Materials Science and Engineering C 69 (2016) 337342

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Materials Science and Engineering C

journal homepage: www.elsevier.com/locate/msec

Deposition of calcium phosphate coatings using condensed phosphates

(P2O47 and P3O510) as phosphate source through induction heating
Huan Zhou a,b, Saisai Hou a,c, Mingjie Zhang a,d, Mengmeng Yang a,d, Linhong Deng a, Xinbo Xiong e, Xinye Ni f,
a
Institute of Biomedical Engineering and Health Sciences, Changzhou University, Changzhou, Jiangsu 213164, China
b
International Research Centre for Translational Orthopedics (IRCTO), Suzhou, Jiangsu 215006, China
c
School of Materials Science and Engineering, Changzhou University, Changzhou, Jiangsu 213164, China
d
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu 213164, China
e
College of Materials Science and Engineering, Shenzhen University, Shenzhen, Guangdong 518060, China
f
Second People's Hospital of Changzhou, Nanjing Medical University, Changzhou, Jiangsu 213003, China

a r t i c l e i n f o a b s t r a c t

Article history: In present work condensed phosphates (P2O4 5

7 and P3O10 ) were used as phosphate source in induction heating
Received 21 April 2016 to deposit calcium phosphate coatings. The phase, morphology, and composition of different phosphate-related
Received in revised form 13 June 2016 coatings were characterized and compared using XRD, FTIR, and SEM analyses. Results showed that P2O4 7 -
Accepted 26 June 2016
formed calcium pyrophosphate hydrate coatings with interconnected cuboid-like particles. The as-deposited cal-
Available online 01 July 2016
cium tripolyphosphate hydrate coating with P3O5 10 was mainly composed of ower-like particles assembled by

Keywords:
plate-like crystals. The bioactivity and cytocompatibility of the coatings were also studied. Moreover, the feasibil-
Induction heating ity of using hybrid phosphate sources for preparing and depositing coatings onto magnesium alloy was
Condensed phosphate investigated.
Coating 2016 Elsevier B.V. All rights reserved.

1. Introduction properties of CaP. In particular, Ca-based components can be modied

Surface modication is essential to enhance the biocompatibility and
osteoconductivity of inert orthopedic devices. Coating a base material
with a thin layer of calcium phosphate (CaP) is a typical and effective
approach used to enhance strong bone bonding to the implant in vivo
[1]. Numerous techniques, such as biomimetic coating, microwave-
assisted wet-chemical deposition, and plasma spraying, can be used to
successfully deposit CaP coatings onto the implant surface [24].
Induction heating (IH) is a convenient and effective method for de-
position of CaP coatings onto conductive materials [5]. CaP coatings
can be successfully deposited within hours through electromagnetic in-
duction heating to substrates immersed in aqueous solutions containing
Ca2+ and PO34 . This technology provides stronger coating binding force
and faster coating rate than biomimetic coating. In our previous study,
we determined the effects of temperature, processing time, and Mg2+
doping on CaP coating properties [6,7]. Increasing the temperature
from 70 C to 100 C and prolonging processing from 0.5 h to 8 h
could increase coating density and adhesion force. Moreover, Mg2 +
doping could induce the formation of micropores and promote cell ad-
hesion and proliferation. CaP contains both Ca- and P-based compo-
nents. Replacement of either Ca or P component can alter the
Corresponding author.
E-mail address: nxy2000@aliyun.com (X. Ni).
through Mg2+ doping.
This paper is inspired by our long-standing efforts to apply con-
densed phosphates for generating CaPs [8,9]. Condensed phosphates
are more important in some biomolecules, such as guanosine-5-tri-
phosphate and adenosine triphosphate, than regular PO3 4 . The phos-
phate chain of these molecules can be broken down by enzyme
hydrolysis to release PO3
4 . Supplementation of condensed phosphates
in bone formation promotes the differentiation and maturation of oste-
oblasts [10,11] but inhibits the activity of osteoclast cells [12]. The prop-
erties of CaP coatings can be controlled by modifying the composition of
aqueous solutions for synthesis [2,13]. However, condensed phosphates
can be degraded under hydrothermal treatment; specically, heat is
generated around the coil during IH. Therefore, the inuences of replac-
ing PO34 with condensed phosphates on CaP coatings deposited
through IH must be investigated.
2. Material and methods
2.1. IH coating system set-up
IH coating deposition was performed using SP-15 high-frequency in-
duction power with a frequency of 15 kHz (Shenzhen SuanPin Power
Co. Lit). The experimental setup for induction heat deposition is similar
to that in reference [5]. This system is composed of an induction current
power, which consists of a copper coil with appropriate water cooling, a

http://dx.doi.org/10.1016/j.msec.2016.06.090
0928-4931/ 2016 Elsevier B.V. All rights reserved.
338 H. Zhou et al. / Materials Science and Engineering C 69 (2016) 337342

Table 2
Compositions of aqueous coating solution using hybrid phosphate sources.

CaCl2 NH4H2PO4 Na4P2O7 H2O

Sample (g) (g) (g) Na5P3O10(g) (mL) Ca/P

\P\
Ca\ \P2 17.4 7.2 4.6 1000 1.67
\P\
Ca\ \P3 17.4 7.2 3.83 1000 1.67

The incubated samples were examined using SEM. Moreover, the

cytocompatibility of the deposited coatings was evaluated by seeding
10,000 MC3T3-E1 osteoblast cells on the coated sample surface. The os-
teoblast cells were provided by Lei Yang, Soochow University. The sam-
ples were further incubated for up to 7 d, and the medium was
refreshed every other day. The number of cells in each group on days
1 and 7 was measured through MTT assay to determine cell density.

Fig. 1. Schematic graph of IH coating system: (1) induction powder, (2) copper coil, (3)
glass tube, (4) sample, (5) agitator, (6) pump, and (7) coating solution.
2.3. IH coating deposition using solution of hybrid phosphate sources

The feasibility of depositing hybrid composite coatings onto the C/C

glass tube of 20 mm diameter inside the coil, a peristaltic pump, a feed-
ing tank, and tubes that connect the feeding tank to the bottom of the
glass tube and the top of the glass tube to the tank for the residual solu-
tion. A narrowed area of 16 mm diameter is inside the glass tube and
supports the samples in a vertical position. The schematic of the coating
system is presented in Fig. 1.
sample surface was studied using solutions containing hybrid phos-
phate sources (Table 2). Coating was conducted in the same way as de-
scribed above. The as-prepared samples were characterized using XRD
and SEM analyses.
2.4. IH coating deposition onto magnesium substrates
Mg alloy (AZ31) square with 15 mm side length was placed into the
IH coating system. Solutions containing PO3 4 5
4 , P2O7 , or P3O10 were

2.2. IH coating deposition onto carbon/carbon substrates used to coat Mg alloys by following the steps described above. The coat-
ed samples were compared visually.
All chemicals were purchased from Aladdin (Shanghai, China). The
coating solution used in this study was prepared by dissolving given 3. Results
amounts of reagent-grade CaCl2 and different phosphate sources into
distilled water (Table 1). The pH of solution was adjusted to 4.5 by 3.1. IH coating deposition onto carbon/carbon substrates
adding 1 M HCl to obtain a transparent solution.
Cylindrical carbon/carbon (C/C) composites with 8 mm diameter The XRD patterns of the coated samples are presented in Fig. 2. After
and 10 mm length were prepared by Northwest Polytechnology Univer- deposition for 2 h, the characteristic diffraction peaks on the surface of
sity in China. Samples were pretreated in high-pressure steam in a hydroxyapatite (HA, Ca10(PO4)6(OH)2, PDF#09-0432) crystal almost
50 mL autoclave with 40 mL of 2 M H2O2 solution at 160 C before coat-
ing. The cylinders were removed from the H2O2 solution, rinsed ultra-
sonically with deionized water, and dried in air. The C/C sample was
subsequently placed into a tube and coated for 4 h by using different so-
lutions. The C/C samples were rewashed using deionized water to re-
move uncoated powder on the surface. The samples were then
immersed in an autoclave containing 20% ammonia water at 130 C
for 6 h to mature the deposited crystals.
The coated samples were characterized using X-ray diffraction (XRD,
D/MAX2500, Rigaku, Japan), Fourier transform infrared spectroscopy
(FTIR, PROTG 460, Nicolet, USA), and scanning electron microscopy
(SEM, SUPRA 55, Zeiss, Germany) analyses. The bioactivity of the coat-
ings was studied in vitro by soaking the samples in simulated body
uid (SBF) for up to 2 weeks; SBF was replaced every other day [14].

Table 1
Compositions of aqueous coating solution using different phosphate source.

Sample CaCl2 (g) Phosphate source (g) H2O (mL) Ca/P

\P
Ca\ 17.4 NH4H2PO4 10.8 1000 1.67
\P2
Ca\ 17.4 Na4P2O7 12.5 1000 1.67
Fig. 2. XRD patterns of the coated C/C composite samples, refers to carbon, refers
\P3
Ca\ 17.4 Na5P3O10 11.5 1000 1.67
to HA, refers to CPPH, and refers to unknown phase.
 H. Zhou et al. / Materials Science and Engineering C 69 (2016) 337342 339

signicantly changed after the ammonia treatment (Fig. 5). Numerous

voids were found in the Ca\\P coating at low magnication, and the
coating was composed of needle-like nanoparticles. The coating layer
of Ca\\P2 was formed by interconnected cuboid-like particles, whereas
that of Ca\\P3 was mainly composed of ower-like particles assembled
by plate-like crystals. Moreover, high-magnication analysis indicated
the presence of signicant amounts of cavities among the particles in
all samples.
The bioactivity difference of the three sample series was observed
and compared after 2 weeks of SBF incubation (Fig. 6). Analysis of the
density of the deposited apatite showed that bioactivity decreased in
the order of Ca\\P, Ca\\P2, and Ca\\P3.
The results of MTT tests revealed that all the synthesized samples
promoted cell proliferation (Fig. 7). However, both Ca\\P2 and Ca\\P3
showed improved cell adhesion than Ca\\P (p b 0.05) on day 1, and
Ca\\P2 showed improved cell proliferation than Ca\\P (p b 0.05) on
day 7. The cell average number in Ca\\P3 was higher than that in Ca\\P.

Fig. 3. Typical FTIR spectrum of the as-deposited coatings.

3.2. IH coating deposition using solutions containing hybrid phosphate
sources

The crystalline structure, morphologies, and compositions of the as-

appeared in the XRD topography of Ca\\P. Strong carbon diffraction
deposited coatings were considerably changed when P2O47 or
peaks were also detected (PDF#656212). By contrast, the intensity of
P3O510 was combined with PO4
3
in the coating solution. As shown in
carbon diffraction peaks was low in samples Ca\\P2 and Ca\\P3. The
Fig. 8, strong carbon diffraction peaks and poor CPPH peaks were detect-
Ca\\P2 coating was mainly composed of calcium pyrophosphate hy-
ed in Ca\\P\\P2. Unknown phase as well as HA and carbon diffraction
drate (CPPH, Ca2P2O72H2O, PDF #41-0488). The coating phase of
peaks were observed in Ca\\P\\P3. The intensity of carbon diffraction
Ca\\P3 was unidentied when searching the JADE database.
peaks was reduced compared with that in Ca\\P\\P2, and the coating
The FTIR spectra of the samples are shown in Fig. 3. Ca\\P within the
was partially amorphous. The SEM analysis showed that the Ca\\P\\P2
wavenumber range of 9001150 cm1 showed one strong absorption
surface was partly covered by deposited bean-like particles (Fig. 9).
band, whereas Ca\\P2 and Ca\\P3 showed clusters of absorption
Moreover, the coating of Ca\\P\\P3 was composed of ower-like parti-
bands. The band at 760 cm1 could be attributed to condensed phos-
cles assembled by different-sized plate-like crystals as well as small
phates. The absorption bands of water were found at 1656 and
spherical particles.
3448 cm1.
The morphologies of the coatings after IH are presented in Fig. 4. All
samples contained a coating layer on the surface. The coatings of Ca\\P, 3.3. IH coating deposition onto magnesium substrates
Ca\\P2, and Ca\\P3 were composed of plate-like particles, cuboid-like
particles embedded in badland-like layer, and self-assembled plate- Mg alloy is another conductive material. Fig. 10 shows that Ca\\P3
like particles, respectively. The morphologies of the coatings were provided the optimal coating density.

Fig. 4. SEM photos of the surface morphologies of achieved coatings on the C/C composites surface: (a and d) Ca\
\P, (b and e) Ca\
\P2, and (c and f) Ca\
\P3.
340 H. Zhou et al. / Materials Science and Engineering C 69 (2016) 337342
Fig. 5. SEM photos of the surface morphologies of ammonia-treated coatings on the C/C composite surface: (a and d) Ca\
4. Discussion
The results of XRD, FTIR, and SEM analyses conrm that condensed
phosphates, namely, P2O4 7 and P3O5
10 , can be used in the IH coating
system to generate coatings with features different from those of
PO3
4 . The XRD patterns indicated that Ca\ \P and Ca\\P2 had HA and
CPPH coating on the C/C surface, respectively. Ca2+ and PO34 are com-
monly formed into plate-like acidic calcium phosphates in an acidic en-
vironment but could be converted into HA after subsequent alkali
treatment [15,16]. SEM observations of the Ca\\P coating morphology
before and after ammoniawater incubation are in agreement with
these reports (Figs. 4d & 5d). For Ca\\P2, although CPPH crystals pre-
served the cuboid-like structure after the treatment, the particle sizes
decreased as shown in Figs. 4 and 5. These cuboid-like crystals were in-
terconnected with one another to form a porous coating layer. For
Ca\\P3, the as-formed unidentied materials were found to be calcium
tripolyphosphate hydrate (Ca5(P3O10)2 nH2O, CTPH). First, PO3 4 and
P2O4
7 reacted with Ca2+ to form CaPs. P3O5 10 then reacted with Ca
2+
to form a complex with both ions. Second, the FTIR spectra (Fig. 3) indi-
cated that Ca\\P2 and Ca\\P3 presented a similar spectrum to that of
Fig. 6. Typical surface morphologies of the coated C/C composites after 2 weeks of SBF incubation: (a) Ca\
\P, (b and e) Ca\
\P2, and (c and f) Ca\
\P3.
Ca\\P. This nding conrms the presence of H2O and condensed phos-
phates in Ca\\P3. Third, condensed phosphates can delay the deposition
of apatite to the substrate surface from SBF [17], and Ca\\P3 showed
stronger inhibitory ability than Ca\\P2. These ndings support the con-
clusion that CTPH is deposited onto Ca\\P3.
The MTT study indicated that Ca\\P2 and Ca\\P3 are biocompatible
biomaterials to osteoblasts. CPP-based ceramics are potential materials
for bone repair because of their higher biodegradation than HA in vivo
and ability to intensify osteoblast differentiation [18,19].
For coating deposition using solutions containing hybrid phosphate
sources, Ca\\P\\P2 could not provide uniform coating to fully cover
the C/C surface. This phenomenon was attributed to the competition
of PO3
4 and P2O7
4
to bond to Ca2+ and the inhibitory effect of P2O47
on calcium orthophosphate crystal formation. The as-deposited CPPH
presented a new morphology relative to that of Ca\\P2 or those report-
ed in other studies [20]. The Ca\\P\\P3 coatings were composed of
small spherical particles and ower-like particles assembled by differ-
ent-sized plate-like crystals. The ower-like particles assembled by
large-sized plate-like crystals are CTPH. Flower-like particles assembled
by smaller-sized plate-like crystals are HA [21], whereas the small
\P, (b) Ca\
\P2, and (c) Ca\
\P3.
 H. Zhou et al. / Materials Science and Engineering C 69 (2016) 337342
Fig. 7. MTT results of cell culture.
Fig. 8. XRD patterns of the coated C/C composite samples using hybrid phosphate sources:
refers to carbon, refers to HA, refers to CPPH, and refers to unknown
phase.
Fig. 9. SEM photos of the surfaces of the coated C/C composite samples using hybrid phosphate sources: (a and b) Ca\
341
spherical particles are amorphous calcium phosphate (ACP). When a
CaP formation inhibitory factor exists, ACP is favored instead of crystal-
lized HA [22]. The inhibitory factor is P3O5
10 .
Both Ca\\P2 and Ca\\P3 showed faster coating deposition rates than
Ca\\P for C/C (Fig. 2), which indicates that condensed phosphate ions
exhibit higher afnity to Ca2+ than PO3 4 in aqueous media. Ca\\P3 is
the recommended material for Mg alloys to generate protective coating
layer within a short time (Fig. 10). Controlling the degradation rate of
Mg alloy is a challenging task. CaP, especially HA coatings, have been
signicantly studied to be deposited as a protective layer on the Mg
alloy surface [20,23]. Unlike stable C/C composites, Mg alloy is biode-
gradable and continuously releases hydrogen. The released Mg2+ ions
can inhibit the growth of crystallized CaPs [24], thereby decreasing coat-
ing density. This study presents an alternative coating in the IH coating
system for the Mg alloy surface, but further studies must be conducted.
5. Conclusions
P2O4
7 and P3O5
10 were used as phosphate sources to generate bio-
compatible calcium condensed phosphate coatings in the IH coating
system. Ca\\P2 and Ca\\P3 generated coatings with higher density
than that of regular Ca\\P. The as-deposited coatings showed different
phase compositions and coating morphologies depending on the phos-
phate source. Only Ca\\P\\P3 formed thick coatings on the C/C surface
when P2O4 5 3
7 or P3O10 ions were combined with PO4 . The generated
coatings were composed of CTPH, HA, and ACP. Comparison with stud-
ies on Mg alloy indicated that Ca\\P3 provided coatings with the
highest density than other compositions. Therefore, condensed phos-
phates can be potentially applied to prepare coatings for biomedical ap-
plications through IH.
Acknowledgements
This work was partially supported by the Natural Science Founda-
tion of Jiangsu Province Research (no. BK20151181), the Scientic Re-
search Foundation for the Returned Overseas Chinese Scholars, State
Education Ministry, 2014 (1685) and High-Level Medical Talents Train-
ing Project of Changzhou (no. 2016CZLJ004).
\P\
\P2 and (c and d) Ca\
\P\
\P3.
342 H. Zhou et al. / Materials Science and Engineering C 69 (2016) 337342
Fig. 10. Mg alloy squares treated using different coating solutions.
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