Scripta Materialia 54 (2006) 131–135

www.actamat-journals.com

Synthesis and characterization of hydroxyapatite using

carbon nanotubes as a nano-matrix
Santosh Aryal a, K.C. Remant Bahadur a, N. Dharmaraj b,c
,
Kawn-Woo Kim a, Hak Yong Kim b,*
a
Department of Bionanosystem Engineering, Chonbuk National University, Chonju 561-756, Republic of Korea
b
Department of Textile Engineering, Chonbuk National University, 664-14 Duckjin-dong, Chonju 561-756, Republic of Korea
c
Department of Chemistry, Government Arts College, Udumalpet 642 126, India

Received 5 August 2005; received in revised form 13 September 2005; accepted 27 September 2005
Available online 26 October 2005

Abstract

Multilayered hydroxyapatite (HA) crystals were chemically synthesized by using carboxyl functionalized multi-walled carbon nano-
tubes (MWNTs) as a matrix and characterized by various physico-chemical techniques. The results showed that MWNTs functionalized
with –COOH group form an efficient matrix for the growth of HA crystals.
Ó 2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Keywords: Biomaterials; Biomineralization; Carbon nanotubes; Calcium compounds; Nanomaterials

1. Introduction resources. However, crystallographic properties of HA

Hydroxyapatite (HA) [Ca10(PO4)6(OH)2], the prime
constituent of teeth and bone minerals, has attracted a
great deal of attention because of its involvement in the
biological calcification process of teeth and bone formation
[1–3]. The development of novel methods for its synthesis
has been the focus of a lot of recent work. Earlier work
mainly focused on controlling the stoichiometry of the
products, whereas with the development of nanotechnolo-
gies, considerable efforts are now focused on controlling
morphology and size also [3–6]. It has been shown that
many clinical capabilities of HA mainly depends on mor-
phology and size of HA particles [7]. Therefore, we believe
that the synthesis of nanoscale HA with suitable morpho-
logy will facilitate its improved clinical application.
HA with characteristic features of a biomaterial, in
particular, a crystallographic similarity with natural bone
minerals, can be derived from both synthetic and natural
*
Corresponding author. Tel.: +82 63 270 2351; fax: +82 63 270 2348.
E-mail address: khy@moak.chonbuk.ac.kr (H.Y. Kim).
prepared from various resources differ widely due to the
synthetic procedures and key ingredients used [8], which
in turn directly affects its bioactivity. Hence, the synthesis
of phase pure HA with controlled morphology is highly
desirable. There is a resurgence of interest in controlling
such crystal nucleation and growth for making composite
materials analogous to those produced by nature [9–11].
Most of the earlier efforts on the biomineralization of bone
involved the deposition of calcium phosphate crystals on a
matrix rich in negatively charged groups, particularly
acidic proteins or macromolecules [12–17], which are
believed to control the nucleation, polymorphism, and
growth of the crystals [18–22]. However, these macromole-
cules do not provide significant control over crystal
morphology. Recently, Oh et al. [23] and Rautary et al.
[24] have demonstrated the use of a nano-matrix for the
synthesis of HA with controlled morphology.
Carbon nanotubes (CNTs) are novel molecular scale
wires which exhibit useful properties for various potential
applications, including in miniature biological devices.
Also, the electrical properties of CNTs are sensitive to
surface charge transfer and changes in the surrounding

1359-6462/$ - see front matter Ó 2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.scriptamat.2005.09.050
132 S. Aryal et al. / Scripta Materialia 54 (2006) 131–135
electrostatic environment, undergoing drastic changes by
simple adsorptions of certain molecules or polymers [25–
28]. The growth of plate-like HA crystals was achieved
by using carboxylate ions of nano gold surface bound
aspartic acid molecules for binding Ca2+ ions, followed
by reaction with (NH4)2HPO4 [24]. Based on the general
concept of using a nano-matrix as a template for the
growth of inorganic crystals and the specific potential of
carboxylate ions to bind Ca2+ for the growth of HA, we
report herein the synthesis of HA with controlled morpho-
logy by employing –COOH functionalized multi-walled
carbon nanotubes (MWNTs) as a matrix.
2. Experimental section
2.1. Material and instrumentation
MWNTs were purchased from CNT Co. Ltd. Yeonsu-
Cu, Incheon, South Korea and were purified by washing
with concentrated hydrochloric acid and nitric acid, respec-
tively, to remove the amorphous carbon particles. All other
reagents used in this experiment were purchased from
Aldrich and used as received. Fourier transform-infrared
(FT-IR) spectra of the samples were recorded on a Bio-
Rad Win spectrometer in KBr pellets and powder X-ray
diffraction (XRD) patterns of the crystals were obtained
from a Shimadzu Lad-X 600 X-ray diffractometer using
Cu Ka radiation. Morphology and elemental analysis of
the HA crystals have been studied by a JEOL GSM scan-
Scheme 1. Schematic representation of the growth of hydroxyapatite on multi-walled carbon nanotubes functionalized with –COOH.
ning electron microscope (SEM) equipped with energy
dispersive X-ray (EDX) analysis. Transmission electron
microscopy (TEM) images were taken using a JEOL JEN
2010 operated at 200 kV accelerating voltage.
2.2. Synthesis of Hydroxyapatite on MWNTs
nano-matrix (MWNTs–HA)
Carboxyl group functionalized MWNTs (MWNTs–
COOH) have been synthesized by refluxing MWNTs with
nitric acid. In a typical experiment, 1 g of MWNTs with
60 ml of 60% nitric acid were sonicated for 20 min in a
round bottom flask to get a homogeneous mixture and
then refluxed for 18 h at 110 °C. Then the mixture was
filtered under vacuum through 0.22 lm millipore polycar-
bonate membrane and washed with distilled water until
the pH of the filtrate became 7. The resulting MWNTs–
COOH was dried under vacuum for 30 h at 50 °C.
MWNTs–COOH thus obtained was used for the synthesis
of HA crystal. MWNTs–COOH (200 mg) was reacted with
5 ml of 0.01 M aqueous solution of calcium chloride and
stirred for 1 h. To this mixture 5 ml of 0.01 M aqueous
solution of disodium hydrogen phosphate was added drop-
wise with continuous stirring. After 1 h, the colour of the
reaction mixture had changed from black to gray indicat-
ing the precipitation of HA. The product formed was
filtered and washed several times with distilled water to
remove any unreacted reagents and by products. Finally
the product was dried in air at room temperature for
 S. Aryal et al. / Scripta Materialia 54 (2006) 131–135
10 h and then under vacuum for 24 h. A possible mecha-
nism for the formation of HA crystals is furnished in
Scheme 1.
3. Results and discussion
HA crystals synthesized by reacting Na2HPO4 with
Ca2+ ions bound to carboxylate ions of MWNTs–COOH
matrix were found to be nanocrystalline in nature with a
multilayered array over the surface of the carbon nano-
tubes. The structure, crystalline phase, and morphology
of the synthesized HA have been investigated by using
various analytical and electron microscopic methods.
The formation of HA using carboxylate functionalized
MWNTs was identified by analyzing its FT-IR spectrum
(Fig. 1). The spectrum showed the m3 and m4 stretching
modes of phosphate group at 1100 and 960 cm 1, respec-
tively. In addition, a band due to excitation of the m4 mode
of PO34 group was observed at 600 cm 1. The absorptions
corresponding to the hydroxyl (–OH) group of HA and the
carboxylate ion of MWNTs were found at 3500 and
1500 cm 1, respectively. These bands and their positions
in the FT-IR spectrum indicated the formation of HA on
the MWNTs matrix [29].
The evolution of HA crystalline phase was studied by
X-ray diffraction (XRD) patterns and given in Fig. 2. It
is seen from this figure that diffraction peaks corresponding
to the HA crystalline phase were found at 2 h values 23, 26,
32, 49, and 53°, which are indexed to the (1 1 1), (0 0 2),
(1 1 2), (2 1 3), and (0 0 4) planes, respectively. The XRD
did not show any peaks corresponding to Ca3(PO4)2 [22],
suggesting that the reaction resulted in the formation of
phase pure HA. The presence of an intense diffraction peak
at the 2 h value of 26° corresponding to (0 0 2) plane
revealed the growth of HA crystals along the c-axis
[8,30,31]. The broad nature of the diffraction peaks which
are seen in Fig. 2 suggests that the HA crystals are very
Fig. 1. FT-IR spectra of carboxyl functionalized MWNTs and MWNTs–
HA.
133
Fig. 2. XRD pattern of hydroxyapatite.
small in size. These XRD results support the conclusion
that HA was formed.
The surface morphology and the elemental composition
of HA synthesized using carboxyl functionalized MWNTs
were studied by SEM equipped with an EDX analyzer
and presented in Figs. 3 and 4. The SEM image at low re-
solution showed unique multilayered morphology resem-
bling that of natural rock formed by biomineralization
(Fig. 3(A)). The same sample showed an interesting stack-
like morphology with a uniform stack width at higher mag-
nification (Fig. 3(B) and (C)). This multilayered plate-like
morphology may be due to the growth of HA crystals in
between the bundles of MWNTs. As we know, the presence
of carboxylate ions capable of binding with Ca2+ ions on
the surface of MWNTs through chelation is mainly respon-
sible for the controlled growth of HA upon the addition of
disodium hydrogen phosphate. Earlier reports also con-
firmed the morphology of HA grown by different routes
[8,23–28]. Further proof regarding elemental composition
was furnished by the EDX spectrum (Fig. 4). The ratio of
Ca/P was found to be 1.58, which is close to that of the ideal
stoichiometry of HA and better than that reported else-
where [32].
TEM images (Fig. 5) of HA crystals were taken by dip-
ping a copper grid in a dispersion of HA in water. From
these images, it was found that multilayered plate-like
HA crystals were formed over the surface of carboxyl func-
tionalized MWNTs. Selected area electron diffraction
(SAED) patterns of the HA crystals showed that the
(0 0 2) plane is aligned with the crystalsÕ axis, indicating that
the crystal growth is along the c-axis direction, which is
consistent with the XRD data discussed above.
4. Conclusion
In this study, the synthesis of multilayered HA crystals
was achieved by using carboxyl functionalized MWNTs.
The structure of HA was confirmed by identifying the
134 S. Aryal et al. / Scripta Materialia 54 (2006) 131–135

Fig. 4. EDX profile of HA grown on MWNTs nano-matrix.

Fig. 3. SEM images recorded from MWNTs–HA at different magni-

fications.

peaks characteristic of the phosphate group in the FT-IR

spectra. XRD studies confirmed its crystalline structure.
The appearance of multilayered plate-like assemblies of
HA crystals with uniform plate width in SEM and TEM Fig. 5. TEM images of MWNTs–HA crystal taken at different magnifi-
images, support the suggestion that there is formation of cations. Inset in (B) shows the SAED pattern of the crystal.
 S. Aryal et al. / Scripta Materialia 54 (2006) 131–135
HA on MWNTs with a stoichiometric value that complies
with natural HA. From our results, we believe that the car-
boxyl functionalized MWNTs could serve as an efficient
nano-matrix for the growth of HA crystals.
Acknowledgements
This research was supported by the Korean Ministry of
Education and Human Resource Development through the
Center for Healthcare Technology Development, Chonbuk
National University, Jeonju 561-756, Republic of Korea.
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