Ceramics International 44 (2018) 17612–17622

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Ceramics International
journal homepage: www.elsevier.com/locate/ceramint

Silicon-doped hydroxyapatite prepared by a thermal technique for hard T

tissue engineering applications
Maryam Tavafoghia, Joseph M. Kinsellab, Cé Guinto Gamysc, Mathilde Gosselinc,
⁎
Yaoyao Fiona Zhaoa,
a
Department of Mechanical Engineering, McGill University, Montreal, QC, Canada
b
Department of Bioengineering, McGill University, Montreal, QC, Canada
c
Materium Innovations Company, Granby, QC, Canada

A R T I C LE I N FO A B S T R A C T

Keywords: Hydroxyapatite (HA, Ca5(PO4)3OH) has been extensively used for bone implantation due to its similarity to the
Si-doped hydroxyapatite mineral component of bone, which makes it strongly osteoconductive. However, HA has low resorbability, and it
Resorbability is difficult to replace by a newly regenerated bone. Si doping can enhance the resorbability of HA by modifying
Mineralization its crystal structure. Here, we developed a simple thermal technique for preparing Si-doped HA from silica (SiO2)
Hard tissue engineering
and HA precursors, both of which are inexpensive and commercially available. This method included the
physical binding of SiO2 and HA particles, followed by pressing and sintering the mixture at an elevated tem-
perature, which enhanced the atomic diffusion of Si into HA unit cells. We also evaluated the simulated body
fluid (SBF) activity of the Si-doped HA prepared by this technique and showed that it significantly had higher
resorbability and mineralizing potential compared to the pure HA. Our experimental design including, the in-
dividual precipitation and resorption assays enabled us to explain the mechanism behind the improved activity
of Si-doped HA in SBF. This was attributed to the formation of new phases, such as β-tricalcium phosphate (β-
TCP) and calcium silicate (Ca2SiO4) with higher solubility than HA on the SiO2-contating HA during the sintering
stage. This can provide some guidelines for designing new calcium phosphate-based materials for hard tissue
engineering applications.

1. Introduction other hand, is shown to enhance the resorbability of HA by modifying

Hydroxyapatite (HA, Ca5(PO4)3OH) is the main inorganic compo-
nent of bone and teeth. Early studies on the use of HA in hard tissue
engineering go back to the 1950s, and since that time, the synthesis of
HA as a potential material for bone implantation and replacement has
been extensively investigated [1–4]. Synthetic HA has been extensively
used for bone grafting due to its similarity to the inorganic component
of bone, which makes it extremely osteoconductive [5,6]. As opposed to
bioglasses especially borate glasses, HA is highly biocompatible, and it
possesses higher mechanical strength compared to many other bioac-
tive glasses and some types of calcium phosphates. This can make HA a
better candidate for load bearing sites [6]. However, the low resorb-
ability of HA compared to the bioglasses and its difficulty to replace by
a newly generated bone in vivo has been increasingly limited the
clinical use of HA in bone scaffolds [7,8]. In fact, clinical investigations
have shown that HA implants are virtually inert and can remain in body
for approximately six years after implantation [6]. Si doping, on the
its morphology and crystal structure [8–10]. The Si-doped HA is also
shown to have enhanced in vitro [10,11] and in vivo [8,12] bone re-
generation properties and cell proliferation capability [13,14] com-
pared to the stoichiometric HA. In addition, the presence of silicon itself
can promote bone formation and calcification, and it is generally noted
that silicon is essential for the growth and development of biological
tissues, such as bone and teeth [15]. Overall, an ideal set of properties,
such as bioactivity and biocompatibility combined with an acceptable
range of mechanical strength can make Si-doped HA a more competent
osteogenic agent compared to bioactive glasses and biopolymers [8].
Aqueous precipitation techniques [15–19] are commonly used for
synthesizing Si-doped HA. However, these chemical techniques are
complex in terms of optimizing the processing conditions, and usually
have poor ability in controlling the crystallinity of the produced Si-
doped HA particles. Also, they are not efficient for producing Si-doped
HA on a large scale. The Si-doped HA can be alternatively produced by
thermal techniques [20–23], where HA and a Si-containing precursor

⁎
Corresponding author.
E-mail address: yaoyao.zhao@mcgill.ca (Y.F. Zhao).

https://doi.org/10.1016/j.ceramint.2018.06.071
Received 29 March 2018; Received in revised form 9 June 2018; Accepted 9 June 2018
Available online 15 June 2018
0272-8842/ © 2018 Elsevier Ltd and Techna Group S.r.l. All rights reserved.
M. Tavafoghi et al. Ceramics International 44 (2018) 17612–17622

are mixed physically and sintered at temperatures high enough to en- Table 1
hance the atomic diffusion of Si into HA. However, the bone re- The list of samples and their preparation conditions.
generation properties of the Si-doped HA material produced by such Name Component (theoretical values) Processing
thermal techniques has been hardly investigated.
In this work, we investigated the in vitro resorbability and miner- HA HA As received powder
HA-E HA HA immersed in
alizing potential of Si-doped HA produced by a simple thermal tech-
ethanol
nique. This method included the physical mixing of HA and silica (SiO2) HA-G HA HA-E after grinding
powder by the aid of an organic binder, followed by pressing and sin- SiO2 SiO2 As received powder
tering the mixture at an elevated temperature, which enhanced the SiO2-E SiO2 SiO2 immersed in
atomic diffusion of Si into HA unit cells. The sintered compacts were ethanol
SiO2-G SiO2 SiO2-E after grinding
then characterized by a variety of techniques to verify the formation of
HA-GP HA (97 wt%)+ PEG (3 wt%) Green powder
Si-doped HA on these samples. Finally, the compacts were immersed in HA-GC Green compact
simulated body fluid (SBF) to investigate their dissolution and miner- HA-CC HA (100 wt%) + SiO2 (0 wt%) Calcined compact
alization behavior at physiological mimicking conditions. Differently HA-SC Si: 0 wt%, Si: 0 at% Sintered compact
HA-(2% SiO2)- HA (1.9 wt%) + SiO2 ( 95.1 wt%) + Green powder
from other studies on Si-doped HA materials, here we provided a pre-
GP PEG (3 wt%), HA:SiO2 = 98:2 wt%,
cise experimental design, including the individual mineralization and HA-(2% SiO2)- Green compact
dissolution assays. This enabled us to explain in more detail the me- GC
chanism behind the improved bone regeneration activity of Si-doped HA-(2% SiO2)- HA (98 wt%) + SiO2 (2 wt%) Calcined compact
HA. Our goal was to develop a simple and cost-effective technique for CC
HA-(2% SiO2)- Si: 0.94 wt%, Si: 0.77 at% Sintered compact
preparing HA-based materials with improved bone regeneration prop-
SC
erties for hard tissue engineering applications. SiO2-GP SiO2 (97 wt%)+ PEG (3 wt%) Green powder
SiO2-GC Green compact
2. Materials and method SiO2-CC HA (0 wt%) + SiO2 (100 wt%) Calcined compact
SiO2-SC Si: 46.76 wt%, Si: 33.33 at% Sintered compact

2.1. Materials

HA powder (Ca5(PO4)3OH, purity ≥ 95%) was purchased from
Berkeley Advanced Biomaterials Inc., and amorphous silica micro-
spheres (SiO2, purity > 95%) were received from Materium
Innovations (MATSPHERE™MAT300). More details about Materium
company can be found in the affiliation. Materium also assisted in
characterizing the initial SiO2 and HA powder. The HA and silica par-
ticle sizes were 50–150 and 5–40 µm, and their surface areas were 10
and > 100 m2/g, respectively. Polyethylene glycol 8000 (PEG, purity
≥ 95%) and ethanol (≥ 99.5%) were purchased from Fisher Scientific.
2.2. Preparing SiO2-containing HA powder
Adequate amounts of as-received silica (3 g) and HA powder (147 g)
with SiO2:HA weight ratio of 2:98 were immersed in 250 ml of ethanol.
4.6 g (resulting in 3 wt% of total weight) of a binder, PEG, was dis-
solved in 30 ml of deionized (DI) water and then added to the HA and
silica mixture to bind the silica and HA particles together. The same
procedure was followed for 150 g of as-received SiO2 and HA powders
as control samples, and the binder was also added to keep consistency
with the SiO2-containing HA powder. The solutions were then kept
stirring on a magnetic stirrer with the rate of 500 rpm until the ethanol
evaporated partially and then they were dried in the oven at 60 °C
overnight. The dried mixtures were ground manually by a mortar and
pestle to obtain green powders (GPs), HA-GP, HA-(2% SiO2)-GP, and
SiO2-GP as shown in Table 1. To investigate the changes in the mor-
phology of powders during this process, the same procedure was fol-
lowed for as-received HA and SiO2 powder without adding any binder.
These powders were called SiO2-EtOH and HA-EtOH after immersion,
and SiO2-G and HA-G after grinding.
2.3. Si-doped HA preparation
Cylindrical compacts (15 mm in diameter, and 6–7 mm in thickness
depending on the loose density of initial powders) were prepared by
uniaxial pressing at 5 metric ton in a steel die using 2 g of each HA-GP,
HA-(2% SiO2)-GP and SiO2-GP powder. The compacts were calcined at
400 °C for 5 h in air to remove the binder and then sintered in an
electrical furnace (ExOne) at 1100 °C for 8 h in air at a heating and
cooling rate of 10 °C/min. The weight of compacts was measured by a
digital analytical balance, and the dimensions were measured by a di-
gital caliper. Absolute (ρ) and relative (ρr) density of compacts were
calculated according to the following equations for at least 3 samples
for each set, and the values were average.
ρ = m/v (1)
ρr = ρ/ρth (2)
ρth = (ρth of SiO2) × (wt% of SiO2) + (ρth of HA) × (wt% of HA) (3)
where, m and v are mass and volume of compacts, respectively; ρth is
theoretical density of compacts, and ρth of HA and SiO2 is 3.156 and
2.648 g/cm3, respectively.
2.4. Mineralization and dissolution experiments
SBF was prepared according to a well-known recipe provided by
Kokubo et al. [24]. Also, to evaluate HA dissolution, a Ca- and P-defi-
cient SBF solution (so called DSBF) was prepared by eliminating Ca-
and P-containing precursors from SBF. It should be noted that Ca
(2.5 mM) and P (1 mM) concentration in SBF is very low [24], and the
elimination of these ions would not have a significant impact on the
ionic strength (0.16 vs 0.15 for SBF and DSBF, respectively) and ki-
netics of calcium phosphate dissolution. The ionic strength in body fluid
is mainly regulated by NaCl, significant amount of which (137.5 mM)
was present in both SBF and DSBF. Also, both SBF and DSBF contained
a buffer reagent (tris, 50.5 mM), which kept the pH constant at about
7.4 during the course of reaction. The sintered compacts prepared by
pellet-pressing (SC samples) were immersed in 40 ml of SBF or DSBF
solution in centrifugation tubes and the tubes were placed in an in-
cubator at 37 °C for 15 days. The solutions were agitated gently every 2
days. The samples were finally removed from the incubator, washed 3
times with DI water, and dried in the oven at 60 °C.

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M. Tavafoghi et al. Ceramics International 44 (2018) 17612–17622

Fig. 1. BSE-SEM (b and c) and SEM (a and d–l) images of HA-(2% SiO2)-GP (a-c), HA (d and e), HA-E (f), HA-G (g), HA-GP (h), SiO2 (i),SiO2-E (j), SiO2-G (k), and
SiO2-GP (l) samples. The red and blue arrows on panel b and c show the HA and SiO2 particles, respectively, that were identified by EDS analysis. (For interpretation
of the references to colour in this figure legend, the reader is referred to the web version of this article.)

2.5. Characterizations
2.5.1. Inductively coupled plasma atomic emission spectroscopy (ICP-AES)
Ca, P, and Si concentration in the SBF and DSBF solutions con-
taining HA-SC, HA-(2% SiO2)-SC, and SiO2-SC samples were measured
at different incubation times using an ICP-AES instrument (ICAP 6500
Duo). 1 ml aliquots were taken from the SBF and DSBF solutions at the
desired incubation times, filtered with a syringe filter with a pore size of
100 nm to remove any trace of precipitates or particles released from
the samples, and then immediately diluted 5 times with 4% nitric acid.
The dilution with 4% nitric acid was to make sure that no more pre-
cipitate formed in the solutions after filtration. The Ca, P, and Si con-
centrations were measured at the wavelength of 317.9, 178.3, and
212.4 nm, respectively. Dissolved amounts of Ca (Cad) and P (Pd) at
different incubation times were measured from DSBF solution that did
not contain any Ca and P-containing precursors. To keep consistency,
the dissolved amounts of Si (Sid) were also measured from DSBF solu-
tion. For this, the initial amounts of Si (SiiDSBF), P (PiDSBF), and Ca
(CaiDSBF) in blank DBSF were subtracted from the measured amounts of
these elements at different incubation times in DSBF. The precipitated

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amounts of Ca (Cap) and P (Pp) were also calculated according to the
following equations,
Cap (mol) = CaiSBF + Ca d − CaSBF (4)
Pp (mol) = PiSBF + Pd − PSBF (5)
where CaiSBF and PiSBF are the initial amounts of Ca and P in blank SBF,
and CaSBF and PSBF are the measured amounts of Ca and P in SBF so-
lution at different incubation times.
Also, the dissolution and mineralization of compacts after 14 day
immersion in DSBF and SBF, respectively, were investigated by calcu-
lating the compacts weight changes according to the following equa-
tions,
(mDSBF − m s)
Dissolution(%) = × 100
mS (6)
mSBF + (ms − mDSBF) − m s
Mineralization(%) = × 100
mS (7)
where ms is the mass of sintered compacts (SCs), and mDSBF, and mSBF
are the mass of SCs after 14 day immersion in DSBF and SBF, respec-
tively.
2.5.2. Fourier transform infrared (FT-IR) spectroscopy
IR spectra were recorded on a Bruker Tensor 27 FT-IR spectrometer
using diffuse reflectance (DRIFT) mode. The samples were diluted with
KBr to approximately 10 wt/wt%. Pure KBr powder was used as a
background. The FT-IR spectra were recorded from 600 to 4000 cm−1
using a deuterated triglycine sulfate (DTGS) detector. The spectra were
collected by averaging 256 scans at 4 cm−1 resolution.
2.5.3. X-ray diffraction (XRD)
XRD spectra were collected using a Bruker AXS XRD instrument
with Cu Kα radiation generated at 40 kV and 40 mA. A quartz and
metallic sample holder was used for the powders and compacts, re-
spectively. Diffraction angles collected (2θ) ranged from 5° to 54°, with
step size of 0.05°.
2.5.4. X-ray photoelectron spectroscopy (XPS)
XPS measurements were performed using a monochromatic X-ray
photoelectron spectrometer K Alpha (Thermo Scientific). The setup was
equipped with an Al Kα X-Ray source (1486.6 eV, 0.834 nm), a micro-
focused monochromator and an ultrahigh vacuum chamber (10−9
Torr). The survey scans were collected with energy steps of 1 eV and
spot size of 400 µm. Scans were taken on at least 3 points on each
sample and the quantitative results were averaged. A flood gun was
used to neutralize electrical built-up charge generated. Peak fitting and
quantitative analysis of survey spectra were performed using the soft-
ware Thermo Avantage (version 4.60).
2.5.5. Scanning electron microscopy (SEM)
The morphology of samples was analyzed with a scanning electron
microscope (SEM) from Hitachi (S-4700 FE-SEM) using variable pres-
sure (VP) mode and an acceleration voltage of 10 kV. Energy dispersive
spectrometer (EDS) was used for the elemental analyses. EDS spectra
were collected on at least 5 points on each sample and the quantitative
results were averaged. The specimens were mounted on a double sided
conductive carbon tape and were not metal coated.
2.6. Statistical analysis
All quantitative results are shown as standard deviation of the
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Ceramics International 44 (2018) 17612–17622
mean. Statistical analyses were performed by Student's T-Test with
p < 0.05 considered significant.
3. Results and discussions
Si was doped into HA by sintering, and the effect of Si doping on the
sinterability and SBF activity of HA was studied. A wide range of SiO2
concentrations from 0 to 100 wt% were investigated, and it was shown
that 2 wt% SiO2 was the optimum concentration, where the SBF activity
in terms of the changes observed in Ca and P concentration was sig-
nificantly different from 0 wt% SiO2 (Fig. S1), while the mechanical
properties of compacts were not yet deteriorated (Fig. S2). Therefore,
this concentration of SiO2 was chosen for the rest of the analyses.
3.1. The homogeneity of SiO2-containing HA powder
SiO2-containing HA powder (HA-(2% SiO2)-GP) was prepared ac-
cording to a technique explained in Section 2.2. SEM images of HA-(2%
SiO2)-GP are shown in Fig. 1a–c. The dark gray particles observed more
clearly on the BSE-SEM images (see blue arrows on Fig. 1b and c) were
identified by EDS (3 spectra at least) to be SiO2 and the lighter particles
(see red arrows on Fig. 1c) were HA. The quantitative EDS analysis was
performed on the circular area of 25 µm in diameter, and the results
indicated that within this range, Si was homogeneously dispersed in HA
at a concentration of 0.8 ± 0.1 at%, which was very close to theore-
tical concentration of Si at 0.77 at%.
Overall, the particles observed on the green powders, HA-(2%
SiO2)-GP (Fig. 1a–c), HA-GP (Fig. 1h), and SiO2-GP (Fig. 1l), were
much smaller (~0.5–12 µm) than those observed on the as-received
SiO2 (~25 µm) (Fig. 1i) and HA (~100 µm) (Fig. 1d and e) powders. To
investigate the changes in the morphology and size of particles during
the preparation of green powders, the pure (no binder was added) HA
and SiO2 powders seperately underwent the same procedure as the
green powders. The results indicated that only immersing and stirring
the HA (Fig. 1f) and SiO2 (Fig. 1j) powders in ethanol was enough to
break down the particles and make them smaller, while no significant
changes were observed in the ground HA (Fig. 1g) and SiO2 (Fig. 1k)
compared to the ethanol-immersed powders. The presence of large
aggregates on these samples could be attributed to electrostatic forces
usually observed between small particles.
3.2. Effect of SiO2 addition on the morphology and sinterability of HA
compacts
The green compacts (GC samples) were made by pressing the HA-
GP, HA-(2% SiO2)-GP, and SiO2-GP powders listed in Table 1. These
compacts were then calcined (CC samples) to remove the organic
binder and sintered (SC samples) at 1100 °C for 8 h to make the com-
pacts stronger. In this section, we show the changes in the morphology
and composition of compacts during these thermal processes, and also
investigate the effect of SiO2 addition on the sinterability of the HA
compacts. Sintering at elevated temperatures can also make the Si
atoms become incorporated into the HA lattice [10,17]. This will be
discussed in detail in Section 3.3.
The SEM images of CC and SC samples containing 0, 2, and 100 wt%
of SiO2 are shown in Fig. 2. While the morphology of particles in the HA
compact did not change considerably during sintering (Fig. 2a and b),
the SiO2 compact underwent significant changes during the sintering
stage (Fig. 2e and f), where closely packed particles and well defined
grain boundaries were observed on SiO2-SC (Fig. 2f). This indicated a
higher sinterability of SiO2 compared to HA powder, which is also
confirmed by the density results shown in Fig. 3: both the absolute
M. Tavafoghi et al. Ceramics International 44 (2018) 17612–17622

Fig. 2. SEM images of HA-CC (a), HA-SC (b), HA-SC immersed in SBF (c), HA-SC immersed in DSBF (d), SiO2-CC (e), SiO2-SC (f), SiO2-SC immersed in SBF (g), SiO2-
SC immersed in DSBF (h), HA-(2% SiO2)-CC (i), HA-(2% SiO2)-SC (j), HA-(2% SiO2)-SC immersed in SBF (k), and HA-(2% SiO2)-SC immersed in DSBF (l). The blue
arrows on panel i show the SiO2 particles that were identified by EDS analysis. (For interpretation of the references to colour in this figure legend, the reader is
referred to the web version of this article.)

(Fig. 3a) and relative (Fig. 3b) density of SiO2 compact increased
drastically after sintering while the density of the HA compact did not
change significantly. Also, adding 2 wt% SiO2 to HA did not help im-
prove the sinterability of this powder (Fig. 3a and b): both the absolute
(2.05 vs 2.16 g/cm3) and relative (65.3 vs 68.4%) density measured on
HA-(2% SiO2)-SC was lower than on HA-SC. This could be attributed to
the low concentration of SiO2, or to the fact that SiO2 was not homo-
genously dispersed in HA. In fact, SiO2 aggregates of ~ 15 µm in size
were observed on HA-(2% SiO2)-CC (Fig. 2i); however, these aggregates
disappeared after sintering on HA-(2% SiO2)-SC (Fig. 2j). This was
investigated in more detail by SEM-EDS (Fig. 4a) and XPS (Fig. 4b)
compositional analyses that were performed on the compacts surfaces.
While Si content in HA-(2% SiO2)-GP was estimated to be
0.8 ± 0.1 at% by EDS, the Si concentration within the same analysis
range was higher at 2.0 ± 0.2 and 2.8 ± 0.3 at% on the surface of HA-
(2% SiO2)-GC and HA-(2% SiO2)-CC, respectively. This indicated the
homogeneity of SiO2 within the HA-(2% SiO2)-GP was disrupted during
the compaction and calcination stage and it became more concentrated
on the surface likely due to the fact that SiO2 has a lower density than
HA (2.65 vs 3.16 g/cm3). However, the Si content on HA-(2% SiO2)

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M. Tavafoghi et al.
decreased from 2.8 ± 0.3 to 1.8 ± 0.1 at% after sintering (Fig. 4a),
which indicated that sintering at an elevated temperature (1100 °C)
could make the SiO2 migrate from the surface into the compacts. To
confirm this, XPS analysis, which is more sensitive to surface compo-
sition than EDS, was performed. As expected, the Si concentration
measured by XPS (Fig. 4b) on HA-(2% SiO2)-CC was significantly
higher than the one measured by EDS (Fig. 4a). However, consistent
with the EDS results, XPS results also confirmed that the Si measured on
HA-(2% SiO2)-SC (4.2 ± 0.1 at%) was lower than on HA-(2% SiO2)-CC
(10.1 ± 0.6 at%) indicating the effect of sintering on improving the Si
homogeneity. These compositional analyses indicated that the homo-
geneity of Si distribution on HA-(2% SiO2)-GP decreased during the
compaction process; however, sintering helped to improve the homo-
geneity of the compacts, but Si was still more concentrated on the
surface than inside the compacts.
Overall, the results reported in the literature indicate that Si addi-
tion has an adverse effect on the sinterability of HA scaffolds
[18,25,26]. Our results as well confirmed this observation; however,
the deteriorating effect of 2 wt% SiO2 on sinterability of HA was neg-
ligible here (Fig. 3). The densities obtained for HA-SC (2.16 ± 0.03 g/
cm3) and HA-(2% SiO2)-SC (2.05 ± 0.02 g/cm3) samples were both
significantly higher than the mineral density of trabecular bone
(~ 0.2 g/cm3) [27], and were more comparable to that of cortical bone
(~ 2.0 g/cm3) [28]. Yet, the relative densities obtained for HA-SC
(68.4 ± 0.8%) and HA-(2% SiO2)-SC (65.3 ± 0.8%) were still lower
than the maximum density of ~ 98% reported for HA-based compacts
prepared under the same conditions [15,18]. This can be attributed to
the low sinterability of our HA-GP powder, which had large particle
sizes and diverse morphologies.
3.3. Si-doping into HA by sintering
The SiO2 and HA powders were mixed physically, so only the
characteristic features related to these two materials were expected to
be present on the HA-(2% SiO2) samples. However, sintering the sam-
ples at an elevated temperature could enhance the atomic diffusion of Si
into the HA lattice, thus resulting in the formation of new crystalline
phases according to the following reactions [10,17],
Ca10 (PO4)6 (OH)2 +xSiO44 − → Ca10 (PO4)6 − x (SiO4 ) x (OH)2 − x + xPO34−
+xOH−
x
Ca10 (PO4)6 − x (SiO4 ) x (OH)2 − x → ⎛1− ⎞ Ca10 (PO4 )6 (OH)2 +xCa3 (PO4 )2
⎝ 2⎠
+xCa2SiO4
The formation of new phases on the samples during sintering was
investigated by IR (Fig. 5a) and XRD (Fig. 5b) techniques. Table 2
shows the assignment of peaks found on FTIR spectra recorded on HA-
CC, HA-SC, HA-(2% SiO2)-CC, HA-(2% SiO2)-SC, SiO2-CC, and SiO2-SC
samples [29–36]. The HA sample (Fig. 5a, i and ii) did not undergo
significant changes during sintering; only a small peak at 716 cm−1
corresponding to OH [35] appeared on HA-SC, which could be attrib-
uted to the increase in crystallinity HA by sintering [37]. However,
SiO2-SC (Fig. 5a, vi) showed more significant changes compared to
SiO2-CC (Fig. 5a, v): overall, the peaks became sharper on SiO2-SC.
Also, the δ Si-O-Si band at 812 cm−1 on SiO2-C shifted to 795 cm−1 on
SiO2-SC, and a small peak at 623 cm−1 corresponding to δ Si-O-Si ap-
peared on SiO2-SC [33]. The δ Si-O-Si bands at 623 and 795 cm−1
observed on SiO2-SC are the features of silicate chains ((SiO3)∞) [34]
and can be attributed to the formation of crystalline silica after sin-
tering.
Ceramics International 44 (2018) 17612–17622
HA-(2% SiO2)-CC (Fig. 5a, iii) showed almost all characteristic
peaks relative to its HA component and it also showed some major
peaks of amorphous silica at 812, 1105, and 1215 cm−1. However,
upon sintering (Fig. 5a, iv), the peak at 812 cm−1 shifted to 795 cm−1
on HA-(2% SiO2)-SC due to the formation of crystalline silica. Also, the
peak at 1215 cm−1 corresponding to ν Si-O-Si [36] in silica dis-
appeared, and the peak at 962 cm−1 originating from ν1 PO4 [29] in HA
became weaker. Instead, some new peaks emerged at 498 and
947 cm−1 on HA-(2% SiO2)-SC, which could be attributed to δ SiO4 and
ν SiO4, respectively [34,38–40]. These IR results indicated the decrease
in the SiO2 content of HA-(2%SiO2)-SC sample due to the Si diffusion
into HA during sintering and the formation of SiO44- groups, which
substituted for PO43- in HA. This could result in the formation of Si-
doped HA (Ca10(PO4)6-x(SiO4)x(OH)2-x) according to Eq. (8)
[10,17,38,39]. The lower concentration of Si on the surface of HA-(2%
SiO2)-SC compared to HA-(2% SiO2)-CC (Fig. 4) can further confirm
this hypothesis. SiO44- observed on HA-(2%SiO2)-SC could also origi-
nate from calcium silicate (Ca2SiO4) that can form following the de-
composition of Si-doped HA (Eq. (9)) [10]. However, IR results did not
evidently show the formation of TCP, which is also expected to form
according to Eq. (9). To further investigate the formation of new
crystalline phases during sintering, XRD analysis (Fig. 5b) was per-
formed.
Consistent with the IR results, the XRD spectra for the HA sample
did not change significantly during sintering (Fig. 5b, i and ii), while
the initially amorphous SiO2 became completely crystalline (Fig. 5b, v
and vi). This was evident from the sharp peaks on SiO2-SC, which
matched well the XRD reference pattern for crystalline silica. HA-(2%
SiO2)-CC (Fig. 5b, iii) only showed the peaks relative to HA because
SiO2 was amorphous at this stage; however, HA-(2% SiO2)-SC (Fig. 5b,
iv) showed both the HA and SiO2 peaks since SiO2 became crystalline
during sintering. HA-(2% SiO2)-SC also showed some relatively strong
peaks corresponding to beta-tricalcium phosphate (β-TCP) and calcium
silicate as expected to form according to Eq. (9) following the decom-
position of Si-doped HA [10,17]. In fact, the XRD peaks of HA are ex-
pected to shift to lower angles upon the incorporation of Si into the HA
lattice [41]. However, since such changes were not evident on our high
resolution XRD spectra (Fig. S3), it can be deduced that the Si-doped
HA was totally decomposed into TCP and calcium silicate during the
relatively long sintering time (8 h).
(8)
3.4. Effect of SiO2 addition on SBF activity of HA
Fig. 6a–g shows the amount of Ca, P, and Si that are dissolved from
the compacts in DSBF (Fig. 6a–d), or are removed from SBF due to the
(9)
calcium phosphate precipitation on the compacts (Fig. 6e–g), and
Fig. 6h shows the Ca/P ratio in the precipitates. These values are
measured by ICP according to a technique that is discussed in detail in
Section 2.5.1. The total dissolution and mineralization were also in-
vestigated by measuring the changes in the weight of compacts after
immersion in DSBF or SBF, respectively (Fig. 6i).
3.4.1. HA dissolution
HA-(2% SiO2)-SC showed significantly higher Ca dissolution than
HA-SC that did not contain any silica (Fig. 6a). However, P dissolution
on HA-(2% SiO2)-SC was less than on HA-SC (Fig. 6b), but it should be
considered that P dissolution for all samples was almost negligible
(< 1 µmol). HA-(2% SiO2)-SC also showed a significantly higher Si
dissolution compared to HA-SC (Fig. 6c). The Si dissolution on HA-(2%
SiO2)-SC was even faster than the pure silica samples, SiO2-SC. Overall,
HA-(2% SiO2)-SC showed the fastest dissolution rate in DSBF, followed
by HA-SC and SiO2-SC (Fig. 6d and i). The faster dissolution rate of HA-
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M. Tavafoghi et al.
Fig. 3. Absolute (a) and relative (b) density of HA, HA-(2% SiO2), and SiO2
compacts after calcination (CC samples) and sintering (SC samples). Asterisk
signs (*) show the values that were statically significantly different with
P < 0.05.
Fig. 4. Si at% measured on HA, HA-(2% SiO2), and SiO2 samples after calci-
nation (CC samples), sintering (SC samples), and immersion in SBF and DSBF by
either SEM-EDS (a) or XPS (b). Asterisk signs (*) show the values that were
statically significantly different with P < 0.05.
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Ceramics International 44 (2018) 17612–17622
(2% SiO2)-SC was also evident from the SEM images (Fig. 2). While the
surface morphology of HA-SC (Fig. 2d) and SiO2-SC (Fig. 2h) immersed
in DSBF was not significantly different from HA-SC (Fig. 2b) and SiO2-
SC (Fig. 2f), the surface of HA-(2% SiO2)-SC (Fig. 2j) became rougher
and pitted after immersion in DSBF (Fig. 2l).
3.4.2. Calcium phosphate precipitation
Precipitation results in Fig. 6e–g and i indicated that the calcium
phosphate precipitate formed on all samples; however, precipitation on
SiO2-SC was almost negligible. This was also evident from the SEM
image of the SiO2-SC immersed in SBF (Fig. 2g). The rate of calcium
phosphate precipitation on HA-(2% SiO2)-SC was significantly higher
than on HA-SC; however, the particles of the newly formed calcium
phosphate were not distinguishable on the SEM images of either HA-
(2% SiO2)-SC (Fig. 2k) or HA-SC (Fig. 2c) immersed in SBF. This is
while, spherulitic HA particles are usually expected to form on biolo-
gical surfaces immersed in SBF [11,17,30]. This could be attributed to
the fact that the precipitates that formed on the samples were removed
during the washing stage, or possibly the precipitating particles here
had a similar morphology to those already present on the compacts,
thus making them undistinguishable. However, the XPS results in
Fig. 4b showed that Si concentration on HA-(2% SiO2)-SC immersed in
SBF was significantly lower than on both the HA-(2% SiO2)-SC, and the
HA-(2% SiO2)-SC immersed in DSBF. This could be attributed to the
presence of a layer of newly formed calcium phosphate on HA-(2%
SiO2)-SC immersed in SBF [42]. Therefore, the latter scenario seems to
be more likely to happen considering the fact that the formation of
similar particles to those on HA-SC and HA-(2% SiO2)-SC is also re-
ported by other authors on the Si-doped HA scaffolds placed in SBF
[10,11,42,43].
3.4.3. How Si doping can improve HA resorbability and mineralization?
It is shown that Si doping, even in a small amount at ~ 0.8 wt%, can
significantly improve the SBF activity of HA [10,11,16,38,42,44].
Consistent with these results, we showed that our Si-doped HA, pro-
duced by sintering the 2 wt% SiO2-containng HA compacts, had higher
SBF activities than the pure HA (Fig. 6). Such SBF activity tests are
widely used for predicting the bioactivity of biomaterials in vivo [16].
However, SBF studies on Si-doped HA have rarely been able to distin-
guish between the mineralizing and dissolving phases [10,11,42].
Moreover, these studies merely reported the changes in [Ca] and [P] in
SBF rather than measuring the amounts of precipitating and dissolving
Ca and P [10,11,38,42]. Introducing a Ca- and P-deficient solution (so
called DSBF) enabled us to investigate the dissolution, which was fol-
lowed by some simple calculations (explained in detail in Section 2.5.1)
to measure the amounts of dissolving and precipitating Ca and P. This
allowed us to better understand the mechanism of calcium phosphate
mineralization or dissolution in the presence of Si-doped HA in SBF. For
example, the higher resorbability of Si-doped HA is usually attributed to
the partial transformation of HA to TCP with a higher resorbability than
HA [10]. However, our results showed that while Ca dissolution was
significantly higher for HA-(2% SiO2)-SC than for HA-SC, the P dis-
solution for both of these samples was comparable and almost negli-
gible. This indicated that the higher solubility of HA-(2% SiO2)-SC
cannot be attributed to the faster dissolution of TCP formed on this
sample during sintering (Fig. 5b). On the other hand, XRD results
(Fig. 5b) showed that other phases, such as Ca2SiO4 were also present
on HA-(2% SiO2)-SC. Ca2SiO4 (5.8 ×10−4 M at 20 °C) has a higher
solubility than both HA (3.2 ×10−5 M at 37 °C) and TCP (1.6 ×10−4 M
at 37 °C) [45], and considering the significantly higher amounts of Ca
and Si dissolution for HA-(2% SiO2)-SC than for HA-SC (Fig. 5a and c),
it can be concluded that the dissolution of Ca2SiO4 was mainly re-
sponsible for the higher resorbability of HA-(2% SiO2)-SC. Also, the Ca/
Si ratio of 1.6 ± 0.3 measured based on the dissolved Ca and P (Fig. 6a
and c) for HA-(2% SiO2)-SC after 14 day incubation was close to that
for Ca2SiO4 (2), which further confirms the dissolution of Ca2SiO4.
M. Tavafoghi et al. Ceramics International 44 (2018) 17612–17622

Fig. 5. FTIR spectra (a) and XRD pat-

terns (b) recorded on HA-CC (i), HA-SC
(ii), HA-(2% SiO2)-CC (iii), HA-(2%
SiO2)-SC (iv), SiO2-CC (v), and SiO2-SC
(vi) samples. The numbers in black and
blue on panel a indicate the peaks ori-
ginating from HA and SiO2, respectively,
and the numbers in red indicate new
phases that were formed during the
sintering stage and are discussed in de-
tail in Section 3.3. The XRD patterns are
compared with reference spectra for HA
(black lines, PDF card n. 00-009-0432),
SiO2 (blue lines, PDF card n. 00-041-
0539), β-TCP (red lines, database code
AMCSD 0007447), and β-Ca2SiO4 (green
lines, database code AMCSD 0020214).
(For interpretation of the references to
colour in this figure legend, the reader is
referred to the web version of this ar-
ticle.)

Table 2 glasses can act as intermediating crystals for the formation of bone like
Assignment of peaks found on FTIR spectra shown in Fig. 5 for HA-CC, HA-SC, crystals [47–49], and this too can be responsible for the faster miner-
HA-(2% SiO2)-CC, HA-(2% SiO2)-SC, SiO2-CC, and SiO2-SC. alization of HA-(2% SiO2)-SC sample. In fact, Si is an essential trace
Peaks Wavenumbers (cm−1) element in many organs of animals, and its presence plays a significant
role in bone mineralization through promoting collagen type 1 synth-
HA-CC HA-SC HA-(2% HA-(2% SiO2-CC SiO2-SC esis and enhancing osteoblast differentiation [6,16,50,51]; therefore, Si
SiO2)-CC SiO2)-SC dissolution can also play a role in improving HA mineralization at in
ν2 PO4 432 474 432 474 – – vivo conditions.
474 474 To identify the precipitating calcium phosphate, the Ca/P ratios
δ Si-O-Si – – – – 476 476 (Fig. 6h) were measured for the precipitates based on the amount of
δ SiO4 – – – 498 – – precipitating Ca and P shown in Fig. 6e and f. The Ca/P for HA-SC was
ν4 PO4 575 575 575 575 – –
almost constant at 1.8 ± 0.1 during the whole incubation period in
604 604 604 604
635 635 635 635 SBF. This Ca/P ratio can be attributed to the formation of carbonated
OH – 716 – 716 – – HA (CHA), which has Ca/P ratios ranging from 1.67 to 2 depending on
δ Si-O-Si – – 812 – 812 – its carbonate content [52]. The Ca/P for HA-(2% SiO2)-SC was
δ Si-O-Si in – – – 795 – 795
2.0 ± 0.1 after 3 day incubation and it increased to 2.2 ± 0.1 after 15
(SiO3)∞ 623
ν1 CO3 891 891 891 891 – – days, which could be attributed to the incorporation of more carbonate
ν SiO4 – – – 947 – – ions into the HA precipitate forming on this sample. The Ca/P was
ν1 PO4 962 962 962 962 – – significantly high for SiO2-SC at 3 day incubation (8.2 ± 0.7) and it
ν3 PO4 1028 1028 1028 1028 – – decreased to 2.7 ± 0.2 after 15 days. The high Ca/P ratio here could be
1057 1057 1057 1057
attributed to the formation of Ca(OH)2 derivatives [53] on SiO2-SC;
1092 1092 1092 1092
ν Si-O-Si – – 1105 1105 1105 1105 however, it should be considered that the precipitation on SiO2-SC was
1215 1215 1215 almost negligible (Figs. 2g and 6g).
To further evaluate the precipitating material, XRD analysis was
performed on the samples after immersion in SBF (Fig. S5). No calcium
However, the XRD patterns of HA-(2% SiO2)-SC immersed in SBF or phosphate phases were detected on the XRD pattern of SiO2-SC im-
DSBF (Fig. S4) did not show any decline in the peaks related to Ca2SiO4 mersed in SBF, which indicates the precipitation on SiO2-SC was not
due to the fact that the main peaks of Ca2SiO4 overlaps with HA. significant. Also, the XRD patterns of HA-SC and HA-(2% SiO2)-SC did
Total calcium phosphate precipitation also was higher for HA-(2% not show significant changes after immersion in SBF. This can indicate,
SiO2)-SC than for HA-SC (Fig. 6g). This can be explained by the higher the newly formed calcium phosphates (as were already confirmed by
dissolution rate of these samples, which could increase the local con- ICP analyses in Fig. 6f and g) were probably HA or TCP, which were
centration of Ca in SBF and favor calcium phosphate mineralization on already present on the samples. However, the precipitation of TCP from
the samples [11,46]. Also, it is shown that the silane groups in bioactive SBF is not favored at physiological conditions [24]. Also, the Ca/P

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M. Tavafoghi et al. Ceramics International 44 (2018) 17612–17622

Fig. 6. Dissolution and mineralization of HA-SC, HA-(2% SiO2)-SC, and SiO2-SC scaffolds immersed in DSBF and SBF, respectively, at different incubation times
evaluated by ICP: (a, b, c, d) Ca (Cad), P (Pd), Si (Sid), and total (Cad + Pd + Sid) dissolution of scaffolds in DSBF; (e, f, g) Ca (Cap), P (Pp), and total calcium phosphate
(Cap + Pp) precipitation from SBF. (h) Ca/P ratio in the precipitated phase. (i) scaffold weight changes after 14 day immersion in DSBF or SBF corresponding to
dissolution or mineralization, respectively. Asterisk (*) and plus (+) signs indicate the values that are statically significantly different from the correspondent values
measured on HA-SC and SiO2-SC, respectively with P < 0.05.

ratios (Fig. 6h) measured for HA-SC (~ 1.8) and HA-(2% SiO2)-SC
(2–2.2) were closer to that for CHA (1.67–2) than for TCP (1.5). This
can indicate the formation of HA on these samples, which is also con-
firmed by other researches to form on HA-based scaffolds immersed in
SBF [54,55].
Overall, HA-(2% SiO2)-SC showed significantly improved both the
resorbability and mineralizing potential compared to the pure HA in
physiological conditions (Fig. 6), and this would make the Si-doped HA
a more ideal candidate than HA for hard tissue engineering applica-
tions, where both the scaffold resorbability and regenerability is re-
quired. The dissolution and mineralization rate of ~ 0.001 wt% h−1
obtained for HA-(2% SiO2)-SC sample was comparable to the values
reported for the Si-doped HA produced by aqueous precipitation tech-
niques and tested in vitro [56]; however, they were approximately 30
times lower than when the bioactivity tests were conducted in in vivo
conditions [16,57]. Also, the resorbability of HA-(2% SiO2)-SC was still
significantly lower than some bioglasses such as borosilicate and borate
bioactive glasses that showed dissolution rate of 0.06 and 0.3 wt% h−1,
respectively, in SBF [58]. However, it should be noted that the low
degree of porosity in our Si-doped HA samples produced by a com-
paction technique could also be responsible for its lower activity at
biological conditions.
4. Conclusion
In this work, we shed some light on the mechanism under which Si
doping can improve the resorbability and mineralizing potential of HA.
This can provide some guidelines for designing new HA-based materials
to be used for hard tissue engineering applications.
We showed the Si-doped HA prepared by sintering 2 wt% SiO2-

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contating HA powder had both the higher mineralization and dissolu-
tion rate in SBF compared to the pure HA. This is while the deterior-
ating effect of 2 wt% SiO2 on sinterability and mechanical properties of
HA compacts was almost negligible. The enhanced SBF activities of 2 wt
% SiO2-contating HA were attributed to the formation of Si-doped HA
on this sample at elevated sintering temperatures. The decomposition of
Si-doped HA consequently led to the formation of new phases, such as
β-TCP and Ca2SiO4, which had higher water solubility than HA. Also,
the presence of silane groups on Si-doped HA could be responsible for
the faster mineralization of this sample.
Acknowledgment
This research project is supported by the Natural Science and
Engineering Research Council of Canada (NSERC) EGP 469410-2014.
Also, we would like to thank Prof. Marta Cerruti from Materials
Engineering, McGill University for providing access to a FT-IR instru-
ment.
Appendix A. Supplementary material
Supplementary data associated with this article can be found in the
online version at http://dx.doi.org/10.1016/j.ceramint.2018.06.071.
References
[1] A. Perloff, A.S. Posner, Preparation of pure hydroxyapatite crystals, Science 124
(3222) (1956) 583–584.
[2] M.I. Kay, R.A. Young, A.S. Posner, Crystal structure of hydroxyapatite, Nature 204
(4963) (1964) 1050–1052, http://dx.doi.org/10.1038/2041050a0.
[3] M.T. Jahromi, G. Yao, M. Cerruti, The importance of amino acid interactions in the
crystallization of hydroxyapatite, J. R. Soc. Interface 10 (80) (2012), http://dx.doi.
org/10.1098/rsif.2012.0906.
[4] M. Tavafoghi Jahromi, M. Cerruti, Amino acid/ion aggregate formation and their
role in hydroxyapatite precipitation, Cryst. Growth Des. 15 (3) (2015) 1096–1104.
[5] J.R. Woodard, et al., The mechanical properties and osteoconductivity of hydro-
xyapatite bone scaffolds with multi-scale porosity, Biomaterials 28 (1) (2007)
45–54.
[6] G. Kaur, O.P. Pandey, K. Singh, D. Homa, B. Scott, G. Pickrell, A review of bioactive
glasses: their structure, properties, fabrication and apatite formation (in Eng), J.
Biomed. Mater. Res A 102 (1) (2014) 254–274.
[7] M.B. Conz, J.M. Granjeiro, G. d.A. Soares, Hydroxyapatite crystallinity does not
affect the repair of critical size bone defects, J. Appl. Oral. Sci. 19 (4) (2011)
337–342 (Jul-Aug 03/22/received 11/20/revised 10/26/accepted).
[8] A.E. Porter, N. Patel, J.N. Skepper, S.M. Best, W. Bonfield, Comparison of in vivo
dissolution processes in hydroxyapatite and silicon-substituted hydroxyapatite
bioceramics (in eng), Biomaterials 24 (25) (2003) 4609–4620.
[9] A.E. Porter, C.M. Botelho, M.A. Lopes, J.D. Santos, S.M. Best, W. Bonfield,
Ultrastructural comparison of dissolution and apatite precipitation on hydro-
xyapatite and silicon-substituted hydroxyapatite in vitro and in vivo (in eng), J.
Biomed. Mater. Res A 69 (4) (2004) 670–679.
[10] Y. Belmamouni, M. Bricha, J. Ferreira, K. El Mabrouk, Hydrothermal synthesis of Si-
doped hydroxyapatite nanopowders: mechanical and bioactivity evaluation, Int. J.
Appl. Ceram. Technol. 12 (2) (2015) 329–340.
[11] E.S. Thian, J. Huang, S.M. Best, Z.H. Barber, W. Bonfield, Novel silicon-doped hy-
droxyapatite (Si-HA) for biomedical coatings: an in vitro study using a cellular si-
mulated body fluid (in eng), J. Biomed. Mater. Res B Appl. Biomater. 76 (2) (2006)
326–333.
[12] N. Patel, et al., A comparative study on the in vivo behavior of hydroxyapatite and
silicon substituted hydroxyapatite granules (in eng), J. Mater. Sci. Mater. Med. 13
(12) (2002) 1199–1206.
[13] J.L. Xu, K.A. Khor, Chemical analysis of silica doped hydroxyapatite biomaterials
consolidated by a spark plasma sintering method, J. Inorg. Biochem. 101 (2) (2007)
187–195.
[14] M. Palard, J. Combes, E. Champion, S. Foucaud, A. Rattner, D. Bernache-Assollant,
Effect of silicon content on the sintering and biological behaviour of Ca10(PO4)(6-
x)(SiO4)x(OH)(2-x) ceramics (in eng), Acta Biomater. 5 (4) (2009) 1223–1232.
[15] S.R. Kim, et al., Synthesis of Si, Mg substituted hydroxyapatites and their sintering
behaviors (in eng), Biomaterials 24 (8) (2003) 1389–1398.
[16] Z.-Y. Qiu, I.-S. Noh, S.-M. Zhang, Silicate-doped hydroxyapatite and its promotive
effect on bone mineralization, Front. Mater. Sci. 7 (1) (2013) 40–50.
[17] K. Nakata, T. Kubo, C. Numako, T. Onoki, A. Nakahira, Synthesis and
17621
Ceramics International 44 (2018) 17612–17622
characterization of silicon-doped hydroxyapatite, Mater. Trans. 50 (5) (2009)
1046–1049.
[18] I.R. Gibson, S.M. Best, W. Bonfield, Effect of silicon substitution on the sintering and
microstructure of hydroxyapatite, J. Am. Ceram. Soc. 85 (11) (2002) 2771–2777.
[19] I.R. Gibson, S.M. Best, W. Bonfield, Chemical characterization of silicon-substituted
hydroxyapatite (in eng), J. Biomed. Mater. Res. 44 (4) (1999) 422–428.
[20] S. Langstaff, M. Sayer, T.J.N. Smith, S.M. Pugh, S.A.M. Hesp, W.T. Thompson,
"Resorbable bioceramics based on stabilized calcium phosphates. Part I: rational
design, sample preparation and material characterization, Biomaterials 20 (18)
(1999) 1727–1741.
[21] J.W. Reid, A. Pietak, M. Sayer, D. Dunfield, T.J.N. Smith, Phase formation and
evolution in the silicon substituted tricalcium phosphate/apatite system,
Biomaterials 26 (16) (2005) 2887–2897.
[22] M. Sayer, et al., Structure and composition of silicon-stabilized tricalcium phos-
phate, Biomaterials 24 (3) (2003) 369–382.
[23] X.W. Li, H.Y. Yasuda, Y. Umakoshi, Bioactive ceramic composites sintered from
hydroxyapatite and silica at 1200 °C: preparation, microstructures and in vitro
bone-like layer growth (in eng), J. Mater. Sci. Mater. Med. 17 (6) (2006) 573–581.
[24] T. Kokubo, H. Takadama, How useful is SBF in predicting in vivo bone bioactivity?
Biomaterials 27 (15) (2006) 2907–2915.
[25] A. Bianco, I. Cacciotti, M. Lombardi, L. Montanaro, Si-substituted hydroxyapatite
nanopowders: Synthesis, thermal stability and sinterability, Mater. Res. Bull. 44 (2)
(2009) 345–354.
[26] S.R. Kim, et al., Synthesis of Si, Mg substituted hydroxyapatites and their sintering
behaviors, Biomaterials 24 (8) (2003) 1389–1398.
[27] Y. Jiang, et al., Trabecular bone mineral and calculated structure of human bone
specimens scanned by peripheral quantitative computed tomography: relation to
biomechanical properties (in eng), J. Bone Miner. Res 13 (11) (1998) 1783–1790.
[28] D.D. Thompson, Age changes in bone mineralization, cortical thickness, and
Haversian canal area, Calcif. Tissue Int. 31 (1) (1980) 5–11.
[29] E. Garskaite, K.-A. Gross, S.-W. Yang, T.C.-K. Yang, J.-C. Yang, A. Kareiva, Effect of
processing conditions on the crystallinity and structure of carbonated calcium hy-
droxyapatite (CHAp), CrystEngComm 16 (19) (2014) 3950–3959, http://dx.doi.
org/10.1039/C4CE00119B.
[30] M. Tavafoghi, N. Brodusch, R. Gauvin, M. Cerruti, Hydroxyapatite formation on
graphene oxide modified with amino acids: arginine versus glutamic acid, J. R. Soc.
Interface 13 (114) (2016), http://dx.doi.org/10.1098/rsif.2015.0986.
[31] S. Roushdey, Defect Related Luminescence in Silicon Dioxide Network: A Review,
2011.
[32] G. Dingemans, C.A.A. Van Helvoirt, D. Pierreux, W. Keuning, W.M.M. Kessels,
Plasma-assisted ALD for the conformal deposition of SiO2: process, material and
electronic properties, J. Electrochem. Soc. 159 (3) (2012) H277–H285.
[33] J.F.B. Barata, A.L. Daniel-Da-Silva, M.G.P.M.S. Neves, J.A.S. Cavaleiro, T. Trindade,
Corrole-silica hybrid particles: synthesis and effects on singlet oxygen generation,
RSC Adv. 3 (1) (2013) 274–280, http://dx.doi.org/10.1039/C2RA22133K.
[34] P. Yu, R.J. Kirkpatrick, B. Poe, P.F. Mcmillan, X. Cong, Structure of calcium silicate
hydrate (C-S-H): near-, mid-, and far-infrared spectroscopy, J. Am. Ceram. Soc. 82
(3) (1999) 742–748.
[35] L. Berzina-Cimdina, N. Borodajenko, Research of Calcium Phosphates Using Fourier
Transform Infrared Spectroscopy, INTECH Open Access Publisher, Croatia, 2012.
[36] G.F. Andrade, D.C.F. Soares, R.K.D.S. Almeida, E. Sousa, S.M. Barros, Mesoporous
silica SBA-16 functionalized with Alkoxysilane groups: Preparation, characteriza-
tion, and release profile study, J. Nanomater. 2012 (2012) 10 (Art. no. 816496.).
[37] P.E. Wang, T.K. Chaki, Sintering behaviour and mechanical properties of hydro-
xyapatite and dicalcium phosphate, J. Mater. Sci.: Mater. Med., vol. 4(2), pp.
150–158.
[38] L.T. Bang, B.D. Long, R. Othman, Carbonate hydroxyapatite and silicon-substituted
carbonate hydroxyapatite: synthesis, mechanical properties, and solubility evalua-
tions, Sci. World J. 2014 (2014) 9 (Art. no. 969876).
[39] D. Arcos, J. Rodrı́guez-Carvajal, M. Vallet-Regı, The effect of the silicon in-
corporation on the hydroxylapatite structure. A neutron diffraction study, Solid
State Sci. 6 (9) (2004) 987–994.
[40] P. Saravanapavan, L.L. Hench, Mesoporous calcium silicate glasses. I. Synthesis, J.
Non-Cryst. Solids 318 (1–2) (4/1/ 2003) 1–13.
[41] A. El Yacoubi, A. Massit, M. Fathi, B.C. El Idrissi, K. Yamni, Characterization of
silicon-substituted hydroxyapatite powders synthesized by a wet precipitation
method, IOSR J. Appl. Chem. 7 (11) (2014) 24–29.
[42] F. Balas, J. Perez-Pariente, M. Vallet-Regi, In vitro bioactivity of silicon-substituted
hydroxyapatites (in eng), J. Biomed. Mater. Res A 66 (2) (2003) 364–375.
[43] M.S. Sadjadi, H.R. Ebrahimi, M. Meskinfam, K. Zare, Silica enhanced formation of
hydroxyapatite nanocrystals in simulated body fluid (SBF) at 37 °C, Mater. Chem.
Phys. 130 (1–2) (2011) 67–71.
[44] G. Munir, G. Koller, L. Di Silvio, M.J. Edirisinghe, W. Bonfield, J. Huang, The
pathway to intelligent implants: osteoblast response to nano silicon-doped hydro-
xyapatite patterning, J. R. Soc. Interface 8 (58) (2011) 678–688, http://dx.doi.org/
10.1098/rsif.2010.0548.
[45] J. Rakovan, Growth and surface properties of apatite, Rev. Mineral. Geochem. 48
(1) (2002) 51–86.
[46] J. Weng, Q. Liu, J.G. Wolke, X. Zhang, K. De Groot, Formation and characteristics of
the apatite layer on plasma-sprayed hydroxyapatite coatings in simulated body
fluid (in eng), Biomaterials 18 (15) (1997) 1027–1035.
M. Tavafoghi et al.
[47] S. Hayakawa, K. Tsuru, C. Ohtsuki, A. Osaka, Mechanism of apatite formation on a
sodium silicate glass in a simulated body fluid, J. Am. Ceram. Soc. 82 (8) (1999)
2155–2160.
[48] M. Vallet-Regí, A.M. Romero, C.V. Ragel, R.Z. LeGeros, XRD, SEM-EDS, and FTIR
studies of in vitro growth of an apatite-like layer on sol-gel glasses, J. Biomed.
Mater. Res. 44 (4) (1999) 416–421.
[49] C.A. Miller, T. Kokubo, I.M. Reaney, P.V. Hatton, P.F. James, Formation of apatite
layers on modified canasite glass–ceramics in simulated body fluid, J. Biomed.
Mater. Res. 59 (3) (2002) 473–480.
[50] E.M. Carlisle, Silicon: a possible factor in bone calcification (in eng), Science 167
(3916) (1970) 279–280.
[51] D.M. Reffitt, et al., "Orthosilicic acid stimulates collagen type 1 synthesis and os-
teoblastic differentiation in human osteoblast-like cells in vitro (in eng), Bone 32 (2)
(2003) 127–135.
[52] J.P. Lafon, E. Champion, D. Bernache-Assollant, Processing of AB-type carbonated
hydroxyapatite Ca10−x(PO4)6−x(CO3)x(OH)2−x−2y(CO3)y ceramics with con-
trolled composition, J. Eur. Ceram. Soc. 28 (1) (2008) 139–147.
17622
Ceramics International 44 (2018) 17612–17622
[53] S. Raynaud, E. Champion, D. Bernache-Assollant, P. Thomas, Calcium phosphate
apatites with variable Ca/P atomic ratio I. Synthesis, characterisation and thermal
stability of powders (in eng), Biomaterials 23 (4) (2002) 1065–1072.
[54] H.M. Kim, T. Himeno, M. Kawashita, T. Kokubo, T. Nakamura, The mechanism of
biomineralization of bone-like apatite on synthetic hydroxyapatite: an in vitro as-
sessment, J. R. Soc. Interface 1 (1) (2004) 17–22.
[55] T. Kokubo, H. Takadama, How useful is SBF in predicting in vivo bone bioactivity?
Biomater. Rev. 27 (15) (2006) 2907–2915.
[56] S. Sprio, et al., Physico-chemical properties and solubility behaviour of multi-sub-
stituted hydroxyapatite powders containing silicon, Mater. Sci. Eng.: C 28 (1) (1/
10/ 2008) 179–187.
[57] A.M. Pietak, J.W. Reid, M.J. Stott, M. Sayer, Silicon substitution in the calcium
phosphate bioceramics, Biomaterials 28 (28) (2007) 4023–4032.
[58] Q. Fu, M.N. Rahaman, H. Fu, X. Liu, Silicate, borosilicate, and borate bioactive glass
scaffolds with controllable degradation rate for bone tissue engineering applica-
tions. I. Preparation and in vitro degradation, J. Biomed. Mater. Res. Part A 95A (1)
(2010) 164–171.