CRYSTAL

GROWTH
Well-Formed One-Dimensional Hydroxyapatite Crystals Grown by & DESIGN
an Environmentally Friendly Flux Method
2009
Katsuya Teshima,*,† SunHyung Lee,‡ Mitsuo Sakurai,† Yoshitaka Kameno,† VOL. 9, NO. 6
Kunio Yubuta,§ Takaomi Suzuki,†,| Toetsu Shishido,§ Morinobu Endo,|,⊥ and Shuji Oishi†,| 2937–2940
Department of EnVironmental Science and Technology, Faculty of Engineering, Shinshu UniVersity,
Nagano 380-8553, Japan, Faculty of Engineering, Shinshu UniVersity, Nagano 380-8553, Japan,
Institute for Materials Research, Tohoku UniVersity, Sendai 980-8577, Japan, Institute of Carbon
Science and Technology, Shinshu UniVersity, Nagano 380-8553, Japan, and Department of Electrical
and Electronic Engineering, Faculty of Engineering, Shinshu UniVersity, Nagano 380-8553, Japan

ReceiVed February 10, 2009; ReVised Manuscript ReceiVed April 13, 2009

ABSTRACT: Highly crystalline, well-developed hydroxyapatite (HAp) crystals were grown successfully for the first time by a
flux cooling method at a temperature less than 500 °C under air pressure. Environmentally friendly growth was induced by heating
a mixture of solute [Ca(NO3)2 · 4H2O, (NH4)2HPO4, and KOH] and flux (KNO3 and LiNO3) at 400-500 °C and holding at this
temperature for 10 h. After that, the mixture was cooled at various rates of 5, 200, and >1000 °C · min-1 (water quenching). The
obtained HAp crystals, which had average sizes of up to 18.0 × 2.1 µm and aspect ratios of up to 8.6, were colorless and transparent.
They had hexagonal prismatic one-dimensional (1-D) forms with pyramidal or truncated pyramidal end faces. The 1-D crystal
forms and sizes were obviously dependent on the growth conditions, such as the holding temperature and the cooling rate of the
high-temperature solution. High-resolution transmission electron microscopy (HR-TEM) images indicated that the grown HAp crystals
were of a very good crystallinity. From the HR-TEM and X-ray diffraction, the 1-D HAp crystals, which were hexagonal cylinders
({101j0} faces), elongated in the 〈0001〉 directions.

Introduction NaCl flux.20,21 The forms of grown crystals were a hexagonal

prism and needle (whisker) with pyramidal end faces. The CAp
Natural calcium apatites [Ca5(F,OH,Cl)(PO4)3] exhibit large
films, which were colorless and transparent, had relatively
variations in F, OH, and Cl contents and extensive substitution
smooth surfaces. Additionally, barium chlorapatite [Ba5Cl(PO4)3]
with the incorporation of Na, K, Co, Cu, Zn, and so on. The
and strontium chlorapatite [Sr5Cl(PO4)3] have also been grown
stoichiometric compositions such as fluorapatite [Ca5F(PO4)3,
by the NaCl flux method.22,23 Furthermore, FAp crystals have
FAp], hydroxyapatite [Ca5(OH)(PO4)3, HAp], and chlorapatite
been grown by the cooling of a CaF2 or KF flux.24,25 However,
[Ca5Cl(PO4)3, CAp] are uncommon in nature. Calcium apatites,
no study has been reported on the fabrication of highly
which are highly biocompatible, are well-known as the main
crystalline, well-developed HAp crystals by flux method. The
component of bones and teeth in the human body, and are also
purpose of this study, therefore, is the environmentally friendly
used as artificial biomaterials.1-3 Among these apatites, HAp
synthesis of highly crystalline, well-developed HAp crystals by
has received much attention as a material for artificial bones,
the cooling of a nitrate system flux. The flux growth is
scaffolds for tissue engineering, protein chromatography ap-
particularly preferred because it readily allows crystal growth
plications, and so on.4-7 In this context, HAp is considered as
at a temperature well below the melting point of the solute.
a model compound to study biomineralization phenomena.8-10
Other important advantages of flux growth are that the grown
Extra pure, well-developed HAp crystals are, therefore, of great
crystals have an idiomorphic or enhedral habit and a reasonably
interest for use in detailed medical applications and surgery.
lower degree of dislocation density. For the flux method, the
The crystals of Ca5(OH)(PO4)3 belong to the hexagonal system
most important thing is to choose a proper flux (solvent). In
with space group P63/m.11 Hydroxyapatite has a melting point
this study, KNO3-LiNO3 was chosen as a flux to grow crystals
of 1614 °C and lattice parameters a ) 0.942 nm and c ) 0.688
of Ca5(OH)(PO4)3 because it has a very low eutectic temperature
nm.11
(about 125 °C).
There have been many studies on the preparation of HAp
crystals and films by various techniques, such as solid-state
Experimental Section
reaction, hydrothermal growth, plasma techniques, pulsed laser
deposition, sol-gel crystallization, wet processes, and biomi- Hydroxyapatite crystals were grown by a cooling method of the
metic processes.5,6,10,12-19 There are, however, several disad- KNO3-LiNO3 flux. Reagent-grade Ca(NO3)2 · 4H2O, (NH4)2HPO4,
vantages in these techniques, such as expensive equipment, high KOH, KNO3, and LiNO3 (Wako Pure Chemical Industries, Ltd.) were
used for the growth of HAp crystals. A mixture of Ca(NO3)2 · 4H2O
total costs, high environmental loads, and poor crystallinity.
(2.557 g), (NH4)2HPO4 (0.858 g), and KOH (0.125 g) powders were
In our previous study, CAp crystals and their films have been, used as a solute. To be accurate, KOH is not a constituent of HAp;
respectively, grown by the cooling and the evaporation of the however, we define a solute as the mixture in this study. In addition,
a mixture of KNO3 (13.002 g) and LiNO3 (5.911 g) was chosen as the
* Corresponding author. E-mail: teshima@gipwc.shinshu-u.ac.jp. flux. The eutectic temperature of flux was approximately 125 °C. The
†
Department of Environmental Science and Technology, Faculty of Engineer- solute and flux powders were weighed out, mixed together, and put
ing, Shinshu University. into a platinum crucible (capacity: 30 cm3). The crucibles were heated
‡
Faculty of Engineering, Shinshu University. to 400-500 °C at rates of 15 °C · min-1 or 45 °C · h-1 and held at this
§
Tohoku University.
|
Institute of Carbon Science and Technology, Shinshu University. temperature for 10 h. After that, they were cooled to 100 °C at a rate
⊥
Department of Electrical and Electronic Engineering, Faculty of Engineering, of 5, 200, or >1000 °C · min-1 (water quenching). The cooling rates of
Shinshu University. 5 and 200 °C · min-1 were controlled by the cooling program, and the

10.1021/cg900159j CCC: $40.75  2009 American Chemical Society

Published on Web 05/14/2009
2938 Crystal Growth & Design, Vol. 9, No. 6, 2009 Teshima et al.

Figure 1. SEM images showing typical 1-D HAp crystals grown at the cooling rate of (a) and (b) 5 °C · min-1 and (c) and (d) 200 °C · min-1. The
holding temperature was fixed at 500 °C. (a) and (c) Low magnification and (b) and (d) high magnification, top of the crystals.

crucibles were allowed to cool to room temperature in the furnace.

The cooling rate of >1000 °C · min-1 was not able to be attained by the
cooling program in the furnace. For >1000 °C · min-1 (quenching in
water), the crucibles were held at the holding temperature of 500 °C
for 10 h, and then removed from the furnace and immersed into water
and cooled extremely rapidly. This cooling rate, >1000 °C · min-1, is
an approximate estimated value. The crystal products were separated
from the remaining flux in warm water. The obtained crystals were
observed by use of an optical microscope and field emission scanning
electron microscope (FE-SEM, JEOL, JSM-7000F). Phases and elon-
gated directions of the crystals were studied by X-ray diffraction (XRD,
SHIMADZU, XRD-6000). The chemical compositions of the 1-D
grown crystals were studied by means of Fourier-transform infrared
spectroscopy (FTIR, JASCO, FT/IR-6100). The high-resolution trans-
mission electron microscopy (HRTEM) and electron diffraction ob-
servations were carried out on JEM-2010 (JEOL) and JEM-2000EXII
(JEOL) instruments operated at 200 kV to analyze the crystallinity and
elongated direction of the grown crystals. The presence of impurities Figure 2. SEM image showing typical 1-D HAp crystals grown at a
from the KNO3-LiNO3 flux and platinum crucible was also observed. holding temperature of 500 °C and a cooling rate of >1000 °C · min-1
The length (L, parallel to the 〈0001〉 directions) and width (water quenching).
(W, perpendicular to the 〈0001〉 directions) of relatively large HAp
crystals (200 samples) were measured, and their average sizes (Lav and
Wav) and aspect ratios (Lav/Wav) were calculated for each growth run.
faces were rarely observed in Figure 1b,d. From the SEM
images, it was clearly found that the number of HAp crystals,
which had pyramidal or truncated pyramidal end faces, grown
Results and Discussion
at the cooling rate of 5 °C · min-1 was much larger than that of
Idiomorphic one-dimensional (1-D; needle- and prism-shaped) 200 °C · min-1. Figure 2 shows a SEM image of HAp crystals
HAp crystals with Lav of 18.0 µm and Wav of 2.1 µm were grown at water quenching (the cooling rate g 1000 °C · min-1).
successfully grown for the first time by the cooling of It seems that the widths of the grown crystals became small
KNO3-LiNO3 flux from all growth runs used in this study. The although the lengths were almost the same. This tendency is
grown 1-D HAp crystals were colorless and transparent. Figure the same as our previous study for the Na2Ti6O13 whisker
1 shows SEM images of HAp crystals grown at a cooling rate growth.26,27 (The columnar structure of HAp resembles the
of 5 (Figure 1a,b) and 200 °C · min-1 (Figures 1c, d). With both tunnel structure of Na2Ti6O13.) In order to examine the effect
cooling rates, well-developed HAp crystals were grown from of thermal condition on crystal averages (Lav and Wav), growth
the KNO3-LiNO3 mixed flux. The tendency of aspect ratios experiments were carried out at various holding temperatures
(or crystal sizes) of the grown crystals to increase with an and cooling rates. The values of Lav and Wav are presented in
increasing cooling rate is clearly observed in Figure 1a,c. It Table 1. When the holding temperature and cooling rate were,
seems that the widths of the grown crystals became small respectively, 500 °C and 5 °C · min-1, the aspect ratio was about
although the lengths are almost the same. The basic form of 7.2 (Lav ) 14.4 µm and Wav ) 2.0 µm). When the holding
the grown crystals was a hexagonal cylinder. Hexagonal temperature was fixed at 500 °C, the aspect ratio gradually
cylindrical crystals with pyramidal or truncated pyramidal end increased with increasing cooling rate, reaching about 12.1
Well-Formed 1-D Hydroxyapatite Crystals Crystal Growth & Design, Vol. 9, No. 6, 2009 2939

Table 1. Average Sizes of 1-D HAp Crystals Grown at Various

Thermal Conditions
run heating rate/ holding cooling rate/ average average average
no. °C · min-1 temp/°C °C · min-1 length/µm width/µm aspect ratio
1 15 500 5 10.8 1 10.8
2 15 500 200 11.4 1 11.4
3 0.75 400 5 14 1.3 10.8
4 0.75 500 5 14.4 2 7.2
5 0.75 500 200 18 2.1 8.6
6 0.75 500 >1000 17 1.4 12.1

(water quenching; Lav ) 17.0 µm and Wav ) 1.4 µm). On the

other hand, when the cooling rate was fixed at 5 °C · min-1, the
aspect ratio drastically increased with a decrease in holding
temperature (400 °C; Lav/Wav ) 14.0 µm/1.3 µm ) 10.8). As
mentioned above (Figure 2), the aspect ratios were significantly
dependent on the widths of the grown HAp crystals. Hydroxya-
patite readily produces anisotropic growth for its unique crystal Figure 4. FTIR spectra obtained from the 1-D HAp crystals grown at
a cooling rate of (a) 5 °C · min-1 and (b) 200 °C · min-1. The holding
structure, that is, columnar structure. Because it has different temperature was fixed at 500 °C.
crystallographic planes ({101j0} and {0001} planes), which have
different surface energies, polarities, and chemical activities.
In the case of Ba5Cl(PO4)3 crystals grown by the flux method,
the surface tension of the {101j0} planes was smaller than that
of the {101j1} planes. This result indicates that the {101j0} planes
are more stable than the {101j1} planes and the crystals elongate
predominantly in the 〈0001〉 directions.
Figure 3 shows XRD profiles for the transparent, colorless
whiskers (Figure 3a), pulverized crystallites (Figure 3b) at a
cooling rate of 5 °C · min-1 and Ca5(OH)(PO4)3 (hydroxyapatite)
ICDD PDF (Figure 3c). Characteristic four diffraction lines
corresponding to Ca5(OH)(PO4)3 between 31.5 and 34.5° were
clearly observed in Figure 3b. In addition, the XRD pattern of
the grown crystals in this study indicated good crystallinity. The
grown crystals were finally identified as Ca5(OH)(PO4)3 by their
XRD pattern (Figure 3b), using data given in the literature
(Figure 3c).11 Figure 4 shows FTIR spectra obtained from the
grown HAp crystals at a cooling rate of 5 (Figure 4a) and 200
°C · min-1 (Figure 4b). Absorption bands at approximately 3570
and 635 cm-1 are, respectively, assigned to stretching mode
(νs) and vibrational mode (νL) of the hydroxyl group, OH.13,17,28
The broad absorption band having the center wavenumber of
3400 cm-1 originates from adsorbed water molecules. It decides
the absorbed water when crystals were separated from flux in
warm water. Absorption bands at about 1090, 1060, 1025, 960,
605, and 560 cm-1 are assigned to vibrations of the phosphate

Figure 5. (a) TEM micrograph, (b) diffraction pattern, and (c) lattice
image of a typical 1-D HAp crystal (holding temperature ) 500 °C
and cooling rate ) 5 °C · min-1).

group, PO43-. Therefore, it was confirmed that the HAp

including OH group could be grown from the temperature of
500 °C.13,17,28
The HAp crystals grown from the KNO3-LiNO3 mixed flux
had well-developed faces. Typical 1-D HAp crystal tops are
shown in Figure 1b,d. The orientated 1-D crystals were
investigated by the XRD measurement to determine the Miller
indices of the crystal faces (Figure 3a). In the diffraction pattern,
the diffraction intensity of the (101j0) and (303j0) planes was
Figure 3. X-ray diffraction patterns (Cu KR) of 1-D HAp crystals. (a)
1-D crystals of well-developed faces were laid in parallel with the holder
plate, (b) pulverized crystallites, and (c) HAp ICDD PDF.11 The 1-D predominant. The indices of side faces were {101j0} and were
HAp crystals were grown at a holding temperature of 500 °C and a a result similar to our previous studies of chlorapatite and
cooling rate of 5 °C · min-1. fluorapatite.23,25 Figure 5a,b show a bright field TEM image of
2940 Crystal Growth & Design, Vol. 9, No. 6, 2009 Teshima et al.

a typical hexagonal cylinder and a selected area electron crystalline, well-formed 1-D HAp crystals. The nitrate system
diffraction pattern (SAED) (heating rate ) 15 °C · min-1, holding flux KNO3-LiNO3 is adequate for an environmentally friendly
temperature ) 500 °C, cooling rate ) 5 °C · min-1). The bright growth of HAp crystals. Since the nitrate system flux has very
field TEM image showed the grown crystal had a well-formed, low eutectic temperatures, it can be applied to grow high-quality,
truncated cylindrical shape (Figure 5a). The diffraction spots functional oxide crystals under much lower temperature
in the SAED pattern were confirmed to be in accordance with conditions.
an apatite structure. As clearly seen in Figures 3a and 5a,b, the
elongated direction clearly corresponded to the 〈0001〉 directions. Acknowledgment. This research was partially supported by
Considering the lattice parameters and HAp crystal structure, CLUSTER Project (the second stage) and a Grant-in-Aid (No.
this result is found to be reasonable to elongate to the 〈0001〉 20350093) of Ministry of Education, Culture, Sports, Science
directions. Furthermore, the lattice image obtained from a HAp and Technology, Japan. A part of this work was supported by
1-D crystal is shown in Figure 5c, taken with the incident beam the TEPCO Research Foundation and The Steel Industry
along the [11j00] direction. The crystal was of very good Foundation for the Advancement of Environmental Protection
crystallinity because no defects were observed in this image. Technology (2007-2008). This research was partially performed
The SAED indicated that the lattice parameters of the 1-D under the interuniversity cooperative research program of
Advanced Research Center Metallic Glasses, Institute for
crystals are a ) 0.947 and c ) 0.686 nm, which are in good
Materials Research, Tohoku University.
agreement with those found in the previous study.11 From these
results of SEM, XRD, and TEM, we conclude that the HAp References
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Finally, we found the flux which is able to make highly CG900159J