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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


[13–16]. It is well known that apatite layers are formed
Synthesis of chemically modified or ion-substituted on the surface of bioactive silicate glass ceramics, such
hydroxyapatite has drawn interest of many scientists as Bioglass and Cerabone A-W, after a few days soaking
since major component of biological tissues such as in Simulated Body Fluid at 371C. The silanol groups in
bone, teeth and some invertebrate skeletons are com- glass ceramics or bioglass have been proposed as
posed of hydroxyapatite containing various kinds of catalysts of the apatite phase nucleation in forming
inorganic substances [1–11]. Especially, incorporation of surface apatite layers [15–16]. It is generally noted that
silicon and magnesium ions into hydroxyapatite struc- silicon is essential to the growth and development of
ture is of great interest since they play an important role biological tissue such as bone, teeth and some inverte-
in developing artificial bone. brate skeletons.
Carlisle et al. have indicated the importance of silicon Magnesium is also undoubtedly one of the most
for bone formation and calcification through electron important bivalent ions associated with biological
microprobe study. It has been shown that silicon is apatites [6–7]. It has been verified that in calcified
localized in active growth areas, such as the osteoid, of tissues, the amount of magnesium associated with the
the young bone of mice and rats, and silicon levels up to apatitic phase is higher at the beginning of the
0.5 wt% were observed in these areas [12]. Further calcification process and decreases with increasing
evidence of the importance of silicon on bone formation calcification. Also, there is growing evidence that
magnesium may be an important factor in the qualita-
*Corresponding author. Fax: +82-2-3282-2430.
tive changes of the bone matrix that determines bone
E-mail address: yhkokim@kicet.re.kr (Y.H. Kim). fragility. Magnesium depletion adversely affects all
1
Also for correspondence. 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- solution was maintained at 10.0 by the addition of
ities, osteopenia and bone fragility [17]. NH4OH solution. The mole ratios of Ca(OH)2 and
Several attempts have been made to prepare Si- or H3PO4 required to prepare a stoichiometric Si-substi-
Mg-substituted hydroxyapatite through a variety of tuted hydroxyapatite are listed in Table 1. Reaction
synthetic routes. For examples, Gibson et al. [1] and temperature was maintained at 251C. Various amounts
Marques et al. [3] prepared Si-substituted hydroxyapa- of 99% TEOS were added to the resulting hydroxyapa-
tite using silicon acetate as a silicon source. Ruys [4] and tite suspension (Table 1). The reaction mixture was
Kim et al. [5] also prepared Si-substituted hydroxyapa- stirred for 2 h and aged for 1 day at 251C. The resulting
tite utilizing tetraethyl orthosilicate (TEOS) as a silicon precipitate was filtered, dried at 901C overnight. The
source. Also, Mg-substituted hydroxyapatite was stu- chemical compositions of the samples were determined
died by many researchers [18–23]. In the previous study, by ICP spectrometer (Perkin–Elmer Optima 3300DV).
it has been revealed that magnesium inhibits apatite
crystallization in solution and destabilizes the structure 2.2. Preparation of Si, Mg-substituted hydroxyapatite
of hydroxyapatite and favors its thermal conversion into (Si, Mg-HA)
b-tricalcium phosphate. However, it is noted that
substitution of silicon or magnesium should not result Similar to the Si-substituted hydroxyapatite, the
in thermal decomposition upon sintering in order to Si, Mg-substituted hydroxyapatites were prepared using
maintain the integrity of the ion-substituted hydroxya- a precipitation method. CaCO3 powder was heated at
patites toward bioactivity. No detailed information of 10001C for 3 h and resultant 5.30 g CaO powder was
sintering behavior of Si- or Mg-substituted hydroxy- hydrated to produce Ca(OH)2 with 1.7 l distilled water
apatite has been presented in the relevant literature. from which CO2 was removed by bubbling with
The purpose of the present study is to synthesize Si- nitrogen and entire reaction was carried out under
substituted hydroxyapatite and the combinated ion of nitrogen atmosphere. Mg(OH)2 was completely dis-
Si- and Mg-substituted hydroxyapatite and to examine solved in 0.3 m H3PO4 solution and added into Ca(OH)2
their sintering behavior to explore optimum sintering suspension 251C using an automatic titrator with
conditions of substituted hydroxyapatites. 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
2. Materials and methods added to the resulting hydroxyapatite suspension and
stirred for 2 h. The mole ratio of each reactant required
2.1. Preparation of Si-substituted hydroxyapatite to prepare a stoichiometric Si, Mg-substituted hydro-
(Si-HA) xyapatite is listed in Table 1. The reaction mixture was
aged for 1 day at 251C. The resulting precipitate was
The Si-substituted hydroxyapatites were prepared filtered, dried at 901C overnight. The chemical composi-
using a precipitation method. CaCO3 powder was tions of the samples were determined by ICP spectro-
heated at 10001C for 3 h and resultant 5.30 g CaO meter.
powder was hydrated to produce Ca(OH)2 with 1.7 l
distilled water. Prior to use, CO2, which dissolved in 2.3. Characterization techniques
distilled water, was eliminated by bubbling with nitro-
gen gas for 30 min and the subsequent reaction In order to examine the sintering behavior, the
proceeded under nitrogen atmosphere. The appropriate synthesized powders of ion-substituted hydroxyapatites
amounts of 0.3 m H3PO4 solution was added into were uniaxially pressed to form a disk-shaped compact
Ca(OH)2 suspension using an automatic titrator with (10 mm diameter and 1.5 mm thick) and the compact
vigorous stirring. During the addition, pH of the was then pressed at 1600 bar using a cold isostatic

Table 1
Quantities of reactants used and the expected wt% of Si, Mg

Samples No. of moles Expected wt%

Ca(OH)2 H3PO4 TEOS Mg(OH)2 Si Mg

Si-HA I 10.5 5.29 0.71 0 2 0


Si-HA II 10.5 4.61 1.39 0 4 0
Si, Mg-HAI 9.90 5.50 0.50 0.60 2 2
Si, Mg-HAII 10.05 5.50 0.50 0.45 1.5 1.5
Si, Mg-HAIII 10.19 5.75 0.25 0.31 1 1

Ca/(P+Si) or (Ca+Mg)/(P+Si) is 1.75 for starting materials.


S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398 1391

pressing (CIP). The green pellets were sintered between 2.4. Evaluation of biocompatibility of ion-substituted
10001C and 15001C in an atmospheric condition, and hydroxyapatite
maintained for 4 h at the peak temperatures. The phase
transformations of sintered samples at different heating Cell proliferation experiment was carried out to
temperatures were observed using X-ray powder dif- evaluate biocompatibility of the pure HA, Si-HA and
fractometer (MAC Science Co., Ltd.). Cu Ka radiation Si, Mg-HA. MG63 osteoblast-like cells, a continuous
was used at the operating condition of 40 kV and 20 mA. line derived by human osteosarcoma [Korean Cell Line
The XRD data were collected over 2y range of 10–601 at Bank, Seoul, Korea], were cultured in Dulbecco’s
a step size of 0.021. The density of sintered samples was modified Eagle’s medium (DMEM) containing 10%
measured using Archimedes’ principles. Transmission fetal bovine serum (FBS; HyClone, Logan, Utah) and
electron microscopy (TEM) (200 kV, JEM-2000EX and 0.5% antibiotics at 371C in a humidified atmosphere of
400 kV, JEM-4010, JEOL) was used to observe particle 95% air and 5% CO2. After subculture of MG63 cells,
shape and to examine the grain boundary and distribu- 2  104 cells/ml were placed in the plates containing pure
tion of Si along grain boundary and inside the grain of and the ion-substituted hydroxyapatite substrates and in
Si-substituted hydroxyapatite. TEM with EDAX was the polystyrene control plates. The number of cells in the
employed for characterization of the microstructure and trypsinized plates was counted by a hemocytometer at
for the distribution of silicon ion. Sample for TEM was the incubation times, 3, 6 and 24 h.
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 3. Results
30 mm thickness. The thinned specimen was further
thinned by Ar-ion sputtering down to electron transpar- 3.1. As-synthesized powders and their chemical analysis
ency. A thin carbon film was coated before observation
to prevent the electron discharge during observation. The TEM micrographs of as precipitated powder of
The IR spectra were recorded between 400 and hydroxyapatites are shown in Fig. 1. The nano-sized
1400 cm1 on a Perkin–Elmer 1600 M-80 spectrometer. powders in needle shape were synthesized of 50 nm for
Chemical analyses were performed by a Jarrell–Ash Si-substituted and 100 nm for Si, Mg co-substituted
Poliscan 61E ICP spectrometer and a Perkin–Elmer hydroxyapatites in average size. The chemical composi-
5000 atomic absorption spectrophotometer. The cell tions determined by ICP of the ion-substituted samples
parameters were refined using the LeBail method in the are shown in Table 2. The silicon and magnesium
generalized structure analysis system (GSAS). Determi- contents are lower than that of the corresponding
nation of the lattice parameters of pure and ion- amount of starting material, which implies that some
substituted samples sintered at 12001C for 4 h with of silicon and magnesium ions remain in the mother
background parameters, zero point, and peak shape liquor solution after precipitation. X-ray diffraction
parameters was made by LeBail techniques of the X-ray data of as-synthesized pure and ion-substituted hydro-
diffraction data collected for powders. The refinement xyapatites show broad diffraction patterns and this is
software GSAS was used and the refinements were attributed to small crystallite size and poor crystallinity
based on the structural data of Kay et al. using the space of as-synthesized forms (Fig. 2). No observable differ-
group P63/m. ence in X-ray diffraction data of as-synthesized form of

(a) (b)

Fig. 1. TEM micrographs of the samples: (a) Si-substituted hydroxyapatite (Si-HA I), and (b) Si, Mg-substituted hydroxyapatite (Si, Mg-HA II).
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

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 cm1.
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 cm1. Substitution of silicon into the ( c ¼ 6:8980 A,
a ¼ 9:4023 A, ( respectively. Meanwhile,
hydroxyapatite framework (Ca10(PO4)6x(SiO4)x the lattice constants for pure HA synthesized under
(OH)2x) 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 cm1 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
S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398 1393

(c)

Intensity (arbitrary)
(b)

(a)

4000 3000 2000 1000

-1
Wavenumber (cm )
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 which means that structure of Si, Mg co-substituted


Lattice constants of synthesized pure and ion-substituted hydroxya- hydroxyapatite maintains its original structures up to
patites
12001C. However, the X-ray diffraction patterns of
Sample Space group Lattice constants above 1 wt% of each Si, Mg co-substituted hydroxya-
HA P63/m ( c ¼ 6:875 A
a ¼ b ¼ 9:413 A, ( patite at the same sintered temperature contain the
Si-HA I P63/m ( c ¼ 6:902 A
a ¼ b ¼ 9:416 A, ( peaks indicating the presence of tricalcium phosphate.
Si, Mg-HA II P63/m ( c ¼ 6:898 A
a ¼ b ¼ 9:402 A, (
3.4. Densification and microstructure

4 h. Below 12001C, all the peaks in the data are indexed The density of sintered hydroxyapatite has been
for hydroxyapatite phase on the basis of JCPDS Card significantly influenced by sintering temperature. The
No. 9-432. The data do not reveal the presence of extra bulk density of pure and ion-substituted samples as a
phases related to silicon oxide species or other calcium function of sintering temperature is shown in Fig. 7. The
phosphate species and indicates that structure of Si- density of sintered samples at each temperature was
substituted hydroxyapatite keeps its original structures measured using Archimedes’ principles. Theoretical
intact up to 12001C. However, the X-ray diffraction density of pure hydroxyapatite calculated from unit cell
patterns of sintered sample containing the same silicon parameter is known as 3.156 g/cm3. The density of pure
content at 13001C or higher temperature for 4 h exhibit hydroxyapatite reaches 3.071 g/cm3 at 10001C. 1.97 wt%
the peaks corresponding to tricalcium phosphate of Si-substituted hydroxyapatite reaches 3.014g/cm3 at
(Ca3(PO4)2) and calcium phosphate silicate 12001C of sintering temperature. Theoretical density of
(Ca10(PO4)4(SiO4)2) phases (Fig. 4). Fig. 5 shows the silicon-substituted hydroxyapatite is expected less than
X-ray diffraction patterns of sintered sample of pure hydroxyapatite’s due to its change of unit cell
3.76 wt% Si-HA in the range 1000–13001C for 4 h. In parameters by Si substitution [1]. At temperatures above
this case, the X-ray diffraction pattern of sintered 12001C, there is a slight decrease in the bulk density due
sample at 11001C for 4 h already shows the peaks to production a-tricalcium phosphate and calcium
corresponding to tricalcium phosphate and calcium phosphate silicate (Ca10(PO4)4(SiO4)2) phases by the
phosphate silicate phases. decomposition Si-substituted hydroxyapatite. It is well
Fig. 6 shows the X-ray diffraction patterns of sintered known in the literature that a bulk density of a-
sample of Si, Mg-HA at 12001C for 4 h with various tricalcium phosphate is 2.863 g/cm3, which is lower than
amounts of silicon and magnesium. At the temperature that of hydroxyapatite [27].
12001C, the XRD data of about 1 wt% of each Si, Mg For 3.76 wt% Si-substituted hydroxyapatite, the
co-substituted hydroxyapatite do not indicate the density higher than 2.525 g/cm3 is never reached. It is
presence of extra phases related to silicon oxide, shown that 3.76 wt% Si-substituted hydroxyapatite
magnesium species or other calcium phosphate species, 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

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

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

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

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
2nm Fig. 10. Cell proliferation result as a function of time.

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. 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
indicated by the arrows. However, any amorphous silicate ion, while magnesium ion substitutes for Ca site
phase or segregation of SiO2 or MgO was not present and exists as a magnesium oxide ion [3].
along the grain boundary. Also, the distribution of It is known that substitution of Mg+2 by Ca+2 in
silicon along the grain boundary and inside the grain the synthetic hydroxyapatites is very limited (o1%)
was qualitatively analyzed with EDAX using beam size [19]. Lack of substitution is due in part to the differences
of 15 nm. There was no increased segregation along in the hydrated properties of the Mg+2 and Ca+2 ions.
grain boundary. Also, it is not observed any other The smaller ionic radius of Mg+2 causes its hydration
phases associated with the silicon addition. It is layer to be bound much more tightly than that of
confirmed that silicon-substitution is to be fully stabi- Ca+2 ion. Therefore, the dehydration of the aqueous
lized inside the lattice structure of the hydroxyapatite as ions, which needs to occur before incorporation
long as the content of the silicon is limited below into a crystalline structure, is much slower for Mg+2
1.97 wt%. ion. In the present study, however, Mg ion is readily
incorporated into the hydroxyapatite structure at
3.5. Evaluation of biocompatibility of Si-substituted levels up to 1 wt% when it co-substitutes with Si
hydroxyapatite ion. The substituted amount of Si in synthesized
Si-HA using our method is also higher than the
The number of cells proliferated in the pure and ion- values reported in the literature [1] within the thermal
substituted hydroxyapatite sample plate and in the stability region. It is attributed to the fact that
control plate was counted as shown in Fig. 10. With our synthetic procedure was keeping away CO2 to
increasing time, the number of cells were gradually prevent producing carbonated hydroxyapatite since Si
increased up to 12  104 cells/ml for the control plate and CO 3 ions are known to replace phosphorus site
and 29  104, 20  104, and 33  104 cells/ml for the competitively [1,11].
pure, Si, Mg-HA II and Si-HA I sample plates, Substitution of silicon into the hydroxyapatite frame-
respectively. Even though the degree of cell proliferation work (Ca10(PO4)6x(SiO4)x(OH)2x) reduces amount of
of Si, Mg-HA II was lower than that of the pure hydroxyl group to compensate for the extra negative
hydroxyapatite, the degree of cell proliferation in the charge of the silicate group. Intensities at 3570 and
sample plates was higher than that in the control one. 630 cm1 of ion-substituted samples drastically decrease
The degree of cell proliferation on the Si, Mg-HA II and with increasing substituted silicon amount compared
Si-HA I gradually increased with time. These results with those of hydroxyapatite spectra. This result clearly
indicate that the ion-substituted hydroxyapatite samples shows that PO4 tetrahedra are replaced by SiO4
have better biocompatibilities than the control one. 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-
4. Discussion 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
Both Ca and P ions in hydroxyapatite structure can agreement with the observed increase in lattice constants
be substituted with various ions and isomorphous for Si-substituted HA. However, the lattice constants
S.R. Kim et al. / Biomaterials 24 (2003) 1389–1398 1397

both for a and c decrease when Ca+2 ions are replaced 5. Conclusion
by Mg+2 in the hydroxyapatite framework. The ionic
radius of Mg+2 (0.69 A) ( is considerably smaller than the To obtain biomaterials having an improved biocom-
one of Ca +2 (
(0.99 A), which is consistent with the patibility, Si, Mg-substituted hydroxyapatites, individu-
observed reduction in lattice constants Si, Mg-substi- ally and in combination, have been prepared and their
tuted HA. The change of the lattice constants of ion- sintering behaviors have been examined. From the XRD
substituted hydroxyapatite clearly demonstrates that analysis, it was confirmed that the single-phase hydro-
Si, Mg ions are structurally incorporated, they do not xyapatites with silicon level 1.97 wt% for singly sub-
just cover the surface of hydroxyapatite. stituted sample kept its original structure with fully
TEM micrographs data show the morphology of the dense sintered body. TEM analysis confirmed that Si-
ion-substituted hydroxyapatites. The crystals of Si, Mg substituted hydroxyapatite consisted of mono-phase
co-substituted hydroxyapatite became larger than those hydroxyapatite grains without any amorphous phase.
of Si mono substituted hydroxyapatite. As previously Co-substitution with magnesium, Si-substitution into
reported [19], Mg inclusion produced larger crystals. hydroxyapatite structure was limited by 1.05 wt%. It
In general, pure hydroxyapatite is known to be was evidenced by XRD analysis at different sintered
thermally stable up to 13001C and above that tempera- temperatures. The results of cell proliferation experi-
ture, it starts to decompose to a- or b-tricalcium ments suggest that the Si, Mg- and Si-substituted
phosphate, CaO, oxyapatite, etc. depending on Ca/P hydroxyapatites have a good biocompatibility and can
ratio of hydroxyapatite [28]. From the XRD analysis, be the useful materials for implants and bone augmen-
1.97 wt% Si-singly substituted hydroxyapatite is ther- tation.
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-
Acknowledgements
tricalcium phosphate (Ca3(PO4)2) and calcium phos-
phate silicate (Ca10(PO4)4(SiO4)2) phases at 11001C.
Financial support of this work was provided by Next
From the microstructural observation and EDAX
Generation Project (Development of Biological Hybrid
analysis of the sample sintered at 12001C using
Materials) of Ministry of Commerce, Industry and
TEM, it was concluded that Si-substituted hydroxyapa-
Energy, Korea The authors would like to thank Dr. Jae-
tite consisted of mono-phase hydroxyapatite grains
hyun Park for her assistance in determination of lattice
without any amorphous phase. Although the sinter-
parameters of the samples using the generalized
ability of Si-substituted hydroxyapatite was significantly
structure analysis system (GSAS).
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- References
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