Bio-inspired self-healing structural color hydrogel

Fanfan Fua,1, Zhuoyue Chena,1, Ze Zhaoa, Huan Wanga, Luoran Shanga, Zhongze Gua, and Yuanjin Zhaoa,2
a
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China

Edited by David A. Weitz, Harvard University, Cambridge, MA, and approved April 24, 2017 (received for review March 3, 2017)

Biologically inspired self-healing structural color hydrogels were enzyme additives. As both the GelMA and protein hydrogels are
developed by adding a glucose oxidase (GOX)- and catalase derived from organisms, the composite materials had high bio-
(CAT)-filled glutaraldehyde cross-linked BSA hydrogel into metha- compatibility and plasticity. It was demonstrated that a series of
crylated gelatin (GelMA) inverse opal scaffolds. The composite new structural color materials with one-dimensional (1D) linear
hydrogel materials with the polymerized GelMA scaffold could microfiber, 2D pattern, and 3D photonic path structures could be
maintain the stability of an inverse opal structure and its resultant developed by assembling and healing the composite structural
structural colors, whereas the protein hydrogel filler could impart color hydrogel elements. These features make our self-healing
self-healing capability through the reversible covalent attachment structural color hydrogels highly promising for different applica-
of glutaraldehyde to lysine residues of BSA and enzyme additives. tions, such as counterfeit prevention, integrated optics, and
A series of unprecedented structural color materials could be cre- biomedical engineering.
ated by assembling and healing the elements of the composite
hydrogel. In addition, as both the GelMA and the protein hydro- Results and Discussion
gels were derived from organisms, the composite materials pre- In a typical experiment, the GelMA hydrogel inverse opal scaf-
sented high biocompatibility and plasticity. These features of self- folds were fabricated by replicating silica colloidal crystal tem-
healing structural color hydrogels make them excellent functional plates. These colloidal crystal templates were prepared by the
materials for different applications. self-assembly of silica nanoparticles in silica capillaries or on
glass slides, which closely packed and finally formed an ordered
colloidal crystal | self-healing | inverse opal | structural color | hydrogel structure during dehydration (Fig. 2A). This ordered packing of
the nanoparticles endowed the colloidal crystals with inter-
connected nanopores throughout the templates, which enabled
S tructural colors, arising from intrinsic periodic nanostructures
and resulting in the interaction of light with these photonic
nanostructures, have attracted much interest because of the fasci-
infiltration of the GelMA pregel solution. After the pregel so-
lution penetrated the nanopores and filled all of the voids of the
nation associated with the display of various brilliant examples (1 templates by capillary action, the solution was polymerized to
6). The creatures displaying brilliant structural colors are prevalent form a hydrogel by UV light. Finally, the inverse opal scaffolds
in nature, and their life, including communication, shielding, and were obtained by etching the silica nanoparticles, leaving an in-
other biological functions, is closely linked with these structural verse opal GelMA hydrogel scaffold (Fig. 2B). This kind of
colors (7). Researchers, marveling at these miracles, have devoted scaffold displays various brilliant structure colors (Fig. S1), which
their work to bio-inspired structure colors materials. Structural color is an important feature of the materials.
hydrogels are one of the most important examples and have been To impart to the structural color hydrogel the capability of
widely studied and used in switches (8, 9), optical devices (10, 11), self-healing, the glutaraldehyde cross-linked BSA hydrogel with
sensing materials (12, 13), and wearable electronics (14), etc. (15 enzyme additives of GOX and CAT was filled into the inverse
opal scaffold. In this process, the pregel was first prepared with a
20). However, because the deterioration and accumulation of
damage of these materials during applications are inevitable, to
achieve next-generation materials for both fundamental research Significance
and practical applications, the creation of bio-inspired structural
color materials with increased survivability is still necessary. Structural color hydrogels have been widely studied and used
Coincidentally, for the purpose of increasing the survivability in different applications, such as in switches, optical devices,
and lifetime of an organism, the function of self-healing that can etc. However, because the deterioration and accumulation of
spontaneously heal injury and recover functionality in creatures is damage of these materials are inevitable during applications,
ubiquitous in nature (2125). Inspired by these creatures, re- the creation of bio-inspired structure color materials with in-
searchers have developed numerous self-healing hydrogel mate- creased survivability is still desired for both fundamental re-
rials that fuse together with their homogeneous hydrogels for search and practical applications. In this study, inspired by
various important applications (2637). However, the fusion pro- creatures in nature with spontaneous healing from injury and
cess of the self-healing hydrogels could also happen in their recovering of functionality, we demonstrated a self-healing
neighboring nanostructures; this would cause the destruction of structural color hydrogel by filling a healable protein hydro-
the periodic photonic scaffolds. Therefore, a self-healing hydrogel gel into an inverse opal scaffold. A series of new structural
with stable structural color has not yet been reported, and its color materials with 1D linear microfiber, 2D pattern, and 3D
construction remains a challenge. photonic path structures could be constructed by assembling
In this paper, we present the desired self-healing structural color and healing the composite structural color hydrogel elements.
hydrogels by constructing them with a composite nanostructure, as
Author contributions: Y.Z. designed research; F.F. and Z.C. performed research; F.F., Z.C.,
indicated in Fig. 1. This nanostructure was composed of a meth- Z.Z., H.W., L.S., Z.G., and Y.Z. analyzed data; F.F. and Y.Z. wrote the paper; Z.Z. contrib-
acrylated gelatin (GelMA) hydrogel inverse opal scaffold and a filler uted to scientific discussion; and H.W. and L.S. contributed to scientific discussion of
of glutaraldehyde cross-linked BSA hydrogel with enzyme additives the article.

of glucose oxidase (GOX) and catalase (CAT). The polymerized The authors declare no conflict of interest.
GelMA hydrogel scaffold in the composite materials could guaran- This article is a PNAS Direct Submission.
tee the stability of both the inverse opal structure and its resultant 1
F.F. and Z.C. contributed equally to this work.
structural colors, whereas the protein hydrogel filler could impart the 2
To whom correspondence should be addressed. Email: [email protected].
materials with self-healing capability through the reversible covalent This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.
attachment of the glutaraldehyde to lysine residues of BSA and the 1073/pnas.1703616114/-/DCSupplemental.

59005905 | PNAS | June 6, 2017 | vol. 114 | no. 23 www.pnas.org/cgi/doi/10.1073/pnas.1703616114

A the imine bonds being oxidized and to support the cyclic reaction
of glucose oxidation. Finally, with the regulation of pH, the
GelMA BSA-gel imine bonds provide the opportunity to heal the protein hydro-
replication infiltration
gel. As a benefit from the reversible binding of the inverse opal
filler protein hydrogel, the GelMA and proteins hybrid structural
color hydrogel system would also be imparted with a self-
B healing function.
To investigate the self-healing property of GelMA and the pro-
Damage Healing teins hybrid structural color hydrogel system, hybrid hydrogel
microfibers with the same inverse opal nanostructures and com-
posite protein materials were fabricated and cut into segments.
Then, the segments of the structural color hydrogels were brought
together slightly to ensure that their surfaces were in full contact. It
was found that by simply connecting two of the segments, they could
not adhere to each other and remained independent. However, with
the addition of glucose, the two segments could adhere tightly to
each other and form an integrated microfiber (Fig. 3A). Although
the repaired traces could not be hidden and the reflection peak
Fig. 1. Schematic diagram of the self-healing structural color hydrogel. width at the fracture increased slightly (Fig. S4), the self-healing
(A) Fabrication of the self-healing structural color hydrogel. It was composed structural color microfiber maintained its vivid structural color
of a GelMA hydrogel inverse opal scaffold and a filler of glutaraldehyde cross- through the whole body. In addition, the self-healing microfibers
linked BSA hydrogel with GOX and CAT additives. (B) Self-healing process of
the structural color hydrogel.
show elasticity as good as their original elasticity. Thus, the enzyme-
mediated hybrid structural color hydrogel exhibits excellent self-

APPLIED PHYSICAL
healing properties with high recovery and reversibility.

SCIENCES
protein concentration of 13.5 wt% (GOX, CAT, and BSA for It is noteworthy that our strategy could even heal the microfiber
0.2%, 0.8%, and 12.5%, respectively). The concentration of segments with different structural colors. To demonstrate this, hy-
glutaraldehyde was 0.5 wt%, and the pH of the solution was brid hydrogel microfiber segments with blue, green, and red struc-
adjusted to 7.0. To fully fill the protein hydrogel into the nano- tural colors were assembled together. The joints of these microfiber
pores of the inverse opal, the structure color hydrogels should be segments were simply treated with glucose. It was found that al-
dehydrated and then immersed in the pregel solution in a vac- though having different structural colors, neighbor segments could
uum environment. After these steps, the structure color hydro- still adhere together tightly and form an integrated microfiber. The
gels were transferred to a closed environment with a certain combined microfiber inherited the multiplex structural colors of
humidity for the polymerization of the infiltrated pregel in the each segment and preserved the good elasticity of the original
inverse opals (Fig. 2C). Finally, a hybrid inverse opal hydrogel microfibers (Fig. 3B). This indicated that the enzyme-mediated
with brilliant structure color was achieved. hybrid structural color hydrogel was suitable for different kinds of
The formation of the structural colors of the hydrogels was inverse opal scaffolds with different nanometer aperture sizes.
ascribed to their orderly arranged nanostructure, which imparts to Therefore, a new assembly strategy for the construction of a mul-
the inverse opal hydrogel and its derived hybrid hydrogel a unique tiplex structural color hydrogel was developed.
photonic band gap (PBG). This PBG leads light with certain To demonstrate the versatility of the self-healing structural
wavelengths or frequencies to be located in and reflected instead color hydrogels in assembling other shapes besides cylindrical,
of propagating through the materials. As a result, the colloidal three pieces of different hybrid hydrogel films were used for
crystal templates and the inverse opal scaffold, together with the structural color pattern construction. By adding glucose to the
intersection line, these films were stitched together to form an
hybrid hydrogel, all showed vivid colors and possessed character-
istic reflection peaks. Under normal incidence, the main reflection
peak position of these materials can be estimated by Braggs
equation, = 2d111naverage, where d111 is the interplanar distance A B
of the (111) diffracting planes, and naverage refers to the average
refractive index of the materials. Although the infiltration of the
protein hydrogel into the inverse opal could increase the naverage of
the hybrid material, the d111-related scaffold has also shrunk
during the polymerization of the filler, and thus the structural
color and the reflection peak of the hybrid material have blue
shifted (Fig. S1). In addition, by using different sizes of silica
nanoparticles for the colloidal crystal templates, a series of
GelMA hydrogel inverse opals and resultant hybrid hydrogels with C D
different diffraction peaks and structural colors could also be
obtained (Fig. 2D and Fig. S2).
In the GelMA and proteins hybrid hydrogel system, the fea-
ture of reversible imine covalent attachment of the glutaralde-
hyde to lysine residues of BSA, GOX, and CAT proteins imparts
to the inverse opal filler protein hydrogel the self-healing func-
tion, which uses GOX and CAT to adjust the pH of the system by
adding extra traces of glucose (Fig. S3). In this process, GOX
assists the glucose to be oxidized to gluconolactone, which is then Fig. 2. (AC) SEM images of (A) colloidal crystal template, (B) inverse opal
hydrolyzed to gluconic acid to decrease the pH value of the scaffold, and (C) hybrid self-healing hydrogel surface. (Scale bars: 500 nm.)
protein hydrogel. The by-product H2O2 of the glucose oxidation (D) Optical images and absolute reflection spectra of six kinds of hybrid self-
will decompose into H2O and O2 by the CAT enzyme to avoid healing hydrogel microfibers.

Fu et al. PNAS | June 6, 2017 | vol. 114 | no. 23 | 5901

A the repaired sample. It was found that the self-healing film could
keep its integrated structures not only in the assembled units but
also in the repaired section (Fig. 4B). Thus, the self-healing
structural color film is sufficiently flexible to resist an external
tensile force (Fig. S5). In each assembled unit, it could be ob-
served that all of them showed an equal ratio of stretching with
the whole film, which caused the blue shift of their structural
colors. These colors blue shift should ascribe to the gradual
decreasing of the interplanar distance d111 of the (111) dif-
fracting planes during the stretching of the inverse opal mate-
rials. In addition to the simple indexed pattern of structural
colors, a much more complex 2D pattern, such as Chinese Taiji
(Fig. 4C), could also be constructed by using the same self-
B healing assembly strategy.
Besides the 2D patterns, the self-healing assembly strategy
could also be used for the development of 3D structural color
materials that have potential values in the areas of art creation,
counterfeit prevention, and 3D integrated optics, etc. To dem-
onstrate these concepts, a GelMA and proteins hybrid yellow
structural color hydrogel film was cut into pieces of different
sizes. These pieces were stacked together from large to small
(Fig. 5A). Because of the complete porous inverse opal structure
of these pieces, the filler self-healing protein hydrogels in the
inverse opal scaffolds from the surface of the pieces could touch
each other. Thus, these pieces could form an integrated 3D
pyramid structure by a self-healing assembly strategy (Fig. 5B).
With the same method, we could also construct 3D structural
Fig. 3. Optical images of the self-healing process of the structural color
color objects in a hydrogel block. In this process, hybrid blue
microfibers. (A) Optical images of the self-healing process of two segments
of the structural color hydrogel microfiber. (B) Optical images of the self- structural color hydrogel pieces with different 2D green triangle
healing process of three different structural color hydrogel segments. (Scale patterns were first prepared by using the above process. Then,
bars: 2 mm.) the pieces were stacked together and treated with glucose (Fig.
5C). Finally, a transparent blue hydrogel block containing a 3D
triangle-stacked structural color object was generated (Fig. 5D).
indexed pattern of an integrated film with blue, green, and red By designing the objects with more complex shapes and struc-
structural colors (Fig. 4A). To investigate the practical value of tural colors, advanced counterfeit prevention tags could be
the self-healing hybrid hydrogel materials, a tensile test was achieved. By using slender structural color microfibers instead of
performed to quantitatively evaluate the mechanical stability of the triangle pattern, a 3D integrated photonic path could be

B
Stretch

Fig. 4. Optical images of the self-healing process of the structural color films. (A) The construction process of a structural color hydrogel film by using three
pieces of hybrid hydrogel with different structural colors. (B) The self-healing structural color hydrogel film from A showed good flexibility to resist external
tensile force. (C) Optical image of a Chinese Taiji pattern assembled by the self-healing structural color hydrogel. (Scale bars for A and C: 5 mm.)

5902 | www.pnas.org/cgi/doi/10.1073/pnas.1703616114 Fu et al.

 A B C D

E F G H

Fig. 5. Construction of 3D structural color objects. (A and B) Schematic diagram and optical image of a yellow pyramid-structured structural color hydrogel.
The hydrogel had an integrated structure and could be hung in the air in B. (C and D) Schematic diagram and optical image of a blue hydrogel block with a 3D
triangle stacked green object inside. (EH) Schematic diagram and optical images of the 3D integrated photonic paths in a hydrogel block. The images were
taken under natural light (F), 550 nm monochromatic light (G), and 450 nm monochromatic light (H), respectively. (Scale bars: 5 mm.)

developed in the hydrogel block (Fig. 5 E and F). Because of the were suitable for cell culture and reproduction. It is noteworthy
existence of the PBGs in the hydrogel block and its encapsulated that in many cases tissue engineering required hydrogels have no
photonic paths, the hybrid hydrogel could show excited 3D green self-healing function, which greatly limits their application.

APPLIED PHYSICAL
or invisible optical paths under green or blue light irradiation, However, with our strategy, both non- and self-healing hydrogels

SCIENCES
respectively (Fig. 5 G and H). This implies that the materials could be combined as a hybrid together through cross-linking
could be used as new carriers for a 3D optical waveguide or nanoscaffolds, and the resultant hydrogels could be endowed
optical communication. with the self-healing function to remedy the restrictions of the
As both the GelMA hydrogel inverse opal scaffold and the hydrogel biomaterials in biomedical applications.
filler of the protein hydrogel were derived from organisms,
the self-healing structural color hybrid hydrogels should have the Conclusion
same high biocompatibility. To demonstrate this, the hybrid In summary, we have developed self-healing structural color
hydrogels before and after self-healing were all used for a he- hydrogels with a GelMA inverse opal scaffold and BSA protein
patocellular carcinoma (HepG2) cell culture, respectively. The filler with GOX and CAT enzyme additives. The GelMA scaffold
inverse opal GelMA hydrogel scaffold was also cultured with in the hybrid hydrogels guaranteed the stability of the inverse opal
cells for the control group. It was found that the HepG2 cells
structure and the resultant structural colors, whereas the protein
could adhere and grow on the surface of the hybrid hydrogels
filler could impart the hydrogels self-healing capability through
irrespective of the repair process. These cells formed tight cell
cell connections both on the hybrid hydrogel film and on the the reversible covalent attachment of the glutaraldehyde to lysine
healing section of the film after 24 h culture, as shown in Fig. 6 residues of proteins. We have demonstrated that a series of un-
AD and Fig. S6. The cell viabilities on the structural color hy- precedented structural color materials with 1D linear structures,
brid hydrogels before and after self-healing, as well as on the 2D patterns, and 3D counterfeit-prevention objects and photonic
commercial multiwell and on the inverse opal GelMA hydrogel path structures could be created by assembling and healing the
scaffold, were also investigated quantitatively by using 3-(4, hybrid hydrogel elements. In addition, the biocompatibility and
5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) biological applicability of the GelMA and proteins hybrid struc-
assays, as presented in Fig. 6E. It could be observed that the tural color hydrogels were also demonstrated. These features of
viability of the HepG2 cells shows no obvious differences on the self-healing structural color hydrogels indicated their versatile
these hydrogel substrates. Thus, the self-healing hybrid hydrogels values in different areas.

A B E

C D

Fig. 6. (AD) CLSM images of the HepG2 cells cultured on hybrid hydrogels before (A and C) and after (B and D) self-healing. A and B are the bright-field
microscopy images, and C and D are the calcein-AM fluorescent images. (Scale bar: 100 m.) (E) Results of the HepG2 cell MTT assays cultured on different
hydrogels for 24 h.

Fu et al. PNAS | June 6, 2017 | vol. 114 | no. 23 | 5903

Methods were recorded at a fixed glancing angle, using an optical microscope
Materials. Eight kinds of SiO2 nanoparticles with sizes 200 nm, 210 nm, 230 nm, equipped with a fiber-optic spectrometer (Ocean Optics; USB2000-FLG).
250 nm, 270 nm, 290 nm, 300 nm, and 320 nm were purchased from Nanjing
Dongjian Biological Technology Co., Ltd. The GelMA hydrogel was self- The Construction Process of Structured Structural Color Hydrogels. The self-
prepared. Gelatin (from porcine skin), methacrylic anhydride, and MTT were healing property of cross-linked protein hydrogel systems was investigated
purchased from Sigma Aldrich. Calcein-AM (molecular probe) was purchased by cutting the hybrid structural color hydrogels into two segments. Then, two
from Life Technologies, and glutaraldehyde was derived from Aladdin. BSA, segments of the hybrid structural color hydrogels were brought together
GOX, and CAT were acquired from Sigma Aldrich. Cellulose dialysis mem- slightly to ensure the two surfaces were fully contacted and stimulated with
branes [molecular weight cutoff (MWCO) = 8,00014,000] were acquired from external glucose (0.1 mg) for 3 h under a closed condition at 4 C. Finally, the
Shanghai Yuanye Biotechnology Corporation. HepG2 cells were from the In- enzyme-mediated hybrid structural color hydrogels, exhibiting excellent
stitute of Biochemistry and Cell Biology, the Chinese Academy of Sciences, self-healing properties, were prepared. In another experiment, three kinds
Shanghai, China. Dulbeccos modified Eagles medium (DMEM) and FBS were of hybrid hydrogel fibers with different structure colors were assembled
purchased from HyClone. Penicillinstreptomycin was obtained from Gibco. together under the same conditions. The 2D pattern and 3D photonic path
Water used in all experiments was purified using a Milli-Q Plus 185 water structures could also be developed by assembling and healing the composite
purification system (Millipore) with resistivity higher than 18 Mcm. structural color hydrogel elements. Obtained by using mask templates of UV
light, these inverse opal scaffolds in different shapes were first dehydrated
for 2 h at 35 C and quickly filled with the prepared solution in a vacuum
Preparation of Inverse Opal Scaffold. The inverse opal scaffolds were fabri-
environment for 20 min. Then, the 2D pattern and 3D photonic path
cated using a sacrificial template method. The colloidal crystal templates
structures could also be assembled together with an external glucose for 3 h
were obtained at invariant temperature and humidity by a vertical deposition
under a closed condition at 4 C. The optical microscopy images of the
method. In brief, the colloidal crystal templates were prepared with the self-
samples were obtained under the same conditions by a digital camera
assembly of silica nanoparticles in silica capillaries or on glass slides. The SiO2
(Canon5D Mark II).
nanoparticles (50 wt%) with a variety of particle sizes (200 nm, 210 nm,
230 nm, 250 nm, 270 nm, 290 nm, 300 nm, and 320 nm) were dispersed in
water, showing a good monodispersity. For the preparation of colloidal Cell Culture. Cells were regularly cultured and passaged with DMEM sup-
crystal fiber templates, the SiO2 solution was injected into silica tubes (d = plemented with 10% FBS and 1% penicillinstreptomycin in a humidified
1.56 mm) and formed an ordered fiber cluster structure during the de- incubator at 37 C with 5% CO2. The structural color hydrogels were first
hydration procedure at 40 C for 15 d. Then the fiber templates were cal- disinfected by exposure to UV light for 2 h and rinsed with sterile PBS so-
cined at 750 C for 5 h to improve their mechanical strength. Finally, the lution three times before cell culture. Then the HepG2 cells, cultured on the
silica tubes were removed and the free templates were obtained. The col- surface of the structure color hydrogels, were treated in traditional ways.
loidal crystal film templates (with thickness of about 0.5 mm) were also The cells were seeded on the surface of hydrogel films (1 cm2) in a six-well
prepared to obtain different patterns under the same condition. The silica tissue culture plate (2 105 cells per well) for 24 h. The viability of
nanoparticles self-assembled on glass slides with a silica solution (ethyl al- HepG2 cells cultured in different structural color hydrogels was analyzed.
cohol) concentration of 20 wt% at 4 C for 3 h, and then the glass were Briefly, cells were first cultured on the surface of the structure color hydrogel
calcined at 450 C for 5 h to improve their mechanical strength. The inverse films for 24 h, then MTT/PBS solution (5 g/mL) was added, and the cells
opal structural color hydrogels scaffold was obtained based on these col- were incubated for another 4 h. Then the cell viability was quantified by the
loidal crystal templates. The GelMA pregel solution (0.2 g/mL) was infiltrated MTT assays according to the manufacturers instructions. To test different
into the silica templates by capillary force, and the solution was polymerized cell viabilities under the same conditions, the cells cultured on the tissue
to form a hydrogel (with refractive index about 1.387) by exposure to UV culture plate were set as control experiments. The mean value and SD of five
light. Finally, the inverse opal scaffold was obtained by etching (4 wt% paralleled assays for each sample were recorded. The morphology of cells
hydrofluoric acid) the silica nanoparticles, leaving an inverse opal GelMA was also observed. After being cultured on the surface of the structure color
hydrogel scaffold. These inverse opal scaffolds with different patterns could hydrogel films for 24 h, the cells were stained with calcein AM (2 g/mL, 2 mL
also be obtained by exposure to UV light with mask templates. per well) for 20 min at 37 C, followed by being rinsed twice with PBS and
fixed with glutaraldehyde (2.5%, 2 mL per well) for 6 h at 4 C. Finally, the
cells were observed using an inverted fluorescence microscope.
Preparation of Bio-Inspired Self-Healing Structural Color Hydrogels. The bio-
inspired self-healing structural color hydrogels were prepared based on
Characterization. Reflection spectra were obtained at a fixed glancing angle,
the enzyme additives of the GOX and CAT. The glutaraldehyde (0.5 wt%)
using an optical microscope equipped with a fiber-optic spectrometer (Ocean
cross-linked BSA (12.5 wt%) hydrogel with GOX (0.2 wt%) and CAT (0.8 wt%)
Optics; USB2000-FLG). SEM images of samples were taken by a scanning
was filled into the inverse opal scaffold. In this process, the pH of the pregel
electron microscope (Hitachi S-3000N). Microscopy images of the samples
solution was adjusted to 7.0. The inverse opal scaffold was dehydrated for 2 h
were obtained with an optical microscope (Olympus BX51) equipped with a
at 35 C and quickly filled with the pregel solution (with refractive index
CCD camera (Media Cybernetics Evolution MP5.0) and a digital camera
about 1.352) in a vacuum environment for 20 min. After these steps, the
(Canon5D Mark II). The stiffness of the hydrogel materials was characterized
structure color hydrogels were transferred into a closed environment with a
by Single Column Table Top Systems (5943; Instron).
certain humidity at 4 C for another 3 h for polymerization of the infiltrated
pregel in the inverse opals. Finally, the hybrid structural color hydrogels with
good visibility and brilliant structural colors were prepared. In addition, by ACKNOWLEDGMENTS. This work was supported by the National Science
Foundation of China (Grants 21473029 and 51522302), the NSAF Foundation
using different sizes of silica nanoparticles, a series of hybrid hydrogels with
of China (Grant U1530260), the National Science Foundation of Jiangsu
different diffraction peaks and structural colors could also be obtained. The (Grant BK20140028), the Program for New Century Excellent Talents in Uni-
optical microscopy images of the colloidal crystal templates, inverse opal versity, the Scientific Research Foundation of Southeast University, and the
scaffold, and hybrid hydrogels were obtained under the same conditions by Scientific Research Foundation of the Graduate School of Southeast Univer-
a digital camera (Canon5D Mark II). The reflection spectra of these samples sity (Grant YBJJ1671).

1. Ge J, Yin Y (2011) Responsive photonic crystals. Angew Chem Int Ed Engl 50: 8. Hou J, et al. (2015) Hydrophilic-hydrophobic patterned molecularly imprinted pho-
14921522. tonic crystal sensors for high-sensitive colorimetric detection of tetracycline. Small 11:
2. Zhao Y, Xie Z, Gu H, Zhu C, Gu Z (2012) Bio-inspired variable structural color materials. 27382742.
Chem Soc Rev 41:32973317. 9. Gu H, et al. (2013) Tailoring colloidal photonic crystals with wide viewing angles.
3. Ye B, et al. (2014) Photonic crystal microcapsules for label-free multiplex detection. Small 9:22662271.
Adv Mater 26:32703274. 10. Hou J, et al. (2014) Bio-inspired photonic-crystal microchip for fluorescent ultratrace
4. Zhao Y, Shang L, Cheng Y, Gu Z (2014) Spherical colloidal photonic crystals. Acc Chem detection. Angew Chem Int Ed Engl 53:57915795.
Res 47:36323642. 11. Zhang P, et al. (2016) Superspreading on immersed gel surfaces for the confined
5. Lee HS, et al. (2014) Magnetoresponsive discoidal photonic crystals toward active synthesis of thin polymer films. Angew Chem Int Ed Engl 55:36153619.
color pigments. Adv Mater 26:58015807. 12. Kim SH, Shim JW, Yang SM (2011) Microfluidic multicolor encoding of microspheres
6. Qin M, et al. (2016) A rainbow structural-color chip for multisaccharide recognition. with nanoscopic surface complexity for multiplex immunoassays. Angew Chem Int Ed
Angew Chem Int Ed Engl 55:69116914. Engl 50:11711174.
7. Shang LR, Gu ZZ, Zhao YJ (2016) Structural color materials in evolution. Mater Today 13. Liu CH, et al. (2016) Tunable structural color surfaces with visually self-reporting
19:420421. wettability. Adv Funct Mater 26:79377942.

5904 | www.pnas.org/cgi/doi/10.1073/pnas.1703616114 Fu et al.

14. Su B, Tian Y, Jiang L (2016) Bioinspired interfaces with superwettability: From ma- 26. Xing LX, et al. (2016) Self-healable polymer nanocomposites capable of simulta-
terials to chemistry. J Am Chem Soc 138:17271748. neously recovering multiple functionalities. Adv Funct Mater 26:35243531.
15. Yu L, Liu X, Yuan W, Brown LJ, Wang D (2015) Confined flocculation of ionic pol- 27. Shang L, et al. (2017) Bioinspired multifunctional spindle-knotted microfibers from
lutants by poly (L-dopa)-based polyelectrolyte complexes in hydrogel beads for three- microfluidics. Small 13:201600286.
dimensional, quantitative, efficient water decontamination. Langmuir 31:63516366. 28. Liu X, Wang S (2014) Three-dimensional nano-biointerface as a new platform for
16. Shang L, et al. (2015) Photonic crystal microbubbles as suspension barcodes. J Am guiding cell fate. Chem Soc Rev 43:23852401.
Chem Soc 137:1553315539. 29. Wang SG, et al. (2016) Phase-changeable and bubble-releasing implants for highly
17. Wang M, He L, Xu W, Wang X, Yin Y (2015) Magnetic assembly and field-tuning of efficient HIFU-responsive tumor surgery and chemotherapy. J Mater Chem B Mater
ellipsoidal-nanoparticle-based colloidal photonic crystals. Angew Chem Int Ed Engl Biol Med 4:73687378.
54:70777081. 30. Gao Y, et al. (2014) Enzymetically regulating the self-healing of protein hydrogels
18. Zhao Y, et al. (2013) Bioinspired multifunctional Janus particles for droplet manipu- with high healing efficiency. Angew Chem Int Ed Engl 53:93439346.
31. Cai G, et al. (2016) Extremely stretchable strain sensors based on conductive
lation. J Am Chem Soc 135:5457.
self-healing dynamic cross-links hydrogels for human-motion detection. Adv Sci
19. Bai S, Nguyen TL, Mulvaney P, Wang D (2010) Using hydrogels to accommodate hy-
4:1600190.
drophobic nanoparticles in aqueous media via solvent exchange. Adv Mater 22:
32. Zhao X, Zhang MY, Guo BL, Ma PX (2016) Mussel-inspired injectable supramolecular
32473250.
and covalent bonds crosslinked hydrogels with rapid self-healing and recovery via a
20. Kim SH, Shum HC, Kim JW, Cho JC, Weitz DA (2011) Multiple polymersomes for
facile approach under metal-free conditions. J Mater Chem B Mater Biol Med
programmed release of multiple components. J Am Chem Soc 133:1516515171.
4:66446651.
21. Cordier P, Tournilhac F, Souli-Ziakovic C, Leibler L (2008) Self-healing and thermor-
33. Huang W, et al. (2016) Strong and rapidly self-healing hydrogels: Potential hemo-
eversible rubber from supramolecular assembly. Nature 451:977980. static materials. Adv Healthc Mater 5:28132822.
22. Griffin DR, Weaver WM, Scumpia PO, Di Carlo D, Segura T (2015) Accelerated wound 34. Dong R, Zhao X, Guo B, Ma PX (2016) Self-healing conductive injectable hydrogels
healing by injectable microporous gel scaffolds assembled from annealed building with antibacterial activity as cell delivery carrier for cardiac cell therapy. ACS Appl
blocks. Nat Mater 14:737744. Mater Interfaces 8:1713817150.
23. Li CH, et al. (2016) A highly stretchable autonomous self-healing elastomer. Nat Chem 35. Zeng Q, et al. (2016) Self-healing elastin-bioglass hydrogels. Biomacromolecules 17:
8:618624. 26192625.
24. Deng GH, Tang CM, Li FY, Jiang HF, Chen YM (2010) Covalent cross-linked polymer 36. Wang S, et al. (2011) Highly efficient capture of circulating tumor cells by using
gels with reversible sol-gel transition and self-healing properties. Macromolecules 43: nanostructured silicon substrates with integrated chaotic micromixers. Angew Chem
11911194. Int Ed Engl 50:30843088.
25. Cheng Y, et al. (2014) Bioinspired multicompartmental microfibers from microfluidics. 37. Zhu Y, Xuan H, Ren J, Ge L (2015) Self-healing multilayer polyelectrolyte composite

APPLIED PHYSICAL
Adv Mater 26:51845190. film with chitosan and poly(acrylic acid). Soft Matter 11:84528459.

SCIENCES

Fu et al. PNAS | June 6, 2017 | vol. 114 | no. 23 | 5905