Biomaterials 24 (2003) 1389–1398

Synthesis of Si, Mg substituted hydroxyapatites and their

sintering behaviors
S.R. Kima,*, J.H. Leeb, Y.T. Kimb, D.H. Riua, S.J. Junga, Y.J. Leea, S.C. Chunga,
Y.H. Kima,1
a
Korea Institute of Ceramic Engineering and Technology, Seoul 153-801, South Korea
b
Department of Orthopedic Surgery, University of Ulsan College of Medicine, ASAN Medical Center, South Korea
Received 1 May 2002; accepted 15 October 2002

Abstract

Si, Mg-substituted hydroxyapatites, alone and co-substituted, have been prepared to obtain biomaterials having an improved
biocompatibility. From FT-IR, XRD and ICP analyses, it was confirmed that single phases of hydroxyapatite substituted by Si
alone or co-substituted by Si, Mg. The XRD data indicated the absence of extra phases related to silicon and magnesium oxide or
other calcium phosphate species. Si-substituted hydroxyapatite of up to 2 wt% for Si and Si, Mg co-substituted hydroxyapatite of
1 wt% for the each ion keep their original structures intact for the sintering temperatures of up to 12001C. However, it is observed
that ion substitutions by an amount higher than the above ratios for each hydroxyapatite lead to destabilization of original
structures of the hydroxyapatite and to the production of tricalcium phosphate and calcium phosphate silicate phases when the
samples were sintered at 11001C or higher.
r 2002 Elsevier Science Ltd. All rights reserved.

Keywords: Silicon; Magnesium; Substitution; Hydroxyapatite; Sintering behavior

1. Introduction was found in bioactive silicate containing glass ceramics

Synthesis of chemically modified or ion-substituted
hydroxyapatite has drawn interest of many scientists
since major component of biological tissues such as
bone, teeth and some invertebrate skeletons are com-
posed of hydroxyapatite containing various kinds of
inorganic substances [1–11]. Especially, incorporation of
silicon and magnesium ions into hydroxyapatite struc-
ture is of great interest since they play an important role
in developing artificial bone.
Carlisle et al. have indicated the importance of silicon
for bone formation and calcification through electron
microprobe study. It has been shown that silicon is
localized in active growth areas, such as the osteoid, of
the young bone of mice and rats, and silicon levels up to
0.5 wt% were observed in these areas [12]. Further
evidence of the importance of silicon on bone formation
*Corresponding author. Fax: +82-2-3282-2430.
E-mail address: yhkokim@kicet.re.kr (Y.H. Kim).
1
Also for correspondence.
[13–16]. It is well known that apatite layers are formed
on the surface of bioactive silicate glass ceramics, such
as Bioglass and Cerabone A-W, after a few days soaking
in Simulated Body Fluid at 371C. The silanol groups in
glass ceramics or bioglass have been proposed as
catalysts of the apatite phase nucleation in forming
surface apatite layers [15–16]. It is generally noted that
silicon is essential to the growth and development of
biological tissue such as bone, teeth and some inverte-
brate skeletons.
Magnesium is also undoubtedly one of the most
important bivalent ions associated with biological
apatites [6–7]. It has been verified that in calcified
tissues, the amount of magnesium associated with the
apatitic phase is higher at the beginning of the
calcification process and decreases with increasing
calcification. Also, there is growing evidence that
magnesium may be an important factor in the qualita-
tive changes of the bone matrix that determines bone
fragility. Magnesium depletion adversely affects all
stages of skeletal metabolism, causing cessation of bone

0142-9612/02/$ - see front matter r 2002 Elsevier Science Ltd. All rights reserved.
PII: S 0 1 4 2 - 9 6 1 2 ( 0 2 ) 0 0 5 2 3 - 9
1390 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398
growth, decreased osteoblastic and osteoclastic activ-
ities, osteopenia and bone fragility [17].
Several attempts have been made to prepare Si- or
Mg-substituted hydroxyapatite through a variety of
synthetic routes. For examples, Gibson et al. [1] and
Marques et al. [3] prepared Si-substituted hydroxyapa-
tite using silicon acetate as a silicon source. Ruys [4] and
Kim et al. [5] also prepared Si-substituted hydroxyapa-
tite utilizing tetraethyl orthosilicate (TEOS) as a silicon
source. Also, Mg-substituted hydroxyapatite was stu-
died by many researchers [18–23]. In the previous study,
it has been revealed that magnesium inhibits apatite
crystallization in solution and destabilizes the structure
of hydroxyapatite and favors its thermal conversion into
b-tricalcium phosphate. However, it is noted that
substitution of silicon or magnesium should not result
in thermal decomposition upon sintering in order to
maintain the integrity of the ion-substituted hydroxya-
patites toward bioactivity. No detailed information of
sintering behavior of Si- or Mg-substituted hydroxy-
apatite has been presented in the relevant literature.
The purpose of the present study is to synthesize Si-
substituted hydroxyapatite and the combinated ion of
Si- and Mg-substituted hydroxyapatite and to examine
their sintering behavior to explore optimum sintering
conditions of substituted hydroxyapatites.
2. Materials and methods
2.1. Preparation of Si-substituted hydroxyapatite
(Si-HA)
The Si-substituted hydroxyapatites were prepared
using a precipitation method. CaCO3 powder was
heated at 10001C for 3 h and resultant 5.30 g CaO
powder was hydrated to produce Ca(OH)2 with 1.7 l
distilled water. Prior to use, CO2, which dissolved in
distilled water, was eliminated by bubbling with nitro-
gen gas for 30 min and the subsequent reaction
proceeded under nitrogen atmosphere. The appropriate
amounts of 0.3 m H3PO4 solution was added into
Ca(OH)2 suspension using an automatic titrator with
vigorous stirring. During the addition, pH of the
Table 1
Quantities of reactants used and the expected wt% of Si, Mg
Samples No. of moles
Ca(OH)2 H3PO4
Si-HA I 10.5 5.29
Si-HA II 10.5 4.61
Si, Mg-HAI 9.90 5.50
Si, Mg-HAII 10.05 5.50
Si, Mg-HAIII 10.19 5.75
Ca/(P+Si) or (Ca+Mg)/(P+Si) is 1.75 for starting materials.
solution was maintained at 10.0 by the addition of
NH4OH solution. The mole ratios of Ca(OH)2 and
H3PO4 required to prepare a stoichiometric Si-substi-
tuted hydroxyapatite are listed in Table 1. Reaction
temperature was maintained at 251C. Various amounts
of 99% TEOS were added to the resulting hydroxyapa-
tite suspension (Table 1). The reaction mixture was
stirred for 2 h and aged for 1 day at 251C. The resulting
precipitate was filtered, dried at 901C overnight. The
chemical compositions of the samples were determined
by ICP spectrometer (Perkin–Elmer Optima 3300DV).
2.2. Preparation of Si, Mg-substituted hydroxyapatite
(Si, Mg-HA)
Similar to the Si-substituted hydroxyapatite, the
Si, Mg-substituted hydroxyapatites were prepared using
a precipitation method. CaCO3 powder was heated at
10001C for 3 h and resultant 5.30 g CaO powder was
hydrated to produce Ca(OH)2 with 1.7 l distilled water
from which CO2 was removed by bubbling with
nitrogen and entire reaction was carried out under
nitrogen atmosphere. Mg(OH)2 was completely dis-
solved in 0.3 m H3PO4 solution and added into Ca(OH)2
suspension 251C using an automatic titrator with
vigorous stirring. During the addition, pH of the
solution was maintained at 10.0 by the addition of
NH4OH solution. Various amounts of 99% TEOS were
added to the resulting hydroxyapatite suspension and
stirred for 2 h. The mole ratio of each reactant required
to prepare a stoichiometric Si, Mg-substituted hydro-
xyapatite is listed in Table 1. The reaction mixture was
aged for 1 day at 251C. The resulting precipitate was
filtered, dried at 901C overnight. The chemical composi-
tions of the samples were determined by ICP spectro-
meter.
2.3. Characterization techniques
In order to examine the sintering behavior, the
synthesized powders of ion-substituted hydroxyapatites
were uniaxially pressed to form a disk-shaped compact
(10 mm diameter and 1.5 mm thick) and the compact
was then pressed at 1600 bar using a cold isostatic
Expected wt%
TEOS Mg(OH)2 Si Mg
0.71 0 2 0
1.39 0 4 0
0.50 0.60 2 2
0.50 0.45 1.5 1.5
0.25 0.31 1 1
 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398
pressing (CIP). The green pellets were sintered between
10001C and 15001C in an atmospheric condition, and
maintained for 4 h at the peak temperatures. The phase
transformations of sintered samples at different heating
temperatures were observed using X-ray powder dif-
fractometer (MAC Science Co., Ltd.). Cu Ka radiation
was used at the operating condition of 40 kV and 20 mA.
The XRD data were collected over 2y range of 10–601 at
a step size of 0.021. The density of sintered samples was
measured using Archimedes’ principles. Transmission
electron microscopy (TEM) (200 kV, JEM-2000EX and
400 kV, JEM-4010, JEOL) was used to observe particle
shape and to examine the grain boundary and distribu-
tion of Si along grain boundary and inside the grain of
Si-substituted hydroxyapatite. TEM with EDAX was
employed for characterization of the microstructure and
for the distribution of silicon ion. Sample for TEM was
prepared by mechanical grinding and polishing the
3 mm disk specimen separated by ultrasonic disk cutter.
The one side of the specimen was dimple ground to the
30 mm thickness. The thinned specimen was further
thinned by Ar-ion sputtering down to electron transpar-
ency. A thin carbon film was coated before observation
to prevent the electron discharge during observation.
The IR spectra were recorded between 400 and
1400 cm
1 on a Perkin–Elmer 1600 M-80 spectrometer.
Chemical analyses were performed by a Jarrell–Ash
Poliscan 61E ICP spectrometer and a Perkin–Elmer
5000 atomic absorption spectrophotometer. The cell
parameters were refined using the LeBail method in the
generalized structure analysis system (GSAS). Determi-
nation of the lattice parameters of pure and ion-
substituted samples sintered at 12001C for 4 h with
background parameters, zero point, and peak shape
parameters was made by LeBail techniques of the X-ray
diffraction data collected for powders. The refinement
software GSAS was used and the refinements were
based on the structural data of Kay et al. using the space
group P63/m.
(a)
Fig. 1. TEM micrographs of the samples: (a) Si-substituted hydroxyapatite (Si-HA I), and (b) Si, Mg-substituted hydroxyapatite (Si, Mg-HA II).
1391
2.4. Evaluation of biocompatibility of ion-substituted
hydroxyapatite
Cell proliferation experiment was carried out to
evaluate biocompatibility of the pure HA, Si-HA and
Si, Mg-HA. MG63 osteoblast-like cells, a continuous
line derived by human osteosarcoma [Korean Cell Line
Bank, Seoul, Korea], were cultured in Dulbecco’s
modified Eagle’s medium (DMEM) containing 10%
fetal bovine serum (FBS; HyClone, Logan, Utah) and
0.5% antibiotics at 371C in a humidified atmosphere of
95% air and 5% CO2. After subculture of MG63 cells,
2  104 cells/ml were placed in the plates containing pure
and the ion-substituted hydroxyapatite substrates and in
the polystyrene control plates. The number of cells in the
trypsinized plates was counted by a hemocytometer at
the incubation times, 3, 6 and 24 h.
3. Results
3.1. As-synthesized powders and their chemical analysis
The TEM micrographs of as precipitated powder of
hydroxyapatites are shown in Fig. 1. The nano-sized
powders in needle shape were synthesized of 50 nm for
Si-substituted and 100 nm for Si, Mg co-substituted
hydroxyapatites in average size. The chemical composi-
tions determined by ICP of the ion-substituted samples
are shown in Table 2. The silicon and magnesium
contents are lower than that of the corresponding
amount of starting material, which implies that some
of silicon and magnesium ions remain in the mother
liquor solution after precipitation. X-ray diffraction
data of as-synthesized pure and ion-substituted hydro-
xyapatites show broad diffraction patterns and this is
attributed to small crystallite size and poor crystallinity
of as-synthesized forms (Fig. 2). No observable differ-
ence in X-ray diffraction data of as-synthesized form of
(b)
1392 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398

Table 2
Chemical analysis data of the samples

Sample Measured wt% (in solid) Measured wt% (residue in mother liquor) Chemical formula of ion-substituted hydroxyapatite

Si Mg Si Mg

Si-HA I 1.97 0 0.02 0 Ca10(PO4)5.53(SiO4)0.47(OH)1.53

Si-HA II 3.76 0 0.04 0 Ca10(PO4)5.11(SiO4)0.89(OH)1.11
Si, Mg-HAI 1.73 1.38 0.27 0.64 Ca9.62Mg0.38(PO4)5.59(SiO4)0.41(OH)1.59
Si, Mg-HAII 1.05 0.99 0.43 0.50 Ca9.73Mg0.27(PO4)5.75(SiO4)0.25(OH)1.75
Si, Mg-HAIII 0.92 0.65 0.08 0.34 Ca9.82Mg0.18(PO4)5.78(SiO4)0.22(OH)1.78

Uncertainty: 72%, Relative Standard deviation.

No. of moles of residue in mother liquor was converted to wt.% in solid fraction.

(c)
Intensity (arbitrary unit)

(b)

(a)

10 20 30 40 50 60
2θ
Fig. 2. X-ray powder diffraction patterns of as-synthesized samples. (a) Pure hydroxyapatite, (b) Si-substituted hydroxyapatite (Si-HA I), and (c)
Si, Mg-substituted hydroxyapatite (Si, Mg-HA II).

pure and ion-substituted hydroxyapatites is detected, hydroxyapatite structure. The FT-IR spectra of both
even though their chemical analysis confirmed calcined pure and ion-substituted hydroxyapatite sam-
the presence of certain amount of ion substitution ples show typical absorption bands of PO4 vibrations at
(Table 2). 800–1100 and 500–700 cm
1.
Lattice parameters of pure and ion-substituted
3.2. FT-IR data and lattice constant refinement hydroxyapatites [24–26] were refined by the LeBail
method in the GSAS using the X-ray powder diffraction
Comparing the FT-IR spectra of the calcined pure data of each sample sintered at 12001C for 4 h. The cell
and ion-substituted hydroxyapatites, the most notable parameters of the samples are shown in Table 3. The
effect of ion substitution on FT-IR spectra of hydro- space group is P63/m. The new lattice constants for Si-
xyapatite is the change in the hydroxyl stretching bands HA and Si, Mg-HA are a ¼ 9:4163 A, ( c ¼ 6:9019 A
( and
at 3570 and 630 cm
1. Substitution of silicon into the ( c ¼ 6:8980 A,
a ¼ 9:4023 A, ( respectively. Meanwhile,
hydroxyapatite framework (Ca10(PO4)6
x(SiO4)x the lattice constants for pure HA synthesized under
(OH)2
x) reduces amount of hydroxyl group to com- ( and c ¼ 6:8751 A.
the same condition are a ¼ 9:4126 A, (
pensate for the extra negative charge of the silicate
group. In Fig. 3, intensities at 3570 and 630 cm
1 of ion- 3.3. Thermal stability of sintered samples and phase
substituted samples drastically decrease with increasing characterization
substituted silicon amount compared with those of
hydroxyapatite spectra. This result clearly shows that Fig. 4 shows the X-ray diffraction patterns of sintered
PO4 tetrahedra are replaced by SiO4 tetrahedra in the sample of 1.97 wt% Si-HA in the range 1000–15001C for

(c)
Intensity (arbitrary)
(b)
(a)
4000
Fig. 3. FT-IR spectra of the samples sintered at 12001C for 4 h: (a) pure hydroxyapatite (HA), (b) Si, Mg-substituted hydroxyapatite (Si, Mg-HA II),
and (c) Si-substituted hydroxyapatite (Si-HA I).
Table 3
Lattice constants of synthesized pure and ion-substituted hydroxya-
patites
Sample Space group
HA P63/m
Si-HA I P63/m
Si, Mg-HA II P63/m
4 h. Below 12001C, all the peaks in the data are indexed
for hydroxyapatite phase on the basis of JCPDS Card
No. 9-432. The data do not reveal the presence of extra
phases related to silicon oxide species or other calcium
phosphate species and indicates that structure of Si-
substituted hydroxyapatite keeps its original structures
intact up to 12001C. However, the X-ray diffraction
patterns of sintered sample containing the same silicon
content at 13001C or higher temperature for 4 h exhibit
the peaks corresponding to tricalcium phosphate
(Ca3(PO4)2) and calcium phosphate silicate
(Ca10(PO4)4(SiO4)2) phases (Fig. 4). Fig. 5 shows the
X-ray diffraction patterns of sintered sample of
3.76 wt% Si-HA in the range 1000–13001C for 4 h. In
this case, the X-ray diffraction pattern of sintered
sample at 11001C for 4 h already shows the peaks
corresponding to tricalcium phosphate and calcium
phosphate silicate phases.
Fig. 6 shows the X-ray diffraction patterns of sintered
sample of Si, Mg-HA at 12001C for 4 h with various
amounts of silicon and magnesium. At the temperature
12001C, the XRD data of about 1 wt% of each Si, Mg
co-substituted hydroxyapatite do not indicate the
presence of extra phases related to silicon oxide,
magnesium species or other calcium phosphate species,
S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398 1393
3000 2000 1000
-1
Wavenumber (cm )
which means that structure of Si, Mg co-substituted
hydroxyapatite maintains its original structures up to
12001C. However, the X-ray diffraction patterns of
Lattice constants above 1 wt% of each Si, Mg co-substituted hydroxya-
( c ¼ 6:875 A
a ¼ b ¼ 9:413 A, ( patite at the same sintered temperature contain the
( c ¼ 6:902 A
a ¼ b ¼ 9:416 A, ( peaks indicating the presence of tricalcium phosphate.
( c ¼ 6:898 A
a ¼ b ¼ 9:402 A, (
3.4. Densification and microstructure
The density of sintered hydroxyapatite has been
significantly influenced by sintering temperature. The
bulk density of pure and ion-substituted samples as a
function of sintering temperature is shown in Fig. 7. The
density of sintered samples at each temperature was
measured using Archimedes’ principles. Theoretical
density of pure hydroxyapatite calculated from unit cell
parameter is known as 3.156 g/cm3. The density of pure
hydroxyapatite reaches 3.071 g/cm3 at 10001C. 1.97 wt%
of Si-substituted hydroxyapatite reaches 3.014g/cm3 at
12001C of sintering temperature. Theoretical density of
silicon-substituted hydroxyapatite is expected less than
pure hydroxyapatite’s due to its change of unit cell
parameters by Si substitution [1]. At temperatures above
12001C, there is a slight decrease in the bulk density due
to production a-tricalcium phosphate and calcium
phosphate silicate (Ca10(PO4)4(SiO4)2) phases by the
decomposition Si-substituted hydroxyapatite. It is well
known in the literature that a bulk density of a-
tricalcium phosphate is 2.863 g/cm3, which is lower than
that of hydroxyapatite [27].
For 3.76 wt% Si-substituted hydroxyapatite, the
density higher than 2.525 g/cm3 is never reached. It is
shown that 3.76 wt% Si-substituted hydroxyapatite
starts to decompose into a-tricalcium phosphate and
1394 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398

α -TCP
Ca 5 (PO4 )2 (SiO4 )

(e)

Intensity (arbitrary unit) (d)

(c)

(b)

(a)

10 20 30 40 50
2θ
Fig. 4. X-ray powder diffraction patterns of 1.97 wt% Si-substituted hydroxyapatite (Si-HA I) sintered at various temperatures: (a) 10001C, (b)
12001C, (c) 13001C, (d) 14001C and (e) 15001C.

Ca 5 (PO4 ) 2 (Si O4 )
α -TCP
CaO

(d)
Intensity (arbitrary unit)

(c)

(b)

(a)

10 20 30 40 50
2θ
Fig. 5. X-ray powder diffraction patterns of 3.76 wt% Si-substituted hydroxyapatite: (Si-HA II) sintered at various temperatures: (a) 10001C, (b)
11001C, (c) 12001C and (d) 13001C.
 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398 1395

(a)
β-TCP

Intensity (arbitrary unit)

(b)

(c)

10 20 30 40 50 60
2θ
Fig. 6. X-ray powder diffraction patterns of Si, Mg-substituted hydroxyapatites: sintered at 12001C for 4 h: (a) Si, 1.73 wt%, Mg, 1.38 wt% (Si, Mg-
HA I); (b) Si, 1.05 wt%, Mg, 0.99 wt% (Si, Mg-HA II); (c) Si, 0.92 wt%, Mg, 0.65 wt% (Si, Mg-HA III).

100

HA
90
Relative Density (%)

80
MgSiHA
70

60

50
SiHA

40

1000 1100 1200 1300

Temperature (°C)

Fig. 7. Changes of relative densities of ion-substituted hydroxyapatites

after sintering: at different temperatures.

calcium phosphate silicate phases at 11001C, which

means that the 3.76 wt% Si-substituted hydroxyapatite
sample is decomposed before it is fully sintered.
Density of 1.05 wt% each of silicon, magnesium co-
substituted hydroxyapatite reaches 3.017 g/cm3 at Fig. 8. Overall microstructure of Si-HA I sample sintered at 12001C
12001C of sintering temperature while the sample for 4 h.
maintains its original structure.
The microstructure of 1.97 wt% Si-substituted hydro-
xyapatite was observed using a transmission electron that no thermal decomposition occurs during sintering
microscope. The overall microstructure of the Si- process at this temperature. Also no pore was observed
substituted hydroxyapatite consisted of equiaxed grains during the TEM analysis, which indicated that the
with an average grain size of 0.5 mm. Fig. 8 shows a materials was fully densified with sintering at 12001C.
typical microstructure of 1.97 wt% Si-substituted hydro- Fig. 9 shows the high-resolution electron micrographs
xyapatite. No other crystalline phase except hydroxya- around the grain boundary of Si-substituted hydro-
patite is detected during TEM analysis, which suggests xyapatite. The grain boundary was wavy in nature as
1396 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398
2nm
Fig. 9. High-resolution electron micrograph that shows the clean grain
boundary between two grains of Si-HA I sample; No amorphous phase
was present along the grain boundary. Wavy nature of the grain
boundary is evident as indicated by arrows.
indicated by the arrows. However, any amorphous
phase or segregation of SiO2 or MgO was not present
along the grain boundary. Also, the distribution of
silicon along the grain boundary and inside the grain
was qualitatively analyzed with EDAX using beam size
of 15 nm. There was no increased segregation along
grain boundary. Also, it is not observed any other
phases associated with the silicon addition. It is
confirmed that silicon-substitution is to be fully stabi-
lized inside the lattice structure of the hydroxyapatite as
long as the content of the silicon is limited below
1.97 wt%.
3.5. Evaluation of biocompatibility of Si-substituted
hydroxyapatite
The number of cells proliferated in the pure and ion-
substituted hydroxyapatite sample plate and in the
control plate was counted as shown in Fig. 10. With
increasing time, the number of cells were gradually
increased up to 12  104 cells/ml for the control plate
and 29  104, 20  104, and 33  104 cells/ml for the
pure, Si, Mg-HA II and Si-HA I sample plates,
respectively. Even though the degree of cell proliferation
of Si, Mg-HA II was lower than that of the pure
hydroxyapatite, the degree of cell proliferation in the
sample plates was higher than that in the control one.
The degree of cell proliferation on the Si, Mg-HA II and
Si-HA I gradually increased with time. These results
indicate that the ion-substituted hydroxyapatite samples
have better biocompatibilities than the control one.
4. Discussion
Both Ca and P ions in hydroxyapatite structure can
be substituted with various ions and isomorphous
4.E+05
4.E+05 0h
3.E+05
Number of cells/n
3h
3.E+05 6h
2.E+05 24 h
2.E+05
1.E+05
5.E+04
0.E+00
Control HA Si, Mg- Si-HAI
HA II
Fig. 10. Cell proliferation result as a function of time.
substitution at each of this site is governed by ionic
radii limits. Due to their ionic sizes, it is generally
accepted that silicon substitutes for P site and exists as
silicate ion, while magnesium ion substitutes for Ca site
and exists as a magnesium oxide ion [3].
It is known that substitution of Mg+2 by Ca+2 in
the synthetic hydroxyapatites is very limited (o1%)
[19]. Lack of substitution is due in part to the differences
in the hydrated properties of the Mg+2 and Ca+2 ions.
The smaller ionic radius of Mg+2 causes its hydration
layer to be bound much more tightly than that of
Ca+2 ion. Therefore, the dehydration of the aqueous
ions, which needs to occur before incorporation
into a crystalline structure, is much slower for Mg+2
ion. In the present study, however, Mg ion is readily
incorporated into the hydroxyapatite structure at
levels up to 1 wt% when it co-substitutes with Si
ion. The substituted amount of Si in synthesized
Si-HA using our method is also higher than the
values reported in the literature [1] within the thermal
stability region. It is attributed to the fact that
our synthetic procedure was keeping away CO2 to
prevent producing carbonated hydroxyapatite since Si
and CO
3 ions are known to replace phosphorus site
competitively [1,11].
Substitution of silicon into the hydroxyapatite frame-
work (Ca10(PO4)6
x(SiO4)x(OH)2
x) reduces amount of
hydroxyl group to compensate for the extra negative
charge of the silicate group. Intensities at 3570 and
630 cm
1 of ion-substituted samples drastically decrease
with increasing substituted silicon amount compared
with those of hydroxyapatite spectra. This result clearly
shows that PO4 tetrahedra are replaced by SiO4
tetrahedra in the hydroxyapatite structure.
The lattice constants for both a and c slightly increase
when P+5 ions are replaced by Si+4 in the hydro-
xyapatite framework. The ionic radius of Si+4 (0.42 A) (
is larger than the one of P+5 (0.35 A), ( which is in
agreement with the observed increase in lattice constants
for Si-substituted HA. However, the lattice constants
 S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398
both for a and c decrease when Ca+2 ions are replaced
by Mg+2 in the hydroxyapatite framework. The ionic
radius of Mg+2 (0.69 A) ( is considerably smaller than the
one of Ca +2 (
(0.99 A), which is consistent with the
observed reduction in lattice constants Si, Mg-substi-
tuted HA. The change of the lattice constants of ion-
substituted hydroxyapatite clearly demonstrates that
Si, Mg ions are structurally incorporated, they do not
just cover the surface of hydroxyapatite.
TEM micrographs data show the morphology of the
ion-substituted hydroxyapatites. The crystals of Si, Mg
co-substituted hydroxyapatite became larger than those
of Si mono substituted hydroxyapatite. As previously
reported [19], Mg inclusion produced larger crystals.
In general, pure hydroxyapatite is known to be
thermally stable up to 13001C and above that tempera-
ture, it starts to decompose to a- or b-tricalcium
phosphate, CaO, oxyapatite, etc. depending on Ca/P
ratio of hydroxyapatite [28]. From the XRD analysis,
1.97 wt% Si-singly substituted hydroxyapatite is ther-
mally stable up to 12001C. However, substitution of an
amount higher than 1.97 wt% of silicon leads to
decomposition of Si-substituted hydroxyapatite to a-
tricalcium phosphate (Ca3(PO4)2) and calcium phos-
phate silicate (Ca10(PO4)4(SiO4)2) phases at 11001C.
From the microstructural observation and EDAX
analysis of the sample sintered at 12001C using
TEM, it was concluded that Si-substituted hydroxyapa-
tite consisted of mono-phase hydroxyapatite grains
without any amorphous phase. Although the sinter-
ability of Si-substituted hydroxyapatite was significantly
reduced with the presence of Si, fully dense sintered
body can be obtained within the thermal stability
region.
It is known that magnesium inhibits apatite crystal-
lization in solution and destabilizes the structure of
hydroxyapatite and favors its thermal conversion into a-
or b-tricalcium phosphate. Our experimental results
show that co-substitution of both magnesium and
silicon leads to destabilization its original structure.
When co-substitution with magnesium is carried out
maintaining its original structure, amount of Si-sub-
stitution into hydroxyapatite is reduced. Si-substitution
singly into hydroxyapatite structure was achieved by
1.97 wt% Si level. However, co-substitution with mag-
nesium, Si-substitution into hydroxyapatite structure
was limited by 1.05 wt%. It was evidenced by XRD
analysis at different sintered temperatures.
To evaluate biocompatibility of the synthesized ion-
substituted hydroxyapatite, cell proliferation experiment
was carried out for the short period of time, up to 24 h at
this stage. The degree of cell proliferation on the Si, Mg-
and Si-substituted hydroxyapatites gradually increased
with the increase of time. Based on these results, the
Si, Mg- and Si-substituted hydroxyapatites are consid-
ered to have a good biocompatibility.
1397
5. Conclusion
To obtain biomaterials having an improved biocom-
patibility, Si, Mg-substituted hydroxyapatites, individu-
ally and in combination, have been prepared and their
sintering behaviors have been examined. From the XRD
analysis, it was confirmed that the single-phase hydro-
xyapatites with silicon level 1.97 wt% for singly sub-
stituted sample kept its original structure with fully
dense sintered body. TEM analysis confirmed that Si-
substituted hydroxyapatite consisted of mono-phase
hydroxyapatite grains without any amorphous phase.
Co-substitution with magnesium, Si-substitution into
hydroxyapatite structure was limited by 1.05 wt%. It
was evidenced by XRD analysis at different sintered
temperatures. The results of cell proliferation experi-
ments suggest that the Si, Mg- and Si-substituted
hydroxyapatites have a good biocompatibility and can
be the useful materials for implants and bone augmen-
tation.
Acknowledgements
Financial support of this work was provided by Next
Generation Project (Development of Biological Hybrid
Materials) of Ministry of Commerce, Industry and
Energy, Korea The authors would like to thank Dr. Jae-
hyun Park for her assistance in determination of lattice
parameters of the samples using the generalized
structure analysis system (GSAS).
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