Materials Science Forum Submitted: 2019-12-17

ISSN: 1662-9752, Vol. 1000, pp 97-106 Accepted: 2020-02-25

doi:10.4028/www.scientific.net/MSF.1000.97 Online: 2020-07-03
© 2020 Trans Tech Publications Ltd, Switzerland

The Coating of Bovine-Source Hydroxyapatite on Titanium Alloy

(Ti-6Al-4V ELI) Using Electrophoretic Deposition for Biomedical
Application
Dian Juliadmi1,a, Nuzul Ficky Nuswantoro1,b , Hidayatul Fajri1,c,
Irma Yulia Indriyani2,d, Jon Affi2,e,Menkher Manjas1,f, Djong Hon Tjong3,g,
and Gunawarman2,h *
1
Biomedical Science, Faculty of Medicine, Andalas University, Indonesia
2
Mechanical Engineering Department, Faculty of Engineering, Andalas University, Indonesia
Biology Department, Faculty of Mathematics and Science, Andalas University, Indonesia
3

dianjuliadmi@gmail.com, bnuswantoro.ratissa@gmail.com, cfhidayatul13@gmail.com,

a
d
irmayuliaindriani93@gmail.com,ejonaffi@gmail.com,fmenkherm@yahoo.com,
g
tjong20@yahoo.com, hgunawarman@eng.unand.ac.id
*corresponding author

Keywords: bovine hydroxyapatite, Ti-6Al-4V ELI, electrophoretic deposition, biomedical application

Abstract. Research about the utilization of titanium alloy (Ti-6Al-4V ELI) as implant material in the
treatment of orthopedic cases had been increasing. Health problems appear due to the drawbacks of
using titanium. The lack of titanium using is bio-inertness characteristic, which decreasing its
bioactivity and results in low bone growth and effect for implant failure. The titanium can be modified
with coating on the surface using a bioactive substance that is natural-source hydroxyapatite. Bovine-
source hydroxyapatite (bovineHA) contains apatite component that is similar to human bone apatite.
The coating process was carried out using particle size variation (25 μm, 63 μm, and 125 μm) of
bovineHA. The electrophoretic deposition (EPD) method was applied to coat hydroxyapatite with 10
volt for 5 minutes onto the titanium surface. The result showed that different size particles have an
effect on coating properties. The coating composed by particle-sized 25 μm has better surface
coverage (95.89%), indicating more particle mass (particle weight 6.97x103 μg) attached to surface
material, thus resulting thick coating. The good coating characteristic using bovine-source
hydroxyapatite with small particle size was expected can be used in biomedical applications due to
fulfill the prerequisite of the bone implant.

Introduction
The challenge in medical application, particularly in orthopedics, about using biomaterial for bone
defects treatment caused by disease or trauma had been rising and aims for increasing the quality of
human [1-3]. The requisite for choosing biomaterial must have good biomechanical properties, and
the pivotal thing is excellent biocompatibility. Thus it can be accepted within the body regardless of
rejection and infection. Inquiry for metal-based implant also must role for support healing process in
the orthopedic application as bone prostheses [4]. Titanium is successful as a clinically used
biomaterial for the treatment of bone defects, which has superior biomechanical properties such as
high resistance corrosion and good biocompatibility in aspect interaction with proteins and cells [5,6].
Using titanium alloy, Ti-6Al-4V ELI as an implant considered, must have integration with bone
tissue (osseointegration) preventing implant failure [7]. Meanwhile, the material property of bio-
inertness titanium results in less bioactive or poor osteoinductive and cannot induce bone healing
around implantation forming chemical bonds with material [8-10]. An approach to exploit
biomechanical properties of titanium, (high strength, low elastic modulus, and high corrosion
resistance) [11], material surface was modified using organic material. The coating by hydroxyapatite
aims for improving the load-bearing ability of titanium combine with bioactivity to achieve healing
process and implantation successful [12-14].The coating on surface materials will interact with the

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98 Advanced Materials Research QiR 16

biological environment involved in new bone formation, and herein will be interfacial formed
chemical bonds between the implant and bone [15,16] for stable fixation avoiding implant loosening.
The existence of hydroxyapatite coating will extend using time of implant to enhance long-term
reliability [17]supposed to be followed by no immune rejection reaction [18].
The titanium materials coated with chemically synthetized-hydroxyapatite (Ca10(PO4)6(OH)2)
reported increasing osseointegration between material and bone tissue due to its biocompatibility and
favorable osteoconductivity [19]. Cause of its similarity with the inorganic component of bone apatite
[20], calcium and phosphate ions in hydroxyapatite compounds would attract proteins and cells,
leading to new bone ingrowth [21,22]. Synthetic hydroxyapatite has a different crystal structure with
natural-source apatite [23]. Pure hydroxyapatite synthesized by chemical process needs complex and
high-cost method; contains a shortage of trace elements that is beneficial in bone metabolism; and
results in non-stoichiometric calcium-dependent hydroxyapatite powders [24-27]. Nevertheless,
natural sources hydroxyapatites are favorable for bioactivity indicated by the formation of new
apatite. Biological sources from animals or biowaste such as xenogenic bone and dental from bovine,
fishbone, eggshells, and seashells can be used for hydroxyapatite extraction, and it is eco-friendly
[23,26-31].
Bovine-source hydroxyapatite (bovineHA) can be coated onto titanium surface,Ti-6Al-4V ELI,
through such methods, the one is electrophoretic deposition (EPD), a feasible coating method for
implant-orthopedic application [13,32]. Principally, the method uses an electric field to migrate and
deposit of charged particles suspended within a certain solution, then performance by a sintering
process to increase the adhesion strength of green coating on the material surface [33,34]. Those
advantages are efficient in technical working, thickness, and morphology of coating can be controlled
according to its parameters and can be applied to complex materials [35]. Therefore, in this research,
using the electrophoretic deposition method for coating bovineHA by variant particle size on Ti-6Al-
4V ELI was conducted in achieving good coating characteristic, which is suitable for biomedical
application.

Materials and Methods

Material titanium alloys, a Ti-6Al-4V ELI in disc shape with size 4x4mm2, was used as a research
substrate. Residual stress from lathe machining removed with solution treatment using vacuum
furnace with temperature 950oC for 1 hour. Substrates were ground using SiC paper in achieving an
even and clean surface after cutting processing. The cleaning process was conducted with some
solutions in steps are ethanol and acetone for every 15 minutes by an ultrasonic bath. Samples
immersed in nitric acid solution 25% (v/v) for 30 minutes and followed by NaOH solution for 1 hour.
All of the samples dried using a hot plate with temperature 50oC for 5 minutes.
Hydroxyapatite extracted from bovine bone (Universitas Andalas, Padang, Indonesia) was
prepared in different particle sizes that are 25 µm, 63 µm, and 125 µmusing a ball milling process.
Bovine bone-source hydroxyapatite (bovineHA) powders dissolved in ethanol with concentration 40
mg/mL and then stirred for 1 hour and pH value 4 adjusted with the addition of nitric acid solution.
The EPD process was carried out at constant voltage 10 volts for 5 minutes using DC power supply
under continuous stirring 100 rpm and cathodic technique (substrate as the cathode and graphite as
the anode in the distance 1 cm apart) [36]. Deposited materials dried in room temperature for 24 hours
and sintered with vacuum furnace at temperature 800oC for 15 minutes with heating rate 10oC/min to
increase binding of coating. Morphology characteristic was observed using an optical microscope and
Scanning Electron Microscope (SEM) Hitachi S 3400 with voltage 10kV. The chemical composition
assessment tested with EMAX x-act-liquid nitrogen less X-Ray Detector connected with SEM.
Analysis for coating thickness was measured using Thickness Gauge Series GM280.
 Materials Science Forum vol. 1000 99

Results and Discussions

Bovine-source hydroxyapatite (bovineHA) powders were obtained by the ball milling process and
shieved to get variant particle size that are 25 μm, 63 μm, and 125 μm. The bovineHA coating on Ti-
6Al-4VELI was successfully produced using electrophoretic deposition methods. The deposition
process by voltage 10 volt for 5 minutes could deposit bovineHA particles with different sizes so
formed particles covering up the material surface in plain view. Low voltage and short deposition
time are sufficient to migrate and deposit hydroxyapatite [37] up to particle size 125 μm. The coating
composed by bovineHA particles size 125 μm revealed homogenous morphology, whereas coating
thickness with particles size 25 μm and 63 μm are thicker than others (Figure 1). Observation using
an optical microscope revealed uneven coating for all over samples but less agglomeration at the
coating layer composed by particle size 125 μm.

A B C

Figure 1.Bovine-source hydroxyapatite cover-up Ti-6Al-4V ELI material surface by different

particle size; (a)25 μm (b)63 μm (c)125 μm after coating using EPD method observed with an
optical microscope

The voltage migrates and deposit of charged hydroxyapatite to material surface acting as a cathode
may affect agglomeration. High voltage will migrate small-sized particles and affect particle
arrangement improperly in short deposition time [38]. However, low voltage (10 volt) still results in
the agglomeration of some particles in this research. Using of dispersant can avoid agglomeration
particles due to its role to stabilize the suspension in achieving good particle distribution on the
material surface [33].

A B C

Figure 2. Morphology of bovine-source hydroxyapatite coating on titanium Ti-6Al-4V ELI formed

by different particle size using SEM; a) 25 μm, b) 63 μm, and c) 125 μm

Morphology observed using SEM in Figure 2 showed that the coating composed by particles size
25 μm more reliable than other coatings. No crack indicated coating formation with even particle
distribution and homogenous thickness. The crack formation was seen on coatings consisted of
particle size 63 μm with heterogeneous layers. The formation of asymmetry multilayer effect for
uneven thickness along with cracking could promote delamination. Releasing of particles cause of
low adhesion coating in the crack region would initiate degradation [39]. Dissolution had
disadvantages that may disturb the metabolism system due to releasing harmful ions. Otherwise,
dissolution may induce healing process in fracture repairing involved osteogenesis [40,41].
100 Advanced Materials Research QiR 16

Unavoidable firing shrinkage process of hydroxyapatite and mismatch thermal expansion

coefficient between Ti-6Al-4V ELI material and HA coating will promote cracking after heat
treatment [39,42]. Karimi et al. [43] also founded crack propagation within bovineHA coatings on
NiTi material using the EPD method and sintering at 660oC with porous particles, which possible
trigger loosening or exfoliating of coating layers. The bovineHA can induce bone-like apatite
formation after immersion in simulated body fluid (SBF), indicating bioactivity characteristics.

250

10
200
9

Coating Thickness (µm)

Coating Weight (x103 μg)

8
7 150
6
5 100
4
3
50
2
1
0 0
25 µm 63 µm 125 µm 25 µm 63 µm 125 µm
Particle Size Particle Size

100 3

98 2.5

96
Surface Coverage (%)

2
Ca/P Ratio

94
1.5
92
1
90

88 0.5

86 0
25 μm 63 μm 125 μm 25 μm 63 μm 125 μm
Particle Size Particle Size

Figure 3. Characterization of bovine-source hydroxyapatite coating on titanium Ti-6Al-4V ELI

formed by different particle size using SEM; 25 μm, 63 μm, and 125 μm based on coating weight,
thickness, surface coverage and Ca/P ratio

Measurement of surface coverage value for bovineHA coating showed that particle sizes 125 μm
covered up material surface preferable rather than others (Figure 3). However, those coatings have
lower thickness even than its particle size. The coating might be composed of smaller than particle
sizes 125 μm. Assessment using atomic force microscopy (AFM) with higher resolution (nanoscale),
Duta et al. [24] discovered the smallest particle sizes ~45nm within natural hydroxyapatite coating
 Materials Science Forum vol. 1000 101

consisted of grain sizes of ~0.87 μm. Particle sizes 125 μm did not exist within coating might be
caused by the exfoliating of particle as impact from lower adhesion strength with the material surface.
Hadidi et al. [44] stated that particle size, substrate preparation, temperature, and sintering time effect
to adhesion strength of coating obtained by the EPD process.

Figure 4. EDX results of bovineHA coating on material Ti-6Al-4V ELI surface with particle size: a)
25 μm, b) 63 μm, and c) 125 μm

The used particle size of hydroxyapatite has an association with deposition of particle mass during
the coating process using the EPD method. Calculation of coating mass was applied for sintered
coatings. The graph in Figure 3 showed that treatment using the same voltage and deposition time
may increase deposition for particle sizes 25 μm rather than large particle size. By using the same
energy, smaller particles migrate and deposit faster on surface material in a short time. Migration of
large particle size needs high energy for migration and deposition [33]. The total of hydroxyapatite
coating mass indicates coating thickness on surface material Ti-6Al-4V ELI and area covered up by
bovineHA coating. The high mass of coating by particle sizes 25 μm revealed that the increasing
thickness and followed by the wide-area covered up by the coating. The more deposited mass
bovineHA coating by particle size 63 μm, the more thickness but decreasing surface coverage. Weak
bonds of between-particles beyond size 63 μm will promote exfoliation. Meanwhile, bovineHA
coating by particle sizes 125 μm had low particle mass and results in high thickness and surface
coverage.
102 Advanced Materials Research QiR 16

The sintering process carried out after the coating method has an effect on coating composition.
Observation using EDX in Figure 4 produced coating by particle sizes 25 μm has Ca/P ratio of 1.95,
and in charged of a prerequisite for biomedical application 1.67 Ca/P ratio. Using hydroxyapatite by
Ca/P ratio 1.0-1.7 has neither toxic nor induction foreign body reaction (immunological process),
therefore lead to the healing process induced by hydroxyapatite [21]. The high Ca/P ratio may be
caused by the sintering process and consequence as composition changes. Ramesh et al. [28] stated
that bovine HA has good thermal stability because no decomposition even though after sintering with
temperature 1000oC. The comparable result obtained by Ramesh et al. [28] having Ca/P ratio 1.58-
1.88 and Herliansyah et al. [45] founding Ca/P ratio of 1.93. Sintering parameters such as holding
temperature and atmospheric conditions are a crucial for affecting transformation in resultant HA
phase [46,47].
Principally, biological apatites are non-stoichiometric compound because it has other ions
substituting Ca2+, OH-, or PO43- on apatite lattice component [48]. Substitution of natural ions like as
Na, Mg, Zn, Cl, CO32- have the capability to increase biological activity by hydroxyapatite from
organic sources [45,49]. Therefore, natural hydroxyapatite with impurities may involve in new bone
growth and biological metabolism [28]. The characteristic of natural-source hydroxyapatite
influenced by the used extraction method, which can be obtained by facile and feasible technical with
using organic residue from living things due to abundant stock [50]. Research by Shi et al. [51] proved
that natural-sources hydroxyapatite has good biochemical and involved in bone metabolisms such as
proliferation, differentiation, cell adhesion, and mineralization. Rincon-Lopez et al. [52] stated that
no restriction in biomedical application for either synthetic or natural hydroxyapatite. In addition, for
the religion aspect, natural hydroxyapatite categorized as the halal product for Moslem people [28].
Although there is a slight change in Ca/P ratio, the EPD method can be used for acquiring
bovineHA coating on material surface Ti-6Al-4V ELI in appropriate with biomedical application.
Therefore, using this coating by the EPD method is one of the utilization solutions for orthopedic
applications, particularly. The consideration is bovineHA up to standard as coating with good
biocompatibility and bioactivity accelerating the healing process in a relatively short time to achieve
implant stability and achieving osseointegration.

Conclusions
Bovine hydroxyapatite (bovineHA) contain apatite component that is similar to human bone apatite
and can be used as source hydroxyapatite. The coating process was carried out using different particle
size variation (25 μm, 63 μm, and 125 μm) of bovineHA. The electrophoretic deposition (EPD)
method was applied to coat hydroxyapatite with 10 volts for 5 minutes parameters onto the titanium
surface. The result showed that different size particles have an effect on coating characteristics. The
coating composed by particle-sized 25 μm has better surface coverage (95.89%), indicating more
particle mass (particle weight 6.97x103 μg) attached to surface material, thus resulting thick coating.
On the contrary, no found particle with the same size on the coating formed particle-sized 125 μm.

Acknowledgment
This research was supported by the Ministry of Research, Technology, and Higher Education of the
Republic of Indonesia for supporting and funding under Penelitian Berbasis Kompetensi (PBK)
program with contract no. T/5/UN.16.17/PT.01.03/ PT-MM/2019.
 Materials Science Forum vol. 1000 103

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