Materials Letters 186 (2017) 1720

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Materials Letters
journal homepage: www.elsevier.com/locate/matlet

Improved mechanical strength of porous chitosan scaold by graphene

coatings
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Shaolin Wena, Zan Wangb, Xianliang Zhenga, Xin Wanga,
a
College of Materials Science and Engineering, Key laboratory of Automobile Materials of MOE, Jilin University, Changchun 130012, PR China
b
The First Hospital of Jilin University, Changchun 130021, PR China

A R T I C L E I N F O A BS T RAC T

Keywords: Chitosan (CS) scaolds coated by reduced graphene oxide (rGO) lm were prepared and characterized using X-
Graphene coating ray diraction, X-ray photoelectron spectroscopy, scanning electron microscopy and a dynamic mechanical
Chitosan scaold analyze. The results exhibited that the rGO coating with wrinkled structure attached on pore wall of CS scaolds
Compressive strength and formed a rough and irregular surfaces. The compressive mechanical strength increased with increasing
Biomaterials
graphene oxide dispersion (0, 0.02, 0.1, and 1.0 mg/ml) and the maximum value of about 0.707 MPa was
Carbon materials
achieved at the concentration of 1.0 mg/ml, 1.26-fold greater compared with CS scaold. The porosity and pore
diameter of the scaold did not change much and the composite scaold might be considered as potential
platform for tissue engineering applications.

1. Introduction coating on the improvement of a composite scaold with good

Scaold or three-dimensional construct is one of the key compo-
nents of tissue engineering for cell adhesion, proliferation and dier-
entiation. A high-performance scaold not only possesses biological
properties, regulating cell behavior for tissue development, but also
provide enough mechanical support for cell and tissue growth [1,2]. As
a natural biopolymer, chitosan (CS) is a high-molecular-weight linear
cationic polysaccharide derived from the extensive deacetylation of
chitin, having been served as conventional three-dimensional scaold
in clinical applications [3]. However, weak mechanical strength of
chitosan scaolds prepared by traditional freeze-drying method has
limited their further commercialization [4]. Therefore, much eort has
been made to enhance their mechanical properties either by optimizing
fabrication method to increase crystallinity of chitosan scaold [5] or
through adding nanollers as a reinforcement such as hydroxyapatite
[6] and carbon-based nanomaterials including carbon nanotubes and
graphene [7]. Graphene and its chemical derivatives (graphene oxide
(GO) and reduced graphene oxide (rGO) sheets) were found to be able
to improve mechanical strength of matrix and support cellular pro-
liferation, adhesion, and even induce the dierentiation of stem cells
[7,8]. Thus, covering graphene lm onto pore wall of relatively soft
polymer scaolds may reinforce the mechanical properties of these
scaolds and change their surface micro-topography. Very recently,
Kanayama et al. reported a positive eect of reduced graphene oxide
bioactivity and enhanced compressive strength [9].
In this letter, graphene oxide coatings were formed by dipping CS
scaolds into GO dispersion and through electrostatic and hydrogen
bonding interaction between the oxygen containing functional groups
of GO and chitosan amino groups. Then, the prepared samples were
heated at 150 to obtain rGO-coated CS scaolds. To our best
knowledge, rGO/CS composite scaolds have been rarely reported up
till now.
2. Experimental section
2.1. Materials
The materials we had used were shown as follows: Chitosan, natural
powder graphite, sulfuric acid, sodium hydroxide, hydrochloric acid,
acetic acid, sodium nitrate, potassium permanganate, hydrogen per-
oxide, ethanol. The details of materials are available in Electronic
Supplementary material.
2.2. Synthesis of GO colloidal solution
Graphite oxide was prepared according to the modied Hummers
method [10]. The particular steps are available in Electronic
Supplementary material. Then, graphite oxide solution was mixed with

Corresponding author.
E-mail address: wang_xin@jlu.edu.cn (X. Wang).

http://dx.doi.org/10.1016/j.matlet.2016.09.040
Received 20 July 2016; Received in revised form 25 August 2016; Accepted 10 September 2016
Available online 12 September 2016
0167-577X/ 2016 Elsevier B.V. All rights reserved.
S. Wen et al.
Fig. 1. (ad) Digital photographs of CS scaold, 0.02 mg - rGO/CS scaold, 0.1 mg -
rGO/CS scaold, and 1.0 mg - rGO/CS scaold; (e) TEM image of graphene oxide sheets;
(f) XRD patterns of GO and rGO coatings.
Fig. 2. XRD patterns of CS and rGO-coated CS scaolds.
ethanol and ultrasonically treated for 1 h to obtain graphene oxide
dispersion (0, 0.02, 0.1, and 1.0 mg/ml).
2.3. Preparation of CS scaolds
A chitosan concentration of 4 wt% was prepared by thorough
dissolution of chitosan powder in 0.2 M acetic acid. After that, the
solution was poured into 24 well plates and frozen under 20 ,
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Materials Letters 186 (2017) 1720
followed by lyophilization in a freeze-dryer to form the CS scaolds.
Lyophilized scaolds were immersed in sodium hydroxide dispersion
and washed with DI water for several times until pH near 7. Finally,
they were frozen and lyophilized again for use.
2.4. Preparation of rGO-coated CS scaolds
CS scaolds were dipped into the GO dispersions for 48 h and dried
in air. Finally, the samples were heated at 150 for 12 h to get
chitosan scaolds coated by rGO lm, which are entitled as CS, 0.02 mg
- rGO/CS, 0.1 mg - rGO/CS, and 1.0 mg - rGO/CS.
2.5. Characterization of materials
Transmission electron microscopy (TEM), X-ray diraction (XRD),
X-ray photoelectron spectroscopy (XPS), scanning electron microscopy
(SEM) and dynamic mechanical analyzer were used to characterize
materials. The method of ethanol substitution were used to measure
the porosity of the scaolds. The details of this part are available in
Electronic Supplementary material.
3. Results and discussion
From the digital photographs shown in Fig. 1(a)(d), we can see
that the yellow-colored CS scaolds (~25 mm thickness and ~15 mm
diameter) get darker with increasing the coated GO concentrations. To
characterize the graphene oxide sheet, transmission electron micro-
scopy (TEM) was used. Fig. 1(e) exhibits a successful exfoliation of
individual graphene oxide sheets oxide sheet that displays as a thin
extended lm with a wrinkled surface and a lateral size of about several
micrometers. Fig. 1(f) gives the XRD patterns of the as-prepared GO
and 150 -heated GO lm coated on circular glasses, which shows the
main diraction peak of 11.98 shifted to 23.50, conrming the
formation of rGO lm.
XRD measurements were performed to analyze the crystal structure
of the prepared samples. It can be seen from Fig. 2 that all the XRD
patterns look similar and two strong diraction peaks around 14.90
and 20.41 indicate high degree of crystallinity of the scaolds [11,12].
The rGO coatings hardly disrupt the crystalline constituent of chitosan.
The XPS was used to further conrm the reduction of GO and the
combination of rGO and chitosan. In the C1s spectrum of GO (Fig. 3a),
ve binding energy peaks at 284.4, 285.4, 286.8, 287.7, and 288.6 eV
are corresponding to sp2 C=C, sp3 CC, COH, COC, and C=O,
respectively. After heating treatment at 150 (Fig. 3b), all of the
oxygen containing functional groups were reduced largely and a new
peak appears at 289.2 eV, which attributes to O=COH. Compared
these two C1s spectra, we can conrm that GO lm coated on chitosan
scaold has been reduced to rGO. In addition, from Fig. 3c and d, the
C1s spectra of pure CS scaold and rGO/CS composite scaold,
respectively, we can deduce the covering of rGO on the surface of the
scaold wall, which agrees well with the result of SEM measurement.
SEM images in Fig. 4(a)(c) reveal a highly porous, relatively
homogeneous and interconnected structure of the pure scaolds. The
pore diameter was in the range of 150300 m for all scaolds
including the composites, implying that the incorporation of GO
coatings did not obviously alter the porous structure of the scaolds
(porosity keeps nearly constant at about 92%,). From Fig. 4(d)(f), it
can be seen that not only did the surface of pore wall of rGO/CS
scaolds appear wrinkles, but also the degree of wrinkles be enhanced
with the increase of GO dispersion concentration due to the covering of
rGO lms. The SEM images suggested that rGO had coated on CS
scaolds and the thickness of rGO coatings could be adjusted by the
concentration of GO dispersion and thus regulate surface topography of
the composite scaold and inuence cell behavior [13].
In addition to be able to add elasticity of the matrix scaolds,
graphene-based coating may also increase their compressive strength
S. Wen et al. Materials Letters 186 (2017) 1720

Fig. 3. XPS spectra of (a) GO, (b) rGO, (c) CS scaolds and (d) 1.0 mg - rGO/CS scaold.

Fig. 4. SEM micrographs of (ac) CS scaolds, (d) 0.02 mg - rGO/CS scaold, (e) 0.1 mg - rGO/CS scaold, (f) 1.0 mg - rGO/CS scaold, and (g) compressive strengths of the scaolds.

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S. Wen et al.
[9]. In this letter, the compressive strength of chitosan scaold was
found dependent on the concentration of GO dispersions, and the
dierences in the strength are statistically signicant (Fig. 4g). The
maximum value of about 0.707 MPa was achieved for sample of rGO/
1.0 mg CS scaold, 1.26-fold greater compared with pure CS scaold
(0.561 MPa). As the strong interfacial interactions between the GO
sheets and the polymer matrix and the increase of rGO thickness due to
increasing GO dispersion concentration, the stress applied to the
composite scaolds could be eciently transferred through the rGO
coatings, leading to high strength.
4. Conclusions
We coated CS scaolds with GO sheets followed by thermal
reduction treatment at low heating temperature. The as-prepared
rGO coatings modied the surface morphology of the CS scaolds
and also increased their compressive strength without changing the
porous structures. The rGO/CS composite scaolds may oer a
powerful platform for stem cell research and tissue regenerative
medicine.
Acknowledgement
We would like to thank the Project sponsored by Jilin Province
Development and Reform Commission (Grant NO. 2015Y032-7), the
China Scholarship Council (CSC, File NO. 201406175003) for a visiting
scholar, and the supporting from Suzhou Institute of Nano-Tech and
Nano-Bionics, Chinese Academy of Sciences (16NBI01).
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.matlet.2016.09.040.
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Materials Letters 186 (2017) 1720
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