducing the immune response. With the
help of SPH, the ratio of generated antibodies against phospho-
rylated peptides increased. Fast clearance is another issue pre-
venting effective immunization, as prolonged stimulation is nec-
essary to maximize antigen presentation. Liu et al. thus created
a peptide hydrogelator (Fbp-GD FD FD YD K(𝛾E)2 -NH2 ), which self-
assembled upon the mixture of the gelator with protein anti-
gens ovalbumin (OVA) and hepatitis B surface antigen (HBsAg).
The formed SPH significantly delayed antigen release and fa-
cilitated their delivery to antigen-presenting cells, allowing an
effective immune response and repression of the, E.G.,7-OVA
tumor growth.[143] Recently, many peptides are found to be po-
tent adjuvants.[171] Nap-GFFY peptide designed by Yang et al.
was one such example and reported to serve as a peptide ad-
juvant to evoke the immune response. The peptide could form
a hydrogel by simple heating-cooling process and the antigens
like OVA could be mixed into the hydrogel through a vortex.[172]
The Nap group could be replaced with flurbiprofen or carprofen,
nonsteroidal anti-inflammatory drugs (NSAID), and the result-
ing peptide hydrogels had both antitumor immunity and anti-
inflammatory properties.[173]
Antibodies have long half-lives in circulation and show a high
affinity for corresponding antigens. However, anticancer antibod-
ies that can recognize antigens universally expressed through the
body should be targeted delivered to the tumor. For instance,
off-tumor accumulation of anti-PD-L1 and anti-PD1 antibodies
cause severe side effects in the lungs. Another potential tar-
get for cancer immunotherapy is the integrin-associated protein
(IAP)—CD47 known as a “don’t eat me” signal.[174–176] Although
CD47 was upregulated in many tumors, it is also regularly ex-
pressed on other normal tissue. To limit anti-CD47 antibody in
the B16F10 murine melanoma, Gu’ group loaded the anti-CD47
antibody in the calcium carbonate nanoparticles and then encap-
sulated them in the fibrous gel, which formed quickly upon the
mixture of fibrinogen and thrombin (Figure 9).[177] This treat-
ment changed the tumor microenvironment (pH and immune
checkpoint), and induced the M1-oriented polarization of tumor-
associated macrophages (TAM). High antitumor immunity was
realized using this SPH design, without systemic side effects.
Nucleic acid-based vaccination has unique advantages because
specific antigens can be easily encoded into the sequence with-
out tedious protein expression and purification. However, nucleic
acid-based vaccine suffers from enzymatic degradation and lim-
ited expression efficiency. As a response to this challenge, Op-
penheim et al. designed a set of cationic peptide hydrogelators
and found that the HLT2 peptide (net charge = +5 under neutral

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www.advancedsciencenews.com
Figure 10. Schematic illustration of the mechanism of action of alkaline phosphatase (ALP). The Nap-D FD FD Yp D EMe2 was cleaved by ALP to generate
Nap-D FD FD YD EMe2 , which self-assembled in the cancer cells to induce cell stress and apoptosis. Reproduced with permission.[180] Copyright 2019,
Elsevier Inc.
pH) could efficiently encapsulate a plasmid DNA (TA) encoding
a melanoma-specific gp100 antigen and adjuvant HMGB1 (high-
mobility group binding protein 1). This hydrogel protected the
DNA from swift enzymatic degradation and fragmentation and
improved DNA retention and expression. The SPH-based vac-
cine induced an acute inflammatory response, improved infiltra-
tion of macrophages and polymorphonuclear cells, and enhanced
the immune response mediated by CD4+ /IFN𝛾 + expressing Th1
cells and gp100-specific antibodies.[178]
3.3. Drug-Free Anticancer SPH
The self-assembly of peptide and its derivatives, not only creates
hydrogels but in some cases endows the molecules with self-
assembly-based anticancer activity. Xu and colleagues linked an L -
amino acid to the C-terminal of a D -tripeptide (Nap-D FD FD YR) to
create a peptide hydrogelator that could assemble into a crescent-
shaped supramolecular structure at the presence of ALP.[179]
They showed that the formed supramolecular structure would
damage cell membrane of HeLa and HS-5 cells and interact with
the endoplasmic reticulum (ER) to induce cell death, which pro-
cess depended on the formation of the crescent-shaped mor-
phology. A similar phenomenon was observed on another ALP-
responsive peptide hydrogelator (Nap-D FD FD Yp D EMe2 ), which im-
proved the survival of mice bearing osteosarcoma tumor (Fig-
ure 10).[180] SPH could also be engineered to target mitochon-
dria other than ER to kill tumor cells. A feasible strategy to
achieve this is conjugating mitochondria-targeting moiety such
as triphenyl phosphonium (TPP) to peptide hydrogelators. For
instance, Xu et al. modified a pair of enantiomeric tripeptides
with TPP to produce phosphorylated tetrapeptides (L-1P or D-
1P).[181] They found that the oligomers could self-assemble on
the surface of human osteosarcoma cells at the presence of ALP,
and the hydrogel was internalized by the cancer cells to interfere
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with the function of mitochondria. Yang and colleagues created
a dual-responsive hydrogelator (NBD-GFFpY-ss-ERGD), which
could transform into NBD-GFFY-ss-ERGD and NBD-GFFY-thiol
subsequently after exposure to extracellular ALP and intracellu-
lar GSH.[39] The intracellular hydrogel caused the death of tumor
cells selectively.
3.4. Combination Delivery
Combination therapy of cancer with small molecular drugs and
biomacromolecules is becoming popular in recent years. This is
because small molecular weight chemotherapeutics are potent
immunogenic cell death (ICD) inducers and biomacromolecules
are usually effective immune modulators, which together could
induce prolonged tumor regression. As we discussed above,
SPHs can provide both hydrophilic network and hydropho-
bic cavity to accommodate small molecular weight drugs and
biomacromolecules either covalently or non-covalently, which
make SPH an ideal platform for combination therapy of
cancer.
3.4.1. Combinational Delivery of Small Molecular and
Biomacromolecular Drugs
As we discussed above, many peptides can serve as po-
tent adjuvants to potentiate antitumor immune response.
Inspired by this, immune active peptides were engineered
onto peptide hydrogelators for co-delivery of small molecu-
lar weight ICD inducers. Melittin, a 26-amino-acid polypep-
tide (GIGAVLKVLTTGLPALISWIKRKRQQ-NH2 ) with the po-
tential to perforate on cell membranes and modulate the
innate immune response,[182,183] was the one most investi-
gated. For instance, Jin et al. created a melittin-containing
© 2020 Wiley-VCH GmbH
www.advancedsciencenews.com
peptide hydrogelator namely melittin-RADA32 ((RADA)8 -GG-
GIGAVLKVLTTGLPALISWIKRKRQQ-NH2 ) for localized deliv-
ery of DOX.[40] The resulting hydrogel showed controlled re-
lease of DOX that suppressed the growth of primary B16F10
tumor. In the meanwhile, it also activated DC maturation in
the draining lymph nodes, eliminated alternatively activated
macrophage, and activated cytotoxic T cells to kill metastatic tu-
mors in vivo. The same group then optimized their molecule de-
sign, and used melittin-(RADA)6 for KN93 (a Ca2+ /calmodulin-
dependent protein kinase II (CAMKII) inhibitor) delivery. In
B16F10 mouse melanoma model, melittin-(RADA)6 facilitated
ICD by improving the uptake of KN93 through creating tunnels
on cell membranes and induced stronger cell-mediated antitu-
mor immunity compared with melittin-(RADA)4 and melittin-
(RADA)8 hydrogels.[184]
Co-delivery of small molecular weight ICD inducer and
chemokines is another widely adopted strategy in hydrogel-
based local cancer therapy. For instance, Chen and colleagues
have developed poly(ethylene glycol)-poly(𝛾-ethyl-l-glutamate) di-
block copolymers (mPEG-b-PELG) and PELG-PEG-PELG tri-
block copolymers for codelivery of ICD inducers (cisplatin or
DOX) and immune modulators (interleukin-15, interleukin-2, or
interferon-𝛾).[185,186] The resulting hydrogels showed potent anti-
tumor activity via a mechanism involving CD8+ T cells and nat-
ural killer (NK) cells.
Enhanced cytotoxic T lymphocyte infiltration triggered by ICD
inducers usually resulted in upregulation of immune check-
points. Thus, combination delivery of ICD inducers and immune
checkpoint inhibitors may be an effective way to enhance anti-
cancer immunity. To test this concept, Cui and colleagues created
an MMP-2 responsive hydrogelator diCPT-PLGLAG-iRGD (Fig-
ure 11a), which could self-assemble into supramolecular nan-
otubes (P-NTs) and then formed hydrogel with anti-PD1 antibod-
ies under physiological conditions (Figure 11b).[80] After intra-
tumoral injection, the diCPT-PLGLAG-iRGD showed better tu-
mor penetration compared with the hydrogel formed by diCPT-
PLGLAG-iRGRD. At the presence of MMP-2 and GSH, free CPT
was released to boost ICD of cancer cells, while at the same time
liberate anti-PD1 antibodies. As a result, the SPH boosted T-cell
mediated immune response, leading to the 100% repression of
tumor growth for all treated mice with either CT 26 colon can-
cer or GL-261 brain cancer and repressed the tumor recurrence
significantly.
3.4.2. Combinational Delivery of Cells/Cell Debris and Other
Ingredients
SPHs have been widely used in 3D cell culture and tissue engi-
neering. Recently, SPHs have also been used for local delivery of
cell debris or live DC for cancer immunotherapy. Li and cowork-
ers designed an Fmoc-KCRGDK (FK) based SPH for encapsula-
tion of JQ1 (a BRD4 inhibitor) and indocyanine green (ICG, a
photosensitizer) loaded dead tumor cells.[187] The resulting per-
sonalized cancer vaccine, namely PVAX, could kill cancer cells
upon NIR irradiation to enhance DC maturation and enhance
tumor infiltration of CD8+ T lymphocytes, the activity of which
could be further improved by the released JQ1. Instead of deliv-
ering cell debris, Wang and colleagues explored the use of SPH
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for live DC delivery.[42] The SPH was prepared by encapsulating
live dendritic cells (DCs), anti-PD-1 antibodies, and tumor anti-
gen ovalbumin (OVA) into RADA16 (Ac-(RADA)4 -CONH2 ) (Fig-
ure 12a) based scaffold. The SPH served as a mild reservoir for ec-
togenous DCs where they retained its bioactivity, interacted with
OVA, and finally migrated out to tumor site sustainably com-
pared with OVA-pulsed DCs. It also helped to recruit endogenous
DCs and activated them by sustained OVA release, thus inducing
much stronger activation of antigen-specific effector T cells to kill
tumor cells (Figure 12b). Therefore, the SPH induced strong pro-
tective and therapeutic antitumor response.
4. Conclusions and Perspectives
SPH is a class of biomaterial that closely mimics ECM both in its
structure and biological functions. It is mainly composed of short
peptides that can be degraded into natural amino acids, which
nature endows it with good biocompatibility and controllable im-
munogenicity. Although a short peptide, a decapeptide, for ex-
ample, can have a sequence space with >1013 possibilities, sur-
passing most synthetic materials. Compared to the natural ECM,
it is self-assembled from simple building blocks through non-
covalent interactions such as 𝜋–𝜋 interaction, hydrogen bonding,
hydrophobic force, and electrostatic interactions. Thus, different
functional moieties can be easily integrated into SPH through a
co-assembly strategy to create novel SPHs. With these unique ad-
vantages, SPH has shown great potential in cancer therapy. SPH
has been used as localized delivery vehicles for small molecular
weight drugs, biomacromolecules, nanoparticles, and even cells
through non-covalent or covalent encapsulation mechanisms.
SPH itself can also directly modulate the function and viabil-
ity of cancer cells and stromal cells in the TME. With increas-
ing advances in cancer biology and oncoimmunology, SPH has
also demonstrated its great potential in cancer immunotherapy.
As a result, SPH can selectively eradicate primary tumors, but at
the same time induce systemic anticancer immunity. Neverthe-
less, some issues must be addressed before the clinical transla-
tion of SPH in cancer therapy. First, the key features that deter-
mine the biological activities of SPH mush be understood. Sim-
ilar to ECM that can exert both protumoral and antitumoral ef-
fects, SPH can also exert biological activities of opposite effects.
Recent studies by Hartgerink and Elisseeff and their colleagues
showed that the composition of hydrogels has a dramatic effect
on the types of recruited immune cells into the hydrogels and
their biological functions,[131,188] revealing that the charge and ori-
gin of hydrogels significantly affected their biological functions.
However, more function-related features must be identified for
rational SPH design. Second, the dynamic feature of the SPH
must be controllable. SPH is characteristic of its dynamics be-
cause of its supramolecular nature. However, the dynamic tran-
sition of SPH is mainly driven by physical and chemical cues,
which is in sharp contrast with ECM the transitions of which
are mainly biological cue driven. Strategies that introduce bio-
logical stimuli-responsive linkers into SPH design has been ex-
tensively explored. Recently, Stupp and colleagues explored other
strategies such as simultaneous covalent and non-covalent hy-
brid polymerization and peptide-DNA hybrid building blocks,
which can be potentially used in the design of more intelligent
SPH. Third, the optimized combination treatment regime shall
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Figure 11. Schematic illustration of the design and mechanism of action of P-NT–aPD1 hydrogel. a) The chemical structure of diCPT-PLGLAG-iRGD.
b) PD1 antibodies could be efficiently entrapped within the network form by diCPT-PLGLAG-iRGD solution. After intratumoral injection, the solution
turned into hydrogel, which released CPT and aPD1 in response to MMP-2 and GSH, exerting combination chemotherapy and immunotherapy against
tumor. Reproduced with permission.[80] Copyright 2020, American Association for the Advancement of Science.

be further pursued. SPH has great potential in co-delivery of
multiple cargoes of different chemical and biological properties.
The strategies to realize controlled release of a single therapeu-
tic component have been vastly explored. However, the drug re-
lease kinetics of the co-delivered drugs in an SPH were usu-
ally not optimized. Since combination therapy is a routine prac-
tice in cancer therapy, it is critical to optimize drug combina-
tion, dosage, and release profiles to achieve maximized efficacy.
Forth, imaging-guided hydrogel delivery is required to expand
the application of SPH. Currently, the SPH is mainly applied
in surgical beds, subcutaneous tissues, and tumors that can be
easily accessed (e.g., breast cancer). To treat tumors of the in-
ternal organs, clinic-compatible and imaging-guided drug deliv-
ery platform must be established. Lastly, peptides may be rela-
tively more expensive compared with polymers. The requirement
of protected amino acids during peptide synthesis and the dif-
ficulty in peptide purification are two major challenges to ob-
tain peptide hydrogelators. Recently, a water/dichloromethane
biphasic system with macroinitiators anchored at the interface
was developed by Cheng and colleagues to simplify the impurity
treatment process.[189] Method to synthesize polypeptides directly
from amino acids has also been reported by the same group.[190]
With these new and more upcoming technologies, a cost reduc-
tion in peptide hydrogelator synthesis is foreseeable in the near

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Figure 12. Schematic illustration of the design and mechanism of action of SPH-based vaccine. a) The chemical structure of RADA16 peptide. b) RADA16
peptide could form hydrogel and entrap antigens, anti-PD-1 antibodies, and DC within its network. After subcutaneous injection, the mature DCs would
migrate to the lymph nodes, while the hydrogel could further recruit naïve DC to further improve antigen presentation and CD8 T cell proliferation.
Reproduced with permission.[42] Copyright 2018, American Chemical Society.

future. Despite these challenges, clinical translation of SPH is Received: July 15, 2020
expected in the near future, given its advantages such as tumor Revised: September 4, 2020
targeted drug delivery, low systemic drug exposure, the capability Published online:
to induce systemic antitumor immunity, and realize combination
therapy, as well as good biocompatibility.

Acknowledgements [1] R. O. Hynes, Science 2009, 326, 1216.

Y.C. and C.Z. contributed equally to this work. The authors thank the
National Natural Science Foundation of China (31870995, 81690265,
81671808, 81630052), the Youth Innovation Promotion Association of CAS
(2017335), and the SA-SIBS Scholarship Program for financial support.
Conflict of Interest
The authors declare no conflict of interest.
Keywords
cancer, hydrogels, local therapy, peptides, supramolecular self-assembly
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www.advancedsciencenews.com www.advhealthmat.de

Yaping Li is a principal investigator and director in the Center of Pharmaceutics, Shanghai Institute
of Materia Medica, Chinese Academy of Sciences. He has made a series of contributions to the basic
research and clinic translation of targeted drug delivery systems, including the design, construction,
and mechanistic study of novel nanosized drug delivery systems with higher efficacy and less toxicity
for the treatment of metastatic and multi-drug resistant cancer.

Pengcheng Zhang is a professor at the Shanghai Institute of Materia Medica, Chinese Academy of
Sciences. His research interest mainly focuses on the development of nanomedicines using self-
assembling and biomimetic strategies.

Adv. Healthcare Mater. 2020, 2001239 2001239 (23 of 23) © 2020 Wiley-VCH GmbH