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Original Article

Effect ofanodization and alkaliheat treatmenton

the bioactivity of titanium implant material
(an invitro study)
Ramy A. Abdelrahim1,2, Nadia A. Badr3, Kusai Baroudi4
Department of Dental Biomaterials, School of Dentistry, AlAzhar University, 3Department of Dental Biomaterials, Faculty
of Oral and Dental Medicine, Cairo University, Egypt, Departments of 2Restorative Dental Sciences and 4Preventive Dental
Sciences, Alfarabi Colleges, Riyadh, Kingdom of Saudi Arabia
Corresponding author(email: <>)
Dr.Kusai Baroudi, Department of Preventive Dental Sciences, Alfarabi Colleges, P. O. Box: 85184, Riyadh 11691,
Kingdom of Saudi Arabia.

Received: 04-01-16 Accepted: 04-04-16 Published: 30-05-16

Objective: This study was aimed to assess the effect of anodized and alkaliheat surface treatment on the bioactivity of
titanium alloy(Ti6Al4V) after immersion in Hanks solution for 7days. Materials and Methods: Fifteen titanium alloy
samples were used in this study. The samples were divided into three groups(five for each), five samples were anodized
in 1M H3PO4 at constant voltage value of 20v and another five samples were alkalitreated in 5 M NaOH solution for
25min at temperature 60C followed by heat treatment at 600C for 1h. All samples were then immersed in Hanks
solution for 7 days to assess the effect of surface modifications on the bioactivity of titanium alloy. The different
treated surfaces and control one were characterized by Xray diffraction, atomic force microscopy, scanning electron
microscopy, energydispersive Xray spectroscopy, and Fourier transformation infrared spectroscopy. Statistical
analysis was performed with PASW Statistics 18.0(Predictive Analytics Software). Results: Anodization of Tialloy
samples(GroupB) led to the formation of bioactive titanium oxide anatase phase and PO43 group on the surface. The
alkaliheat treatment of titanium alloy samples (Group C) leads to the formation of bioactive titania hydrogel and
supplied sodium ions. The reaction between the Ti sample and NaOH alkaline solution resulted in the formation of
a layer of amorphous sodium titania on the Ti surface, and this layer can induce apatite deposition. Conclusions: The
surface roughness and surface chemistry had an excellent ability to induce bioactivity of titanium alloy. The
anodization in H3PO4 produced anatase titanium oxide on the surface with phosphate originated from electrolytes
changed the surface topography and allowed formation of calciumphosphate.

Key words: Alkaliheat treatment, anodization, bioactivity, surface roughness

INTRODUCTION their superior biocompatibility due to their excellent

corrosion resistance, and bioinertness of the oxide
Titanium and its alloys widely used as implant layer.[1,2] Therefore, titanium(Ti) and its alloys
biomaterials because of their relatively superior mainly(Ti6Al4V) became the most suitable metal for
physical properties such as lower modulus, low density, endosseous implants fabrication.[3,4]
and relatively low elastic modulus and because of
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How to cite this article: Abdelrahim RA, Badr NA, Baroudi K.

DOI: Effect of anodization and alkali-heat treatment on the bioactivity
10.4103/2231-0762.183107 of titanium implant material (an in vitro study). J Int Soc Prevent
Communit Dent 2016;6:189-95.

189 2016 Journal of International Society of Preventive and Community Dentistry | Published by Wolters Kluwer -Medknow
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Abdelrahim, etal.: Bioactivity of titanium implant materials

One of the most serious and prevalent mechanisms, Among surface treatments, the anodizing exhibited
for obtaining biofunctional and biocompatible material effectiveness in modifying surface properties of
for biomedical use, is surface modification. The surface metals,[19] particularly in inducing the bioactivity of
modification technique is a process that changes the titanium.[20] Anodic oxidation of titanium implants
material surface through changes in their chemical involved several changes in oxide characteristic increases
composition, structure, and morphology, but the bulk not only the thickness of native oxide layer but also the
mechanical properties of the material not affected.[5,6] surface texture such as the ability of fabricating porous
Ti oxide films, the changeability of the crystalline
The surface of a biomaterial plays a key role in structure, chemical composition of oxide film, and
determining biocompatibility.[7,8] Therefore, through formation of rough and homogeneous TiO2
biomaterials surface in contact with the biological tissue layer with numerous nanoscale pores.[20] The surface
or fluid is named biointerface.[9] The biointerface is modification of titanium implants by anodic oxidation
the place where the modifications are commonly carried in phosphoric/sulfuric acid electrolyte systems showed
out to make biomaterials more compatible, and these an increase in oxide thickness,[20] which significantly
modifications are fall under three categories, namely, improves the bone response for better osseointegration
morphologic, physicochemical, and biological. Surface between Ti surface and surrounding bone.[21]
of biomaterials represents a crucial value for biomedical
field and determine biocompatibility of the biomaterial This study aimed to investigate the effect of different
because only the uppermost molecular layers are in surface treatment on the bioactivity of titanium alloy
intimate contact with the biological environment.[10] (Ti6Al4V) and their ability to form calcium phosphate
Thus, any interaction between the implant materials apatite crystals on their surfaces after immersion in a
and the biological environment occurs at or through the simulated body fluid.
level of narrow zone termed as the interface. Therefore,
the biomaterial scientists have a special concern with the MATERIALS AND METHODS
interfacial chemistry that determined by the uppermost
few nanometers that composed the materials surface.[10] This study was performed in the Department of
Biomaterials, Faculty of Dental Medicine, AlAzhar
For an optimally bonding between the implant material University. Ethical approval of AlAzhar University was
and living bone in the human body, it is essential that taken(under number 207/2009).
the implant can form bonelike apatite layer on its
surface. The nucleation of apatite on the surface of A total of fifteen samples made of Ti6Al4V alloy(extra
a material is induced by the functional groups on its low interstitials F 13084 alloy, USA) were used in
surface.[11,12] The surface modification of titanium this study. The sample dimensions were rectangular in
implants is an interested issue for investigation aiming shape of 15mm in length, 7mm in width, and 2mm
to induce acceleration of normal bone healing allowing in thickness. The titanium samples were silky polished
immediate loading of the implant.[11,12] using sandpaper starting from gravel #400 to #1200,
and then ultrasonically cleaned in deionized water for
The treatment of titanium surfaces by alkaline solution 5min before dried by air dryer.
followed by heat treatment was described by some
investigators,[1316] who successfully proved the bioactive Then, the samples were classified into three groups
induction both in vitro[17] and in vivo.[18] Hydroxyapatite according to the following treatments: Five sample
formation onto Ti surface can be accelerated by treatment each, according to the received surface treatment. The
titanium surface by NaOH solution followed by heat surface treatment involved anodization(GroupB) and
treatment to produce layer of sodium titanate on titanium alkaliheat treatment(GroupC). Five samples were
surface. Titanium surface that treated with alkaline kept untreated and served as control(GroupA). Ten
solution followed heat treatment is thought to bond to samples of GroupsB and C were etched in acid blend
living bone via formation of apatite layer on its surface of 80ml/l HNO3, 60ml/l HF, and 150ml/l H2O2 for
while the titanium surface that treated with alkaline 5min at room temperature before the surface treatment.
solution only did not.[13,14] Recently, invitro studies showed
that bonelike apatite could be formed on the titanium Five samples were subjected to anodization process,
surface that treated with alkaline followed by heat GroupB, after acid etching to form an oxide porous
treatment when immersed in simulating body fluid(SBF) film on their surfaces. The anodizing processes were
via formation of titana hydrogel on titanium surface.[13,14] performed in electrochemical cell that has an anodizing

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Abdelrahim, etal.: Bioactivity of titanium implant materials

solution as electrolyte and two electrodes using a direct the precipitation. Fourier transformation infrared
current source with value of 20v as constant voltage. spectroscopy (FTIR) (FT/6300 Type A. Jasco, Japan)
Acathode of platinum is connected to the negative was used to analyze qualitatively the chemical structure
terminal of a voltage source and placed in the solution, of biomimetic coat which precipitated on the immersed
and Tialloy sample is connected to the positive surface.
terminal of the voltage source and also place in the
solution as anode with about 4cm as a fixed distance Statistical analysis
between cathode and anode. An electrolyte bath consists
of 98g/L(1 M H3PO4) aqueous solution(1L total Statistical analysis was performed using oneway
ANOVA followed by Tukeys post hoc test at the
volume) having pH 1.5 that was stirred with magnetic
significance level of = 0.05.
stirrer with 180rpm. The anodizing process was
carried out at room temperature for 15min, and then
the samples were ultrasonically cleaned for 5min in RESULTS
deionized water. The XRD analysis of the anodized group revealed
the presence of anatase TiO2 phase while the
The samples of GroupC were alkalitreated by
alkaliheattreated group reveals the presence of rutile
immersion in 5 M NaOH solution for 25min at TiO2 and TiOH phases as shown in Figure1.
temperature 60C and washed with distilled water, and
dried in the air. Then, the samples were heat treated Atomic Force Micrograph of the alkaliheat treated
at 600C for 1h in a furnace at heating rate 30C/min. group shows the highest surface roughness among
Then, they gradually cooled to room temperature. the all investigated group with multiple deep valises
in threedimensional(3D) image, followed by the
The surfaces of each group were characterized after anodized group, which shows severe undulation of the
surface treatment using Xray diffraction (XRD) surface; however, no sharp peaks were evident in the 3D
(Philips Analytical Xray B.V.), scanning electron images and the least surface roughness was the control
microscopy (SEM) (Technics Inc., JEOL JSM 5410, group, and its 3D image shows slight irregularities and
Japan production), and atomic force microscopy serration of the metal surface due to polishing as shown
(Thermomicroscopes Autoprobe CP research, scanner in Figure2.
100m, USA).
The statistical analysis of the surface roughness
Before immersion in Hanks balanced salt solution of the different surface treated groups is shown
(HBSS), the samples were rinsed with distilled in Figure3. ANOVA test showed that there was a
water, washed further with ethanol in an ultrasonic statistically significant difference among mean(Ra)
bath for 20min, and then dried in air. All the values of the investigated groups with P<0.05.
investigated samples were immersed for 7days in the Pairwise comparisons among the groups revealed that
HBSS which was used as SBF at pH7.4 and 37C. alkaliheattreated group showed the highest statistically
HBSS was renewed every 3days to maintain the significantly mean(Ra) value.
level of ion concentration allowing for biomimetic
coat precipitation on the sample surfaces. After the
immersion of the Tialloy samples in HBSS, the
specimens were removed from the solution, gently
rinsed with distilled water and then dried at 60C for

To assess the biomimetic coat on the differently treated

samples, microstructural/morphological changes of the
specimens surfaces were characterized. Meanwhile,
the surface morphology was examined using SEM.
Energy dispersive X-ray analysis (EDXA)(Model: Link
ISISOXFORDEngland. The device is attached
to JEOL5400 Scanning Microscope) assembled on
the SEM was used to determine the atomic % of
calcium, phosphorous elements, and Ca/P ratio of Figure1: Xray diffraction pattern of all investigated groups

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Abdelrahim, etal.: Bioactivity of titanium implant materials

Figure2: Atomic force micrographs of GroupA, B, and C(threedimensional)

The scanning electron micrograph of the control group

showed that polishing marks of Tialloy obviously seen
on the sample surface, and the anodized group revealed
presence of numerous and unattached, welldefined,
nanosized pores while the alkaliheattreated group
revealed the presence of numerous, unattached,
welldefined, nanosized pores that arranged in an
ordered pattern and equally distributed all over the
structure on the surface spread among microsized
particles. It was prominent that the pores are deeper
with darker contrast[Figure4].

After immersion in Hanks solution, the SEM showed

Figure3: Mean values of surface roughness(Ra) of the investigated
discrete small apatite crystals formed on the surface of groups
the control samples and the EDXA spectrum showed
phosphorus atomic% of 53.94 and calcium atomic%
of 46.05 and Ca/P ratio of 0.85, which is much lower
than stoichiometric value of hydroxyapatite(1.67). It
indicates the synthesis of monocalcium phosphate
monohydrate of chemical composition of Ca
(H2PO4)2.H2O and/or dicalcium phosphate dehydrate
(brushite) Ca HPO4.2H2O[Figures4IIand 5].

The SEM of the anodized group showed contiguous

layer of apatite nuclei formed on the surface of
the anodized samples such that the substrate is Figure4: Scanning electron micrographs of GroupA, B, and C before
not obviously seen and EDXA spectrum showed and after immersion in Hanks solution
phosphorus atomic% of 43.72 and calcium atomic%
of 56.28 and Ca/P ratio of 1.28, which is less than or occasionally named precipitated HA of chemical
stoichiometric value of hydroxyapatite(1.67). This formula Cax Hy(PO4) z.nH2O[Figures4II and 5].
ratio indicates the synthesis of octacalcium phosphate
of chemical composition Ca8(HPO4)2(PO4)45H2O The FTIR analysis of control group, GroupA, is
[Figures4II and 5]. shown in Figure6. The spectrum revealed that the
presence of vibration band at 606 cm1 is for symmetric
While the SEM of the alkaliheattreated group stretching vibration of PO43(4). The bands at 865 and
exhibited layer composed of an interconnected spherical 985 cm1 are bands for symmetric stretching vibration
particle, this layer is to some extent thicker, and the of PO43(1) while band at 1019 cm1 for asymmetric
particles are larger in size. The EDXA spectrum showed stretching vibrations of PO43(3). The symmetric
phosphorus atomic% of 37.96 and calcium atomic% stretching vibrations of PO43(3) are represented
of 62.04 and Ca/P ratio of 1.67. This ratio suggests by bands at 1058 and 1090 cm1 while the band at
the formation of calcium deficient hydroxyapatite 1075 cm1 is absent. The symmetric stretching vibration

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Abdelrahim, etal.: Bioactivity of titanium implant materials

Figure6: Fourier transformation infrared spectroscopy spectra of

investigated GroupsA, B, and C after immersion in Hanks solution

1455 cm1. The reflectance bands characteristic for H2O

are observed at 629 and 3230 cm1 which denoting OH
group of HA. Abroad and not very intense band in HAp
at 3230 cm1 may be due to the presence of lattice water
B in the solid or it caused by the Hbonding between the
adsorbed H2O and the OH group of the apatite. The
carbonyl groupC=O is identified from bands at 1693
and 1760 cm1.

The FTIR analysis of alkaliheattreated samples,

GroupC, is shown in Figure6. The spectrum revealed
that the presence of vibration bands at 561 and
at 602 cm1 is for symmetric stretching vibration of
C PO43(4) while bands at 871, 960, and 985 cm1 are
Figure5: Energydispersive Xray spectrum of group A, B and C after
characteristic bands for symmetric stretching vibration
immersion in Hanks solution of PO43(1) while band at 1019 cm1 is characteristic
for asymmetric stretching vibrations of PO43(3).
bands of HPO42 group are observed at 1113, 1141 and The symmetric stretching vibrations of PO43(3) are
1226 cm1. The symmetric stretching vibration bands of represented by band 1093 cm1 while the band at 1048
CO32 are captured at 873, 1414, and 1453 cm1. The and 1075 cm1 is absent. The symmetric stretching
reflectance bands characteristic for H2O were observed vibration bands of HPO42 group are observed at 1111
at 625, 3083, 3249, 3333, and 3546 cm1 which denoting and 1220 cm1. The symmetric stretching vibration
OH group of HA. The carbonyl groupC=O is bands of CO32 are captured at 1416 and 1455 cm1. The
identified from bands at 1698 and 1712 cm1. reflectance bands characteristic for H2O were observed
at 626, 3001, 3195, and 3550 cm1 which denoting OH
The FTIR analysis of anodized samples, GroupB, is group of HA. The carbonyl GroupC=O is identified
shown in Figure6. The spectrum revealed that the from bands at 1712, 1726 cm1.
presence of vibration bands at 563 and at 604 cm1 is
for symmetric stretching vibrations of PO43(4). The DISCUSSION
bands at 862 and 965 cm1 are bands for symmetric
stretching vibration of PO43(1) while band at The anodization in H3PO4 produced anatase titanium
1020 cm1 is for asymmetric stretching vibrations of oxide on the surface with phosphate originated from
PO43(3). The symmetric stretching vibrations of electrolytes and changed the surface topography.[22,23]
PO43(3) are represented by band 1076 cm1 while In addition, crystal structures of titanium oxide found
the band at 1047 and at 1096 is absent. The symmetric on titanium surface after anodization[24,25] allowed
stretching vibration bands of HPO42 group are observed formation of calciumphosphate in Hanks solution
at 1114, 1143, and 1222 cm1. The symmetric stretching with ion concentrations approximating those of human
vibration bands of CO32 are captured at 875, 1416, and blood plasma.[26]

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It is reported that in aqueous body fluid, the OH apatite nucleation. On accumulation of Ca2+ions, the
bonds easily to the Ti4+of TiO2 and subsequently calcium titanate changed to be positively charged and
TiOH groups is formed.[27] The isoelectric point combined with negatively charged phosphate to form
of titanium oxide surface at electrolyte pH nearly calcium phosphates and then transform to apatite.[32]
56 shows zero net charge. Apositively charged
surface will be created by production of(TiOH2)+ The difference between the surface activities of
in the surface layer at electrolyte pH<5. The TiOH chemically and electrochemically treated Ti6Al4V is
will be deprotonated at electrolyte pH>6, and due to the different surface morphology and structure,
becomes(TiOH) so the surface will be a negatively i.e.,increased surface roughness lead to increased
charged. Therefore, the surface will be negatively bioactivity.[33,34]
charged at the physiological pH of~7.4, due to the
existence of deprotonated acidic hydroxides.[28] That Limitations
negatively charged surface can attract cations such as
calcium that subsequently could react with HPO42 or Although this research was carefully prepared, I am still
H2PO4 to form calcium phosphates.[29] aware of its limitations and shortcomings.

The titanium cations can react with phosphoric acid to First of all, the research was concerned only with invitro
form phosphates as explained in the following reactions: and have not any invivo study.

Ti4+ + 2H3PO4Ti(HPO4)2 +4H+(1) Second, the study did not cover different surface
modification methods.
Ti4+ + Ti(HPO4)2+H3PO4Ti2(PO4)2
(HPO4) + 4H+(2) Third, the study was done using titanium alloy discs
and not titanium implants.
In turn, the nucleation of calcium phosphate
nanocrystals within nanopores could be facilitated by CONCLUSIONS
the presence of these phosphate ions.[30]
Electrochemical treatment of anodic oxidation in
Ca9(HPO4)(PO4)5OH3 Ca3(PO4)2+H2O(3) H3PO4 solution converted titanium oxide to the
favorable anatase phase and provided PO43 which
The reaction between the Ti sample and NaOH acted as nuclei for developing HA crystals. Immersion
alkaline solution will lead to the formation of a layer of in NaOH solution before alkaliheat treatment of
amorphous sodium titania on the Ti surface. Alkaline titanium alloy supplied sodium ions and titania
treatment for 25min would provide a titania layer of hydrogel which are beneficial substrate to precipitate
considerable thickness with mature submicrometer calcium and phosphorus in a ratio comparable to that of
porosity. Further, heat treatment at temperature(600C) stoichiometric HA.
for extended time(1h) resulted in a cracked layer
with an increased thickness. The thickness and gradual Financial support and sponsorship
transformation of the titania layer from an amorphous
to a crystalline state depended almost linearly on
the period and the temperature of the chemical
treatment.[31] Conflicts of interest
There are no conflicts of interest.
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