Bioactive Materials 6 (2021) 1375–1387

Contents lists available at ScienceDirect

Bioactive Materials
journal homepage: www.sciencedirect.com/journal/bioactive-materials

Adaptable hydrogel with reversible linkages for regenerative medicine:

Dynamic mechanical microenvironment for cells
Zongrui Tong a, Lulu Jin a, Joaquim Miguel Oliveira b, c, d, Rui L. Reis b, c, d, Qi Zhong e,
Zhengwei Mao a, *, Changyou Gao a, **
a
MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou, 310027,
China
b
3B’s Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of
Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, 4805-017, Barco GMR, Portugal
c
ICVS/3B’s - PT Government Associate Laboratory, Braga/Guimarães, Portugal
d
The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017, Barco, Guimarães, Portugal
e
Key Laboratory of Advanced Textile Materials & Manufacturing Technology, Ministry of Education, National Base for International Science and Technology
Cooperation in Textiles and Consumer-Goods Chemistry, Zhejiang Sci-Tech University, 310018, Hangzhou, China

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

Keywords: Hydrogels are three-dimensional platforms that serve as substitutes for native extracellular matrix. These ma­
Adaptable hydrogel terials are starting to play important roles in regenerative medicine because of their similarities to native matrix
Dynamic mechanical microenvironment in water content and flexibility. It would be very advantagoues for researchers to be able to regulate cell behavior
Supramolecular chemistry
and fate with specific hydrogels that have tunable mechanical properties as biophysical cues. Recent de­
Dynamic covalent chemistry
Yes-associated protein
velopments in dynamic chemistry have yielded designs of adaptable hydrogels that mimic dynamic nature of
extracellular matrix. The current review provides a comprehensive overview for adaptable hydrogel in regen­
erative medicine as follows. First, we outline strategies to design adaptable hydrogel network with reversible
linkages according to previous findings in supramolecular chemistry and dynamic covalent chemistry. Next, we
describe the mechanism of dynamic mechanical microenvironment influence cell behaviors and fate, including
how stress relaxation influences on cell behavior and how mechanosignals regulate matrix remodeling. Finally,
we highlight techniques such as bioprinting which utilize adaptable hydrogel in regenerative medicine. We
conclude by discussing the limitations and challenges for adaptable hydrogel, and we present perspectives for
future studies.

1. Introduction (ECM) that goes beyond immutable physical support. We need ECM that
provides chemical and physical dynamsism, in order to control cell
The limited innate capacity for self-regeneration of human beings behavior, cell fate, and cell growth [12].
caused wide attention to tissue regeneration, and propelled the advances Hydrogel is an efficient three-dimensional material for tissue engi­
of regenerative medicine which could replace or repair diseased tissue or neering because of its high water content and flexibility, which allows it
organs. Despite the inherent challenges, in recent decates, successes in to mimic native ECM. Benefiting from the aqueous state close to phys­
cell and biomateirals for regenerative medicine has led to practical ap­ iological condition, hydrogel could provide biomimetic microenviron­
plications like artificial skin [1], ear-shaped cartilage [2], bone recon­ ment [13]. The similar stiffness of hydrogel and soft tissue is also
struction [3,4], nerve regeneration [5], etc. Furthermore, some beneficial to tissue regeneration [14]. At the early-stage studies of
commercialized products for stem cell therapy [6–8] and numerous regenerative medicine, covalent hydrogels were dominant; these
candidates in clinical trials [9–11] have been developed. However, included glutaraldehyde (GA) crosslinked poly (vinyl alcohol) (PVA)
further advances in tissue regeneration will require extracellular matrix and chitosan [15], photo-crosslinked methacrylated gelatin (GelMA)

* Corresponding author.
** Corresponding author.
E-mail addresses: [email protected] (Z. Mao), [email protected] (C. Gao).

https://doi.org/10.1016/j.bioactmat.2020.10.029
Received 24 August 2020; Received in revised form 14 October 2020; Accepted 28 October 2020
Available online 10 November 2020
2452-199X/© 2020 The Authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

[16], and transglutaminases crosslinked fibrinogen [17], etc. Such co­ has provided plenty of non-covalent interactions to obtain reversible
valent crosslinked hydrogels are stable but lack of the necessary dy­ linkages, including macrocyclic host–guest interactions, hydrogen
namic for cell expansion, ECM remodeling and other cell behaviors. bonds, electrostatic interactions, and hydrophobic interactions. Also,
Thus, researchers turned their attention to degradable polymers which dynamic covalent chemistry provides several options, including
could impart dynamic changes for cell proliferation and tissue regen­ reversible Diels–Alder reactions, hydrazone bonds, thioester exchanges,
eration [18,19]. Such hydrogels based on degradable polymers can boronate bonds, etc.
adapt to cell culture in terms of ECM chemistry, mechanical properties,
cell type, crosslinking density, and adhesion ligand density [20]. How­ 2.1. Non-covalent interactions
ever, the tradeoff between mechanical loss due to degradation and
required physical support decides the limitation of irreversible change of 2.1.1. Macrocyclic host–guest interactions
degradable covalently crosslinked hydrogel. Macrocyclic host-guest complexation is a kind of non-covalent
In recent decades, adaptable hydrogels with reversible linkages have interaction which consists of macrocyclic host such as cyclodextrin
become able to mimic the dynamic changes of native ECM. Native ECM (CD) or cucurbit[n]uril (CB[n]s), and paired guests including specific
is dynamic with stress relaxation and be remodeled and to provide dy­ small molecules and linear polymers. The combination form of macro­
namic mechanics to cells [21,22]. However, conventional hydrogels cyclic host-guest complex could be “pendant” or “threading” architec­
with irreversible linkages are static which own finite stress relaxation ture (Fig. 1a and b).
and could hardly be remodeled without degradation. Advances in su­ The “pendant” means the host or guest molecule was decorated with
pramolecular chemistry and dynamic covalent chemistry provide op­ functional groups on the polymer chains. The host and guest could be
portunities to develop reversible linkages for hydrogels with paired, according to specific combinations, and typical equilibrium
self-healing, shear thinning, and fast stress relaxation properties to binding constants of specific pairs have been summarized in some re­
supply dynamic physical cues [23,24]. Further studies focus on spatial views [28,29]. Because of the relatively weak interaction and rapid
dynamics (plasticity that enables remodeling) and mechanical dynamics formation, hydrogels with “pendant” host-guest interactions are prone
(stress relaxation that provides biophysical signals) of matrices with to shear thinning and can sometimes be injected. Specific pairs present
reversible linkages [25]. For the purposes of our review, we define various stimulus responses (thermal, pH, light, ion and redox) because of
adaptable hydrogel for tissue engineering as polymer networks with the mutative formation and deformation [30].
adaptable linkages that can be broken and re-formed in a reversible Cyclodextrin is one kind of macrocyclic saccharide with a cavity, and
manner without external triggers [26]. it could be divided into three kinds (α, β and γ) according to the number
Previous reviews have outlined the types of reversible linkages of units. Because of the poor solubility, β-cyclodextrin can be more easily
which could be utilized to fabricate adaptable hydrogels. Previous re­ purified after modification compared to α-, and γ-cyclodextrin.
views have also discussed the applications of adaptable hydrogels in bio- β-cyclodextrin is widely studied because it has good equilibrium asso­
printing and tissue engineering [26,27]. However, the literature has ciation constants and widely-available derivatives [31]. Rodell and
lacked a systematic summary of how to utilize those reversible linkages Burdick et al. have reported their systematic research on hyaluronic acid
to build dynamic networks to accomplish specific influences on cell (HA) hydrogel based on host-guest interactions modified with host
behavior. In this review, we summarizes recent advances in (1) revers­ (β-cyclodextrin) and guest (adamantine or azobenzene) [32,33]. Such
ible linkages which could be utilized to construct adaptable hydrogels; host-guest hydrogels have properties such as shear-thinning, self-healing
(2) the influences of hydrogel stress relaxation and spatial plasticity on and injectability. Furthermore, to make the hydrogels more tunable, a
cell behavior and fate; (3) practical applications of adaptable hydrogels secondary network with covalent crosslink was developed (Fig. 1c) [34,
in the field of tissue regeneration. We discuss the rational design of 35]. Additionally, limited chemical cross-linking was adopted to address
adaptable networks that provide biophysical cues to achieve hydrogels the reduced stability of adaptable hydrogels with reversible linkages.
for regeneration of different tissues/organs. Finally, we conclude the Feng et al. reported preassembled host-guest complexation based on
limitation of adaptable hydrogels and perspective developments. gelation and acrylate β-cyclodextrin to fabricate stable adaptable
hydrogels [36,37]. The tailored degradation and molecule release
2. Reversible linkages for adaptable hydrogels respond to proteases, and they are ensured by dithiol peptide crosslinker
[38]. Such peptide was able to cleave the matrix metalloproteinase and
Adaptable hydrogels are designed to provide mechanical and spatial accomplish cell invasion in the process of tissue regeneration. Practical
dynamic, which plays an important role during cell proliferation, dif­ applications including gene therapy, stem cell therapy and 3D print were
ferentiation, and immigration as well as tissue regeneration. Thus also attempted [39,40].
adaptable hydrogels must provide a physical scaffold to maintain the Cucurbit[n]uril (CB[n], n = 5–8, 10) has a similar structure to
structural integrity of multicellular organisms, and also serve as a cyclodextrin, but it has been limited by several factors: it is insoluble, it
reservoir for biochemical and biophysical signals to support cell sur­ lacks a homologous series of different-sized hosts, and it is unable to
vival, organization, and differentiation. Three characteristics of adapt­ access CB[n] derivatives [41]. So far, only a few hydrogels consisting of
able hydrogels are required for these hydrogels: they must be easy to CB [6] and CB [8] have been reported for stem cell therapy or drug
remodel, instantaneously reversible, and they must feature fast stress delivery, even though CB[n] has high affinity to specific guest mole­
relaxation. . The remodeling of the matrix is important for cell expan­ cules, high selectivity, and strong binding interactions [42,43]. As
sion, and migration, which cannot be achieved by traditional hydrogel shown in Fig. 1a, host-guest interactions based on cucurbit[n]uril could
networks. The instantaneous reversibility could allow for injectability, include functional group as cyclodextrin, but also linkers for more than
and it could maintain hydrogel stability as physical support even with one guest moiety.
multiple deformations. Lastly, fast stress relaxation is a newly discov­ The “threading” architecture, also called rotaxane, means the linear
ered regulator to cell behavior that could reflect specific mechanosignals polymer, such as poly (ethylene oxide) (PEO), poly (ε-caprolactone)
to cells, and this makes adaptable hydrogels different from conventional (PCL), or poly (vinyl alcohol) (PVA) serves as a guest molecule that
irreversible hydrogels. passes through the cavity of host molecule. When the linear polymer
In the design of adaptable materials, in addition to the common passes through the cavity of macrocyclic host molecules, the latter could
molecular weight, stoichiometry of reactive functional groups and slide along the guest polymer chain causing flexibility. Such a structure
crosslink density, reversible linkage between molecular chains is exhibit flexible dynamism without breaking, which is essentially
essential. The degree of equilibrium reversibility is the key to balance different from most reversible hydrogels.
stability and mechanical dynamic. In the past, supramolecular chemistry Highlighted in dynamic hydrogels, sliding hydrogels with rotaxane

1376
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

Fig. 1. Macrocyclic host–guest design for adaptable

hydrogel. a) Illustration of “pendant” architecture; b)
Illustration of “threading” architecture; c) Network
architectures examined included a host-guest (GH)
hydrogel, covalently crosslinked (MeHA) hydrogel,
GH double network (DN), and MethGH DN. Local
stress under loading (red) dissipated through revers­
ible GH complex rupture [34]. Copyright (2016)
Wiley. d) Schematic illustrations of sliding hydrogel
(right) and covalent crosslinked chemical hydrogel
(left). And the confocal microscopy images of cells
cultured in these two kinds of hydrogel [44]. Copy­
right (2016) Wiley.

architecture represent both stability and mobility. Even though sliding formed without chemical modification [47]. As shown in Fig. 2a, TA
hydrogels are ideal strengthen material with complex dynamic, most plays a role as a “molecular glue”, a new mode of reversible action in
reported ones were not biocompatible. Yang et al. [44] made sliding phosphodiester bonds, which is different from the crosslinking that uses
hydrogels for constructing synthetic stem cell niches (Fig. 1d). Molec­ complementary sequences. Shin et al. [48] reported gallol-derived
ular mobilities of sliding hydrogel render the hydrogel network more hydrogel; such reversible hydrogen bonds would transform to covalent
adaptable, giving stem cells the freedom to respond to soluble factors bonds after oxidation, in a process that may last several hours. As a
and to adjust their behavior accordingly. result, such hydrogels could only be adaptable in a short time for cell
encapsulation and 3D bio-printing. The self-healing of supramolecular
2.1.2. Hydrogen bonds hydrogels also provides opportunities to print multiple materials and
Hydrogen bonds are ubiquitous secondary interactions which could complex structures at high resolutions [49].
be destroyed easily due to weak bond energy. Multiple hydrogen bonds Nucleotides, famous for Watson Crick base pairing, are the basic
could form dynamic reversible hydrogels. The large overall association components of genetic substrates. DNA hydrogels could be crosslinked
constant ensures such hydrogels remain reliable and adaptable for tissue under physiological conditions via efficient ligase-mediated reactions
engineering. In the proceeding of regenerative materials, substrates with [50]. As shown in Fig. 2b, the functional group (single-stranded DNA or
multiple hydrogen bonds, including polyphenols, nucleotides, and specific base pairing) modified on the polymer chains could be available
β-sheets of peptides, were adopted to fabricate adaptable hydrogels. to design adaptable hydrogels [51,52].
Polyphenols, also called vegetable tannins, have more than one hy­ Ureido pyrimidinone (UPy) could form self-complementary dimers
droxyl group on a benzene ring. They can form hydrogen bonds with via quadruple hydrogen bonds. However, the strong affinity of UPy di­
themselves, protein, or DNA [45]. Plenty of works have reported mers for this kind of bonding can only be retained in hydrophobic mi­
bio-adhesive polyphenols in tissue engineering [46]. With reversible yet croenvironments, which are not biocompatible; thus, the application of
strong interaction between DNA and tannic acid (TA), hydrogel was UPy based hydrogel in tissue engineering was a challenge [50,53]. Block

Fig. 2. a) Schematic illustrations of the formation

and hydrolysis of DNA/TA hybrid hydrogel [47].
Copyright (2015) Wiley. b) (i) Polypeptide–DNA
conjugate forms hydrogel based on hydrogen bonds
between double-stranded DNA [52]. Copyright
(2015) Wiley. (ii) Schematic illustrations of four-arm
PEG hydrogel network based on hydrogen bonds be­
tween thymine and adenine functionalities [51].
Copyright (2012) The Royal Society of Chemistry. c)
(i) Schematic illustrations of PEG-UPy chain-extended
(co)polymers with segmented multiblock architec­
tures [55]. Copyright (2014) American Chemical So­
ciety. (ii) Schematic illustrations of non-swellable
hydrogel based on quadruple hydrogen bonds.
Copyright (2019) American Chemical Society.

1377
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

copolymerization or graft with hydrophilic polymer (Fig. 2c i) may supramolecular interactions [71]. Additionally, as shown in Fig. 3b,
overcome the limited application of UPy in aqueous condition [54,55]. Mg2+ could stabilize hydrogels by modified with bisphosphonate, which
Another strategy is to crosslink the micelle of amphiphilic copolymer could adapt cell culture and promote a physiological response [72].
with UPy units (Fig. 2c ii) [56]. Polyelectrolyte hydrogels have been formed via electrostatic in­
β-Sheets are common motifs of protein secondary structure. They teractions, which have advantages including solubility, quick gelation,
consist of β-strands with more than two backbone hydrogen bonds. protection for bioactive moluecles, and response stimulation [73–75].
Among biomaterials in tissue engineering, several natural proteins and The quick gelation needs to be utilized skillfully such as through layer by
programmable peptides could form hydrogels via β-sheets. Mechanical layer self-assembly; random mixture would cause unexpected precipi­
properties and gelation of natural materials (silk and collagen I) are tation. As shown in Fig. 3c, polyelectrolyte complexes consist of alginate
largely dictated by domains of β-sheet peptide assemblies. Also, syn­ and polylysine were built stepwise with a DNA template [76].
thetic peptides, including MAX1 (β-hairpin peptide), RADA16 and
(FEFEFKFK)2-OH, etc., have been designed and programmed to fabri­ 2.1.4. Hydrophobic interaction
cate hydrogels for tissue engineering [57–59]. Due to the weak inter­ Derived by the entropic increase, amphiphilic molecules could
action of β-sheets, composition of such synthetic peptide hydrogels assemble by burying the hydrophobic faces away from aqueous envi­
based on β-sheets and supporting matrix are reliable in practical appli­ ronment. Amphiphilic block polymers and coiled-coil peptides are
cation [60]. Researchers describe general rheological analysis [61] and common candidates for adaptable hydrogels assembled via hydrophobic
tutorials [62] which highlights the impact of the conditions used for interaction.
peptide assembly on the formation and mechanical properties of the Amphiphilic block polymers could consist of synthetic polymers,
corresponding hydrogels. polypeptides, or grafted derivatives of former two compounds. As shown
in Fig. 4a, amphiphilic polypeptide KmLn could be synthesized by a block
2.1.3. Electrostatic interaction of hydrophilic polylysine (K) and hydrophobic polyleucine (L), in which
Electrostatic interaction is a molecular interaction between cationic m and n represent unit number of polylysine and polyleucine [77].
and anionic electric charges. Based on electrostatic interaction, hydro­ Synthetic polymers like hydrophilic polyethylene glycol (PEG) are often
gels could be formed between polyelectrolytes, or between multivalent blocked with hydrophobic polymers like polyphenylene sulfide (PPS) to
cationic and anionic polymers. Most natural macromolecules (e.g. build amphiphilic block polymers [78]. Besides, amphiphilic polymers
alginate, hyaluronic acid, chondroitin sulphate, etc.) are anionic, and poly (N-isopropylacrylamide) are popular for fabricating adaptable
they could assemble with cationic polyelectrolyte (e.g. chitin, chitosan, hydrogels for cell culture, release, and separation [79,80].
polylysine or synthetic polymers) or crosslinks with multivalent cations Coiled-coil peptides are structural sequences common in fibrous
(Ca2+, Mg2+, Fe3+, Zn2+, etc.) [63,64]. proteins, which consists of more than two α-helices [81]. Typically, the
Alginate-Ca2+ is widely applied in tissue engineering because it is primary structure of the α-helices consists of (a-b-c-d-e-f-g) n, where “a”
easy to form and control (Fig. 3a) [65,66]. The mechanical properties of
alginate hydrogels have been studied according to the influences of
mechanical signals on cell behavior and fate (see 3.1 for detail). In the
progress of additive manufacturing, 3D bioprinting can combine stem
cells, and other materials to create complex bio-hybrid structures for
various applications [67,68]. The easy and rapid access to digital design
and fabrication propels practical application in tissue engineering such
as bone/cartilage regeneration, and cardiovascular diseases. Adaptable
hydrogels were also developed as bio-inks, because of their dynamic and
shear-thinning features. In recent decades, adaptable hydrogels have
been printed by extrusion technique. Such technique allows high cell
density with low cost, but it is hindered by slow speed and moderate
resolution compared to other techniques (inkjet, laser-assisted, and
stereolithography) [69]. Alginate expansion lattices for neural progen­
itor cells were extrusion-printed via stepwise crosslinking. The
first-stage crosslink gave pre-printed hydrogels proper stiffness that
could support cells and allow for injection, and second-stage crosslink
formed a robust and stable lattice [70]. Another more common strategy Fig. 4. Schematic of a shear-thinning hydrogel formed by coiled-coil self-as­
for bio-printing is photo-crosslink extruded bio-ink with UV light. Stable sembly and reinforced through temperature-responsive aggregation of poly (N-
isopropylacrylamide) [84]. Copyright (2013) Wiley.
and precise structure was achieved by covalent cross-linking or

Fig. 3. a) Schematic illustration of the fabrication of

the self-assembled HA-BP-Mg nanocomposite hydro­
gel. i) The in situ self-assembly of Ac-BP-Mg NPs
around the grafted BP groups of the HA-BP macromer
via BP-Mg2+ coordination. ii) The self-assembled cell-
laden hydrogel are stabilized by the Ac-BP-Mg NPs.
iii) UV-initiated radical polymerization of the acrylate
groups at the surface of Ac-BP-Mg NPs leads to the
secondary crosslinking and a further increase in the
mechanical property of the self-assembled HA-BP-Mg
nanocomposite hydrogel. iv) The cell-laden hydrogel
can be injected and rapidly remolded to conform to
the geometry of the injection sites [72]. Copyright
(2017) Wiley b) DNA-templated crosslinking of algi­
nate and polylysine via polyelectrolyte complexation
[76]. Copyright (2019) Springer Nature.

1378
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

and “d” are usually occupied by hydrophobic amino acids, such as group and hydrazine could be formed rapidly under physiological con­
leucine and valine, “e” and “g” are occupied by charged residues [82]. ditions. Under neutral pH-, the formation and hydrolysis rate of aliphatic
Similar to functional groups, adaptable hydrogels could be formed by aldehyde-derived hydrazones are much faster than that of aromatic
self-assembly of polymers grafted with coiled-coil peptides, or by aldehyde-derived hydrazones. Hdrogels crosslinked completely with
crosslink of micelles that consist of peptide-grafted polymers with aliphatic aldehyde-derived hydrazones have rapid mechanical relaxa­
reactive groups [83]. As shown in Fig. 4b. elastin-like peptide/protein is tion, which is more conducive to cell spreading and migration [91].
another kind of peptide that could form adaptable hydrogels by Hydrogels based on hydrazone bonds feature rapid geletion and
self-assemble via hydrophobic interaction [84]. pH-responsive degradation. They have been widely studied in different
applications including cell delivery and drug release [92–96].
2.2. Dynamic covalent linkages
2.2.3. Schiff base bonds
Dynamic covalent bonds are a class of reversible chemical bonds that The Schiff base is produced from the reaction between aldehyde
can be broken and reformed under equilibrium conditions. From the groups or ketone and amines. Due to the harsh reaction conditions of
safety consideration, toxic catalysts or relatively extreme conditions ketone and amine, only aldehyde groups could be applied for biological
such as temperature and pH reversible reactions are not available for material. The self-healing property of Schiff base is based on rapid
adaptive tissue regenerative materials. Table 1 shows the dynamic co­ dissociation and recombination without external stimuli. Aliphatic
valent bonds that have been studied, including Diels-Alder reactions, Schiff bases are typically more suitable for cell growth because of the
hydrazine bonds, Schiff bases, hydrazine bonds, thioesters, borate es­ easier reversible breakage [97,98]. Because of the wide range of sub­
ters, and disulfide bonds. stances which have aldehyde groups or ketone and amines, hydrogels
based on Schiff base are common. Furthermore, with ε-polylysine and
2.2.1. Reversible Diels–Alder reaction aldehyde groups modified Pluronic F127, Wang et al. reported a method
The Diels-Alder (DA) reaction is a click chemistry with high selec­ to fabricate adapatble hydrogels based on micelles and ε-polylysine
tivity and good yield, without any by-products. DA reactions can occur assembled nanoparticles [99]. Similarly, Zhou et al. reported
in a liquid physiological environment, and the equilibrium of the reac­ polypeptide-based hybrid nanosystems with mutifunctions based on
tion can be controlled by temperature. In past studies, reversible DA Schiff base reaction between -polylysine and aldehyde groups modified
reactions required high temperatures (above 100 ◦ C) which are un­ Pluronic F127 [100].
suitable for the physiological environment [85]. However, recent
studies have found that furan and maleimide can undergo reversible DA 2.2.4. Thioester
reaction at 37 ◦ C, which can be used for tissue engineering [86]. With Thioesters (e.g. coenzyme A) are widely used in biosynthesis, and
disulfide bonds that contain norborene and methylphenyltetrazine, their exchange rate in aqueous solution depends on the nucleophilicity
Delplace et al. designed inverse electron-demand DA reactions which and protonation state of sulfide. The formation of alkyl thioesters ex­
could happen under physiological conditions (pH and temperature) ceeds the competitive hydrolysis reaction at physiological pH. Changes
[87]. However the gelation based on inverse electron-demand DA re­ in pH and reactant metering can modulate thioester exchange to yield
actions is still slow (over 30 min) with pH 5.5 which is not sutiable for thioester-based adaptable hydrogel [101,102].
most cells. Two alternative strategies could be adpoted: (1) replacing the
furan diene with an even more electron-rich cyclic diene, such as a 2.2.5. Boronate bonds
fulvene, would further enhance the reaction rate of the DA reaction [88, The reaction between boric acid and diol compounds could produce
89], or (2) adopting additional weak linkages like thermosensitive dynamic boronate bonds, which are common in biofilms of bacteria that
furyl-modified hydroxypropyl chitin (FGE-HPCH) to stablize the combine carbohydrates with lectin. Phenylboronic acid (PBA), also
hydrogels before DA reaction is accomplished [90]. called “synthetic lectin”, was applied to mimic chemical structure of
lectins in bacteria and obtain self-healing materials [103]. However, its
2.2.2. Hydrazone bonds primary/secondary amines are toxic, and PBA needs to be modified to a
The hydrazone bonds produced from the reaction between aldehyde polymer backbone at a pH lower than the physiological environment. By

Table 1
Dynamic covalent bonds applied for adaptable hydrogel.
Kind Chemical Reaction Ref

C-C
Reversible Diels–Alder Reaction [85–90]

C-N
Hydrazone Bonds [91–96, 111, 122, 172, 173]

Schiff Base Bonds [97,98]

C-S
Thioester [101,102]

B-O
boronate bonds [104]

S-S
Disulfide Bonds [107]

1379
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

synthesizing monomers with PBA, adaptable hydrogels could be formed interface structure [118,119]. However, how to maintain stemness in 3D
under physiological conditions [104]. Additionally, numerous attempts culture is still difficult.
have been made to improve the arylboronic acid/diol binding affinity by Besides of stiffness and degradability, stress relaxation of adaptable
lowering the pKa of the boronic acid. Wu et al. reported nopoldiol, a hydrogels with reversible linkages has inspired researchers to study the
rigid unnatural cisdiol that provides fast, stable, and bioorthogonal influence of dynamic mechanical environments on stem cells [111,120].
binding with arylboronic acids to build novel type of network with Different from the constant elastic modulus of hydrogels with irrevers­
acid/polyol resistance, fast gelation [105]. The cis-diol moiety on fura­ ible linkages, the dynamic elastic modulus of adaptable hydrogels with
nose ring of guanosine can also react with borates or boronates via reversible linkages is enabled by fast stress relaxation. Max et al. utilized
boronic acid-diol complexation, and this property is usually used to alginate hydrogels crosslinked with electrostatic interaction to set a
improve the stability of guanosine-based hydrogels [106]. series of hydrogels with different stress relaxation rates to verify their
influences on the maintenance of stemness [121]. As a result, gene
2.2.6. Disulfide bonds expression of human cortical neuron progenitors (hNPC) and MSCs
In living organisms, disulfide bonds formed by the thiol side chains of showed sensitvity to the stress relaxation of alginate hydrogel.
cysteine residues play a key role in protein folding and assembly. Furthermore, fast stress relaxation of adaptable hydrogels is consistant
Although the thiol is not active in its reduced state and the formation of with native ECM, which caused hydrogels with reversible linkages are
the disulfide requires oxidant, the mild reaction condition is biocom­ more similar to native ECM than those hydrogels with irreversible
patible. Potential limitations of this cross-linking mechanism involve an linkages. The reversible and dynamic network of adaptable hydrogels
off-target reaction with a protein that may interfere with its biological was also proven to be helpful for ECM remodeling like native ECM, and
activity. In addition, it may exhibit poor stability in the presence of a would permit cell spreading and cell-cell contact [122,123].
reductant like glutathione [107].
3.2. Cell secretome regulation
3. Adaptable hydrogel’s effect on cells
In the progress of cell proliferation and tissue regeneration, secretion
Protection is the major aim of hydrogel for cell encapsulation, mostly plays an important role in cell behavior and cell fate [124,125]. Stem
in terms of immunoisolation. Immuno-privilege is an essential difficulty cells could influence tissue regeneration not only by cell proliferation
that cell therapy must face, which requires a protective barrier to ensure but also by cell secretome regulation [126,127]. Secretome from
high cell viability in vivo. Additional multiple functions have also been mesenchymal stem cells (MSCs) includes a great variety of cytokines and
proposed for cell encapsulation systems, such as exchange bioactive chemokines, which have been shown to play important roles in tissue
substances [108,109]. Hydrogels are common matrix for 3D cell repair and regeneration. Therapies with the application of secretome
encapsulation because of their hydrophilicity, injectability, etc (Fig. 5c). have a superior safety profile compared to live cell administration [128,
In comparison to conventional hydrogels, most adaptable hydrogels are 129].
injectable and self-healing through reversible linkages, which provide The constant stiffness of materials was hypothesized to facilitate the
better protection of encapsulated cells through dissipating shear/com­ regulation of stem cell secretome [130]. With engineered recombinant
press force. As shown in Fig. 5a and b, adaptable hydrogel could dissi­ proteins, which crosslinked with tunable stiffness, the stem cell secre­
pate force when sheared or pressed [110,111]. Additionally, the tome was regulated in situ [124]. Such an adaptable design of “shear-­
successful and easy access to cell encapsulation was the basement of thinning hydrogels for injectable encapsulation and long-term delivery”
further bio-printing, tissue regeneration and other applications. In terms was named “SHIELD”, and applied to culture human adipose-derived
of bio-printing, the shear-thinning hydrogel with reversile linkages stem cells (hASCs). Two stages of gelation were adopted to form
could prevent cell sediment, which is common in conventional covlant SHIELD: (1) weak gelation of peptide assemble, ands (2) strong gelation
hydrogel such as GelMA, PEGDA, etc. [112]. of a thermoresponsive PNIPAM. The series of hydrogels were named as
In past decades, strategies including adding bioactive factors and “SHIELD-n”, in which n represnets content of PNIPAM. As shown in
designing materials surface and interface, have been developed to Fig. 6a, gene expression and secreted paracrine factors of cells cultured
regulate or guide cell behavior and fate for tissue engineering. However, SHIELD- 1.25 and SHIELD-2.5 hydrogels were proven in higher level. As
in recent years, dynamic physical cues were discovered to have essential a result, enhanced secretome of hASCs cultured in SHIELD hydrogels
effects in maintenance of stemness, cell secretome regulation, and cell could lead to better angiogenic function of hMVECs. As shown in Fig. 6b.
spread, expansion, migration, and differentiation. Significantly more network junctions and loops were observed in hASCs
cultured in SHIELD than those cultured in basal media.
3.1. Maintenance of stemness
3.3. Cell spreading, expansion and migration
Maintenance of stemness and further differentiation potential of
stem cells is critical for clinical application [113,114]. Substrates with As a basic physical quality, stress relaxation is a matrix biophysical
high stiffness were proven to be helpful for directional differentiation of signal related to cell behaviors such as cell spreading, fiber remodeling,
stem cells, which would cause loss of stemness [115–117]. Previous focal adhesion formation, volume expansion, and migration [131,132].
studies focus on matrix stiffness, topological morphology, and other Both physical and dynamic chemical crosslinked adaptable networks

Fig. 5. Shear-thinning and self-healing ELP–HA

hydrogel can protect cells during syringe needle flow.
a) Live/dead (green/red) confocal projection images
of MSCs exposed to syringe needle flow (28 gauge,
0.5 in., 4 mL min− 1) within PBS or ELP–HA hydrogel.
b) Quantification of cell viability immediately post­
injection for multiple flow rates. Error bars represent
mean ± SD. *p < 0.05, n = 5. c) Confocal 3D re­
constructions of live/dead MSCs (green/red) post­
injection (28 gauge, 0.5 in., 4 mL min− 1) within PBS
and ELP–HA hydrogel [111]. Copyright (2017) Wiley.

1380
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

Fig. 6. a) i. Confocal 3D projection images of hASCs within Matrigel sandwiches stained with DAPI (blue) for cell nuclei and rhodamine phalloidin (red) for F-actin
cytoskeleton after 2 days exposure to various media treatments. ii. Relative mRNA expression of hASCs within SHIELD at day 14 post injection. b). Relative growth
factor concentration of collected media conditioned by hASCs within SHIELD at day 14 post injection. *p < 0.05, n = 4 [124]. Copyright (2016) Wiley.

were studied as regulators of cell spreading. Compared with constant responses (activation/inhibition). Such responses result from ECM
elastic modulus and static spatial architecture of hydrogels with irre­ remodeling and biophysical signals, and further decide cell fates. Spe­
versible linkages, hydrogels with reversible linkages show stress relax­ cifically speaking, native ECM mainly consists on collagen which shows
ation and plasticity [132]. The stress relaxation of adaptable hydrogels fast stress relaxing and could be easily remodeled (Fig. 7a). Adaptable
contributes to biophysical signals to integrin that guides cell behavior hydrogels with reversible linakges present similar stress relaxing with
including spreading and expansion. On the contrary, the static me­ collegan which could provide dynamic mechanical environment for
chanical environment is responsible for inhibited focal adhesion for­
mation and further cell spreading and expansion [122]. As a result,
restriction of expansion by hydrogels with slow stress relaxation with
diminishes in osteogenesis. Beyond the knowledge of
protease-dependent and pores or channels required migration, the
adaptable network allows cells to migrate through confining matrix
without protease [133].

3.4. Cell differentiation

Tunable properties of ECM could signal cells to influence their dif­

ferentiation, with which hydrogels could be designed to induce cells
differentiation [134]. For instance, with viscoelastic alginate-Ca2+
hydrogels that present stress relaxation and creep, chondrogenesis dif­
ferentiation was found to be related to faster relaxation [135]. The slow
stress relaxation restricts cell volume expansion, which further increases
the secretion of IL-1 (cell factor associated with cartilage degradation
and cell death), and finally triggers chondrogenesis differentiation. Also,
when encapsulated in amphiphilic PNIPAM based hydrogel, hMSC
differentiated into nucleus pulposus like cells with undesirable bone
formation under hypoxic conditions [136]. Benefitting from the con­
ductivity of polyelectrolytes, self-assembly hydrogels consist of cationic
PEG-peptide and anionic poly(3,4-ethylenedioxythiophene): poly­
styrene sulfonate (PEDOT: PSS) could cause MSCs to differentiate into
myocardiocyte-like cells [137].
As above mentioned, matrix cues have been demonstrated as potent
regulators of epigenetics and stem cells function. In the past, most effects
of adaptable hydrogels were ascribed to the regulator response to me­
chanical and spatial dynamic [138]. Additionally, pathways activated
by these cues including matrix stiffness, degradability and stress relax­ Fig. 7. a. Stress relaxation of elastic hydrogel based on irreversible covalent
ation; this illustrates how cells functions are regulated were discovered bonds, stress relaxing hydrogels based on reversible linkages and native ECM
[139]. Commonly, biophysical signals of extracellular matrix are collagen. b. Illustration of static mechanical environment and fixed architecture
of elastic hydrogels, and dynamic mechanical environment and remodelable
received by integrins, then affect cell by pathways e.g. Rho/ROCK (Rho
architecture of stress relaxing hydrogels. c. Inhibited YAP/TAZ of cell incubated
protein/Rho-associated kinase), which is necessary for the activity of the
in elastic hydrogels, and activated YAP/TAZ in stress relaxing hydrogels. Also,
YAP (Yes-associated protein) and TAZ (WW domain containing tran­ the stress relaxing rate impacts maturation of FA and F-actin remodeling. d.
scription regulator) transcription factors [140]. Representative immunofluorescence staining for F-actin (orange), nucleus
In recent years, YAP/TAZ were studied as two highly related (blue), and YAP/TAZ (magenta) in hMSCs cultured in elastic and stress-relaxing
mechanosensitive transcriptional regulators for cells [141]. Matrices gels for 1 and 7 d. Images are shown as one cross-sectional slice of a z-stack with
with varied mechanical behaviors affect cells via different YAP/TAZ a step size of 1.5 μm. Scale bar = 5 μm [104]. Copyright (2018) The Authors.

1381
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

cells. However, non-degradable covalent networks are elasctic and host-guest interaction, and slowly released in vivo. After several weeks,
provide constant mechnical environment which is not adaptable for obvious degradation could be seen in microgel morphology. As shown in
cells. Also, spatial ECM remodeling is important for cells. As shown in Fig. 8a, the cleavable microgel was cleared in the MI condition, while
Fig. 7b and c,.the activation of YAP/TAZ, maturation of FA and F-actin large aggregates of partially degraded microgels remained in the healthy
remodeling are impacted by the mechnical environment. Compared to condition. Across all conditions, stable microgels remained present with
elastic hydrogels, cells cultured in stress relax hydrogels present acti­ spherical, nondegraded morphologies throughout the duration of the
vated YAP/TAZ, and further cell volume expansion and spread (Fig. 7d) three weeks study. As shown in Fig. 8b, quantified microgel presence at
[104]. the two weeks time point by measuring the fluorescence intensity and
Additionally, more pathways have been studied. For instance, vol­ area of both microgel types in order to provide a metric of material mass
ume expansion controls nuclear localization of RUNX2 (Runt-related and volume in the injection site. Cleavable material normalized fluo­
transcription factor 2), but not YAP, was demonstrated to promote rescence intensity in the MI condition was ~40% in the no MI condition
osteogenesis in 3D culture [132]. and cleavable material normalized area was ~20% of the normalized
area in the no MI condition. This indicated preferential degradation of
4. Application of adaptable hydrogel cleavable microgels in the MI condition. There were no significant dif­
ferences in stable microgel fluorescence intensity between conditions.
4.1. Cardiac regeneration after myocardial infarction Relative to the no MI condition, there was a small decrease in the
amount of stable normalized microgel area in the MI group, possibly
Myocardial infarctions are the leading cause of mortality in heart indicating increased dispersion of the stable microgels in the tissue after
diseases. These infarctions result in permanent loss of contractile car­ MI, likely due to increased cellular invasion as the cleavable microgels
diomyocytes (native muscle cells in the heart). The major challenge to were degraded.
heart regeneration is the limited capacity for renewal of cardiomyocytes
in the adult heart [142]. In the past, various hydrogels have been
4.2. Bone regeneration
developed for treating myocardial infarction, which still could not meet
the complex demands of heart tissue repair. Most covalent crosslinked
Bone defect is a common clinical disease which raised numerous
hydrogel could not cover the entire surface area of tissues with irregular
strategies including implantation, transplantation and cellular therapy,
surfaces or those that are heavily folded. The gelation of conventional
etc. Bian and his colleagues [144,145] have reported several works that
hydrogels with irreversible linkages is uncontrollable, which means fast
applied adaptable hydrogel to bone regeneration. Bone regeneration is
gelation brings difficulty to operation and slow gelation would cause
an important kind of tissue regeneration, but the bones must be very
failed retention of cells or drugs. Furthermore, the brittle and/or unable
hard. One way to meet this strength requirement through hydrogels is by
to accommodate the dynamic movement of tissues in the body. Staple­
dual crosslinking [145].
ton et al. reported supramolecular polymeric hydrogels composed of
Feng et al. reported their work on gelatin hydrogels based on host-
dodecyl (C12)-modified hydroxypropylmethylcellulose and poly
guest interactions to assist bone regeneration [36]. As shown in
(ethylene glycol)-b-poly(lactic acid) nanoparticles as an effective
Fig. 9a, the host-guest macromer (HGM) hydrogels are crosslinked by
post-operative pericardial adhesion barrier [143]. Such supramolecular
supramolecular interaction between cyclodextrin and aromatic residues
polymeric hydrogels could be injected and self-heal rapidly to reversibly
of gelatin. Injetablility and plasticity could be attributed to the revers­
transition from viscous flow back to a solid-like barrier, allowing the
ible linkages, which enable minimally invasive therapy and cell infil­
hydrogels to quickly adhere and settle on the target tissue. Compared to
tration. To compare the difference of cell infiltration in reversible and
irreversible cardiac adhesions, supramolecular polymeric hydrogels
irreversible linkages. HGM hydrogels and methacrylated gelatin
showed better retention and remodeling that suit soft tissue. Addition­
(MeGel) hydrogels were adpoted to culture hMSCs (Fig. 9b) with SDF-1α
ally, adaptable hydrogels might be helpful to accomplish heart regen­
gradient. Fig. 9c revealed that HGM hydrogels could enable cell infil­
eration by cellular therapy or gene delivery. Burdick and his colleagues
tration and migration because of the reversible nature while MeGel
[38] developed an adaptable hydrogel with shear-thinning and
inhibited cell infiltration.
self-healing properties based on macrocyclic host-guest complexation.
Bisphosphonate-magnesium (Mg2+) coordination was also utilized to
Such an adaptable hydrogel is composed of adamantine modified hy­
obtain adaptable hydrogels which could be infiltrated by cells or regu­
aluronic acid (HA) microgel and cyclodextrin modified HA with gran­
late differentiation [72]. Dexamethasone phosphate-laden hydrogels
ular structure. Besides of injectablity, stress relaxation, and plasticity,
based on pamidronate-grafted hyaluronic acid crosslinked with mag­
the granular structure could be designed to control porosity of hydrogels
nesium ion (HA-Pam-Mg-DexP) were formed by
for cell invasion.
bisphosphonate-magnesium coordination. As a result, self-healing and
Furthermore, they applied such guest–host HA hydrogels to sus­
improved mechanical properties could be found in their adaptable
tained miRNA delivery to promote cardiomyocyte proliferation and
hydrogels. Further design for osteogenic differentiation regulation was
functional regeneration after ischemic injury [39]. MicroRNA mimic
related to release of magnesium ions and dexamethasone, which was
(miR-302) modified with cholesterol was assembled into hydrogel via
catalyzed by activation of alkaline phosphatase (ALP). Such injectable

Fig. 8. a) Two-component granular hydrogels

labeled with FITC for cleavable microgels and
rhodamine for stable microgels were injected into rat
hearts either without myocardial infarction (no MI)
or with MI. Representative images of microgels and
cell nuclei (blue) at one, two, and three weeks after
injection (scale bar = 500 μm). b) Quantification of
remaining microgels at two weeks, as fluorescence
intensity normalized to the injection site area (n = 3).
Statistical significance was assessed relative to
healthy controls with p < 0.05 [38]. Copyright
(2018) Wiley.

1382
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

Fig. 10. Schematic illustration of self-assembling peptide hydrogel-coated PCL

scaffolds in osteochondral defect model and the interactions between the pep­
tide chains of SAPH and the possible interactions between SAPH and PCL [60].
Copyright (2018) Wiley.

4.5. Other applications

Fig. 9. a) The schematic illustration of the injection of hMSC-laden HGM
hydrogels: the shear stress of the injection coverts the solid HGM hydrogel into The applications of adaptable hydrogels have been expanded into
the “sol” state. Cell viability staining of the hMSC-laden HGM hydrogels after 3 wider fields including wound healing, adhesion, and drug delivery. Li
days of in vitro culture following the injection of the HGM hydrogels. b) et al. [153] attempted to build double crosslinked networks with dy­
Schematic illustration of cell migration experiment. The hMSCs seeded on the namic covalent and physical linkages to achieve rapid gelation as well as
surface of the HGM hydrogels infiltrate into the hydrogels, but few seeded cells shear thinning properties. Such hydrogel was constituted with hydrazide
infiltrate into the MeGel hydrogels. c) The 3D distribution of DAPI-stained modified hyaluronic acid (HAAD) and benzaldehyde terminated F127
hMSC nuclei clusters within the MeGel and HGM hydrogels. The confocal mi­ triblock copolymers (BAF127). The rapid gelation satisfied the inject­
crographs show the 3D distribution of DAPI-stained hMSC nuclei clusters within
ability of hydrogel to fit irregular burn wounds. The transplantation of
the MeGel hydrogelsand HGM hydrogels after 2 h of exposure to a chemo­
endothelial cells with adaptable hydrogel can overcome the limitation of
attractant gradient. Scale bar: 100 mm (a) and 100 μm (c) [36]. Copyright
(2016) Elsevier.
transplanted cell death, which was confirmed as a promising therapeutic
strategy for peripheral arterial disease [154].
Additionally, the adaptable network could include post-operative
hydrogel combined with drugs and cells could effectively promote in
adhesions which should satisfy the injection to organs, viscoelasticity
situ bone regeneration via minimally invasive procedures.
that enables organs and tissues to move freely, and effectively preven­
tion of adhesions of tissues. Stapleton et al. [143] reported a
4.3. Cartilage regeneration
dynamically-crosslinked supramolecular polymer–nanoparticle (PNP)
hydrogel for post-operative adhesions. These PNP hydrogels are formed
Cartilage defect caused by osteoarthritis, trauma and other reasons is
by simple mixing of aqueous solutions of dodecyl (C12)-modified
an important health issue but difficult to be repaired [146,147]. Various
hydroxypropylmethylcellulose (HPMC-C12) with biodegradable poly­
strategies including structures [149,150] (fibrous, porous, etc), me­
meric nanoparticles comprising poly(ethylene glycol)-b-poly(lactic
chanical stimulation, and degradability [151] have been attempted and
acid) (PEG–PLA). The simplicity of their preparation through a
achieved only limited success [125]. The differentiation and prolifera­
self-assembly process enables PNP hydrogel manufacturing to be easily
tion of chondrogenic cells are key points limiting clinical effect of
scaled up, producing materials with identical mechanical properties on
cartilage regeneration [152]. Richardson et al. [132] reported an
any scale.
adaptable hydrazone covalent network for cartilage regeneration with
In terms of brain disease, conventional drug delivery strategies are
tunable viscoelastic properties and time-dependent properties, which
typically limited by the blood-brain barrier. Local delivery could be an
impacted chondrocyte proliferation and matrix deposition.
impossible way to ideal therapy which required vehicles should be
mouldable, injectable, and mechanically softer than surrounding brain
4.4. Osteochondral regeneration
tissue, and readily tailorable cargo loading and delivery properties.
Rowland et al. [43] developed physical hydrogel cross-linked via the
Clinically, osteochondral regeneration remains a great challenge
host-guest interactions of cucurbit [8]uril (CB8) as drug-delivery vehicle
because of the concurrence of subchondral bone injuries and cartilage
for brain. Such hyaluronic acid cysteine-phenylalanine/cucurbit [8]uril
damage. The integrative repair of osteochondral regeneration requires
hydrogel can be tuned to appropriately complement the surrounding
mechanical support which should be provided by subchondral bone or
environment, ensuring continuous structural and shape remodeling that
its substitute and nutrient exchange between cartilage and bone marrow
would be coherent with tissue healing. Such hydrogels provide an
cavities. As shown in Fig. 10, the combination of strong scaffolds and
effective alternative to carmustine loaded wafers in order to bypass the
adaptable peptide hydrogel was adopted to meet mechanical support
blood-brain barrier and provide synergistic, personalized therapies
and 3D microenvironment for cell culture [60]. The hydrophobic
locally for patients with glioblastoma.
interaction between β-sheet of peptide and repeated unit -[O
(CH2)5CO]n-of polycaprolactone ensured the stable adhesive for a long
5. Conclusions and perspectives
time. Prolonged therapy was achieved by the separate deposition of
chondrogenic and osteogenic ECM proteins.
In summary, numerous adaptable networks based on chemical or
physical linkages have been developed by researchers, which has pro­
vided tools for further study. The effects of adaptable hydrogel with

1383
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

spatiotemporal and mechanical dynamic on cell behavior has been a Appendix A. Supplementary data
rewarding object of inquiry. Two especially important factors for desi­
gining these adaptable hydrogels are fast stress relaxation and easy Supplementary data related to this article can be found at https://doi
remodeling capacity. Ease of remodeling is ensstional for maintenance .org/10.1016/j.bioactmat.2020.10.029.
of stemness and cell migrations, applications that require networks to
break and reform rapidly. Various reversible linkages could be adopted References
with consideration of gelation rate, and physiological conditions (like
pH, redox, etc). Additionally, fast stress relaxation should be inten­ [1] J.F. Burke, I.V. Yannas, W.C. Quinby, C.C. Bondoc, W.K. Jung, Successful use of a
physiologically acceptable Artificial skin in the treatment of extensive burn
tionally controlled for specific tissue engineering to induce targeted cell injury, Ann. Surg. 194 (4) (1981) 413–428.
differentiation. Plenty of practical applications have been proposed, and [2] J. Xue, B. Feng, R. Zheng, Y. Lu, G. Zhou, W. Liu, et al., Engineering ear-shaped
evidence has been provided to support the effectiveness of some. cartilage using electrospun fibrous membranes of gelatin/polycaprolactone,
Biomaterials 34 (11) (2013) 2624–2631.
However, some problems continue to exist, including the insufficient [3] N. Huebsch, E. Lippens, K. Lee, M. Mehta, S.T. Koshy, M.C. Darnell, et al., Matrix
dynamic due to limited reversible linkage and balance between stability elasticity of void-forming hydrogels controls transplanted-stem-cell-mediated
and dynamic, and the lack of reliable quantitative evaluation and bone formation, Nat. Mater. 14 (12) (2015) 1269–1277.
[4] X. Bai, M. Gao, S. Syed, J. Zhuang, X. Xu, X. Zhang, Bioactive hydrogels for bone
necessary biological knowledge. Even though the reversible linkages regeneration, Bioactive Materials 3 (4) (2018) 401–417.
could benefit cell behavior as a better dynamic ECM network and soft [5] S. Jekhmane, M. Prachar, R. Pugliese, F. Fontana, J. Medeiros Silva, F. Gelain, et
stiffness adapt cells, reversible linkages that can break and form even al., Design parameters of tissue-engineering scaffolds at the atomic scale, Angew.
Chem. 131 (47) (2019) 17099–17107.
more quickly should be developed. Besides, the poor stability and me­
[6] G. Pellegrini, D. Ardigò, G. Milazzo, G. Iotti, P. Guatelli, D. Pelosi, et al.,
chanical strength of single reversible linkages limit practical applica­ Navigating market authorization: the path holoclar took to become the first stem
tions. Most single reversible linkages could not satisfy practical cell product approved in the European union, Stem Cell. Transl. Med. 7 (1)
mechanical support or long term gelation. More strategies, including (2018) 146–154.
[7] M. Allison, Hemacord approval may foreshadow regulatory creep for HSC
double network or composition with strong scaffold should be devel­ therapies, Nat. Biotechnol. 30 (4) (2012) 304.
oped. Additionally, with ideal injectable property, adaptable hydrogels [8] M. Allison, Hemacord approval may foreshadow regulatory creep for HSC
would be important bio-ink for 3D bio-printing. Beyond the present therapies, Nat. Biotechnol. 30 (4) (2012) 304.
[9] C. Willyard Timeline, Regrowing the body, Nature 540 (7632) (2016) S50–S51.
polymer/monomer solutions ink, more styles like granular hydrogels, [10] B.A. Syed, J.B. Evans, Stem cell therapy market, Nat. Rev. Drug Discov. 12 (3)
nanoparticle based hydrogels, microgels etc, could be developed to (2013) 185–186.
satisfy more functions like complex cell coculture or cargo delivery. [11] R. Langer, J. Vacanti, Tissue engineering, Science 260 (5110) (1993) 920–926.
[12] A.M. Rosales, K.S. Anseth, The design of reversible hydrogels to capture
Several papers have reported how material matrices affect cell extracellular matrix dynamics, Nat. Rev. Mater. 1 (2) (2016).
behavior, but conclusive knowledge requires more evidence and dis­ [13] D. Seliktar, Designing cell-compatible hydrogels for biomedical applications,
coveries. For instance, the capability of modulating the behaviors of the Science 336 (6085) (2012) 1124–1128.
[14] G.S. Hussey, J.L. Dziki, S.F. Badylak, Extracellular matrix-based materials for
immune cells (such as macrophage polarization, cell infiltration) with regenerative medicine, Nat. Rev. Mater. 3 (7) (2018) 159–173.
hydrogels which could be helpful to tissue regeneration was reported [15] W.E. Hennink, C.F. van Nostrum, Novel crosslinking methods to design hydrogels,
[155,156]. Macrophage polarization could be induced by adaptable Adv. Drug Deliv. Rev. 64 (2012) 223–236.
[16] J.W. Nichol, S.T. Koshy, H. Bae, C.M. Hwang, S. Yamanlar, A. Khademhosseini,
hydrogels which could be verified by more M1 and M2 macrophage
Cell-laden microengineered gelatin methacrylate hydrogels, Biomaterials 31 (21)
biomarkers expression in the regeneration process compared to con­ (2010) 5536–5544.
ventional hydrogels [157,158]. Further study needs to be carried to [17] N. Annabi, S.M. Mithieux, P. Zorlutuna, G. Camci-Unal, A.S. Weiss,
figure out what factor (stiffness, dynamic mechanical, or spatial cues) A. Khademhosseini, Engineered cell-laden human protein-based elastomer,
Biomaterials 34 (22) (2013) 5496–5505.
should be responsible for the difference. Furthermore, quantitative [18] S.J. Hollister, Porous scaffold design for tissue engineering, Nat. Mater. 4 (7)
analysis and precise model should be set up for universal design of (2005) 518–524.
adaptable matrices for tissue engineering. At present, quantitative [19] P. Tayalia, D.J. Mooney, Controlled, Growth factor delivery for tissue
engineering, Adv. Mater. 21 (32–33) (2009) 3269–3285.
analysis has been considered from chemical equilibrium view or com­ [20] L. Jongpaiboonkit, W.J. King, G.E. Lyons, A.L. Paguirigan, J.W. Warrick, D.
bined view of viscoelasticity and corresponding cell behavior. On the J. Beebe, et al., An adaptable hydrogel array format for 3-dimensional cell culture
one hand, we can understand how to obtain a fast gelation according to and analysis, Biomaterials 29 (23) (2008) 3346–3356.
[21] T. Lecuit, P. Lenne, Cell surface mechanics and the control of cell shape, tissue
chemical equilibrium even though the stability and mechanical prop­ patterns and morphogenesis, Nat. Rev. Mol. Cell Biol. 8 (8) (2007) 633–644.
erties could not be predicted. On the other hand, by adjusting the ma­ [22] F. Burla, Y. Mulla, B.E. Vos, A. Aufderhorst-Roberts, G.H. Koenderink, From
terials, we can obtain materials with different stress relaxation mechanical resilience to active material properties in biopolymer networks,
Nature Reviews Physics 1 (4) (2019) 249–263.
properties and corresponding cell behaviors. But to meet the practical [23] X. Ding, Y. Wang, Weak bond-based injectable and stimuli responsive hydrogels
application, both views need to be considered and carried out for for biomedical applications, J. Mater. Chem. B 5 (5) (2017) 887–906.
follow-up research. The former can provide researchers with a basis for [24] X. Tong, F. Yang, Recent progress in developing injectable matrices for enhancing
cell delivery and tissue regeneration, Adv. Healthc. Mater. 7 (7) (2018) 1701065.
material design, while the latter provides a good way to verify and
[25] C.M. Madl, S.C. Heilshorn, H.M. Blau, Bioengineering strategies to accelerate
evaluate materials. stem cell therapeutics, Nature 557 (7705) (2018) 335–342.
[26] H. Wang, S.C. Heilshorn, Adaptable hydrogel networks with reversible linkages
Declaration of competing interest for tissue engineering, Adv. Mater. 27 (25) (2015) 3717–3736.
[27] P. Heidarian, A.Z. Kouzani, A. Kaynak, M. Paulino, B. Nasri-Nasrabadi, Dynamic
hydrogels and polymers as inks for three-dimensional printing, ACS Biomater.
The authors declare no conflict of interest. Sci. Eng. 5 (6) (2019) 2688–2707.
[28] E.A. Appel, J. Del Barrio, X.J. Loh, O.A. Scherman, Supramolecular polymeric
hydrogels, Chem. Soc. Rev. 41 (18) (2012) 6195–6214.
Acknowledgments [29] A. Harada, Y. Takashima, M. Nakahata, Supramolecular polymeric materials via
cyclodextrin–guest interactions, Accounts Chem. Res. 47 (7) (2014) 2128–2140.
We thank the financial support of the National Key Research and [30] G. Liu, Q. Yuan, G. Hollett, W. Zhao, Y. Kang, J. Wu, Cyclodextrin-based
host–guest supramolecular hydrogel and its application in biomedical fields,
Development Program of China (2016YFE0132700), National Natural Polym. Chem.-Uk 9 (25) (2018) 3436–3449.
Science Foundation of China (51822306, 51673171), Science Technol­ [31] G. Chen, M. Jiang, Cyclodextrin-based inclusion complexation bridging
ogy Department of Zhejiang Province (2020C03042), and the Funda­ supramolecular chemistry and macromolecular self-assembly, Chem. Soc. Rev. 40
(5) (2011) 2254–2266.
mental Research Funds for the Central Universities of China. [32] C.B. Rodell, A.L. Kaminski, J.A. Burdick, Rational design of network properties in
guest–host assembled and shear-thinning hyaluronic acid hydrogels,
Biomacromolecules 14 (11) (2013) 4125–4134.

1384
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

[33] A.M. Rosales, C.B. Rodell, M.H. Chen, M.G. Morrow, K.S. Anseth, J.A. Burdick, [62] J. Raeburn, A. Zamith Cardoso, D.J. Adams, The importance of the self-assembly
Reversible control of network properties in azobenzene-containing hyaluronic process to control mechanical properties of low molecular weight hydrogels,
acid-based hydrogels, Bioconjugate Chem. 29 (4) (2018) 905–913. Chem. Soc. Rev. 42 (12) (2013) 5143.
[34] C.B. Rodell, N.N. Dusaj, C.B. Highley, J.A. Burdick, Injectable and cytocompatible [63] X. Yan, Z. Tong, Y. Chen, Y. Mo, H. Feng, P. Li, et al., Bioresponsive materials for
tough double-network hydrogels through tandem supramolecular and covalent drug delivery based on carboxymethyl chitosan/poly(γ-glutamic acid) composite
crosslinking, Adv. Mater. 28 (38) (2016) 8419–8424. microparticles, Mar. Drugs 15 (5) (2017) 127.
[35] C. Loebel, A. Ayoub, J.H. Galarraga, O. Kossover, H. Simaan-Yameen, D. Seliktar, [64] Z. Tong, J. Yang, L. Lin, R. Wang, B. Cheng, Y. Chen, et al., In situ synthesis of
et al., Tailoring supramolecular guest–host hydrogel viscoelasticity with covalent poly (γ- glutamic acid)/alginate/AgNP composite microspheres with antibacterial
fibrinogen double networks, J. Mater. Chem. B 7 (10) (2019) 1753–1760. and hemostatic properties, Carbohydr. Polym. 221 (2019) 21–28.
[36] Q. Feng, K. Wei, S. Lin, Z. Xu, Y. Sun, P. Shi, et al., Mechanically resilient, [65] J. Sun, X. Zhao, W.R.K. Illeperuma, O. Chaudhuri, K.H. Oh, D.J. Mooney, et al.,
injectable, and bioadhesive supramolecular gelatin hydrogels crosslinked by Highly stretchable and tough hydrogels, Nature 489 (7414) (2012) 133–136.
weak host-guest interactions assist cell infiltration and in situ tissue regeneration, [66] Z. Tong, Y. Chen, Y. Liu, L. Tong, J. Chu, K. Xiao, et al., Preparation,
Biomaterials 101 (2016) 217–228. characterization and properties of alginate/poly(γ-glutamic acid) composite
[37] Q. Feng, J. Xu, K. Zhang, H. Yao, N. Zheng, L. Zheng, et al., Dynamic and cell- microparticles, Mar. Drugs 15 (4) (2017) 91.
infiltratable hydrogels as injectable carrier of therapeutic cells and drugs for [67] R.L. Truby, J.A. Lewis, Printing soft matter in three dimensions, Nature 540
treating challenging bone defects, Acs Central Sci 5 (3) (2019) 440–450. (7633) (2016) 371–378.
[38] J.E. Mealy, J.J. Chung, H. Jeong, D. Issadore, D. Lee, P. Atluri, et al., Injectable [68] A. Lee, A.R. Hudson, D.J. Shiwarski, J.W. Tashman, T.J. Hinton, S. Yerneni, et al.,
granular hydrogels with multifunctional properties for biomedical applications, 3D bioprinting of collagen to rebuild components of the human heart, Science
Adv. Mater. 30 (20) (2018) 1705912. 365 (6452) (2019) 482–487.
[39] L.L. Wang, Y. Liu, J.J. Chung, T. Wang, A.C. Gaffey, M. Lu, et al., Sustained [69] I. Donderwinkel, J.C.M. van Hest, N.R. Cameron, Bio-inks for 3D bioprinting:
miRNA delivery from an injectable hydrogel promotes cardiomyocyte recent advances and future prospects, Polym. Chem.-Uk 8 (31) (2017)
proliferation and functional regeneration after ischaemic injury, Nat. Biomed. 4451–4471.
Eng. 1 (12) (2017) 983–992. [70] C.D. Lindsay, J.G. Roth, B.L. LeSavage, S.C. Heilshorn, Bioprinting of stem cell
[40] C. Loebel, C.B. Rodell, M.H. Chen, J.A. Burdick, Shear-thinning and self-healing expansion lattices, Acta Biomater. 95 (2019) 225–235.
hydrogels as injectable therapeutics and for 3D-printing, Nat. Protoc. 12 (8) [71] L. Ouyang, C.B. Highley, C.B. Rodell, W. Sun, J.A. Burdick, 3D printing of shear-
(2017) 1521–1541. thinning hyaluronic acid hydrogels with secondary cross-linking, ACS Biomater.
[41] J. Lagona, P. Mukhopadhyay, S. Chakrabarti, L. Isaacs, The Cucurbit[n]uril Sci. Eng. 2 (10) (2016) 1743–1751.
Family, Angew. Chem. Int. Ed. 44 (31) (2005) 4844–4870. [72] K. Zhang, Q. Feng, J. Xu, X. Xu, F. Tian, K.W.K. Yeung, et al., Self-assembled
[42] J. Yeom, S.J. Kim, H. Jung, H. Namkoong, J. Yang, B.W. Hwang, et al., injectable nanocomposite hydrogels stabilized by bisphosphonate-magnesium
Supramolecular hydrogels for long-term bioengineered stem cell therapy, Adv. (Mg2+) coordination regulates the differentiation of encapsulated stem cells via
Healthc. Mater. 4 (2) (2015) 237–244. dual crosslinking, Adv. Funct. Mater. 27 (34) (2017) 1701642.
[43] M.J. Rowland, C.C. Parkins, J.H. McAbee, A.K. Kolb, R. Hein, X.J. Loh, et al., An [73] D. Cross, X. Jiang, W. Ji, W. Han, C. Wang, Injectable hybrid hydrogels of
adherent tissue-inspired hydrogel delivery vehicle utilised in primary human hyaluronic acid crosslinked by well-defined synthetic polycations: preparation
glioma models, Biomaterials 179 (2018) 199–208. and characterization in vitro and in vivo, Macromol. Biosci. 15 (5) (2015)
[44] X. Tong, F. Yang, Sliding hydrogels with mobile molecular ligands and crosslinks 668–681.
as 3D stem cell niche, Adv. Mater. 28 (33) (2016) 7257–7263. [74] H. Li, W. Zhang, W. Tong, C. Gao, Enhanced cellular uptake of bowl-like
[45] S. Quideau, D. Deffieux, C. Douat-Casassus, L. Pouységu, Plant polyphenols: microcapsules, Acs Appl. Mater. Inter. 8 (18) (2016) 11210–11214.
chemical properties, biological activities, and synthesis, Angew. Chem. Int. Ed. 50 [75] Y. Chen, F. Wang, N. Zhang, Y. Li, B. Cheng, Y. Zheng, Preparation of a 6-OH
(3) (2011) 586–621. quaternized chitosan derivative through click reaction and its application to
[46] S.K. Madhurakkat Perikamana, J. Lee, Y.B. Lee, Y.M. Shin, E.J. Lee, A.G. Mikos, et novel thermally induced/polyelectrolyte complex hydrogels, Colloids Surf. B
al., Materials from mussel-inspired chemistry for cell and tissue engineering Biointerfaces 158 (2017) 431–440.
applications, Biomacromolecules 16 (9) (2015) 2541–2555. [76] P. Shi, N. Zhao, J. Coyne, Y. Wang, DNA-templated synthesis of biomimetic cell
[47] M. Shin, J.H. Ryu, J.P. Park, K. Kim, J.W. Yang, H. Lee, DNA/Tannic acid hybrid wall for nanoencapsulation and protection of mammalian cells, Nat. Commun. 10
gel exhibiting biodegradability, extensibility, tissue adhesiveness, and hemostatic (1) (2019).
ability, Adv. Funct. Mater. 25 (8) (2015) 1270–1278. [77] A.P. Nowak, V. Breedveld, L. Pakstis, B. Ozbas, D.J. Pine, D. Pochan, et al.,
[48] M. Shin, J.H. Galarraga, M.Y. Kwon, H. Lee, J.A. Burdick, Gallol-derived ECM- Rapidly recovering hydrogel scaffolds from self-assembling diblock copolypeptide
mimetic adhesive bioinks exhibiting temporal shear-thinning and stabilization amphiphiles, Nature 417 (6887) (2002) 424–428.
behavior, Acta Biomater. 95 (2019) 165–175. [78] J. Zhang, T. Tokatlian, J. Zhong, Q.K.T. Ng, M. Patterson, W.E. Lowry, et al.,
[49] C.B. Highley, C.B. Rodell, J.A. Burdick, Direct 3D printing of shear-thinning Physically associated synthetic hydrogels with long-term covalent stabilization
hydrogels into self-healing hydrogels, Adv. Mater. 27 (34) (2015) 5075–5079. for cell culture and stem cell transplantation, Adv. Mater. 23 (43) (2011)
[50] S.H. Um, J.B. Lee, N. Park, S.Y. Kwon, C.C. Umbach, D. Luo, Enzyme-catalysed 5098–5103.
assembly of DNA hydrogel, Nat. Mater. 5 (10) (2006) 797–801. [79] L. Cai, R.E. Dewi, S.C. Heilshorn, Injectable hydrogels with in situ double network
[51] H. Tan, C. Xiao, J. Sun, D. Xiong, X. Hu, Biological self-assembly of injectable formation enhance retention of transplanted stem cells, Adv. Funct. Mater. 25 (9)
hydrogel as cell scaffold via specific nucleobase pairing, Chem. Commun. 48 (83) (2015) 1344–1351.
(2012) 10289. [80] B.L. Ekerdt, C.M. Fuentes, Y. Lei, M.M. Adil, A. Ramasubramanian, R.
[52] C. Li, A. Faulkner-Jones, A.R. Dun, J. Jin, P. Chen, Y. Xing, et al., rapid formation A. Segalman, et al., Thermoreversible hyaluronic acid-PNIPAAm hydrogel
of a supramolecular polypeptide-DNA hydrogel for InSitu three-dimensional systems for 3D stem cell culture, Adv. Healthc. Mater. 7 (12) (2018) 1800225.
multilayer bioprinting, Angew. Chem. 127 (13) (2015) 4029–4033. [81] P.B. Rapp, A.K. Omar, J.J. Shen, M.E. Buck, Z. Wang, D.A. Tirrell, Analysis and
[53] P.Y.W. Dankers, T.M. Hermans, T.W. Baughman, Y. Kamikawa, R.E. Kieltyka, M. control of chain mobility in protein hydrogels, J. Am. Chem. Soc. 139 (10) (2017)
M.C. Bastings, et al., Hierarchical formation of supramolecular transient networks 3796–3804.
in water: a modular injectable delivery system, Adv. Mater. 24 (20) (2012) [82] V. Conticello, S. Hughes, C. Modlin, Biomaterials made from coiled-coil peptides,
2703–2709. Sub-cellular biochemistry 82 (2017) 575.
[54] G. Zhang, Y. Chen, Y. Deng, T. Ngai, C. Wang, Dynamic supramolecular [83] Y. Liu, B. Liu, J.J. Riesberg, W. Shen, In situ forming physical hydrogels for three-
hydrogels: regulating hydrogel properties through self-complementary quadruple dimensional tissue morphogenesis, Macromol. Biosci. 11 (10) (2011) 1325–1330.
hydrogen bonds and thermo-switch, ACS Macro Lett. 6 (7) (2017) 641–646. [84] M.J. Glassman, J. Chan, B.D. Olsen, Reinforcement of shear thinning protein
[55] M. Guo, L.M. Pitet, H.M. Wyss, M. Vos, P.Y.W. Dankers, E.W. Meijer, Tough hydrogels by responsive block copolymer self-assembly, Adv. Funct. Mater. 23 (9)
stimuli-responsive supramolecular hydrogels with hydrogen-bonding network (2013) 1182–1193.
junctions, J. Am. Chem. Soc. 136 (19) (2014) 6969–6977. [85] S. Kirchhof, F.P. Brandl, N. Hammer, A.M. Goepferich, Investigation of the
[56] Z. Qin, X. Yu, H. Wu, J. Li, H. Lv, X. Yang, Nonswellable and tough Diels–Alder reaction as a cross-linking mechanism for degradable poly(ethylene
supramolecular hydrogel based on strong micelle cross-linkings, glycol) based hydrogels, Journal of materials chemistry. B, Materials for biology
Biomacromolecules 20 (9) (2019) 3399–3407. and medicine 1 (37) (2013) 4855.
[57] L.M. De Leon Rodriguez, Y. Hemar, J. Cornish, M.A. Brimble, [86] M.H. Ghanian, H. Mirzadeh, H. Baharvand, In situ forming, cytocompatible, and
Structure–mechanical property correlations of hydrogel forming β-sheet peptides, self-recoverable tough hydrogels based on dual ionic and click cross-linked
Chem. Soc. Rev. 45 (17) (2016) 4797–4824. alginate, Biomacromolecules 19 (5) (2017) 1646–1662.
[58] L.A. Castillo Diaz, M. Elsawy, A. Saiani, J.E. Gough, A.F. Miller, Osteogenic [87] V. Delplace, A.J. Pickering, M.H. Hettiaratchi, S. Zhao, T. Kivijärvi, M.S. Shoichet,
differentiation of human mesenchymal stem cells promotes mineralization within Inverse electron-demand diels–alder methylcellulose hydrogels enable the Co-
a biodegradable peptide hydrogel, J. Tissue Eng. 7 (2016) 1013652270. delivery of chondroitinase ABC and neural progenitor cells, Biomacromolecules
[59] J.D. Tang, C. Mura, K.J. Lampe, Stimuli-responsive, pentapeptide, nanofiber 21 (6) (2020) 2421–2431.
hydrogel for tissue engineering, J. Am. Chem. Soc. 141 (12) (2019) 4886–4899. [88] C.M. Madl, S.C. Heilshorn, Rapid diels–alder cross-linking of cell encapsulating
[60] L. Li, J. Li, J. Guo, H. Zhang, X. Zhang, C. Yin, et al., 3D molecularly hydrogels, Chem. Mater. 31 (19) (2019) 8035–8043.
functionalized cell-free biomimetic scaffolds for osteochondral regeneration, Adv. [89] L.J. Smith, S.M. Taimoory, R.Y. Tam, A.E.G. Baker, N. Binth Mohammad, J.
Funct. Mater. 29 (6) (2019) 1807356. F. Trant, et al., Diels–alder click-cross-linked hydrogels with increased reactivity
[61] C. Yan, D.J. Pochan, Rheological properties of peptide-based hydrogels for enable 3D cell encapsulation, Biomacromolecules 19 (3) (2018) 926–935.
biomedical and other applications, Chem. Soc. Rev. 39 (9) (2010) 3528. [90] B. Bi, M. Ma, S. Lv, R. Zhuo, X. Jiang, In-situ forming thermosensitive
hydroxypropyl chitin-based hydrogel crosslinked by Diels-Alder reaction for three
dimensional cell culture, Carbohydr. Polym. 212 (2019) 368–377.

1385
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

[91] B.M. Richardson, D.G. Wilcox, M.A. Randolph, K.S. Anseth, Hydrazone covalent [117] D. Lü, C. Luo, C. Zhang, Z. Li, M. Long, Differential regulation of morphology and
adaptable networks modulate extracellular matrix deposition for cartilage tissue stemness of mouse embryonic stem cells by substrate stiffness and topography,
engineering, Acta Biomater. 83 (2019) 71–82. Biomaterials 35 (13) (2014) 3945–3955.
[92] L. Koivusalo, M. Kauppila, S. Samanta, V.S. Parihar, T. Ilmarinen, S. Miettinen, et [118] R. Randriantsilefisoa, Y. Hou, Y. Pan, J.L.C. Camacho, M.W. Kulka, J. Zhang, et
al., Tissue adhesive hyaluronic acid hydrogels for sutureless stem cell delivery al., Interaction of human mesenchymal stem cells with soft nanocomposite
and regeneration of corneal epithelium and stroma, Biomaterials 225 (2019) hydrogels based on polyethylene glycol and dendritic polyglycerol, Adv. Funct.
119516. Mater. 30 (1) (2020) 1905200.
[93] J.T. Koivisto, C. Gering, J. Karvinen, R. Maria Cherian, B. Belay, J. Hyttinen, et [119] C.M. Madl, B.L. LeSavage, R.E. Dewi, C.B. Dinh, R.S. Stowers, M. Khariton, et al.,
al., Mechanically biomimetic gelatin–gellan gum hydrogels for 3D culture of Maintenance of neural progenitor cell stemness in 3D hydrogels requires matrix
beating human cardiomyocytes, Acs Appl. Mater. Inter. 11 (23) (2019) remodelling, Nat. Mater. 16 (12) (2017) 1233–1242.
20589–20602. [120] C.M. Madl, B.L. LeSavage, R.E. Dewi, C.B. Dinh, R.S. Stowers, M. Khariton, et al.,
[94] A. Suo, W. Xu, Y. Wang, T. Sun, L. Ji, J. Qian, Dual-degradable and injectable Maintenance of neural progenitor cell stemness in 3D hydrogels requires matrix
hyaluronic acid hydrogel mimicking extracellular matrix for 3D culture of breast remodelling, Nat. Mater. 16 (12) (2017) 1233.
cancer MCF-7 cells, Carbohydr. Polym. 211 (2019) 336–348. [121] M. Darnell, A. O Neil, A. Mao, L. Gu, L.L. Rubin, D.J. Mooney, Material
[95] F. Xu, I. Gough, J. Dorogin, H. Sheardown, T. Hoare, Nanostructured degradable microenvironmental properties couple to induce distinct transcriptional programs
macroporous hydrogel scaffolds with controllable internal morphologies via in mammalian stem cells, Proc. Natl. Acad. Sci. Unit. States Am. 115 (36) (2018)
reactive electrospinning, Acta Biomater. 104 (2020) 135–146. E8368–E8377.
[96] P.K. Sharma, S. Taneja, Y. Singh, Hydrazone-linkage-based self-healing and [122] J. Lou, R. Stowers, S. Nam, Y. Xia, O. Chaudhuri, Stress relaxing hyaluronic acid-
injectable xanthan–poly(ethylene glycol) hydrogels for controlled drug release collagen hydrogels promote cell spreading, fiber remodeling, and focal adhesion
and 3D cell culture, Acs Appl. Mater. Inter. 10 (37) (2018) 30936–30945. formation in 3D cell culture, Biomaterials 154 (2018) 213–222.
[97] H. Tan, J.P. Rubin, K.G. Marra, Injectable in situ forming biodegradable chitosan- [123] O. Chaudhuri, L. Gu, D. Klumpers, M. Darnell, S.A. Bencherif, J.C. Weaver, et al.,
hyaluronic acid based hydrogels for adipose tissue regeneration, Organogenesis 6 Hydrogels with tunable stress relaxation regulate stem cell fate and activity, Nat.
(3) (2014) 173–180. Mater. 15 (3) (2016) 326–334.
[98] B. Yang, Y. Zhang, X. Zhang, L. Tao, S. Li, Y. Wei, Facilely prepared inexpensive [124] L. Cai, R.E. Dewi, A.B. Goldstone, J.E. Cohen, A.N. Steele, Y.J. Woo, et al.,
and biocompatible self-healing hydrogel: a new injectable cell therapy carrier, Regulating stem cell secretome using injectable hydrogels with in situ network
Polym. Chem.-Uk 3 (12) (2012) 3235. formation, Adv. Healthc. Mater. 5 (21) (2016) 2758–2764.
[99] S. Wang, H. Zheng, L. Zhou, F. Cheng, Z. Liu, H. Zhang, et al., Nanoenzyme- [125] E. Oliveira, R.C. Assunção-Silva, O. Ziv-Polat, E.D. Gomes, F.G. Teixeira, N.
reinforced injectable hydrogel for healing diabetic wounds infected with A. Silva, et al., Influence of different ECM-like hydrogels on neurite outgrowth
multidrug resistant bacteria, Nano Lett. 20 (7) (2020) 5149–5158. induced by adipose tissue-derived stem cells, Stem Cell. Int. 2017 (2017) 1–10.
[100] L. Zhou, Y. Xi, Y. Xue, M. Wang, Y. Liu, Y. Guo, et al., Injectable self-healing [126] N.A. Silva, J. Moreira, S. Ribeiro-Samy, E.D. Gomes, R.Y. Tam, M.S. Shoichet, et
antibacterial bioactive polypeptide-based hybrid nanosystems for efficiently al., Modulation of bone marrow mesenchymal stem cell secretome by ECM-like
treating multidrug resistant infection, skin-tumor therapy, and enhancing wound hydrogels, Biochimie 95 (12) (2013) 2314–2319.
healing, Adv. Funct. Mater. 29 (22) (2019) 1806883. [127] M.C. Schneider, C.A. Barnes, S.J. Bryant, Characterization of the chondrocyte
[101] H. Shih, T. Greene, M. Korc, C. Lin, Modular and adaptable tumor niche prepared secretome in photoclickable poly(ethylene glycol) hydrogels, Biotechnol. Bioeng.
from visible light initiated thiol-norbornene photopolymerization, 114 (9) (2017) 2096–2108.
Biomacromolecules 17 (12) (2016) 3872–3882. [128] V.V. Rao, M.K. Vu, H. Ma, A.R. Killaars, K.S. Anseth, Rescuing mesenchymal stem
[102] T.E. Brown, B.J. Carberry, B.T. Worrell, O.Y. Dudaryeva, M.K. McBride, C. cell regenerative properties on hydrogel substrates post serial expansion, Bioeng
N. Bowman, et al., Photopolymerized dynamic hydrogels with tunable Transl Med 4 (1) (2019) 51–60.
viscoelastic properties through thioester exchange, Biomaterials 178 (2018) [129] F. Liu, S. Hu, H. Yang, Z. Li, K. Huang, T. Su, et al., Hyaluronic acid hydrogel
496–503. integrated with mesenchymal stem cell-secretome to treat endometrial injury in a
[103] S.H. Hong, M. Shin, E. Park, J.H. Ryu, J.A. Burdick, H. Lee, Alginate-boronic acid: rat model of asherman’s syndrome, Adv. Healthc. Mater. 8 (14) (2019),
pH-triggered bioinspired glue for hydrogel assembly, Adv. Funct. Mater. 30 (26) e1900411.
(2020) 1908497. [130] K.T. Weber, Y. Sun, S.K. Bhattacharya, R.A. Ahokas, I.C. Gerling, Myofibroblast-
[104] S. Tang, H. Ma, H. Tu, H. Wang, P. Lin, K.S. Anseth, Adaptable fast relaxing mediated mechanisms of pathological remodelling of the heart, Nat. Rev. Cardiol.
boronate-based hydrogels for probing cell-matrix interactions, Adv. Sci. 5 (9) 10 (1) (2013) 15–26.
(2018) 1800638. [131] K.M. Wisdom, K. Adebowale, J. Chang, J.Y. Lee, S. Nam, R. Desai, et al., Matrix
[105] D. Wu, W. Wang, D. Diaz-Dussan, Y. Peng, Y. Chen, R. Narain, et al., In situ mechanical plasticity regulates cancer cell migration through confining
forming, dual-crosslink network, self-healing hydrogel enabled by a microenvironments, Nat. Commun. 9 (1) (2018) 4144.
bioorthogonal nopoldiol–benzoxaborolate click reaction with a wide pH range, [132] H.P. Lee, R. Stowers, O. Chaudhuri, Volume expansion and TRPV4 activation
Chem. Mater. 31 (11) (2019) 4092–4102. regulate stem cell fate in three-dimensional microenvironments, Nat. Commun.
[106] J. Hu, Q. Hu, X. He, C. Liu, Y. Kong, Y. Cheng, et al., Stimuli-responsive hydrogels 10 (1) (2019) 529.
with antibacterial activity assembled from guanosine, aminoglycoside, and a [133] K.M. Wisdom, K. Adebowale, J. Chang, J.Y. Lee, S. Nam, R. Desai, et al., Matrix
bifunctional anchor, Adv. Healthc. Mater. 9 (2) (2020) 1901329. mechanical plasticity regulates cancer cell migration through confining
[107] Q. Xu, C. He, Z. Zhang, K. Ren, X. Chen Injectable, Biomolecule-responsive microenvironments, Nat. Commun. 9 (1) (2018) 4144.
polypeptide hydrogels for cell encapsulation and facile cell recovery through [134] W.L. Murphy, T.C. McDevitt, A.J. Engler, Materials as stem cell regulators, Nat.
triggered degradation, Acs Appl. Mater. Inter. 8 (45) (2016) 30692–30702. Mater. 13 (6) (2014) 547–557.
[108] C.R. Correia, R.L. Reis, J.F. Mano, Design principles and multifunctionality in cell [135] H. Lee, L. Gu, D.J. Mooney, M.E. Levenston, O. Chaudhuri, Mechanical
encapsulation systems for tissue regeneration, Adv. Healthc. Mater. 7 (19) (2018) confinement regulates cartilage matrix formation by chondrocytes, Nat. Mater. 16
1701444. (12) (2017) 1243–1251.
[109] O. Hasturk, D.L. Kaplan, Cell armor for protection against environmental stress: [136] A.A. Thorpe, G. Dougill, L. Vickers, N.D. Reeves, C. Sammon, G. Cooper, et al.,
advances, challenges and applications in micro- and nanoencapsulation of Thermally triggered hydrogel injection into bovine intervertebral disc tissue
mammalian cells, Acta Biomater. 95 (2019) 3–31. explants induces differentiation of mesenchymal stem cells and restores
[110] X. Chen, C. Dong, K. Wei, Y. Yao, Q. Feng, K. Zhang, et al., Supramolecular mechanical function, Acta Biomater. 54 (2017) 212–226.
hydrogels cross-linked by preassembled host–guest PEG cross-linkers resist [137] Y. Xu, X. Yang, A.K. Thomas, P.A. Patsis, T. Kurth, M. Kräter, et al., Noncovalently
excessive, ultrafast, and non-resting cyclic compression, NPG Asia Mater. 10 (8) assembled electroconductive hydrogel, Acs Appl. Mater. Inter. 10 (17) (2018)
(2018) 788–799. 14418–14425.
[111] H. Wang, D. Zhu, A. Paul, L. Cai, A. Enejder, F. Yang, et al., Covalently adaptable [138] Y. Ma, M. Lin, G. Huang, Y. Li, S. Wang, G. Bai, et al., 3D spatiotemporal
elastin-like protein-hyaluronic acid (ELP-HA) hybrid hydrogels with secondary mechanical microenvironment: a hydrogel-based platform for guiding, Stem Cell
thermoresponsive crosslinking for injectable stem cell delivery, Adv. Funct. Fate 30 (49) (2018), e1705911.
Mater. 27 (28) (2017) 1605609. [139] L.G. Major, Y.S. Choi, Developing a high-throughput platform to direct
[112] K. Dubbin, A. Tabet, S.C. Heilshorn, Quantitative criteria to benchmark new and adipogenic and osteogenic differentiation in adipose-derived stem cells, J. Tissue
existing bio-inks for cell compatibility, Biofabrication 9 (4) (2017) 44102. Eng. Regen. M. 12 (2018) 2021–2028.
[113] Q. Xu, S. A, Y. Gao, L. Guo, J. Creagh-Flynn, D. Zhou, et al., A hybrid injectable [140] S.W. Crowder, V. Leonardo, T. Whittaker, P. Papathanasiou, M.M. Stevens,
hydrogel from hyperbranched PEG macromer as a stem cell delivery and Material cues as potent regulators of epigenetics and stem cell function, Cell Stem
retention platform for diabetic wound healing, Acta Biomater. 75 (2018) 63–74. Cell 18 (1) (2016) 39–52.
[114] D. Dong, T. Hao, C. Wang, Y. Zhang, Z. Qin, B. Yang, et al., Zwitterionic starch- [141] G. Brusatin, T. Panciera, A. Gandin, A. Citron, S. Piccolo, Biomaterials and
based hydrogel for the expansion and “stemness” maintenance of brown adipose engineered microenvironments to control YAP/TAZ-dependent cell behaviour,
derived stem cells, Biomaterials 157 (2018) 149–160. Nat. Mater. 17 (12) (2018) 1063–1075.
[115] F. Chowdhury, Y. Li, Y. Poh, T. Yokohama-Tamaki, N. Wang, T.S. Tanaka, Soft [142] M.A. Laflamme, C.E. Murry, Heart regeneration, Nature 473 (7347) (2011)
substrates promote homogeneous self-renewal of embryonic stem cells via 326–335.
downregulating cell-matrix tractions, PloS One 5 (12) (2010), e15655. [143] L.M. Stapleton, A.N. Steele, H. Wang, H.L. Hernandez, A.C. Yu, M.J. Paulsen, et
[116] P.M. Gilbert, K.L. Havenstrite, K.E.G. Magnusson, A. Sacco, N.A. Leonardi, al., Use of a supramolecular polymeric hydrogel as an effective post-operative
P. Kraft, et al., Substrate elasticity regulates skeletal muscle stem cell self-renewal pericardial adhesion barrier, Nat. Biomed. Eng. 3 (8) (2019) 611–620.
in culture, Science 329 (5995) (2010) 1078–1081. [144] L. Shi, F. Wang, W. Zhu, Z. Xu, S. Fuchs, J. Hilborn, et al., Self-healing silk fibroin-
based hydrogel for bone regeneration: dynamic metal-ligand self-assembly
approach, Adv. Funct. Mater. 27 (37) (2017) 1700591.

1386
Z. Tong et al. Bioactive Materials 6 (2021) 1375–1387

[145] K. Zhang, Z. Jia, B. Yang, Q. Feng, X. Xu, W. Yuan, et al., Adaptable hydrogels [153] Z. Li, F. Zhou, Z. Li, S. Lin, L. Chen, L. Liu, et al., Hydrogel cross-linked with
mediate cofactor-assisted activation of biomarker-responsive drug delivery via dynamic covalent bonding and micellization for promoting burn wound healing,
positive feedback for enhanced tissue regeneration, Adv. Sci. 5 (12) (2018) Acs Appl. Mater. Inter. 10 (30) (2018) 25194–25202.
1800875. [154] A.A. Foster, R.E. Dewi, L. Cai, L. Hou, Z. Strassberg, C.A. Alcazar, et al., Protein-
[146] M. Brittberg, A. Lindahl, A. Nilsson, C. Ohlsson, O. Isaksson, L. Peterson, engineered hydrogels enhance the survival of induced pluripotent stem cell-
Treatment of deep cartilage defects in the knee with autologous chondrocyte derived endothelial cells for treatment of peripheral arterial disease, Biomater.
transplantation, N. Engl. J. Med. 331 (14) (1994) 889–895. Sci.-Uk 6 (3) (2018) 614–622.
[147] J. Runhaar, Development and Prevention of Knee Osteoarthritis: the Load of [155] C. Loebel, C.B. Rodell, M.H. Chen, J.A. Burdick, Shear-thinning and self-healing
Obesity, 2013. hydrogels as injectable therapeutics and for 3D-printing, Nat. Protoc. 12 (8)
[149] Y. Zhang, X. Liu, L. Zeng, J. Zhang, J. Zuo, J. Zou, et al., Polymer fiber scaffolds (2017) 1521–1541.
for bone and cartilage tissue engineering, Adv. Funct. Mater. 29 (36) (2019) [156] X. Tong, F. Yang, Sliding hydrogels with mobile molecular ligands and crosslinks
1903279. as 3D stem cell niche, Adv. Mater. 28 (33) (2016) 7257–7263.
[150] I.L. Kim, S. Khetan, B.M. Baker, C.S. Chen, J.A. Burdick, Fibrous hyaluronic acid [157] M. Shin, J.H. Ryu, J.P. Park, K. Kim, J.W. Yang, H. Lee, DNA/Tannic acid hybrid
hydrogels that direct MSC chondrogenesis through mechanical and adhesive cues, gel exhibiting biodegradability, extensibility, tissue adhesiveness, and hemostatic
Biomaterials 34 (22) (2013) 5571–5580. ability, Adv. Funct. Mater. 25 (8) (2015) 1270–1278.
[151] A.M. Kloxin, A.M. Kasko, C.N. Salinas, K.S. Anseth, Photodegradable hydrogels [158] C. Li, A. Faulkner-Jones, A.R. Dun, J. Jin, P. Chen, Y. Xing, et al., rapid formation
for dynamic tuning of physical and chemical properties, Science (American of a supramolecular polypeptide-DNA hydrogel for in situ three-dimensional
Association for the Advancement of Science) 324 (5923) (2009) 59–63. multilayer bioprinting, Angew. Chem. Int. Ed. 54 (13) (2015) 3957–3961.
[152] D.J. Huey, J.C. Hu, K.A. Athanasiou, Unlike bone, cartilage regeneration remains
elusive, Science 338 (6109) (2012) 917–921.

1387