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Thin Solid Films 516 (2008) 6145 – 6150

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Preparation and characterization of hydroxyapatite coatings on human

enamel by electrodeposition
Ying-Min Liao, Zu-De Feng ⁎, Si-Wei Li
Department of Materials Science and Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China
State Key Laboratory of Physical Chemistry at the Solid Surfaces, Xiamen University, Xiamen 361005, China
Received 9 February 2007; received in revised form 17 August 2007; accepted 12 November 2007
Available online 19 November 2007

Abstract

Thin hydroxyapatite (HA) coatings, approximately from 1 μm to 10 μm in thickness, had been prepared on human enamel by electrodeposition
with the current density of 0.5 mA/cm2 at 55 °C in 1 h. They exhibited an acicular morphology and had a tight contact with the substrate by
scanning electron microscope and transmission electron microscope observation. The comparison was made between the enamel specimens with
HA coatings in this study and the sound enamel. In this comparison, while the Vickers micro-hardness showed a similar value, the antibacterial
activities improved significantly after the formation of HA coatings. In conclusion, electrodeposition was proved to be an effective method for
preparing HA coatings on human enamel and could be considered as a promising method to restore the initial enamel lesions in clinic.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Electrochemistry; Hydroxyapatite (HA); Coatings; Enamel

1. Introduction such as spreading the enamel surfaces with special medicine

Enamel, the exterior coating of human teeth, was a biom-
ineral with remarkable hardness and resistance to physical and
biochemical attack. The major inorganic component of enamel
was hydroxyapatite (Ca10(PO4)6(OH)2, HA) that was prone to
dissolve under acid condition. In addition, the dissolution of
HA would easily lead to the appearance of initial enamel lesion
that was observed clinically as subsurface white spot lesions
and had no obvious caries [1]. As patients had no obvious
response when suffered from the outside cold or hot stimulation,
the initial enamel lesions were easy to be ignored and would
develop to the deep caries lesions. With the improvement of
people's living standard, increasing attention was paid to the
restoration of the initial enamel lesions in order to prevent the
deep caries lesions.
In previous study, several methods had been developed
to restore the initial enamel lesions. The conservative methods
⁎ Corresponding author. Department of Materials Science and Engineering,
College of Chemistry and Chemical Engineering, Xiamen University, Xiamen
361005, China. Tel.: +86 0592 8778331; fax: +86 0592 2183937.
E-mail address: zdfeng@xmu.edu.cn (Z.-D. Feng).
0040-6090/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.tsf.2007.11.022
were reported. As fluoride could not only induce the growth of
HA crystal, but also help to form the fluorapatite that had
stronger stability, fluoride was introduced into a special med-
icine called fluoridated mouthrins [2]. However, the risks of
fluorosis had also been reported [3]. Also, phosphoric acid
polypeptide was introduced into the special medicine, as it
could promote the remineralization of enamel, and played an
important role in preventing caries and restoring the enamel
lesions [4,5]. For well restoration of the initial enamel lesions,
the teeth should be treated with these special medicines for quite
a long duration, such as 1 h per day for successional 7 days. As a
result, these methods were not suitable for clinical treatment.
Neither was the method adopted by K. Yamagishi because of
the low pH, though the rate of the restoration was rapid (within
minutes) [6]. However, the results of the research by Yamagishi
et al. shown that the HA nanocrystals seamlessly grew in the
interface between the paste and tooth enamel, and arranged their
(0001) face parallel to the tooth surface same as enamel apatite.
In addition, this HA coatings could help to prevent reoccurrence
of caries.
Consequently, some other techniques used for preparing
HA coatings on the various substrates might be taken into
6146 Y.-M. Liao et al. / Thin Solid Films 516 (2008) 6145–6150
consideration, such as hydrothermal technique [7], plasma
spray method [8], sol–gel method [9], electrophoresis method
[10], biomimetic method [11] and electrodeposition [12].
Among these techniques, the electrodeposition had exhibited
much more interest as it could be carried out under a mild
experimental condition, i.e. at relatively lower temperature and
under atmospheric pressure. In addition, electrodeposition
would possibly improve the bond strength of the substrate/
coatings and also control the thickness, chemical composition
and microstructure of the coatings. Although studies about
depositing the HA coatings onto the metals, especially on
titanium substrate, were well documented [12–16], there had
been no report about preparing HA coating on the enamel
surfaces by electrodeposition up to now. However, studies had
been carried out to judge effectively whether teeth were carious
through electrical resistance [17,18] and teeth were semicon-
ductors that could be served as electrode. Therefore, there was a
feasibility of preparing HA coatings on the surfaces of human
enamel using electrodeposition technique.
As shown in the previous studies, the surfaces properties
of human enamel would be changed, with the solubility and
penetration increased significantly after being treated with 35–
50% phosphorus for 20–60 s [19]. In addition, the treatment of
30% hydrogen peroxide might cause the alteration in the chem-
ical structures of the teeth and result in the dissolution of HA
crystals [20]. So in this article, the acid solution which was
prepared by mixing aqueous solution of 30% hydrogen
peroxide (H2O2) with 85% phosphoric acid (H3PO4) was used
to produce the initial enamel lesions and change the perfor-
mances of enamel surfaces. The detailed preparation and char-
acterization of the HA coatings on the enamel surfaces by
electrodeposition were described and a possible mechanism
of HA crystallization was proposed. The aim of the study was
to prove that electrodeposition was an effective method for
preparing HA coatings on human enamel surfaces and restoring
the initial enamel lesions.
2. Materials and methods
2.1. Preparation of enamel specimens
Six fresh human third molars extracted without visible evi-
dence of caries were used in this study. They were stored in
physiological solution after their roots and pulps were removed.
Subsequently, they were made into slabs (5 mm length × 4 mm
width × 3 mm height) using a low speed diamond saw (SYJ-150,
Shenyang Kejing, China). In order to remove the bacterium
speckle and pigment, the enamel surfaces of each specimen was
mechanically polished using silicon carbide papers and 0.25 μm
aluminium oxide powders. Then the specimens were washed
with de-ionized water and dried in air.
2.2. Acid pretreatment
Initial enamel lesions were produced on human enamel using
acid solution. The acid solution was prepared by mixing aque-
ous solution of 30% hydrogen peroxide (H2O2) with 85%
phosphoric acid (H3PO4) at a volume ratio of 4:1. The spec-
imens were then washed with de-ionized water and dried in air
after immersed into this acid solution for 1 min.
2.3. Preparation of coatings by electrodeposition
After acid pretreatment, the enamel specimen was treated for
1 h with the current density of 0.5 mA/cm2 at 55 °C. As shown
in Fig. 1, it was performed with a two-electrode electrochem-
ical cell at the open circuit potential in the electrolyte. The
galvanostat was provided by Model 263A (EG&G Instruments,
Inc, USA). A platinum plate was served as counter electrode
and a slice of stainless steel as working electrode. The speci-
men of enamel was put entirely close to the stainless steel, and
the enamel surfaces subjected to coatings faced to the counter
electrode. To enhance the conductivity of the electrolyte, NaNO3
was added and the electrolyte that contained 4.175 × 10− 4 M Ca
(NO3)2, 2.4 × 10− 4 M NH4H2PO4 and 0.1 M NaNO3 was ad-
justed to pH 6.5 by NaOH at 55 °C.
2.4. Apparatus and methods
2.4.1. Contact angle and Vickers micro-hardness measurements
The contact angle was determined using a contact angle
analyzer (Powereach JC2000A, Zhongchen Co., Shanghai,
China) at room temperature. A distilled water droplet (1 μL) on
the surfaces of specimen was observed using a video camera
coupled to a light microscope, and the contact angle was deter-
mined automatically using image analysis software. The hard-
ness of specimens at different state was measured using a HV-
1000 Vickers micro-hardness tester (Shanghai Materials Tester
Machine Co., China) with a load of 10 g for a dwell time of
30 s. In order to avoid the lag effect, each specimen was at
least subjected to three measurements under the same testing
condition.
Paired t-test was carried out to analyze the influence of acid
pretreatment and formation of coatings on the contact angles as
well as the Vickers micro-hardness of the specimens.
2.4.2. X-ray diffraction measurements
Crystal structures of the specimens' surfaces were character-
ized and analyzed by an X'Pert X-ray diffractometer (XRD,
X'pert PRO, Panalytical, Netherlands) and attached software.
XRD measurements were carried out using Cu Kα radiation
Fig. 1. The device of electrochemistry experiment.
 Y.-M. Liao et al. / Thin Solid Films 516 (2008) 6145–6150 6147

(λ = 0.15405 nm) at 40 kV and 30 mA. Data were collected from

20° to 80° with a step size of 0.008 and a scanning speed
of 0.0217 s per step. The testing procedure was X'celerator_
grazing_angle (Omega = 0.2) which was suitable for analyzing
thin film.

2.4.3. Scanning electron microscope and transmission electron

microscope observations
The morphologies and structures of cross-sectional speci-
mens were observed through a field emission scanning electron
microscope (SEM, LEO-1530, Germany) and field emission
gun transmission electron microscope (TEM, Tecnai F30, USA)
with acceleration voltage of 300 kV. SEM specimens were
obtained after being embedded in epoxy resin entirely, cut in
half and mechanically polished using silicon carbide papers
and 0.25 μm aluminium oxide powders. TEM cross-sectional
specimens were prepared using the precision ion polishing
system (PIPS, Model 691, Gatan, USA). Before ion thinning,
the specimens were made into slices along the cross-section
using a low speed diamond saw (SYJ-150, Shenyang Kejing,
China) and then embedded in epoxy resin. After being polished
to 0.2 mm in thickness, they were cut into disks of 3 mm in Fig. 3. Vickers micro-hardness of the specimens' surfaces at different states.
diameter using ultrasonic disc cutter (Model 601, Gatan, USA)
and then pre-thinned and polished mechanically using dim- measured directly using a pH meter (PHB-4, Shanghai Leici,
ple grinder (Model 656, Gatan, USA). In addition, the chemi- China). The calibration experiments showed that the micro-
cal composition of the coatings was analyzed by an energy sensor responded well to pH changes in the buffered solutions.
dispersion spectroscopy (EDS, Oxford INCA300, England) The potential decreased linearly with a slope of 0.065123 at
attached to SEM. 55 °C. Then the pH values at the enamel/solution interface were
measured in situ using this pH microsensor.
2.4.4. pH measurements
The sensing material for the pH microprobe was iridium 3. Results
oxide prepared by electrodeposition on platinum wire [21]. To
minimize the potential drift, the pH microsensor was aged for 3.1. Contact angle measurements
24 h in the electrolyte. Then it was calibrated by measuring the
potential of the iridium oxide microsensor referred to a saturated Fig. 2 showed the contact angles of the specimens' surfaces
calomel electrode in a series of buffered solution, pH of which at different state. A slight decrease in contact angle was ob-
ranged from 1 to 12. The pH value of buffered solution was served after acid pretreatment (P b 0.05). And the contact angles
values ranged from 136.4° ± 10.18° to 160.4° ± 6.79° after the
formation of coatings, and were also significantly higher than
those of enamels before or after acid pretreatment (P b 0.001).

Fig. 4. X-ray diffraction patterns of specimen's surfaces at different state: (a)

before acid pretreatment; (b) after acid pretreatment; (c) after formation of
Fig. 2. Contact angles of the specimens' surfaces at different states. coating.
6148 Y.-M. Liao et al. / Thin Solid Films 516 (2008) 6145–6150
Fig. 5. SEM image showed the surface morphology of the specimen's surfaces
after the coating was prepared.
3.2. Vickers micro-hardness measurements
Vickers micro-hardness as a function of the specimens' sur-
faces at different state was shown in Fig. 3. According to the
statistical analyses, the Vickers micro-hardness values exhibited
a significant decrease (P b 0.001) after acid pretreatment and an
obvious increase (P b 0.05) after formation of coatings. The
Fig. 6. (a) SEM image showed the cross-sectional morphology of the specimen's
surfaces after the coating was prepared; (b) EDS spectra of the coating on the
cross-sectional specimen.
Fig. 7. TEM image showed the cross-sectional of the specimen after the coating
was prepared.
Vickers micro-hardness of the enamel specimens ranged from
270.00 ± 15.81 to 339.16 ± 13.21 HV before acid pretreatment
and ranged from 227.32 ± 11.35 to 286.96 ± 11.19 HV after
formation of coatings. The result of paired t-test indicated that
there was no significant difference between them in the Vickers
micro-hardness (P N 0.05).
3.3. Crystallinity of the specimens' surfaces
Fig. 4 showed the XRD patterns of the specimens' surfaces
at different state. A set of peaks matched well with the standard
pattern of the hexagonal HA (PDF #9-432) [22]. And no
extraneous peaks were found in the XRD patterns. In addition,
the intensity of (002) which could reflect the percentage of HA
was diminished after acid pretreatment and greatly intensified
after formation of coatings.
3.4. Microstructure and morphology analysis
While Fig. 5 showed that the HA crystals were orientated
and developed into acicular crystals, Fig. 6(a) showed that
Fig. 8. Variation of the interfacial pH values with the immersed time.
 Y.-M. Liao et al. / Thin Solid Films 516 (2008) 6145–6150
the coatings were about 2 μm thick and were tightly combined
to the enamel surfaces. As shown in Fig. 7, the HA coatings
preferentially grew along the orientation of the HA crystals of
the enamel rod. The EDS analysis of the coatings indicated that
the Ca/P molar ratio was about 1.67 (Fig. 6b) and sodium was
excluded.
3.5. pH measurement at the substrate/solution interface
Fig. 8 showed the changes of the pH values in the vicinity of
the enamel with current density of 0.5 mA/cm2 for different
immersed time at 55 °C. As soon as the experiment began, pH
increased immediately and reached to 7.64. During the process
of the experiment, pH increased to 7.98 by 40 min, and then
decreased slowly to 7.91 when the experiment ended.
4. Discussion
The gaps between the enamel rods, which might act as the
channels of the mineral ions, appeared after acid pretreatment.
With the breakaway of the calcium and phosphorus ions, the
gaps were broadened, and then the rate of the ions escape
accelerated. As a result, the HA crystals were dissolved (Fig. 4a
and b) and the initial enamel lesions were formed. Then as
shown in Fig. 3, the Vickers micro-hardness of the enamel
surfaces decreased significantly (P b 0.001). However, acid pre-
treatment had no effect on the crystal structure and purity of
HA which had highly preferential (002) orientation (Fig. 4a
and b). Owing to the inhomogeneous composition of enamel
and uneven dissolution of HA, the enamel surfaces turned
rougher and the contact angles values (P b 0.05) decreased
which indicated that the enamel surface energy increased. All
these factors mentioned above facilitated the formation of HA
coatings.
Numerous studies had been made on the precipitation of
calcium phosphates from the solution containing Ca2+ and PO43−
with respect to the four kinds of calcium phosphates in order
of increasing solubility: Ca10(PO4)6(OH)2, Ca2(PO4)3·nH2O,
Ca8H2(PO4)6·5H2O [23], and CaHPO4·2H2O. During the
precipitation of calcium phosphates from aqueous solution,
different phases might be formed depending mainly on the
pH value of solution [24,25]. So the pH value at the cathode/
solution interface was considered to be a critical parameter
for determining structure and composition of the coatings
products on the cathode surfaces. When the interfacial pH was
higher than 7.4, it provided an appropriate chemical environ-
ment in the vicinity of the cathode to form HA coatings on the
Ti6Al4V alloy [21]. In this study, the pH microsensor was used
to measure the interfacial pH value at the cathode/electrolyte
interface and the electrodeposition had been applied to prepare
the HA coatings on human enamel with the current density of
0.5 mA/cm2 at 55 °C. Based on the experimental results, the
mechanism of the formation of HA coatings on human enamel
surfaces were explained as follows. As the specimen of enam-
el was regarded as a semiconductor and put entirely close to
the working electrode, the whole of them might work as a
cathode. When the cathodic current was applied, the relevant
6149
electrochemical reactions that were shown as follows took
place.
2H2 O þ 2e− →H2 ↑ þ 2OH− ð1Þ
OH− þ H2 PO−4 →HPO2−
4 þ H2 O ð2Þ
−
4 þ OH →PO4 þ H2 O
HPO2− ð3Þ
3−
−
5Ca2þ þ 3PO3−
4 þ OH →Ca5 ðPO4 Þ3 ðOHÞ2 ð4Þ
The reducing of water resulted in the emission of H2 in the
vicinity of the enamel surfaces and the increase of hydrox-
ide ions, as shown in reaction (1). Then the pH values at the
cathode/electrolyte interface were higher than 7.64 throughout
the electrodeposition (Fig. 8) and the rates of the reactions (2)
and (3) were accelerated. In addition, the electric field was
helpful for the enrichment of Ca2+ ions on the surface of the
enamel surfaces like the metal electrode [26]. Then it needed
only 1 h to prepare 2 μm-thick coatings on human enamel
surfaces by electrodeposition (Fig. 6a), which was regarded to
be faster than that by conservative methods according to the
reaction (4). The primary component of the coatings was HA as
revealed by EDS (Fig. 6b) and XRD (Fig. 4) measurements. The
HA crystals seamlessly grew in the interface between the elec-
trolyte and enamel, and arranged their (002) face parallel to the
tooth surface same as enamel apatite (Figs. 4 and 7). Then the
HA coatings with acicular morphology (Fig. 5) could tightly
combine to the enamel surfaces (Fig. 6a).
In this study, the performances of enamel surfaces were
changed after the formation of HA coating. The surface energy
of enamel decreased and the enamel surfaces became smooth
reflected from the results of the contact angles measurement
(Fig. 2). This indicated that the antibacterial activities of the
enamel surfaces were improved as Kawai et al. had reported that
the more porcelain surface was rough, the more bacteria would
adhere on it [27]. In addition, the Vickers micro-hardness values
of the enamel specimens with HA coatings were significantly
higher than those after acid pretreatment (P b 0.05) (Fig. 3).
However, the Vickers micro-hardness showed a similar value,
ranged from 250 to 360 HV [28], in the comparison between the
enamel specimens with HA coatings in this study and the sound
enamel. This indicated that the restorations of the initial enamel
lesions were effective at 55 °C. For oral cells would be destroyed
to some degree at about 55 °C, studies were being carried on at
the temperatures below 55 °C. Preliminary result showed that
preferential (002) orientation of HA coatings could grow on
human enamel within 1 h at 40 °C by electrodeposition.
In conclusion, electrodeposition was proved to be an effec-
tive method for restoring the initial enamel lesions in 1 h with
the current density of 0.5 mA/cm2 at 55 °C. In addition, several
appliances had been invented to increase the retention or inhibit
the diffusion of a dental therapeutic agent. With the inspiration
of the previous inventions such as the mouth guard [29] for
dental therapy, we could imagine redesigning a mouth guard as
6150 Y.-M. Liao et al. / Thin Solid Films 516 (2008) 6145–6150
the two-electrode electrochemical cell with a suitable mouth
shape. While the inner peripheral wall would be served as the
cathode, the outer peripheral wall would be served as anode.
Then the electrodeposition would show a remarkable potential
application on the restoration of the enamel with HA coating in
clinic with a low cost of equipment.
5. Conclusions
In summary, preferential (002) orientation HA coatings
could grow on human enamel within 1 h by electrodeposition.
The HA coatings combined with the substrate tightly and
showed an acicular morphology. The Vickers micro-hardness
increased and approached to that of the sound enamel, and
the antibacterial activities of the enamel were also improved.
Electrodeposition was proved to be an effective method for
preparing HA coatings on human enamel and could be con-
sidered as a promising method to restore the initial enamel
lesions in clinic.
Acknowledgments
The teeth were provided by Xiamen stomatological hospital
and Xiamen 174 hospital. This study was financially supported
by the National Natural Science Foundation of China (NSFC
30470434) and Key Scientific Project of Fujian Province of
China (2005HZ01-3).
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