View Article Online / Journal Homepage / Table of Contents for this issue

Nanoscale Dynamic Article Links < C

Cite this: Nanoscale, 2012, 4, 2937

www.rsc.org/nanoscale PAPER
Graphene oxide versus functionalized carbon nanotubes as a reinforcing agent
in a PMMA/HA bone cement
Gil Gonçalves,a Sandra M. A. Cruz,a A. Ramalho,b Jos
e Gr
acioa and Paula A. A. P. Marques*a
Received 8th February 2012, Accepted 23rd February 2012
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E

DOI: 10.1039/c2nr30303e

Graphene oxide (GO) and functionalized carbon nanotubes (f-CNTs) (each in the concentration range
of 0.01–1.00 wt/wt%) were investigated as the reinforcing agent in a poly(methyl methacrylate)
(PMMA)/hydroxyapatite (HA) bone cement. Mixed results were obtained for the changes in the
Downloaded by University of Sussex on 14 January 2013

mechanical properties determined (storage modulus, bending strength, and elastic modulus) for the
reinforced cement relative to the unreinforced counterpart; that is, some property changes were
increased while others were decreased. We postulate that this outcome is a consequence of the fact that
each of the nanofillers hampered the polymerization process in the cement; specifically, the nanofiller
acts as a scavenger of the radicals produced during polymerization reaction due to the delocalized p-
bonds. Results obtained from the chemical structure and polymer chain size distribution determined,
respectively, by nuclear magnetic resonance and size exclusion chromatography analysis, on the
polymer extracted from the specimens support the postulated mechanism. Furthermore, in the case of
the 0.5 wt/wt% GO-reinforced cement, we showed that when the concentration of the radical species in
the PMMA bone cement was doubled, mechanical properties markedly improved (relative to the value
in the unreinforced cement), suggesting suppression of the aforementioned scavenger activity.

Introduction addressed in many studies.6,7,21,22 The results show that the

Single-layer graphene has received a lot of research attention
because of its remarkable thermal, electronic, and mechanical
properties; moreover, graphene can significantly improve the
physical properties of many polymers at low loading.1–5 Gra-
phene has been referred to as a credible alternative to carbon
nanotubes (CNTs) for mechanical reinforcement of polymeric
matrixes.6,7 Graphene powder can be produced in a way scalable
to mass production,2 which, in combination with low production
costs, makes graphene-powder-based composite materials
attractive for a variety of uses. In addition to their excellent
physical and electrical properties and outstanding performance
in composite materials,8–12 CNTs have proven to be highly bio-
compatible, a fact that has led them to be used for biomedical
applications.13,14 In particular, the use of CNTs in biomaterials
applied to bone is also expected to act as scaffolds to promote
and guide bone tissue regeneration as well as improve mechanical
properties.13,15,16 One example is the use of CNTs as mechanical
reinforcement of PMMA bone cements.17–20
The question as to which of these two nanofillers—graphene
or CNTs—is the better reinforcing agent in polymeric nano-
composites, in terms of mechanical properties, has been
a
Nanotechnology Research Division, Center for Mechanical Technology &
Automation, University of Aveiro, 3810-193 Aveiro, Portugal. E-mail:
paulam@ua.pt; Fax: +351234370953; Tel: +351234370830
b
CEMUC, Department of Mechanical Engineering, University of Coimbra,
Portugal
answer depends on many factors, such as the polymer, the
method of preparation of the nanofiller, and the nanofiller
content. Also, improvement of the physicochemical properties of
the nanocomposite depends on the distribution of nanofiller in
the polymer matrix, which, in turn, depends on the chemical
affinity between them.23
Since its discovery in 1960, PMMA bone cement remains one
of the most widely used biomedical materials, being employed to
anchor total joint replacements.24 One of the principal draw-
backs of this cement is that it is not bioactive and, hence,
provides no osseointegration with periprosthetic bone.17,25–27 To
improve this situation, cements containing a bioactive agent,
usually, hydroxyapatite (HA) (which is both osseoconductive
and osseoinductive28), have been prepared. To limit the potential
for adversely affecting the mechanical properties of the cement
when HA is employed, only low loadings (<10 vol./vol.%) have
been used.29,30 On the other hand, studies reveal that, in order for
a bioactive cement to display osseoconductivity after setting, the
bioactive filler loading should be >60 wt/wt% (
35 vol./vol.%).31
Thus, Singh et al.32 prepared a PMMA bone cement formulation
with 67 wt/wt% (42 vol./vol.%) of HA and 0.1 wt/wt% of CNTs
and showed that it was bioactive, possessed suitable mechanical
properties, and in vitro and in vivo tests showed accelerated cell
maturation.32,33
The aforementioned advantages of graphene that make it
more attractive than CNTs and the lack of a critical comparison
between these two nanofillers as the reinforcing agent in

This journal is ª The Royal Society of Chemistry 2012 Nanoscale, 2012, 4, 2937–2945 | 2937

a PMMA/high load HA bone cement were the motivation for the
present study, which had two primary purposes. The first was
to critically compare graphene oxide (GO) and functionalized
(f-CNTs) as the reinforcing agent PMMA/high-loading HA bone
cement in terms of a large array of physical and mechanical
properties of the cement. The second was to propose a mecha-
nism for the influence of these carbon nanostructures on the
properties determined. We selected graphene in its oxidized state,
that is, GO, because, in previous studies, it was shown that the
presence of oxygen functional groups at carbon nanostructure
surfaces simultaneously facilitates their dispersion in the polymer
matrix and reduces their toxicity.32–35 The combination of
nanofillers with in situ polymerization is considered an excellent
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E
method for producing truly synergetic composite materials with
carbon structures.36
Experimental section
Downloaded by University of Sussex on 14 January 2013
1. Materials
The PMMA bone cement used was a commercially available
brand, CMW1 (DePuy, Warsaw, IN, USA). For its composition,
see Table 1.
2. Synthesis of graphene oxide (GO)
The method used for the exfoliation of graphite was based on
that detailed in our previous work.37 Briefly, 2 grams of graphite
(99.99%; Sigma-Aldrich) was dispersed into a flask containing 50
mL of concentrated H2SO4 and 7 g of KMnO4 and the solution
was magnetically stirred for 2 h. After that, the solution was
treated with H2O2 until the gas evolution ceased and then the
resultant suspension was intensively washed, first with HCl
solution (0.1 mol dm3) and then with distilled water by filtration
and centrifugation until a pH of 7 was achieved before drying by
lyophilisation.
3. Functionalization of multiwalled carbon nanotubes
Multiwalled CNTs (purity > 95%; Nanocyl-3150; Belgium) were
chemically oxidised using a mixture of concentrated acids
H2SO4/HNO3 (3 : 1) for 24 h, with further details as given by Jia
et al.38 The resulting f-CNTs were intensively washed by centri-
fugation with distilled water until the pH of their aqueous
dispersion was neutral.
4. Preparation of a PMMA/high-load HA/nanofiller cement
powder mixture (solid component)
Three steps were used in the preparation of the cement powder
mixture (herein designed solid component) (Fig. 1). In the first
Table 1 Composition of CMW1 PMMA bone cement
Bone Cement Powder:
Polymethyl methacrylate (wt/wt%) 88.85
Benzoyl peroxide (BPO) (wt/wt%) 2.05
Barium sulfate (wt/wt%) 9.10
Bone Cement Liquid:
Methyl methacrylate (wt/wt%) 98.5
N,N-Dimethyl-p-toluidine (DMT) (wt/wt%) #1.50
Hydroquinone/ppm 75
2938 | Nanoscale, 2012, 4, 2937–2945
View Article Online
step, the nanofiller (GO or f-CNTs) was suspended in water and
tip-sonicated for 30 min by using a probe-tip sonicator (Sonics
Model VC 130, 20 kHz, probe tip (6 mm diameter), amplitude
80%, 2.2 kW). After that, the nanofiller, the cement powder, and
the HA (particle size 2–3 mm; purity > 98%; Agoramat-Advanced
Materials, Portugal) were thoroughly blended. In the second
step, a freeze-granulation technique (Power Pro Freeze-granu-
lator L5-2, Sweden) was used to increase the homogeneity of the
blended powder.32 The suspension was then dried by lyophili-
sation. After this final treatment, the nanocomposite was kept in
a desiccator.
Nine sets of solid component compositions were prepared:
PMMA/67 wt/wt% HA (unreinforced cement) herein referred to
as PMMA/HA and PMMA/HA reinforced by 0.01, 0.1, 0.5 and
1 wt/wt% of GO or f-CNTs.
5. Preparation of PMMA/high load HA/nanofiller bone cement
specimens
These specimens were fabricated by mixing a given solid
component with the liquid monomer of the PMMA bone cement
under ambient conditions (temperature: 22  1  C; relative
humidity: $40%). Hand mixing was performed in a suitable,
clean, dry mixing ceramic bowl when the liquid component was
added to the solid component. The dough was mixed carefully to
minimize the air entrapment, and then it was taken into gloved
hands and kneaded thoroughly according to the CMW1
instruction leaflet. The dough was then poured into poly(tetra-
fluorethylene) (Teflon) moulds designed according to each test,
and pressed at 9.8 MPa using a uniaxial press for 10 min. The
specimens were then stored under ambient laboratory conditions
for
24 h prior to testing.
6. Characterization of GO and f-CNTs
The morphology of each of these nanofillers was determined
using a high-resolution field-emission scanning electron micro-
scope (HRFE-SEM) (Hitachi SU-70; Japan) and a high resolu-
tion transmission electron microscope (HR-TEM) (JEOL 2200F;
Japan) operated at 200 kV. The TEM specimens were prepared
by depositing an aliquot of the solid component onto a carbon
grid and then letting the solvent evaporate.
The thickness and size of GO sheets deposited on SiO2
substrate were determined using an atomic force microscope
(AFM) (VEECO Multimode; USA).
The elemental composition and atomic concentration of
each constituent in each of the nanofillers were determined using
an X-ray photoelectron spectroscope (XPS) (VG Scientific
ESCALAB 200 A; UK).
7. Determination of cement properties
Dynamic mechanical analysis (DMA) (Tritec 2000; UK) tests
were performed on the specimens (20.0 mm  4.0 mm  2.2 mm)
in single cantilever bending mode, at a frequency of 1 Hz, an
amplitude of 0.010 mm and with a specimen heating rate of 2  C
min1 from room temperature (
22  C) to 150  C. The properties
determined were the storage modulus (E0 ), loss modulus (E00 )
and, hence, damping factor (tan d), as a function of specimen
temperature (T). The temperature corresponding to the peak in
This journal is ª The Royal Society of Chemistry 2012

Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E
Fig. 1 Schematic representation of the three steps used in preparing the solid component of the PMMA/HA/nanofiller bone cement. The morphology
Downloaded by University of Sussex on 14 January 2013
of the solid component is shown in the large SEM micrograph, while the morphology of the PMMA bead in the component, with nanofiller particles on
its surface, is shown in the inset.
the tan d vs. T plot is the glass transition temperature of the
cement (Tg). For each study group, 5 specimens were tested.
Four point bending tests were performed on the specimens
(25.0 mm  10.0 mm  3.3 mm) using a universal tensile tester
machine (AG-X; 5 kN load cell; Japan) controlled with
a commercially available computer software (Trapezium X;
Shimadzu; Japan), at a crosshead displacement rate of 5 mm
min1. For each study group, 5 specimens were tested. SEM
images were obtained from the fractured surfaces of the
specimens.
The modulus of elasticity (E) was determined using the
impulse excitation technique (ASTM C1259-96). The specimen
(25.0 mm  10.0 mm  3.3 mm) was set in free flexural vibration
by a light mechanical impulse. A strain gage was used to assess
the vibration. Fast Fourier transform was applied to analyse the
vibration signals to obtain the frequency of the first flexural
vibrational mode. E was calculated using the expression:
! !
mft2 l3
E ¼ 0:9465 T1 $
d t3
where m is the mass of the specimen, ft is the fundamental
frequency, d is the width of the specimen, l the length of the
specimen, t is the specimen thickness, and T1 is a correction
factor that takes into account the finite dimensions of the spec-
imen. For the calculation of T1, a constant Poisson ratio of 0.3
was used. For each study group, 5 specimens were tested.
The maximum exothermic temperature reached during the
polymerization stage (Tmax) was determined per ISO 5833, using
a Ni–Cr–Al k-type thermocouple and a commercially available
data acquisition system (PicoLog Data Acquisition Software;
Pico Tecknology Ltd., UK). For each study group, the tests were
run in triplicate.
Preparation of specimens (1.5 g each) for the nuclear magnetic
resonance (NMR) and size exclusion chromatography (SEC)
analysis needed a preliminary Soxhlet extraction of the polymer
from the specimen. This was carried out for 8 h using 150 mL of
dichloromethane. For the 1H NMR analysis, the specimen was
This journal is ª The Royal Society of Chemistry 2012
View Article Online
dissolved in deuterated chloroform and then the spectra were
acquired with a spectrometer (DRX 500, Bruker; Germany),
operated at 300.13 MHz. For the SEC analysis, 5 mg of polymer
fraction extracted from the specimen was dissolved in 1 mL of
dimethylacetamide (DMAc) solution, at 20  C, for 30 min and
then filtered through a 0.3 mm filter, yielding a sample concen-
tration of
0.5% (5 mg mL1). The columns, injector system, and
detector were each maintained at 70  C during the analysis. The
eluent (DMAc) was pumped at a flow rate of 0.9 mL min1. The
analytical columns were calibrated with polystyrene standards
(Polymer Laboratories; UK) in the range 1.7–100.0 kDa. The
injected volume was 100 mL. Average molecular weight (Mw),
number average molecular weight (Mn) and polydispersity
(PDI ¼ Mw/Mn) were determined from these analysis using
Cirrus software.
8. Preparation and characterization of cements with different
concentrations of radicalars
Four varieties of the nanocomposite cements that contained
0.1 wt/wt% f-CNTs loading were prepared, the difference being
in the additional concentration of the radicalars, namely, BPO
and DMT (RIC), that were mixed in the solid component and
liquid monomer during preparation of the cements (Table 2). For
each cement, the bending strength and E were determined
(in each case, sample size ¼ 5).
Also, using RIC ¼ 2X, nanocomposite cements that contained
0.5 wt/wt% of either GO or f-CNTs were prepared. For each
cement, bending strength, E, and Tg were determined (in each
case, sample size ¼ 5).
For each of the above described cements, the steps in the
preparation and the methods of determination of the properties
were the same as given in the previous sub-sections.
9. Statistical analysis
In the case of each of the mechanical properties and Tg, the
results were evaluated for statistical significance using a one-way
Nanoscale, 2012, 4, 2937–2945 | 2939
 View Article Online

Table 2 Additional quantities of BPO and DMT added to the solid

component (5 g) and the liquid component (1.8 mL), respectively

Radicalar agents

Concentration BPO (solid component) DMT (liquid component)

0 Initial Initial
1 40 mg (0.165 mmol) 26.25 mL (0.182 mmol)
2 80 mg (0.330 mmol) 52.5 mL (0.364 mmol)
3 120 mg (0.495 mmol) 78.75 mL (0.546 mmol)
10 400 mg (1.65 mmol) 262.5 mL (1.819 mmol)

(ANOVA) analysis of variance, with p < 0.05 being considered

Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E

significant. Post-hoc tests were conducted using Duncan

methods (IBM SPSS Statistics; USA).

Results and discussion

Downloaded by University of Sussex on 14 January 2013

1. Characteristics of the carbon nanostructures

The as-synthesized GO consisted of thin flakes, with a well-
defined few-layer structure (Fig. 2a and b). Note the wrinkled
surface, which could play an important role in enhancing
Fig. 3 Topographic view of contact-mode AFM scan of GO deposited
mechanical interlocking and load transfer to the matrix in
on SiO2 glass and height profile through the line.
a GO-reinforced bone cement. The f-CNTs were
1.5 mm long,
with tube diameter ranging from
30 nm to
50 nm (Fig. 2c and
d), and hollow in structure (Fig. 2c). The plate size distribution in Table 3 Results of XPS analysis of the C1s content of the surfaces of
GO ranged from
200 nm to
3 mm (Fig. 3) and the mean GO and f-CNTs
thickness of GO nanosheets was
1.2 nm, corresponding to
GO f-CNT
nanosheets with two or three layers.39
Two clear trends are seen in the XPS results (Table 3). First, Binding Atomic conc. Binding Atomic conc.
the oxidation level of the GO surface (48.57%) is markedly higher energy/eV (at%) energy/eV (at%)
than that of f-CNTs surface (13.96%). This difference is consis- C–C 285 15.67 285.4 12.89
tent with the differences in the methods used to prepare these C–O 286.9 42.55 286.4 6.57
nanofillers. Second, while the GO surface is mainly functional- C¼O 288.2 4.14 287.4 3.51
ized with epoxy groups, the f-CNTs surface presents a higher C(O)O 289.2 1.88 288.6 3.88
amount of carboxylic groups.

2. Characteristics of the PMMA/HA/nanofiller cements

Fig. 2 HRFSEM and HRTEM images of graphene oxide respectively
(a) and (b) and carboxyl functionalized MWCNTs respectively (c) and
(d).
2.1 Mechanical properties. The results for the mechanical
properties (Table 4 and Fig. 4) showed that, relative to the
unreinforced cement, (1) neither of the filler enhanced E0 ; (2) each
nanofiller led to a broadening of the tan d plot (Fig. 4); (3) while
GO enhanced bending strength, f-CNTs did not; and (4) GO
markedly enhanced E but f-CNTs did not.
The trends in the mechanical properties are consistent with the
morphologies of the GO sheets and the f-CNTs. The GO sheets
display a high specific area and wrinkled surface, contributing to
a better integration of the filler in cement matrix and high matrix
adhesion/interlocking. On the other hand, the cylindrical shape
of the f-CNTs does not contribute to identical interlocking with
the micron sized elements present in the solid component
(PMMA beads and HA). The aforementioned trends in
mechanical properties are also consistent with the morphologies
of the surfaces of fractured bending test specimens (Fig. 5).
Specifically, fewer and smaller pores were seen in the GO-rein-
forced cements, regardless of the loading (for example, Fig. 5a
and b), whereas a porous structure was observed in f-CNTs
reinforced cements, especially at high loadings (Fig. 5c and d). In

2940 | Nanoscale, 2012, 4, 2937–2945 This journal is ª The Royal Society of Chemistry 2012
 View Article Online

Table 4 Mechanical properties of the PMMA/HA nanocomposites prepared with different loadings of GO and f-CNTs

Nanofiller loading (wt/wt%)

Nanofiller Properties 0 0.01 0.1 0.5 1

0
GO E /GPa 2.67  0.31 a
2.49  0.23a
2.26  0.14 a
2.20  0.37 a
2.36  0.19a
Tg/ C 118.0  0.8 118.1  2.3a 98.2  0.4 98.0  0.9 98  1.2
Bending strength/MPa 20.00  1.46 23.01  0.37 22.34  0.74 24.89  1.19 28.28  1.09
E/GPa 6.09  0.42 9.53  0.46 9.84  0.47 9.63  0.50 10.22  0.67
Tmax/ C 82.4  1.3 71.3  1.4 64.7  0.9 56.3  1.1 38.6  2.1
f-CNTs E0 /GPa 2.67  0.31 1.66  0.23 2.13  0.34 1.35  0.21 0.99  0.27
Tg/ C 118.0  0.8 95.3  1.1 91.2  1.5 69.9  1.3 68.0  1.5
Bending strength/MPa 20.00  1.46 14.22  0.63 15.11  0.90 12.57  0.63 11.81  1.69
E/GPa 6.09  0.42a 6.89  0.68a 8.47  0.38a 6.93  0.65a 6.80  0.68a
Tmax/ C 82.4  1.3 61.9  1.2 53.8  0.7 28  1.1 22.4  1.4
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E

a
Indicates no statistically significant difference between control and reinforced cements (ANOVA at 0.05 level of significance); all the other values show
statistically significant difference between control and reinforced cement (ANOVA at 0.05 level of significance).
Downloaded by University of Sussex on 14 January 2013

Fig. 4 Summary of the tan delta results for PMMA/HA reinforced with different percentages of GO (a) and f-CNTs (b).

attributed to the release of the unreacted methyl methacrylate

(MMA) monomer (which is volatile) during the polymerization
of the cement. In other words, each of the nanofillers, but
particularly f-CNTs, captures the radicals produced during the
cement polymerization process, thereby hampering their normal
course.

2.2 Tg results. Besides the already mentioned broadening of

the tan d plot in the presence of both nanofillers (Fig. 4), f-CNTs
had a pronounced effect on the Tg values: Tg decreased
substantially with f-CNTs increase on the cement composition
(Fig. 4b). This is indicative of a decrease in the polymer chain size
and the formation of some oligomeric species.

2.3 Tmax results. The trend in the results, of a decrease of

Fig. 5 Morphologies of fracture surfaces of bending test specimens of
nanocomposite PMMA bone cements reinforced with 1 wt/wt% GO (a)
and (b) and 1 wt/wt% of f-CNTs (c) and (d).
each type of carbon nanostructure-reinforced cement, pores
occur in spite of the homogeneous distribution of the nanofiller
cement matrix (Fig. 5a and d). The origin of these pores may be
Tmax with the presence of either GO or f-CNTs (Table 3), is the
same as reported by Ormsby et al.40 for PMMA-CNT bone
cement. The decrease is more pronounced when f-CNTs were
used compared to when GO was used. These results may be
interpreted as evidence of the nanofiller acting as a radical
scavenger, with f-CNTs having a more pronounced scavenging
effect. The higher oxygen density of GO (Table 2) and the
presence of lattice defects (localized sp3 defects within the sp2

This journal is ª The Royal Society of Chemistry 2012 Nanoscale, 2012, 4, 2937–2945 | 2941
 View Article Online

analysis of their relative peaks in the spectra showed that the

former and latter end groups predominate in the case of the
polymer grown in the presence of GO and f-CNTs,
respectively.

2.5 SEC results. These results (Table 5) confirm that GO had

a less detrimental effect on cement polymerization compared
to f-CNTs. Specifically, the organic fraction extracted from the
GO-reinforced specimen displayed only one main peak whereas
two main peaks were displayed in the case of f-CNT-reinforced
cement specimen (corresponding to two main-sized fractions).
These results suggest a more pronounced detrimental effect of
f-CNTs on the MMA polymerization, with the appearance of an
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E

oligomeric fraction.

3. Influence of radicalar concentration on properties of

Fig. 6 Results of the 1H NMR analysis of PMMA with (a) GO and (b)
PMMA/HA/nanofiller bone cement
Downloaded by University of Sussex on 14 January 2013

f-CNTs.
When 0.1 wt/wt% f-CNTs were used as the reinforcing agent,
the highest value of each of the two mechanical properties
carbon network)6,41 lead to a lower density of p-bonds when determined for the cement (relative to both the unreinforced
compared with f-CNTs, thereby reducing GO’s scavenging cement and the reinforced cement when the liquid monomer in
action. CMW1 cement was used (that is, RIC ¼ 0X)) was obtained with
RIC ¼ 2X (Fig. 7).
2.4 1H NMR results. Two key trends are seen in these With RIC ¼ 2X, there were significant increases in both
results (Fig. 6). First, the tacticity is dependent on the nanofiller bending strength and E for cements in which GO was used as the
used; specifically, when the polymer was grown in the presence reinforcing agent (relative to the corresponding values for the
of GO, isotactic conformation increases significantly, whereas unreinforced cement), regardless of the loading of the nanofiller
heterotactic and syndiotactic conformations were dominant for (Table 6). Furthermore, for a cement reinforced with 0.5 wt/wt%
PMMA grown in the presence of f-CNTs. Further work is GO, Mw was 79 100 (PDI ¼ 1.38) compared to 47 700
needed to explain this finding. Second, there are two terminal
end groups, namely benzoyloxy and unsaturated, and careful
Table 6 Mechanical properties of PMMA/HA reinforced with different
loadings of GO and with RIC ¼ 2X
Table 5 Results of SEC analysis of the polymer fraction extracted from GO (wt/wt%) E0 /GPa Bending strength/MPa E/GPa
the nanocomposites prepared with 0.5 wt/wt% of GO and 0.5 wt/wt% of
f-CNT 0 2.67  0.31a 20.0  01.46 6.09  0.42
0.01 3.20  0.36a 21.17  0.37 12.57  0.46
Cement Peak 1 Peak 2 0.1 2.96  0.21a 31.45  0.74 11.52  0.52
0.5 3.15  0.43a 37.53  1.19 12.00  0.49
0.5GO Mw, 47 700 — 1.0 2.77  0.52a 29.50  1.90 12.24  0.67
Mn, 22 800 —
a
PDI, 2.09 — Indicates no statistically significant difference between control and
0.5CNT Mw, 40 400 Mw, 870 reinforced cements (ANOVA at the 0.05 level of significance); all the
Mn, 28 230 Mn, 570 other values show statistically significant difference between control
PDI, 1.43 PDI, 1.51 and reinforced cement (ANOVA at he 0.05 level of significance).

Fig. 7 Variation in bending strength (a) and modulus of elasticity (b) values obtained for the PMMA/HA matrix and for PMMA/HA reinforced with
0.1 wt/wt% of f-CNTs, with cements prepared using additional quantities of radicalar agents (RIC ¼ 0X, 1X, 2X, 3X and 10X).

2942 | Nanoscale, 2012, 4, 2937–2945 This journal is ª The Royal Society of Chemistry 2012

Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E
Downloaded by University of Sussex on 14 January 2013
Fig. 8 Schematic representation of a possible mechanism of the radical polymerization of PMMA under the influence of carbon nanostructures.
(PDI ¼ 2.09) for a cement with the same GO loading but
synthesized using RIC ¼ 0X (Table 5). This indicates a less broad
range of molecular weights in these samples showing that the
increase of RIC compensates for the radical scavenger activity of
the nanofillers.
4. Postulated mechanism
Carbon nanostructures, in particular carbon nanotubes, are
large arrays of conjugated double bonds; they are expected to
have large electron donor–acceptor capability. This specific
property makes them remarkably reactive toward free radi-
cals.42,43 We, therefore, postulate that both GO and f-CNTs are
active participants in the polymerization of the nanocomposites
cement (Fig. 8). In this scheme, we distinguish between two
different types of action assigned to carbon nanostructures that
can restrict the PMMA radicalar polymerization. The first is
polymerization retardation (partial loss of initiation activity). In
this case, the primary radicals formed after the split of BPO
molecules, when they contact the aromatic surface of carbon
nanostructures, lose their unpaired electron, giving rise to
non-reactive species. The second is polymerization inhibition
(chain termination). Here, during the growth of macromolec-
ular radicals, an electron transference may also occur to the
carbon aromatic surface, ending the polymer chain growth,
resulting in the formation of small-sized macromolecules. These
two types of actions may occur simultaneously and/or
competitively.
Jia W. et al.38 were probably the first workers to propose the
participation of CNTs in the MMA radical polymerization,
pointing out that CNTs could be attacked by AIBN, thereby
opening their p-bonds. These opened p-bonds of CNTs could
link with the PMMA, resulting in obstruction of the growth of
PMMA and, possibly, producing a C–C bond between the CNT
and the PMMA. A strong interface between the CNT and the
PMMA was thus claimed, without convincing evidence, to be
produced through that bonding.38 This explanation has been put
forward by other workers.18,19,36,40 In the present work, however,
This journal is ª The Royal Society of Chemistry 2012
View Article Online
since a different initiator was used, namely one that is activated
at room temperature, it is highly unlikely that the p-bonds of
carbon nanostructures could establish covalent bonds with the
polymer chains. Instead, the p-bonds are more likely to act as
scavengers of primary and macromolecular radicals, as discussed
above, because of the high electron affinity of carbon
nanostructures.44–46
We suggest two approaches to counter the negative
influence of a nanofiller on the polymerization of the cement
and, hence, its mechanical properties. The first involves
modification of the surface of the nanofiller with PMMA
chains via atom transfer radical polymerization, as proposed by
us in our previous study.47 In that work, we showed that when
this method was applied to GO-reinforced PMMA/HA, there
was good integration of GO in the matrix and enhanced
mechanical properties. One limitation of this approach,
however, is that it is difficult to scale up. The second approach
involves increasing the concentration of the radicalar species
during the polymerization, thereby attempting to compensate
for a subsequent loss due to a scavenging activity. Results
obtained in the present study (Fig. 7 and Table 6) show the
feasibility of the second approach, particularly in the case of
a GO-reinforced bone cement. These results are consistent with
an improvement in the polymerization, as reflected in the
increase of Mw.
5. Future work
Future work will be conducted on the determination of other key
properties of GO-reinforced PMMA bone cements prepared
with additional radical initiators, such as curing time, setting
time, fracture toughness, fatigue life, and fatigue crack propa-
gation rate. In addition, since the in vitro toxicity of graphene
towards human cells appears to be lower than that of CNTs and
does not hamper the proliferation of human mesenchymal stem
cells and their differentiation into bone cells,48–50 another future
study will be to determine the in vitro and in vivo biocompatibility
of GO-reinforced PMMA bone cements.
Nanoscale, 2012, 4, 2937–2945 | 2943

Conclusions
In the present study, we compared the influence of two carbon
nanostructures (GO and f-CNTs) when blended with the powder
of a PMMA/high-load HA bone cement on a large collection of
properties of the cements. In the case of GO-reinforced cements,
we found higher values of some of the mechanical properties
determined, compared to the corresponding values for the
unreinforced cement. This is attributed to GO’s high specific
area, high degree of surface functionalization, and wrinkled
surface, all of which lead to high adhesion/interlocking of the
nanofiller with the cement matrix. In the case of the other
mechanical properties determined, there is depreciation no
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E
matter which of the two nanofillers was used. We postulate that
this is a consequence of the fact that the carbon nanostructure
acts as a scavenger of the radicals that are produced during the
polymerization process, causing the inhibition and retardation of
the growth of polymer chains. This phenomenon is more
Downloaded by University of Sussex on 14 January 2013
pronounced when f-CNTs are used compared to when GO is
used because of the former’s higher density of p-bond areas.
Based on this explanation, we proposed that one approach that
may be taken to counter the negative impact of a carbon nano-
structure on mechanical properties of the cement is to substan-
tially increase the concentration of the radicalar initiators (RICs)
present in the cement during polymerization. We demonstrated
the feasibility of this approach for both GO- and f-CNT-rein-
forced cements. Taken as a whole, the present results should
contribute to the development of novel PMMA-based nano-
composite bone cements.
Acknowledgements
This work was co-financed by International Iberian Nanotech-
nology Laboratory (INL)—Foundation of Science and Tech-
nology (FCT—Portugal—FCOMP-01-0124-FEDER-008439).
P. Marques thanks the Ciencia 2007 Program and G. Gonclaves
thanks INL for a PhD grant. We thank Prof. Dmitry Evtyugin
(CICECO, University of Aveiro) for help with SEC analysis and
valuable discussions. We especially thank Prof. Alessandro
Gandini for valuable and fascinating discussions regarding
polymer chemistry.
References
1 K. S. Novoselov, A. K. Geim, S. V. Morozov, D. Jiang, Y. Zhang,
S. V. Dubonos, I. V. Grigorieva and A. A. Firsov, Science, 2004,
306, 666–669.
2 S. Stankovich, D. A. Dikin, G. H. B. Dommett, K. M. Kohlhaas,
E. J. Zimney, E. A. Stach, R. D. Piner, S. T. Nguyen and
R. S. Ruoff, Nature, 2006, 442, 282–286.
3 B. G. Choi, J. Hong, Y. C. Park, D. H. Jung, W. H. Hong,
P. T. Hammond and H. Park, ACS Nano, 2011, 5, 5167–5174.
4 Y. Li, Polymer, 2011, 52, 2310–2318.
5 L. L. Tian, P. Anilkumar, L. Cao, C. Y. Kong, M. J. Meziani,
H. J. Qian, L. M. Veca, T. J. Thorne, K. N. Tackett, T. Edwards
and Y. P. Sun, ACS Nano, 2011, 5, 3052–3058.
6 T. Kuilla, S. Bhadra, D. Yao, N. H. Kim, S. Bose and J. H. Lee, Prog.
Polym. Sci., 2010, 35, 1350–1375.
7 M. A. Rafiee, J. Rafiee, Z. Wang, H. Song, Z.-Z. Yu and N. Koratkar,
ACS Nano, 2009, 3, 3884–3890.
8 M. K. Yeh, T.-H. Hsieh and N.-H. Tai, Mater. Sci. Eng., A, 2008,
483–484, 289–292.
9 Y. Jia, K. Peng, X.-L. Gong and Z. Zhang, Int. J. Plast., 2011, 27,
1239–1251.
2944 | Nanoscale, 2012, 4, 2937–2945
View Article Online
10 M. Mazaheri, D. Mari, Z. R. Hesabi, R. Schaller and G. Fantozzi,
Compos. Sci. Technol., 2011, 71, 939–945.
11 P.-C. Ma, N. A. Siddiqui, G. Marom and J.-K. Kim, Composites, Part
A, 2010, 41, 1345–1367.
12 Z. Spitalsky, D. Tasis, K. Papagelis and C. Galiotis, Prog. Polym.
Sci., 2010, 35, 357–401.
13 K. Sahithi, M. Swetha, K. Ramasamy, N. Srinivasan and
N. Selvamurugan, Int. J. Biol. Macromol., 2010, 46, 281–283.
14 D. A. Gomez-Gualdr
on, J. C. Burgos, J. Yu and P. B. Balbuena,
Nanoparticles in Translational Science and Medicine, Academic
Press, 2011, vol. 104, pp. 175–245.
15 E. Hirata, M. Uo, H. Takita, T. Akasaka, F. Watari and
A. Yokoyama, Carbon, 2011, 49, 3284–3291.
16 T. R. Nayak, L. Jian, L. C. Phua, H. K. Ho, Y. P. Ren and
G. Pastorin, ACS Nano, 2010, 4, 7717–7725.
17 D. Renteria-Zamarron, D. A. Cortes-Hernandez, L. Bretado-Aragon
and W. Ortega-Lara, Mater. Des., 2009, 30, 3318–3324.
18 R. Ormsby, T. Mcnally, C. Mitchell, P. Halley, D. Martin,
T. Nicholson and N. Dunne, Carbon, 2011, 49, 2893–2904.
19 R. Ormsby, T. Mcnally, C. Mitchell and N. Dunne, J. Mater. Sci.:
Mater. Med., 2010, 21, 2287–2292.
20 B. Marrs, R. Andrews, T. Rantell and D. Pienkowski, J. Biomed.
Mater. Res., Part A, 2006, 77, 269–276.
21 T. Chatterjee, K. Yurekli, V. G. Hadjiev and R. Krishnamoorti, Adv.
Funct. Mater., 2005, 15, 1832.
22 J. Du, L. Zhao, Y. Zeng, L. Zhang, F. Li, P. Liu and C. Liu, Carbon,
2011, 49, 1094–1100.
23 J. M. Wernik and S. A. Meguid, Appl. Mech. Rev., 2010, 63,
050801.
24 M. R. S. Chandler, N. D. Watkins, A. Briscoe and A. M. New, Proc.
Inst. Mech. Eng., Part H, 2006, 220, 321–331.
25 T. W. Bauer and J. Schils, Skeletal Radiol., 1999, 28, 483–497.
26 R. L. Barrack, J. Arthroplasty, 2000, 15, 1036–1050.
27 P. E. Sinnett-Jones, M. Browne, A. J. Moffat, J. R. T. Jeffers,
N. Saffari, J. Y. Buffiere and I. Sinclair, J. Biomed. Mater. Res.,
Part A, 2009, 89, 1088–1097.
28 P. K. Stephenson, M. A. R. Freeman, P. A. Revell, J. Germain,
M. Tuke and C. J. Pirie, J. Arthroplasty, 1991, 6, 51–58.
29 M. J. Dalby, L. Di Silvio, E. J. Harper and W. Bonfield, Biomaterials,
2001, 22, 1739–1747.
30 M. J. Dalby, L. Di Silvio, E. J. Harper and W. Bonfield, Biomaterials,
2002, 23, 569–576.
31 T. Tsukeoka, M. Suzuki, C. Ohtsuki, A. Sugino, Y. Tsuneizumi,
J. Miyagi, K. Kuramoto and H. Moriya, Biomaterials, 2006, 27,
3897–3903.
32 M. K. Singh, T. Shokuhfar, J. J. D. A. Gracio, A. C. M. De Sousa,
J. M. D. F. Fereira, H. Garmestani and S. Ahzi, Adv. Funct.
Mater., 2008, 18, 694–700.
33 M. K. Singh, J. Gracio, P. Leduc, P. P. Goncalves,
P. A. A. P. Marques, G. Goncalves, F. Marques, V. S. Silva,
F. Capela, E. Silva, J. Reis, J. Potes and A. Sousa, Nanoscale, 2010,
2, 2855–2863.
34 S. K. Smart, A. I. Cassady, G. Q. Lu and D. J. Martin, Carbon, 2006,
44, 1034–1047.
35 H. Hu, Y. Ni, V. Montana, R. C. Haddon and V. Parpura, Nano
Lett., 2004, 4, 507–511.
36 C. Velasco-Santos, A. L. Mart
ınez-Hern
andez, F. T. Fisher, R. Ruoff
and V. M. Casta~ no, Chem. Mater., 2003, 15, 4470–4475.
37 G. Goncalves, P. A. A. P. Marques, C. M. Granadeiro,
H. I. S. Nogueira, M. K. Singh and J. Gr
acio, Chem. Mater., 2009,
21, 4796–4802.
38 Z. Jia, Z. Wang, C. Xu, J. Liang, B. Wei, D. Wu and S. Zhu, Mater.
Sci. Eng., A, 1999, 271, 395–400.
39 H. C. Schniepp, J.-L. Li, M. J. Mcallister, H. Sai, M. Herrera-Alonso,
D. H. Adamson, R. K. Prud’homme, R. Car, D. A. Saville and
I. A. Aksay, J. Phys. Chem. B, 2006, 110, 8535–8539.
40 R. Ormsby, T. Mcnally, C. Mitchell and N. Dunne, J. Mech. Behav.
Biomed. Mater., 2010, 3, 136–145.
41 C. Soldano, A. Mahmood and E. Dujardin, Carbon, 2010, 48, 2127–
2150.
42 A. Galano, Nanoscale, 2010, 2, 373–380.
43 M. Francisco-Marquez, A. Galano and A. Mart
ıNez, J. Phys. Chem.
C, 2010, 114, 6363–6370.
44 P. J. Krusic, E. Wasserman, P. N. Keizer, J. R. Morton and
K. F. Preston, Science, 1991, 254, 1183–1185.
This journal is ª The Royal Society of Chemistry 2012

45 P. C. P. Watts, P. K. Fearon, W. K. Hsu, N. C. Billingham,
H. W. Kroto and D. R. M. Walton, J. Mater. Chem., 2003, 13,
491–495.
46 I. Fenoglio, M. Tomatis, D. Lison, J. Muller, A. Fonseca, J. B. Nagy
and B. Fubini, Free Radical Biol. Med., 2006, 40, 1227–
1233.
47 G. Goncalves, P. Marques, A. Barros-Timmons, I. Bdkin,
M. K. Singh, N. Emami and J. Gracio, J. Mater. Chem., 2010, 20,
9927–9934.
Published on 29 February 2012 on http://pubs.rsc.org | doi:10.1039/C2NR30303E
Downloaded by University of Sussex on 14 January 2013
This journal is ª The Royal Society of Chemistry 2012
View Article Online
48 Z. M. Markovic, L. M. Harhaji-Trajkovic, B. M. Todorovic-
Markovic, D. P. Kepic, K. M. Arsikin, S. P. Jovanovic,
A. C. Pantovic, M. D. Dramicanin and V. S. Trajkovic,
Biomaterials, 2011, 32, 1121–1129.
49 Y. Zhang, S. F. Ali, E. Dervishi, Y. Xu, Z. Li, D. Casciano and
A. S. Biris, ACS Nano, 2010, 4, 3181–3186.
50 T. R. Nayak, H. Andersen, V. S. Makam, C. Khaw, S. Bae, X. F. Xu,
P. L. R. Ee, J. H. Ahn, B. H. Hong, G. Pastorin and B. Ozyilmaz,
ACS Nano, 2011, 5, 4670–4678.
Nanoscale, 2012, 4, 2937–2945 | 2945