Received: 20 March 2019 Revised: 14 April 2019 Accepted: 15 April 2019

DOI: 10.1002/pat.4654

RESEARCH ARTICLE

Injectable poloxamer/graphene oxide hydrogels with well‐

controlled mechanical and rheological properties

Elaheh Rohani Rad1 | Henri Vahabi2,3 | Krzysztof Formela4 | Mohammad Reza Saeb2,3 |

Sabu Thomas5

1
Faculty of Health and Medical Sciences, The
University of Adelaide, Adelaide, South Although significant progress has been made in the design and application of inject-
Australia, Australia
able hydrogels for biomedical applications, concurrent control of rheological and
2
CentraleSupélec, LMOPS, Université de
Lorraine, Metz, France mechanical properties of injectable hydrogels has remained as an open challenge to
3
Laboratoire Matériaux Optiques, the researchers. In this work, we introduce and put into practice a photo‐curable
Photoniques et Systèmes, CentraleSupélec,
poloxamer (also known as Pluronic)/graphene oxide (Plu/GO) injectable hydrogel with
Université Paris‐Saclay, Metz, France
4
Department of Polymer Technology, Gdańsk
well‐controlled rheological and mechanical properties. Acrylate group was anchored
University of Technology, Gdansk, Poland to hydrogel structure to endow photo‐crosslinking ability through decelerating degra-
5
School of Chemical Sciences, MG University, dation rate of poloxamer hydrogels after injection. It was found that the modified Plu
Kottayam, Kerala, India
remains stable in biological media for a long‐term period without significant weight
Correspondence loss. Rheological properties of hydrogels were also carried out as essential prerequi-
M.R. Saeb, Dr.,Laboratoire Matériaux
Optiques, Photoniques et Systèmes, site for an ideal injectability via frequency sweep, flow curve, recovery, and yield
CentraleSupélec, Université Paris‐Saclay, stress before and after modification, signifying shear‐thinning behavior of Plu/GO
57070, Metz, France.
Email: mrsaeb2008@gmail.com hydrogels with high recoverability. The viscosity of shear‐thinning‐like hydrogels
S. Thomas, Department of Polymer dropped at higher shear stress, which facilitated injection process. Moreover,
Technology, Faculty of Chemistry, G.
mechanical behavior of Plu was optimized by manipulating the content of Plu, degree
Narutowicza Str. 11/12, Gdańsk University of
Technology, 80‐233 Gdansk, Poland. of modification with reactive precursor, curing, and particularly incorporation of GO
Email: sabuthomas@mgu.ac.in
without deteriorating effects on rheological behavior of Plu.

K E Y W OR D S

injectable hydrogel, mechanical properties, photo‐crosslinking, Pluronic, rheological properties

1 | I N T RO D U CT I O N regenerate tissue.17-19 Although considerable progress was made in

Hydrogels reveal remarkable features such as high swelling ratio, sol‐
gel transition, similarity to native tissue and provide acceptable perme-
ability when subjected to oxygen and water,1-5 making them potential
candidate for biomedical applications.6-10 For tissue engineering uses,
hydrogels should additionally provide a set of appropriate characteris-
tics, where invasive surgical operation underlines the need for
implanted hydrogels in lesion site to regenerate damaged tissue.10-14
Bearing such requirements in mind, the use of injectable hydrogels
with ability to form stable network is a prerequisite to avoid invasive
therapeutics.13,15,16 Injectable hydrogels serve as carrier for cells or
drugs to the target tissue, and even a scaffold in damaged site to
manufacturing injectable hydrogels, designing injectable hydrogel with
tunable rheological and mechanical properties has remained a chal-
lenging matter.20-22 Highly thixotropic hydrogels offer a great oppor-
tunity in the development of injectable hydrogels as shield cells
against high shear stress during injection. Moreover, thermal gelation
of injectable hydrogels such as N‐isopropylacrylamide23 and methyl-
cellulose24,25 at body temperature preserves a faithful cage for cells
or drugs once they are injected by the hydrogel.
Polxamer, also known as Pluronic, is a synthetic terpolymer com-
posed of poly (propylene oxide) (PPO) block in the middle and poly
(ethylene oxide) (PEO) blocks at two opposite sides.26,27 PEO and
PPO blocks play the role of hydrophilic and hydrophobic segments,

Polym Adv Technol. 2019;1–11. wileyonlinelibrary.com/journal/pat © 2019 John Wiley & Sons, Ltd. 1
2 ROHANI RAD
respectively, giving the macromolecule a dual yet controllable charac-
ter for various circumstances.27 Pluronic forms low‐viscosity solution
at room temperature, which allows easy processing and handling.
However, it undergoes physical gelation at higher temperatures as a
consequence of hydrophobic aggregation of PPO blocks.28 This
amphiphilic feature has been exploited in different biomedical areas
including drug delivery,29 tissue engineering,30 and 3D bioprinting.31
Typically, properties of Pluronic are dependent on temperature and
chain architecture. Pluronic reveals shear‐thinning and recovery for
encapsulated cells.32,33 However, highly deformable microstructure
of Pluronic in biological media because of its low‐molecular weight
may raise questions about its long‐term practical usage.34 To tackle
this issue, Pluronic has been modified with acrylate moieties to impart
photo‐crosslinking ability under UV‐irradiation.35,36 It was also
reported that addition of hyaluronic acid (HA) improves the mechani-
cal properties of Pluronic and allows for sustained release of drug
thanks to the improved intermolecular interaction between HA and
Pluronic.37 Another solution would be preparation of injectable
electroactive hydrogels through chemical reaction between carboxyl
groups of tetraaniline and hydroxyl end groups of Pluronic, so that
high cell viability was reported for Pluronic/tetraaniline electroactive
hydrogel. 38-40
Graphene and its derivatives are ranked as promising carbon allo-
tropes for their unique characteristics such as electrical and thermal
conductivity, mechanical properties, and biocompatibility.41-44
Graphene is a two‐dimensional monolayer structure of carbon atom
with high surface area, which gives it a very suitable carrier character
for many biomedical applications. 45-48
Graphene oxide (GO), as the
most commonly used derivative of graphene, have hydroxyl and car-
boxyl groups on its surface, which add hydrophilic character to
graphene structure with improved interaction with hydrophilic com-
pounds. A great deal of work has been reported regarding the rein-
forcing effect of GO in biopolymer matrices. 49-51
Won et al found
that incorporation of small amount of GO could significantly improve
the stability of Pluronic as well as its mechanical stiffness through
strong hydrophobic interaction between GO surfaces and PPO blocks
of Pluronic backbone.52 In addition to hydrophobic interaction,
hydroxyl groups of GO and Pluronic are able to form hydrogen bond-
ing.52 The large surface area together with superior electrical conduc-
tivity give graphene a pathway for charge transfer. It was proved that
its electrical stimulation potential could promote the growth and pro-
liferation of stem cells. 53
In this work, we developed Pluronic/GO nanocomposite as an
injectable hydrogel with shear‐thinning and recovery properties. We
knew that Pluronic is a thermos‐sensitive copolymer, which is able
to form temporary physical crosslinking at body temperature. Modifi-
cation of Pluronic with methacrylate anhydride could also provide a
secondary permanent crosslinking ability. Fourier transform infrared
spectroscopy (FTIR) and hydrogen‐1 magnetic resonance spectros-
copy (1H‐NMR) were used for chemical structure analyses. Moreover,
thixotropic behavior of Pluronic cause a viscosity drop at high shear
rates to expedite recovery to initial stage after injection‐what facili-
tates injection of Pluronic/GO hydrogel at room temperature. On
ET AL.
the other hand, GO nanosheets serve as reinforcing nanoparticles that
enhance mechanical properties of hydrogel. To evaluate the perfor-
mance of the hydrogel, rheological properties of hydrogel were stud-
ied in terms of frequency sweep, flow curve, recovery, yield stress,
and temperature sweep. In addition, swelling ratio and degradation
rate of the prepared hydrogels were evaluated to track the role of
modification of Pluronic.
2 | EXPERIMENTAL
2.1 | Materials and synthesis
Pluronic F127, methacrylate anhydride (MA), triethylamine (TEA), 2‐
hydroxy‐1‐(4‐(hydroxyethoxy) phenyl)‐2‐methyl‐1‐propanone
(Irgacure 2959) were all purchased from Sigma‐Aldrich Co. Pluronic
was modified according to a published procedure.54 In brief, Pluronic
(10 g) was dissolved in 20 mL chloroform at room temperature under
vigorous stirring before cooling to 0°C in an ice bath. Afterwards, TEA
was pipetted to the solution. MA (300 μL or 1000 μL, denoted 3X and
10X, respectively) was diluted in 5 mL chloroform and then was
poured to the solution. Next, the solution was left for 24 hours at
50°C. To separate the modified Pluronic (PluMA), solution was added
to the diethylether and filtered by glass funnel and dried in vacuum
oven for 48 hours. To synthesis GO according to the modified Hum-
mer's method,55 graphite (2 g) and potassium permanganate (12 g)
were added to the mixture sulfuric acid/phosphoric acid (240 mL/
25 mL) at room temperature under vigorous and heated up to 50°C
stirring for 24 hours. Afterwards, the resulting solution was mixed
with ice and hydrogen peroxide was added to the solution until the
color of solution change to bright brown. Next, solution was washed
with water (three times) and hydrochloric acid (two times) to remove
impurities. Figure 1 visualizes synthesis procedure of PluMA and GO.
2.2 | Fabrication of hydrogel
First, Irgacure 2959 (1 wt.%) was dissolved in deionized water for
10 minutes at 70°C. Next, GO powder was added to the solution
and probe sonicated for 30 minutes to obtain well‐dispersed GO
nanosheets. Afterwards, the untreated and modified Pluronic were
separately added to the solution and stirred for 1 hour in ice bath
and stored in fridge at 4°C to allow complete dissolution of Pluronic.
Thin films of hydrogel were fabricated by transferring 125 μL of pre‐
solution between two hydrophobic glass slides, which was separated
by two spacers with 1 mm thickness and underwent UV‐irradiation
for predetermined time.
2.3 | Raman spectroscopy
Raman spectroscopy (WITEC Alpha300) was used to confirm the suc-
cessful preparation of GO. Spectra were recorded in the range of 800
to 2000 cm−1 wavelength with a 537‐nm laser. For all experiments, sam-
ples were drop casted on glass slide and spectra recorded in a solid form.
ROHANI RAD ET AL.
FIGURE 1 Visualization of synthesis procedure of PluMA and GO [Colour figure can be viewed at wileyonlinelibrary.com]
2.4 | Fourier transform infrared
FTIR spectrometry was conducted on a Nicolet 6700 Thermo Fisher in
the wavelength range of 4000 to 600 cm−1. Samples were dried for
24 hours before experiment to remove moisture.
2.5 | Proton magnetic resonance spectroscopy (1H‐
NMR)
To confirm the successful modification of Pluronic, 1
H‐NMR
(400 MHz Varian) analysis was conducted in which about 20 mg of
untreated and modified Pluronic were dissolved in 1 mL of deuterium
oxide at 4°C and the appearance of new peaks were measured.
2.6 | X‐ray diffraction
XRD analysis was carried out by means of Rigaku MiniFlex 600, Japan
under predefined condition (40 kV and 15 mA, 2θ = 5–75o). To get a
better resolution in XRD pattern, GO was lyophilized for 2 days to
remove solvents.
2.7 | Thermogravimetric analysis
To characterize the loss weight of GO over temperature sweep, 5 mg
of GO was placed in the chamber of Q500 TA instrument Q500 (ther-
mogravimetric analysis [TGA]) and underwent 2°C/min heating regime
from room temperature to 900°C.
2.8 | Rheological analysis
Physica MCR301 rheometer (Anton Paar, Ostfildern, Germany) was
used to measure the rheological behavior of prepared hydrogels.
Before performing the test, hydrogels were transferred on the base
plane and conditioned for 10 minutes to eliminate the memory of
samples. In the frequency test, storage (G′) and loss modulus (G″) of
samples were recorded over 0.01 to 100 Hz. Recoverability of
hydrogels was examined by variation of G′ and G″ at low and high
3
strains. The viscosity of samples were traced varying shear rate from
0.01 to 1000 s−1. Yield stress was also determined from cross section
of G′ and G″ over shear stress sweep.
2.9 | Swelling ratio
The prepared hydrogels were soaked in deionized water for 24 hours
at 37°C to obtain the swelling equilibrium. They were weighted right
before and after they were lyophilized in freeze‐dryer. Swelling ratio
of hydrogels was measured according to following Equation (1):
W ðswellÞ − W ðdryÞ
× 100 (1)
W ðdryÞ
where W (swell) and W (dry) denote the weight of hydrogel in swell
and dry state.
2.10 | Degradation rate
To examine the degradation rate of the prepared hydrogels, identical
concentration of the untreated and modified Pluronic hydrogel were
prepared in a cylindrical shape (D = 20 mm, H = 10 mm) and immersed
in water for 1 month and then weighted in predetermined times to
study their weight loss as a function of time.
3 | RESULTS AND DISCUSSIONS
3.1 | Preparation of Pluronic/GO hydrogel
Pluronic is an amphiphilic commercial copolymer, which offers unique
rheological properties suitable for injectable hydrogels.56-59 However,
fast degradation of Pluronic in biological media restrict its practical
application in clinical usage. We modified Pluronic with MA to give it
permanent photo‐crosslinking ability. Figure 2 shows FTIR spectra of
the untreated and modified Pluronic. The new peak appearing at
1720 cm−1 in FTIR spectra of Pluronic/GO signifies C═O bond of
the modified Pluronic.54
4 ROHANI RAD
FIGURE 2 Fourier transform infrared (FTIR) spectra of Pluronic
before and after modification with methacrylate anhydride [Colour
figure can be viewed at wileyonlinelibrary.com]
In 1H‐NMR spectra (Figure 3), two new peaks are centered at
5.5 ppm and 6 ppm in the PluMA hydrogel, which refer to vinyl proton
of methacrylate group.60
GO nanosheets were prepared according to the modified Hum-
mer's method, as characterized by XRD, Raman, and TGA pattern in
Figure 4. As can be seen, characteristic peak of GO appeared at 2θ
FIGURE 3 1H‐NMR spectra of Pluronic before and after
modification with methacrylate anhydride [Colour figure can be
viewed at wileyonlinelibrary.com]
ET AL.
of 11° in XRD pattern, indicative of GO single layers. The presence
of large number of functional groups and water molecules between
GO sheets cause d spacing between GO nanosheets become
0.79 nm. In the Raman spectroscopy pattern, two peaks appeared at
around 1350 and 1590 cm−1 related to D and G bands, respectively.
The G/D band is greater than one in Raman spectroscopy pattern,
which provides further concrete evidence for formation of GO struc-
ture. Finally, in TGA experiment, initially GO lost weight at around
100°C, related to the absorbed moisture, followed by a considerable
weight loss at around 150 to 200°C related to the degradation of
functional groups.
3.2 | Flow behavior and yield stress of Pluronic/GO
Once hydrogel moves through nozzle during injection, it experiences
high shear stress applied from walls, which can damage cells. Thus,
an injectable hydrogel should possess shear‐thinning behavior, which
allows eliminating the shear stress applied during injection. Figure 5
provides flow curves of the untreated and modified Pluronic with dif-
ferent concentrations. The viscosity of Pluronic declines constantly
over shear rate, regardless of concentration of MA modifier, which
indicates that shear‐thinning behavior of Pluronic does not disrupted
upon modification by MA. Moreover, flow curves of Pluronic shifted
toward higher values by increasing concentration for micelles probably
located at closer distance to each other to form more hydrophilic
interactions. Power‐law model can quantify the shear‐thinning behav-
ior of Pluronic as the following Equation (2):
η ¼ K γ_ 1−n ; (2)
where η is viscosity; n is power‐law index, K is the consistency
index, and γ_ is the shear rate. For Newtonian fluid, power‐law index
is one, while for shear‐thinning and shear‐thickening solutions, n is
lower and greater than one, respectively. The viscosity value at low
shear rate can be detected by K value. Table 1 summarizes the calcu-
lated value of corresponding n and K. Accordingly, for all cases, power‐
law index is lower than one, signifying dominance of shear‐thinning
behavior. Moreover, K value, as a sign of viscosity at low shear rate,
shows upward trend upon increasing Pluronic concentration. It is
interesting to note that the presence of GO does not negatively affect
the shear‐thinning behavior of Pluronic.
In addition to viscosity, yield stress of injectable hydrogel deter-
mines the spreading rate of hydrogel after injection. Typically, a higher
yield stress leads to shape integrity of hydrogel after injection. In
another words, yield stress of injectable hydrogel indicates the resis-
tance of hydrogel against flow before and after injection. Yield stress
and viscosity at low shear concurrently play critical roles in controlling
the drug release and shape fidelity of injectable hydrogels. Cross sec-
tion of G′ and G″ over shear stress sweep is generally considered as
the yield point, where sol‐gel transition occurs (Figure 6). It is apparent
that for all cases G′ and G″ are stable before yield point, where G′
goes under G″ and hydrogel shows sol behavior. Moreover, the yield-
ing point of Pluronic moves to higher values upon increasing Pluronic
ROHANI RAD ET AL. 5

FIGURE 4 Characterization of GO prepared by the modified Hummer's method in term of (a) X‐ray diffraction (XRD), (b) Raman spectroscopy,
and (c) thermogravimetric analysis (TGA)

FIGURE 5 Flow curve of (a) unmodified and (b) modified Pluronic with different concentrations [Colour figure can be viewed at
wileyonlinelibrary.com]

concentration, which agrees well with flow curve results. In addition,

TABLE 1 Calculated value of n and K from power‐law model introduction of GO also improves yield point, which allows injectable
hydrogels to keep their integrity after injection. Modified Pluronic also
Sample n K
follow the same pattern without any noticeable decline in yield stress
Plu‐20 0.17 197 values.
Plu‐25 0.15 242
Plu‐30 0.18 489
Plu‐25/GO‐0.5 0.16 235 3.3 | Recoverability of Pluronic/GO
PluMA10X‐20 0.16 184
Injectable hydrogel should recover fast to its initial state once hydro-
PluMA10X‐25 0.17 253
gel is injected in the damaged site; otherwise, encapsulated cells or
PluMA10X‐30 0.18 502
drugs will release quickly after injection. To measure the
6 ROHANI RAD ET AL.

FIGURE 6 Variation of G′ and G″ over frequency sweep for (a) Plu‐20, (b) Plu‐25, (c) Plu‐30, (d) Plu‐25/GO‐0.5 hydrogels, (e) PluMA‐20, (f)
PluMA‐25, and (g) PluMA‐30 [Colour figure can be viewed at wileyonlinelibrary.com]

recoverability, the prepared hydrogels went under repetitive low and
high strains for several cycles. Figure 7 depicts the variation of G′
and G″ over applying different strains. Accordingly, G′ stands over G
″ at low strain as a signature of gel state. However, G′ plunged sud-
denly at high strain and got lower than G″, which suggests sol‐gel
transition. After removal of high strain, G′ surged in a very short time
and recovered fully to its initial state. This behavior was observed for
all cycles. It is interesting to note that Pluronic is able to recover at all
studied concentrations, where addition of GO only causes G′ to stand
at higher values. Recoverability of the modified Pluronic follows the
same trend, confirming that modification of Pluronic cannot disturb
the inherent rheological behavior of Pluronic.
3.4 | Mechanical behavior of Pluronic/GO
Figure 7 shows the variation of G′ and G″ over frequency sweep for
the untreated and modified Pluronic hydrogels before UV‐curing.
Storage modulus is directly related to the elastic modulus of hydrogel.
Accordingly, for all the prepared concentrations, G′ is above G″ over
ROHANI RAD ET AL. 7

FIGURE 7 Variation of G′ and G″ over time at low and high strain for (a) Plu‐20, (b) Plu‐25, (c) Plu‐30, (d) Plu‐25/GO‐0.5 hydrogels, (e) PluMA‐
20, (f) PluMA‐25, and (g) PluMA‐30 hydrogels [Colour figure can be viewed at wileyonlinelibrary.com]

the whole frequency range implying the gel behavior of Pluronic.
Moreover, G′ shifted toward higher values upon increasing the con-
centration of the untreated or modified Pluronic, as expected form vis-
cosity trend. This behavior proves that a stiffer hydrogel is achieved at
higher concentration of Pluronic. Of note, modification of Pluronic has
no adverse effect on G′ value (Figure 8).
To explore the effect of modification on mechanical properties of
hydrogel after curing, Pluronic was modified with two different
degrees of substitution. Figure 9 reveals the mechanical behavior of
the modified Pluronic with different degrees of substitution after cur-
ing under UV‐irradiation for 2 minutes. With comparing the G′ for
PluMA10X and PluMA3X, it is found that higher degree of substitu-
tion of Pluronic leads to higher value of G′, which means higher elastic
modulus. Moreover, mechanical properties of hydrogel improve upon
increasing the PluMA concentration. Furthermore, addition of GO
enhances the G′ of Pluronic. Curing parameter also could affect the
8 ROHANI RAD ET AL.

FIGURE 8 Variation of G′ and G″ over frequency sweep for (a) unmodified and (b) modified Pluronic before ultraviolet (UV) curing [Colour figure
can be viewed at wileyonlinelibrary.com]

FIGURE 9 Variation of G′ and G″ over frequency sweep for (a) PluMA3X and (b) PluMA3X Pluronic after curing under ultraviolet (UV) for
2 minutes [Colour figure can be viewed at wileyonlinelibrary.com]

ultimate mechanical properties. For example, it was observed that lon-
ger UV‐irradiation led to stiffer hydrogel (Figure 9C). Temperature was
another indicator, which strongly affected rheological properties of
Pluronic hydrogel. As can be seen in Figure 9D, both G′ and G″ of
Pluronic hydrogel jumped to higher values at higher temperature, sig-
nifying change in the micelle hydrophilicity of Pluronic hydrogel.
Swelling and degradation behavior of the prepared hydrogels was
investigated in deionized water (Figure 10). Accordingly, swelling ratio
of hydrogel continuously decreased upon increasing the concentration
of Pluronic due to the higher crosslink density of 3D network at higher
concentration of Pluronic. Moreover, it was found that swelling ratio
of PluMA10X was lower than PluMA3X because higher modification
degree means higher crosslink density and consequently lower
swelling ratio. In the case of degradation rate, although untreated
Pluronic went under degradation after 1 week, the modified Pluronic
remained stable for 1 month, which was more pronounced for
PluMA10X as it almost remained unchanged over 1 month. Improved
stability of Pluronic stemmed from permanent crosslinked network
caused by acrylate moieties.
Figure 11 provides overview of Plu/GO injectable hydrogel.
Pluronic forms thermal gelation at body temperature because of
micelle formation, as reflected in rheological and mechanical proper-
ties. However, thermal gelation is not strong enough to keep the
shape fidelity for long‐term. Modification with MA gives the samples
photo‐crosslinking ability, which itself provides permanent 3D net-
work after injection and, consequently, stability at biological media.
ROHANI RAD ET AL. 9

FIGURE 10 (a) Swelling ratio and (b) degradation behavior of modified and unmodified Pluronic [Colour figure can be viewed at
wileyonlinelibrary.com]

FIGURE 11 Proposed mechanism for modulation of mechanical and rheological properties of Pluronic [Colour figure can be viewed at
wileyonlinelibrary.com]

Notably, modification of Pluronic does not deteriorate rheological
behavior of Pluronic and only equip it with photo‐crosslinking ability.
The GO nanosheets were well‐dispersed in the Pluronic because of
the hydrophobic and hydrophilic interactions, which improved ulti-
mate mechanical properties. Overall, mechanical behavior of Pluronic
hydrogel could be adjusted by manipulating the degree of modifica-
tion, concentration of Pluronic, incorporation of GO nanosheets, and
curing parameter. In other words, the modified Pluronic benefitted
from identical rheological behavior of neat Pluronic and tunable
mechanical properties after curing.
10
4 | C O N CL U S I O N
In this work, an injectable Pluronic/GO hydrogel was introduced with
enhanced mechanical properties without deteriorating rheological
characteristics of Pluronic. Unmodified Pluronic hydrogel showed
rapid dissociation due to the lack of post‐crosslinking ability. However,
modification of Pluronic with MA and crosslinking under UV‐
irradiation led to stabilization of Pluronic in biological media. Pluronic
with different modification degrees were synthesized and assessed
by the appearance of one new peak at 1720 cm−1 in the FTIR spectra,
and two new peaks centered at 5.5 ppm and 6 ppm in the HNMR
spectra, respectively. GO nanosheets were prepared according to
the modified Hummer's method and characterized in terms of XRD,
TGA, and Raman spectroscopy to confirm the structures. Rheological
properties of the untreated and modified Pluronic hydrogels were ana-
lyzed in terms of frequency sweep, flow curve, yield stress, and recov-
ery. Accordingly, Pluronic hydrogels revealed shear‐thinning behavior
with approximately power‐law index of 0.1. In addition, it was found
that both untreated and modified Pluronic possess 100% recoverabil-
ity, which suits injectability. The yield stress and storage modulus of
Pluronic were improved upon increasing its concentration. Incorpora-
tion of GO enhanced yield stress and storage modulus of Pluronic
hydrogels before and after curing. The modification of Pluronic did
not disrupt its rheological properties, but gave it post‐crosslinking
potential. Eventually, it was found that modification degree of Pluronic
plays the main role in tuning the ultimate properties of Pluronic hydro-
gel. Swelling ratio of the modified Pluronic shows downward trend
upon increasing degree of modification because of higher crosslinking
density. Moreover, unlike untreated Pluronic, which was dissociated
very fast in 1 week, the modified Pluronic remained almost unaffected
over a month without weight loss because of the permanent
crosslinking.
ORCID
Henri Vahabi https://orcid.org/0000-0003-0419-7368
Mohammad Reza Saeb https://orcid.org/0000-0001-9907-9414
RE FE R ENC E S
1. Ahmed EM. Hydrogel: preparation, characterization, and applications: a
review. J Adv Res. 2015;6(2):105‐121.
2. Macaya D, Spector M. Injectable hydrogel materials for spinal cord
regeneration: a review. Biomed Mater. 2012;7(1):012001.
3. Zarrintaj P, Urbanska AM, Gholizadeh SS, Goodarzi V, Saeb MR,
Mozafari M. A facile route to the synthesis of anilinic electroactive col-
loidal hydrogels for neural tissue engineering applications. J Colloid
Interface Sci. 2018;516:57‐66.
4. Zarrintaj P, Manouchehri S, Ahmadi Z, et al. Agarose‐based biomate-
rials for tissue engineering. Carbohydr Polym. 2018;187:66‐84.
5. Zarrintaj P, Moghaddam AS, Manouchehri S, et al. Can regenerative
medicine and nanotechnology combine to heal wounds? The search
for the ideal wound dressing. Nanomedicine. 2017;12(19):2403‐2422.
6. Ali A, Ahmed S. Recent advances in edible polymer based hydrogels as
a sustainable alternative to conventional polymers. J Agric Food Chem.
2018;66(27):6940‐6967.
ROHANI RAD ET AL.
7. Song M‐M, Wang Y‐M, Wang B, et al. Super tough, ultrastretchable
hydrogel with multistimuli responsiveness. ACS Appl Mater Interfaces.
2018;10(17):15021‐15029.
8. Zarrintaj P, Bakhshandeh B, Saeb MR, et al. Oligoaniline‐based conduc-
tive biomaterials for tissue engineering. Acta Biomater. 2018;72:16‐34.
9. Mohebbi S, Nezhad MN, Zarrintaj P, et al. Chitosan in biomedical engi-
neering: a critical review. Curr Stem Cell Res Ther. 2018;14(2):93‐116.
10. Manouchehri S, Bagheri B, Rad SH, et al. Electroactive bio‐epoxy incor-
porated chitosan‐oligoaniline as an advanced hydrogel coating for
neural interfaces. Prog Org Coat. 2019;131:389‐396.
11. Dimatteo R, Darling NJ, Segura T. In situ forming injectable hydrogels
for drug delivery and wound repair. Adv Drug Deliv Rev.
2018;127:167‐184.
12. Nilforoushzadeh MA, Zare M, Zarrintaj P, et al. Engineering the niche
for hair regeneration–a critical review. Nanomedicine: Nanotechnology
Bio Med. 2019;15(1):70‐85.
13. Farokhi M, Mottaghitalab F, Fatahi Y, et al. Silk fibroin scaffolds for
common cartilage injuries: possibilities for future clinical applications.
Eur Polym J. 2019;115:251‐267.
14. Zarrintaj P, Bakhshandeh B, Rezaeian I, Heshmatian B, Ganjali MR. A
novel electroactive agarose‐aniline pentamer platform as a potential
candidate for neural tissue engineering. Sci Rep. 2017;7(1):17187.
15. Thambi T, Li Y, Lee DS. Injectable hydrogels for sustained release of
therapeutic agents. J Control Release. 2017;267:57‐66.
16. Liu M, Zeng X, Ma C, et al. Injectable hydrogels for cartilage and bone
tissue engineering. Bone Res. 2017;5:17014.
17. Sivashanmugam A, Kumar RA, Priya MV, Nair SV, Jayakumar R. An
overview of injectable polymeric hydrogels for tissue engineering. Eur
Polym J. 2015;72:543‐565.
18. Bakhshandeh B, Zarrintaj P, Oftadeh MO, et al. Tissue engineering;
strategies, tissues, and biomaterials. Biotechnol Genet Eng Rev.
2017;33(2):144‐172.
19. Bagher Z, Atoufi Z, Alizadeh R, et al. Conductive hydrogel based on
chitosan‐aniline pentamer/gelatin/agarose significantly promoted
motor neuron‐like cells differentiation of human olfactory ecto‐
mesenchymal stem cells. Mater Sci Eng C. 2019;101:243‐253.
20. Maia J, Ferreira L, Carvalho R, Ramos MA, Gil MH. Synthesis and char-
acterization of new injectable and degradable dextran‐based
hydrogels. Polymer. 2005;46(23):9604‐9614.
21. Rahmati M, Milan PB, Samadikuchaksaraei A, et al. Ionically crosslinked
thermoresponsive chitosan hydrogels formed in situ: a conceptual
basis for deeper understanding. Macromol Mater Eng.
2017;302(11):1700227.
22. Nilforoushzadeh MA, Amirkhani MA, Zarrintaj P, et al. Skin care and
rejuvenation by cosmeceutical facial mask. J Cosmet Dermatol.
2018;17(5):693‐702.
23. Liao W, Zhang Y, Guan Y, Zhu X. Gelation kinetics of thermosensitive
PNIPAM microgel dispersions. Macromol Chem Phys.
2011;212(18):2052‐2060.
24. Shirata Y, Wakasa A, Miura K, Nakamura H, Matsumoto Y, Miyada T.
Body heat responsive gelation of methylcellulose formulation contain-
ing betaine. Biosci Biotechnol Biochem. 2017;81(9):1829‐1836.
25. Vahabi H, Shabanian M, Aryanasab F, Mangin R, Laoutid F, Saeb MR.
Inclusion of modified lignocellulose and nano‐hydroxyapatite in devel-
opment of new bio‐based adjuvant flame retardant for poly (lactic
acid). Thermochim Acta. 2018;666:51‐59.
26. Chun KW, Lee JB, Kim SH, Park TG. Controlled release of plasmid
DNA from photo‐cross‐linked pluronic hydrogels. Biomaterials.
2005;26(16):3319‐3326.
ROHANI RAD ET AL.
27. Zarrintaj P, Ahmadi Z, Saeb MR, Mozafari M. Poloxamer‐based stimuli‐
responsive biomaterials. Mater Today: Proc. 2018;5(7, Part
3):15516‐15523.
28. Lee Y, Chung HJ, Yeo S, et al. Thermo‐sensitive, injectable, and tissue
adhesive sol–gel transition hyaluronic acid/pluronic composite
hydrogels prepared from bio‐inspired catechol‐thiol reaction. Soft Mat-
ter. 2010;6(5):977‐983.
29. Batrakova EV, Kabanov AV. Pluronic block copolymers: evolution of
drug delivery concept from inert nanocarriers to biological response
modifiers. J Control Release. 2008;130(2):98‐106.
30. Gioffredi E, Boffito M, Calzone S, et al. Pluronic F127 hydrogel charac-
terization and biofabrication in cellularized constructs for tissue
engineering applications. Procedia CIRP. 2016;49:125‐132.
31. Müller M, Becher J, Schnabelrauch M, Zenobi‐Wong M. Nanostruc-
tured Pluronic hydrogels as bioinks for 3D bioprinting. Biofabrication.
2015;7(3):035006.
32. Guvendiren M, Lu HD, Burdick JA. Shear‐thinning hydrogels for bio-
medical applications. Soft Matter. 2012;8(2):260‐272.
33. Zarrintaj P, Mostafapoor F, Milan PB, Saeb MR. Theranostic platforms
proposed for cancerous stem cells: a review. Curr Stem Cell Res Ther.
2019;14(2):137‐145.
34. Wu C‐J, Gaharwar AK, Chan BK, Schmidt G. Mechanically
tough pluronic F127/laponite nanocomposite hydrogels from
covalently and physically cross‐linked networks. Macromolecules.
2011;44(20):8215‐8224.
35. Lee S‐Y, Tae G. Formulation and in vitro characterization of an in situ
gelable, photo‐polymerizable Pluronic hydrogel suitable for injection.
J Control Release. 2007;119(3):313‐319.
36. Missirlis D, Hubbell J, Tirelli N. Thermally‐induced glass formation from
hydrogel nanoparticles. Soft Matter. 2006;2(12):1067‐1075.
37. Y‐s J, Park W, Park H, Lee D‐K, Na K. Thermo‐sensitive injectable
hydrogel based on the physical mixing of hyaluronic acid and Pluronic
F‐127 for sustained NSAID delivery. Carbohydr Polym.
2017;156:403‐408.
38. Jin E, Zhang Z, Lian H, et al. Injectable electroactive hydrogels based on
Pluronic® F127 and tetraaniline copolymer. Eur Polym J.
2017;88:67‐74.
39. Zarrintaj P, Ahmadi Z, Vahabi H, Ducos F, Saeb MR, Mozafari M.
Polyaniline in retrospect and prospect. Mater Today: Proc.
2018;5(7):15852‐15860.
40. Atoufi Z, Zarrintaj P, Motlagh GH, Amiri A, Bagher Z, Kamrava SK. A
novel bio electro active alginate‐aniline tetramer/agarose
scaffold for tissue engineering: synthesis, characterization, drug
release and cell culture study. J Biomater Sci Polym Ed.
2017;28(15):1617‐1638.
41. Gaharwar AK, Arpanaei A, Andresen TL, Dolatshahi‐Pirouz A. 3D bio-
material microarrays for regenerative medicine: current state‐of‐the‐
art, emerging directions and future trends. Adv Mater.
2016;28(4):771‐781.
42. Gaharwar AK, Peppas NA, Khademhosseini A. Nanocomposite
hydrogels for biomedical applications. Biotechnol Bioeng.
2014;111(3):441‐453.
43. Nonahal M, Rastin H, Saeb MR, et al. Epoxy/PAMAM dendrimer‐
modified graphene oxide nanocomposite coatings: nonisothermal cure
kinetics study. Prog Org Coat. 2018;114:233‐243.
44. Nonahal M, Saeb MR, Hassan Jafari S, et al. Design, preparation,
and characterization of fast cure epoxy/amine‐functionalized
graphene oxide nanocomposites. Polym Compos. 2018;39(S4):
E2016‐E2027.
11
45. Shao Y, Wang J, Wu H, Liu J, Aksay IA, Lin Y. Graphene based electro-
chemical sensors and biosensors: a review. Electroanalysis: An Int J
Devoted Fund Prac Aspects Electroanal. 2010;22(10):1027‐1036.
46. Sanchez VC, Jachak A, Hurt RH, Kane AB. Biological interactions of
graphene‐family nanomaterials: an interdisciplinary review. Chem Res
Toxicol. 2011;25(1):15‐34.
47. Yarahmadi E, Didehban K, Sari MG, et al. Development and curing
potential of epoxy/starch‐functionalized graphene oxide nanocompos-
ite coatings. Prog Org Coat. 2018;119:194‐202.
48. Haghdadeh P, Ghaffari M, Ramezanzadeh B, Bahlakeh G, Saeb MR.
The role of functionalized graphene oxide on the mechanical and
anti‐corrosion properties of polyurethane coating. J Taiwan Inst Chem
Eng. 2018;86:199‐212.
49. Sahu A, Choi WI, Tae G. A stimuli‐sensitive injectable graphene oxide
composite hydrogel. Chem Commun. 2012;48(47):5820‐5822.
50. Yarahmadi E, Didehban K, Shabanian M, Saeb MR. High‐performance
starch‐modified graphene oxide/epoxy nanocomposite coatings: a
glimpse at cure kinetics and fracture behavior. Prog Color Colorants
Coat. 2018;11:55‐62.
51. Choolaei M, Goodarzi V, Khonakdar HA, et al. Influence of graphene
oxide on crystallization behavior and chain folding surface free energy
of poly (vinylidenefluoride‐co‐hexafluoropropylene). Macromol Chem
Phys. 2017;218(19):1700103.
52. Won D‐A, Kim M, Tae G. Systemic modulation of the stability of
pluronic hydrogel by a small amount of graphene oxide. Colloids Surf
B Biointerfaces. 2015;128:515‐521.
53. Li N, Zhang Q, Gao S, et al. Three‐dimensional graphene foam as a bio-
compatible and conductive scaffold for neural stem cells. Sci Rep.
2013;3(1):1604.
54. Hong K‐H, Jeon Y‐S, Kim J‐H. Preparation and properties of modified
PHEMA hydrogels containing thermo‐responsive pluronic component.
Macromol Res. 2009;17(1):26‐30.
55. Tran DN, Kabiri S, Losic D. A green approach for the reduction of
graphene oxide nanosheets using non‐aromatic amino acids. Carbon.
2014;76:193‐202.
56. Yu L, Ding J. Injectable hydrogels as unique biomedical materials. Chem
Soc Rev. 2008;37(8):1473‐1481.
57. Puglia D, Rastin H, Saeb MR, Shojaei B, Formela K. Cure kinetics of
epoxy/MWCNTs nanocomposites: isothermal calorimetric and rheo-
logical analyses. Prog Org Coat. 2017;108:75‐83.
58. Karahrodi MH, Jazani OM, Paran SMR, Formela K, Saeb MR. Modifica-
tion of thermal and rheological characteristics of bitumen by waste
PET/GTR blends. Construct Build Mater. 2017;134:157‐166.
59. Rastin H, Jafari SH, Saeb MR, Khonakdar HA, Wagenknecht U,
Heinrich G. Mechanical, rheological, and thermal behavior assessments
in HDPE/PA‐6/EVOH ternary blends with variable morphology. J Poly-
mer Res. 2014;21(2):352.
60. Lee BH, Shirahama H, Cho N‐J, Tan LP. Efficient and controllable syn-
thesis of highly substituted gelatin methacrylamide for mechanically
stiff hydrogels. RSC Adv. 2015;5(128):106094‐106097.
How to cite this article: Rohani Rad E, Vahabi H, Formela K,
Saeb MR, Thomas S. Injectable poloxamer/graphene oxide
hydrogels with well‐controlled mechanical and rheological
properties. Polym Adv Technol. 2019;1–11. https://doi.org/
10.1002/pat.4654