Sutton E.

Wheelis Effects of decontamination solutions

Izabelle M. Gindri
Pilar Valderrama
on the surface of titanium:
Thomas G. Wilson Jr investigation of surface morphology,
Jessica Huang
Danieli C. Rodrigues
composition, and roughness

Authors’ affiliations: Key words: AFM, corrosion, detoxification, oxide layer, titanium
Sutton E. Wheelis, Izabelle M. Gindri, Danieli C.
Rodrigues, Department of Bioengineering,
University of Texas at Dallas, Richardson, TX, USA Abstract
Pilar Valderrama, Thomas G. Wilson Jr, Jessica Aim: To investigate the impact of treatments used to detoxify dental implants on the oxide layer
Huang, Private Practice of Periodontics, Dallas, TX,
morphology and to infer how changes in morphology created by these treatments may impact
USA
re-osseointegration of an implant.
Corresponding author: Materials and methods: Pure titanium (cpTi) and the alloy Ti6Al4V were subjected to a series of
Danieli C. Rodrigues
800 W. Campbell Road
chemical treatments and mechanical abrasion simulating surface decontamination of dental
Richardson, Texas 75080-3021, USA implants. The morphology and roughness of the surface layer before and after treatment with
Tel.: 972 883 4703 these solutions were investigated with optical and atomic force microscopy (OM, AFM). The
Fax: 972 883 4653
e-mail: Danieli@utdallas.edu solutions employed are typically used for detoxification of dental implants. These included citric
acid, 15% hydrogen peroxide, chlorhexidine gluconate, tetracycline, doxycycline, sodium fluoride,
peroxyacetic acid, and treatment with carbon dioxide laser. The treatments consisted of both
immersions of samples in solution and rubbing with cotton swabs soaked in solution for 1, 2, and
5 min. Cotton swabs used were analyzed with energy dispersive spectroscopy (EDS).
Results: The microscopy investigation showed that corrosion and pitting of the samples were
present in both metal grades with immersion and rubbing methods when employing more acidic
solutions, which had pH <3. Mildly acidic solutions caused surface discoloration when coupled with
rubbing but did not cause corrosion with immersion. Neutral or basic treatments resulted in no
signs of corrosion with both methods. EDS results revealed the presence of titanium particles on all
rubbing samples.
Conclusion: It was demonstrated in this study that acidic environments coupled with rubbing are
able to introduce noticeable morphological changes and corrosion on the surface of both titanium
grades.

Titanium has been placed at the forefront for
use in dental implants because of its mechan-
ical strength and protective oxide layer, which
ensures the bulk material of an implant is
protected against corrosion. This oxide layer
(e.g., TiO2, TiO) is naturally formed and
regenerated in the presence of air and/or aque-
ous media (Gilbert 2012; Swaminathan & Gil-
bert 2013). Besides providing protection
against corrosive environments, oxide layers
also aid in providing porosity to the surface.
Porosity and roughness are elements that
Date: allow bone surrounding an implant to osseo-
Accepted 4 December 2014 integrate, providing stability for the implant.
To cite this article: Titanium is unique in the fact that bone
Wheelis SE, Gindri IM, Valderrama P, Wilson TG Jr, Huang J, finds the oxide layer of the metal favorable
Rodrigues DC. Effects of decontamination solutions on the
surface of titanium: investigation of surface morphology, for integration. This is critical in dental
composition, and roughness.
implant applications because a well-osseoin-
Clin. Oral Impl. Res. 00, 2015, 1–12
doi: 10.1111/clr.12545 tegrated implant can survive the mechanical
requirements of the oral environment, mini-
mizing micromotion and the potential for
fretting-crevice corrosion associated with
occlusal forces (Mouhyi et al. 2012). To
obtain a favorable environment for osseointe-
gration to occur and continue, routine main-
tenance after placement has been suggested
(Wilson et al. 2014). These procedures are
designed to diagnose and treat inflammatory
responses known as peri-implant mucositis
and peri-implantitis (Mouhyi et al. 2012;
Wilson et al. 2014). Peri-implantitis is char-
acterized by inflammation and continued loss
of integrated bone around the implant. If
peri-implantitis does occur, the clinician has
the option to either remove the infected
implant or perform debridement and detoxifi-
cation of the implant surface to remove the
biofilm (Valderrama et al. 2014). The goal of

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd 1
Wheelis et al
Effects of decontamination on the surface of titanium
debridement is the reintegration of bone on
previously infected implant surfaces (Yuan
et al. 2014).
When peri-implantitis occurs, chemical,
mechanical, and laser treatments can be
employed for removal of attached biofilms on
the surface of the implant (Subramani and
Wismeijer; Renvert et al. 2009; Tastepe et al.
2012; Kamel et al. 2013). Chemical treat-
ments typically employed for debridement of
contaminated surfaces include citric acid,
tetracycline, saline, chlorhexidine, hydrogen
peroxide, tetracycline, and doxycycline
(Finnegan et al. 2010; Gosau et al. 2010; De
Waal et al. 2014; Valderrama et al. 2014).
These chemicals may be applied with various
mechanical means to facilitate biofilm
removal. Er:YAG and CO2 lasers have also
been employed to remove biofilms (Thierry
et al. 2000; Schwarz et al. 2003a,b, 2005,
2011, 2013). Mechanical means including cu-
rettes and powder blasting have been typi-
cally used. Although some studies have
indicated that detoxification and debridement
can be effective in removing bacterial bio-
film, which may lead to re-osseointegration
(Ericsson et al. 1996; Persson et al. 2001;
Schou et al. 2003a), no human studies have
demonstrated reintegration of implant sur-
faces previously covered with biofilms. A few
studies, including that published by Alhag
et al. (2008), have found reintegration in the
oral environment in dogs after various detoxi-
fication treatments (H€urzeler et al.; Subrama-
ni and Wismeijer; Ericsson et al. 1996;
Table 1. Concentrations of various treatments
Treatment Concentration
Citric acid 40%
Hydrogen peroxide 3%
Hydrogen peroxide 15%
Chlorhexidine 0.12%
Chlorhexidine 1%
Sodium fluoride 0.12%
Fig. 1. pH value measurements before contact with immersion medium and after 8 min of contact with immersion medium.
2 | Clin. Oral Impl. Res. 0, 2015 / 1–12
Persson et al. 1999, 2001; Kolonidis et al.
2003; Schou et al. 2003a,b; Persson et al.
2004; Takasaki et al. 2007; Alhag et al. 2008;
Parlar et al. 2009).
While the focus of many detoxification
studies is the removal of bacterial biofilms,
there has been little emphasis placed on the
changes in morphological surface properties
that can occur when an implant is subjected
to these treatments. Many of the treatments
used to detoxify titanium implants are either
acidic, contain high concentrations of fluo-
ride, or employ lasers (Gosau et al. 2010; Val-
derrama et al. 2014). Solutions with a low pH
(<3) and/or high fluoride concentrations
(>0.2%) are often shown to disrupt the oxide
layer of titanium, resulting in discoloration
and dissolution of metal ions and debris into
the surrounding medium (K€ on€
onen et al.
1995; Huang 2002; Rodrigues et al. 2009; Sar-
tori et al. 2009; Bhola et al. 2011; Muguruma
et al. 2011; Mathew et al. 2012; Noguti et al.
2012; Toniollo et al. 2012; Duarte et al. 2013;
Suito et al. 2013). Er:YAG lasers have been
shown to alter surface morphology melting
the surface of titanium from the heat pro-
duced during treatment (Kamel et al. 2013).
It has also been demonstrated that these
acidic treatments may inhibit repassivation
of the oxide layer, which can result in corro-
sion, localized dissociation of the bulk tita-
nium (pitting attack), etching, and
discoloration (Rodrigues et al. 2013; Schwarz
et al. 2013). It has been suggested that the
dissolution of metal ions may encourage
Treatment Concentration
Sodium fluoride 0.2%
Sodium fluoride 1.1%
Doxycycline 50%
Tetracycline 50%
Peroxyacetic acid 35%
CO2 and ultraspeed CO2 laser NA
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
peri-mucositis and peri-implantitis (Rodri-
gues et al. 2013; Wilson et al. 2014). This
may be a cause for concern because these
detoxification treatments thought to elimi-
nate the bacterial biofilms from the surface
and create an environment suitable for re-
osseointegration have the possibility of
inducing inflammation. In addition, studies
have shown that along with providing bone
cells a more favorable surface with which to
integrate, an increase in surface roughness
can facilitate bacteria adhesion and coloniza-
tion, also making more difficult complete
removal of biofilms (Zhao et al. 2014).
The goal of this study was to do a qualita-
tive analysis of the effects of decontamina-
tion treatments on the morphology of the
titanium oxide layer. The rationale for this
investigation was based on the hypothesis
that the synergy between acidic medium and
mechanical forces employed in these proce-
dures may cause permanent disruption of the
oxide layer protecting the surface of titanium
implants. This is critical to investigate
because if the oxide layer is disrupted by the
decontamination therapy, re-osseointegration
may be hindered, which could result in
greater incidence of implant failure. In this
study, the effects of a series of decontamina-
tion solutions are investigated on the surface
of titanium, both commercially pure, and the
alloy Ti6Al4V, using microscopy, spectros-
copy, and roughness measurements.
Material and methods
Materials
A series of solutions typically employed in
the clinical setting were investigated in this
study. The solution treatments were hydro-
gen peroxide (CVS Pharmacy, Woonsocket,
RI, USA) prepared at 3% in purified water;
hydrogen peroxide (Ricca Chemical, Arling-
ton, TX, USA) prepared at 15% in reagent
Wheelis et al
Effects of decontamination on the surface of titanium
(a) (b)
(c) (d)
Fig. 3. AFM images obtained for peroxyacetic immersion treatment: (a,b) Ti6Al4V treated 2D; (c,d) Ti6Al4V treated
3D; and (e) OM images for Ti6Al4V treated.
multiple points pre- and post-treatment.
Images were taken at low and high magnifica-
tions (up to 1000x) using a Keyence VHX 2000
Optical Microscope (Keyence, Osaka, OSK,
Japan). Optical images were taken using the
high dynamic range (HDR) setting on the
microscope and composite image function.
The supernatant and rubbing swabs were
reserved for further analysis after coming in
contact with each of the specimens after the
last immersion procedure. After optical
microscopy images were analyzed, three treat-
ments were selected with observed varying
effects on the surface: the most damaging, the
least damaging, and the treatment that dam-
aged the surface moderately (peroxyacetic
acid, citric acid, and sodium fluoride, 0.12%
respectively). The treatments that were
selected for further analysis were performed
again. Both immersion and rubbing were
repeated as above, and only the chemical treat-
ments selected were continuously performed
for 8 min. The control and treated surfaces
were analyzed with atomic force microscopy
(Biocope Catalyst, Bruker Instruments, Billeri-
ca, MA, USA) and scanning electron micros-
copy (SEM, JEOL JSM 6010-LA, Tokyo, Japan)
equipped with energy dispersive X-ray spec-
troscopy. Energy dispersive X-ray spectroscopy
(EDS) was used to verify the composition of
metal particles/debris on the swabs that could
have been generated during the rubbing and/or
immersion procedures. Swabs that were used
in the rubbing procedure were allowed to dry
and then rubbed on a piece of double-sized
tape for fixation on the SEM stub. EDS scans
were performed six times on each sample to
verify the composition of each swab. From the
4 | Clin. Oral Impl. Res. 0, 2015 / 1–12
(e)
six scans per sample, a mean and standard
deviation was calculated for the percent tita-
nium found on the swab, and this was
repeated for each treatment.
Roughness determination
The treatments that were selected for AFM
analysis (peroxyacetic acid, citric acid, and
sodium fluoride) were performed again, as pre-
viously described. AFM was used for verifica-
tion of specific morphological features and
determination of surface roughness. The AFM
was operated in PeakForce QNM (Quantita-
tive NanoMechanics) mode in air by silicon
cantilevers with spring constants of 200 N/m.
Typically, the image scan size was obtained
within the scale of 5 lm 9 5 lm and
50 lm 9 50 lm and with a scan speed of
3.00 Hz. The average surface roughness was
assessed by averaging the absolute value of
the standard deviation of the mean plane of
the scan using the NanoScope analysis tool
(version 1.4 Bruker Corporation, Billerica,
MA, USA) to quantify a roughness that best
expressed the average for a treatment. The
NanoScope analysis tool was used over a vari-
ety of areas on each treated face.
Results
This study qualitatively evaluated the surface
of treated specimens using microscopy tech-
niques (SEM and AFM) and quantification of
roughness using AFM experiments. For each
titanium grade, experimental condition (rub-
bing versus immersion) and chemical treat-
ment, only one sample was evaluated.
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
pH measurements
In general, no notable variations in solution
pH were detected with specimen immersion
for the total net treatment time (8 min). Fig. 1
shows slight variations in pH with each treat-
ment performed. Thus, it was not possible to
correlate pH values pre- and post-treatment
with the level of surface change. The pH
change within treatments did not vary largely
whether the treatment was mild or acidic.
Immersion
Peroxyacetic acid, citric acid, hydrogen perox-
ide 15%, tetracycline, and doxycycline were
observed to have damaging effects on the
surface when immersion was performed.
Samples treated with peroxyacetic acid were
imaged before and after treatment through
OM and AFM (Figs 2 and 3). Different mor-
phological features were observed in treated
samples in comparison with control samples.
Fig. 2 shows AFM images obtained for per-
oxyacetic immersion treatment for both cpTi
and TiAl4V. It is possible to observe that the
surface of control specimens of both titanium
grades (Fig. 2a,e) is smoother. However, after
treatment, their surfaces presented more
irregularities characterized by the appearance
of peaks and valleys as shown in Fig. 2b,d,f,
h. The presence of pits and discoloration was
more prominent with Ti6Al4V specimens
(Fig. 3), while cpTi specimens did not show
these specific characteristics. With this speci-
men, an interesting feature (with the shape
of a cavity with depth of around 80 nm) was
observed post-treatment (Fig. 3b,d). This fea-
ture could have resulted from localized metal
dissolution.
Citric acid caused some degree of surface
change caused by this treatment in compari-
son with control surfaces (Fig. 4a,c,e,g). The
changes introduced on the surface by this
treatment were observed to occur to a lesser
extent in comparison with the peroxyacetic
acid treatment (Figs 2 and 3). Specifically,
only discoloration was apparent on specimens
of both grades (cpTi and Ti6Al4V) as illus-
trated in Fig. 4. Hydrogen peroxide, tetracy-
cline, and doxycycline all inflicted similar
degree of damage on the specimens. In OM
images, discoloration was similar to that
observed for citric acid treatments, which can
be indicative of potential damage if these
acidic treatments were coupled with rubbing.
The treatments that exhibited no damage to
the surface of both titanium grades in the
form of discoloration, pitting, or delamination
were the three different concentrations of
sodium fluoride, both concentrations of
chlorhexidine, and hydrogen peroxide 3%.
Wheelis et al
Effects of decontamination on the surface of titanium
(a) (b)
(c) (d)
Fig. 3. AFM images obtained for peroxyacetic immersion treatment: (a,b) Ti6Al4V treated 2D; (c,d) Ti6Al4V treated
3D; and (e) OM images for Ti6Al4V treated.
multiple points pre- and post-treatment.
Images were taken at low and high magnifica-
tions (up to 1000x) using a Keyence VHX 2000
Optical Microscope (Keyence, Osaka, OSK,
Japan). Optical images were taken using the
high dynamic range (HDR) setting on the
microscope and composite image function.
The supernatant and rubbing swabs were
reserved for further analysis after coming in
contact with each of the specimens after the
last immersion procedure. After optical
microscopy images were analyzed, three treat-
ments were selected with observed varying
effects on the surface: the most damaging, the
least damaging, and the treatment that dam-
aged the surface moderately (peroxyacetic
acid, citric acid, and sodium fluoride, 0.12%
respectively). The treatments that were
selected for further analysis were performed
again. Both immersion and rubbing were
repeated as above, and only the chemical treat-
ments selected were continuously performed
for 8 min. The control and treated surfaces
were analyzed with atomic force microscopy
(Biocope Catalyst, Bruker Instruments, Billeri-
ca, MA, USA) and scanning electron micros-
copy (SEM, JEOL JSM 6010-LA, Tokyo, Japan)
equipped with energy dispersive X-ray spec-
troscopy. Energy dispersive X-ray spectroscopy
(EDS) was used to verify the composition of
metal particles/debris on the swabs that could
have been generated during the rubbing and/or
immersion procedures. Swabs that were used
in the rubbing procedure were allowed to dry
and then rubbed on a piece of double-sized
tape for fixation on the SEM stub. EDS scans
were performed six times on each sample to
verify the composition of each swab. From the
4 | Clin. Oral Impl. Res. 0, 2015 / 1–12
(e)
six scans per sample, a mean and standard
deviation was calculated for the percent tita-
nium found on the swab, and this was
repeated for each treatment.
Roughness determination
The treatments that were selected for AFM
analysis (peroxyacetic acid, citric acid, and
sodium fluoride) were performed again, as pre-
viously described. AFM was used for verifica-
tion of specific morphological features and
determination of surface roughness. The AFM
was operated in PeakForce QNM (Quantita-
tive NanoMechanics) mode in air by silicon
cantilevers with spring constants of 200 N/m.
Typically, the image scan size was obtained
within the scale of 5 lm 9 5 lm and
50 lm 9 50 lm and with a scan speed of
3.00 Hz. The average surface roughness was
assessed by averaging the absolute value of
the standard deviation of the mean plane of
the scan using the NanoScope analysis tool
(version 1.4 Bruker Corporation, Billerica,
MA, USA) to quantify a roughness that best
expressed the average for a treatment. The
NanoScope analysis tool was used over a vari-
ety of areas on each treated face.
Results
This study qualitatively evaluated the surface
of treated specimens using microscopy tech-
niques (SEM and AFM) and quantification of
roughness using AFM experiments. For each
titanium grade, experimental condition (rub-
bing versus immersion) and chemical treat-
ment, only one sample was evaluated.
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
pH measurements
In general, no notable variations in solution
pH were detected with specimen immersion
for the total net treatment time (8 min). Fig. 1
shows slight variations in pH with each treat-
ment performed. Thus, it was not possible to
correlate pH values pre- and post-treatment
with the level of surface change. The pH
change within treatments did not vary largely
whether the treatment was mild or acidic.
Immersion
Peroxyacetic acid, citric acid, hydrogen perox-
ide 15%, tetracycline, and doxycycline were
observed to have damaging effects on the
surface when immersion was performed.
Samples treated with peroxyacetic acid were
imaged before and after treatment through
OM and AFM (Figs 2 and 3). Different mor-
phological features were observed in treated
samples in comparison with control samples.
Fig. 2 shows AFM images obtained for per-
oxyacetic immersion treatment for both cpTi
and TiAl4V. It is possible to observe that the
surface of control specimens of both titanium
grades (Fig. 2a,e) is smoother. However, after
treatment, their surfaces presented more
irregularities characterized by the appearance
of peaks and valleys as shown in Fig. 2b,d,f,
h. The presence of pits and discoloration was
more prominent with Ti6Al4V specimens
(Fig. 3), while cpTi specimens did not show
these specific characteristics. With this speci-
men, an interesting feature (with the shape
of a cavity with depth of around 80 nm) was
observed post-treatment (Fig. 3b,d). This fea-
ture could have resulted from localized metal
dissolution.
Citric acid caused some degree of surface
change caused by this treatment in compari-
son with control surfaces (Fig. 4a,c,e,g). The
changes introduced on the surface by this
treatment were observed to occur to a lesser
extent in comparison with the peroxyacetic
acid treatment (Figs 2 and 3). Specifically,
only discoloration was apparent on specimens
of both grades (cpTi and Ti6Al4V) as illus-
trated in Fig. 4. Hydrogen peroxide, tetracy-
cline, and doxycycline all inflicted similar
degree of damage on the specimens. In OM
images, discoloration was similar to that
observed for citric acid treatments, which can
be indicative of potential damage if these
acidic treatments were coupled with rubbing.
The treatments that exhibited no damage to
the surface of both titanium grades in the
form of discoloration, pitting, or delamination
were the three different concentrations of
sodium fluoride, both concentrations of
chlorhexidine, and hydrogen peroxide 3%.
 Wheelis et al
Effects of decontamination on the surface of titanium

(a) Rubbing
(b)
Morphological damage characterized by pits,
etching, and delamination on the surface was
monitored as well as discoloration. The
images obtained for treated specimens were
compared with their respective controls. It
was observed that after treatments with
hydrogen peroxide (3%), sodium fluoride
(0.12%, 0.2%. 1.1%), and CO2 Superspeed
Laser, both cpTi and Ti6Al4V were not sub-
jected to any particular surface change or dam-
age in comparison with control samples.
(c) (d) There was no evidence of discoloration or
etching, with the control nearly indistinguish-
able from the treated surface in OM. The laser
treatments exhibited no damage to the surface
as well. The acidic treatments, on the other
hand, were observed to cause remarkable fea-
tures on the surface (pH <5.5) post-rubbing.
The rubbing surfaces of samples treated with
citric acid, hydrogen peroxide (15%), chlorh-
exidine (0.12%, 1%), doxycycline (50%), tetra-
(f) cycline (50%), and peroxyacetic acid (35%)
(e)
exhibited severe discoloration and pitting.
Fig. 5a,e illustrates the appearance of the
surfaces of cpTi and Ti6Al4V samples before
and after the rubbing treatment with hydrogen
peroxide 3% and 15%. After treatment with
hydrogen peroxide 3%, the surface of the sam-
ple exhibited the same characteristics than
those observed for controls (Fig. 5b,f). Discol-
oration, pits, and surface damage were not ver-
ified for both titanium grades investigated.
The cpTi samples showed a few inclusions
(g) (h) (Fig. 5a); however, these features were similar
to those observed after sample polishing and
were not necessarily related to the rubbing
process. On the other hand, after treatment
with hydrogen peroxide 15%, the surface of
both titanium grades showed characteristic
features. The surface of cpTi samples showed
a combination of discoloration, severe pitting
attack, and scratches, which could be related
to the corrosive process and the softer nature
of the material in comparison with the alloy.
(i) (j)
Less degree of pitting attack and violet color
were more prominent with Ti6Al4V samples,
as shown in Fig. 5d,h.
The chlorhexidine treatments, although
relatively high in pH compared with the
other treatments (~5), showed signs of oxide
layer damage when coupled with mechanical
abrasion. Tetracycline and doxycycline exhib-
ited the same effects on the surface as the
strong acids, more so in the rubbing than the
immersion side of the samples. Although dis-
Fig. 4. AFM images obtained for citric acid immersion treatment: (a) CpTi control 2D image; (b) CpTi treated 2D; coloration was evident, it was less severe,
(c) CpTi control 3D; (d) CpTi treated 3D; (e) Ti6Al4V control 2D; (f) Ti6Al4V treated 2D; (g) Ti6Al4V control 3D; (h) with less of the total surface discolored in
Ti6Al4V treated 3D; and OM images of (e) CpTi and (j) Ti6Al4V treated.
comparison with the degree of discoloration
produced by the more acidic treatments.

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd 5 | Clin. Oral Impl. Res. 0, 2015 / 1–12
Wheelis et al
Effects of decontamination on the surface of titanium
(a) (b)
(e) (f)
Fig. 5. Pitting and discoloration observed in the (a) CpTi rubbing control for H2O2 3%; (b) CpTi after rubbing with H2O2 3%; (c) CpTi rubbing control for H2O2 15%; (d) CpTi
after rubbing with H2O2 15%; (e) Ti6Al4V rubbing control for H2O2 3%; (f) Ti6Al4V after rubbing with H2O2 3%; (g) Ti6Al4V rubbing control for H2O2 15%; and (h) Ti6Al4V
after rubbing with H2O2 15%.
Other interesting features were observed
for the supernatant from the tetracycline
immersion treatment and the cotton swabs
from the doxycycline rubbing treatment. The
immersion treatment exhibited a change in
color from the original treatment liquid.
After a week, the chemical had visibly dark-
ened in color in both CpTi and Ti6AlV4 su-
pernatants and swabs used in the rubbing
treatment. The darkening was more apparent
in both treatments that came in contact with
cpTi samples. This darkening may suggest
the presence of dissociated Ti ions or debris
in the liquid and swab. To investigate this
further, the cotton swabs that were soaked in
doxycycline and used for the rubbing treat-
ment were subjected to EDS analysis for eval-
uation of the composition of the residue
collected. The analysis confirmed the pres-
ence of trace amounts of Ti and aluminum
(Al) in the swabs, which suggested there was
metal loss from the Ti6Al4V specimens that
came in contact with the treatment.
Regarding the three treatments chosen for
further investigation, peroxyacetic acid
showed the most aggressive attack on the
metal surface where both cpTi and Ti6Al4V
exhibited a high degree of pitting attack and
disruption of the top-layer surfaces as illus-
trated in Figs 6 and 7. Localized metal disso-
lution resulted in structural features similar
to large pits in both grades of titanium as
shown in Fig. 6b,d,f,h. A variety of pit sizes
suggest multiple severities of attack as illus-
trated in Fig. 7.
6 | Clin. Oral Impl. Res. 0, 2015 / 1–12
(c)
(g)
The rubbing treatment with citric acid also
resulted in pits and discoloration corroborat-
ing the previous observations. Fig. 8 suggests
disruption of the normal surface morphology
from etching and pits in both titanium
grades, but to a lesser extent in comparison
with peroxyacetic acid. This is concluded
from both the roughness measurements and
the frequency at which pits and discoloration
could be observed from AFM and OM
images. An example of pitting attack result-
ing from the use of citric acid is shown in
Fig. 8b,d,i,j. The effect of this treatment
when mechanical force was present only
exaggerated surface features.
On the contrary, samples treated with
sodium fluoride did not show any apparent
features associated with wear and/or corro-
sion. Control and treated samples from both
grades (cpTi and Ti6Al4V) exhibited the
same surface appearance and features after
immersion and rubbing procedures.
Roughness
The results showed that almost all the
selected treatments caused an increase in
surface roughness of the samples, as observed
in Fig. 9. According to Zhao et al. (2014), sur-
face roughness of dental implants can be
classified as minimally rough (<0.5 lm),
moderately rough (between 1 and 2 lm), and
rough (>0.5 lm) (Zhao et al. 2014). Therefore,
all the samples analyzed in this study can be
considered minimally rough both before and
after treatment. The most pronounced
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
(d)
(h)
change in surface roughness caused by the
several treatments employed was observed
for the citric acid treatment performed on
cpTi samples (Fig. 9). In further investiga-
tions of surface composition, EDS analysis
was performed and revealed the presence of
trace amounts of titanium on the swabs on
all rubbing treatments (Fig. 10), which sug-
gests that the mechanical force exhibited by
the rubbing procedure was enough to remove
some of the oxide layer. The metal concen-
tration (Ti) found in the swabs ranged from
of 0.06% to 0.85%.
Discussion
The goal of this study was to investigate the
synergistic effects of chemical treatments
and mechanical forces on the surface of tita-
nium simulating detoxification treatments
used clinically. Although a large number of
relevant studies have investigated the effi-
cacy of detoxification procedures, there is a
lack of information about how these treat-
ments interact and change the surface of a
titanium dental implant. Interesting features
were revealed by optical microscopy (OM)
and atomic force microscopy (AFM) in this
study, which resulted from solution pH,
mechanical factors, and the synergistic
effects of both on the surface of the material.
Features such as pits, discoloration, and
scratches were observed for both titanium
grades with the treatments investigated.

(a)
(c)
(e)
(g)
Fig. 6. AFM images obtained for peroxyacetic rubbing treatment: (a) CpTi control 2D image; (b) CpTi treated 2D;
(c) CpTi control 3D; (d) CpTi treated 3D; (e) Ti6Al4V control 2D; (f) Ti6Al4V treated 2D; (g) Ti6Al4V6 control 3D;
and (h) Ti6Al4V treated 3D.
These features were enhanced when the sam-
ples were subjected to abrasion conditions.
The AFM analysis elucidated features on the
microscale, which are considered important
given the osseointegrative process can be
assisted by such surface features.
Previous studies discussed the influence of
solution pH in the corrosion behavior of
metallic implants (Rodrigues et al. 2009,
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
Wheelis et al
Effects of decontamination on the surface of titanium
(b)
(d)
(f)
(h)
2013; Mathew et al. 2012). Particularly, Ti
and its alloys are highly susceptible to
changes in surface structure and potential in
the presence of acidic conditions. The
increase in H+ concentration favors dissolu-
tion of the oxide layer and hence weakens its
protective properties (Sato 1989). Although
strong acids are discussed to cause dissolu-
tion of the oxide layer, it was observed from
the pH measurements (Fig. 1) that any ions
or debris that may have leached from the sur-
face of the specimens did not alter the pH of
the immersion solution (Rodrigues et al.
2009; Suito et al. 2013). Therefore, no corre-
lation was found between variation of pH
and different treatments.
Pitting and discoloration were the main
morphological features investigated to evalu-
ate corrosion of both cpTi and Ti6Al4V in
the presence of the several treatments and
methods employed. In particular, discolor-
ation was a feature of interest because it is
well established that oxidized titanium dis-
plays a characteristic violet color, which is
an indication of the trivalent oxidized state
(Ti3+), which can be further converted to the
pale yellow Ti4+ ion (Gilbert 2012). Thus, dis-
coloration may indicate the occurrence of
corrosion (Gilbert 2012; Swaminathan &
Gilbert 2013). Pitting attack was another fea-
ture of interest. Pitting is characterized by a
localized dissolution of the bulk metal,
which can lead to the generation of metal
ions and debris to the surrounding environ-
ment. Titanium has been reported to be
highly resistant to pitting attack (Velten
et al. 2002). Previous studies, however, have
demonstrated that the material is susceptible
to the formation of pits under acidic condi-
tions and/or highly fluoridated mediums
when abrasion of the oxide layer occurs (Nog-
uti et al. 2012; Suito et al. 2013). Further-
more, the corrosion triggered by mechanical
forces (fretting-corrosion, fretting-crevice cor-
rosion, and tribocorrosion) has been investi-
gated in previous studies (Mathew et al.
2009; Rodrigues et al. 2009; Gilbert 2012;
Swaminathan & Gilbert 2013). Factors such
as cyclic loads and fretting motion coupled
with acidic environments can result in etch-
ing and delamination, as discussed in previ-
ous retrieval studies of both orthopedic and
dental implants (Renvert et al. 2009; Gilbert
2012; Swaminathan & Gilbert 2012 and Ro-
drigues et al.2013). Mathew et al. (2012) eval-
uated the combined effects of corrosion and
tribological stresses in the degradation of
dental implant materials (cpTi) under varying
pHs, to simulate the different conditions of
the oral environment. In that study, surface
damage with cracking and delamination char-
acteristics were observed with scanning elec-
tron microscopy. More pronounced weight
loss and damage of cpTi surfaces were
observed to occur at a mild pH of 6, which is
the pH of the oral environment (Mathew
et al. 2012). Although the above-mentioned
studies investigated the emergent effect of
chemical environments coupled with
7 | Clin. Oral Impl. Res. 0, 2015 / 1–12
Wheelis et al
Effects of decontamination on the surface of titanium
(a)
(b)
(c) (d)
Fig. 7. AFM images obtained for peroxyacetic rubbing treatment: (a) Ti6Al4V control 2D; (b) Ti6Al4V treated 2D;
(c) Ti6Al4V control 3D; (d) Ti6Al4V treated 3D; and (e) optical microscope for Ti6Al4V treated.
mechanical force, there are no reported
results regarding surface damage due to these
combined factors after decontamination and
cleaning maintenance treatments in the oral
environment.
In immersion experiments, the acidic treat-
ments including peroxyacetic acid, citric
acid, hydrogen peroxide 15%, tetracycline,
and doxycycline were observed to cause dis-
coloration, increase in the depth of peaks and
valleys and pits on the surface of the sam-
ples. These observations corroborate with
previous results which reported that acids
have the ability to cause delamination and
etching of the oxide layer (Suito et al. 2013).
Peroxyacetic acid has been used in steriliza-
tion techniques in the final step of implant
device manufacturing (Thierry et al. 2000;
Shabalovskaya 2002). However, very little is
known about the effect of this sterilization
step on the modification of the outermost
surface sublayers of titanium and its alloys.
This compound is an oxidant and can damage
cells by oxidation and disruption of cell
membranes via the hydroxyl radical agent
(Finnegan et al. 2010). It is also the most
acidic of the treatments used in this study.
The presence of discoloration revealed by
OM (Fig. 3e) is an indication of oxide layer
degradation. The depressions with concen-
trated areas of discoloration shown in OM
are hypothesized to be pits. AFM scans
helped gaining better resolution of these
depressions to confirm that they were pits
and not just polishing imperfections or dust,
as observed in Fig. 3b,d. According to the
results of this study, this treatment may have
8 | Clin. Oral Impl. Res. 0, 2015 / 1–12
(e)
the potential to penetrate the oxide layer
even if no mechanical abrasion is introduced.
The data reported (Figs 2 and 3) are in agree-
ment with the results previously reported by
Thierry et al. (2000). The authors observed
modifications induced by sterilization with
peroxyacetic acid on the surface of nitinol
alloys.
Citric acid is a widely applied agent in
detoxification of implants affected by peri-
implantitis (Alhag et al. 2008). The increase
in sharpness of peaks and valleys on the
surface of immersed specimens (Fig. 4b,d,f,h)
could indicate a change in the structure of
the oxide layer, although no apparent pit-
ting was detected on the surface of both
cpTi and Ti6Al4V samples. This indicates
that localized bulk dissolution was not
induced by immersion with citric acid. Fur-
thermore, the discoloration observed from
OM indicates the possibility of oxide layer
penetration and etching (Fig. 4i,j). In com-
parison with results obtained for the per-
oxyacetic acid treatment (Figs 2 and 3), it is
reasonable to infer that the lower pH had
less effect on the surface (Suito et al. 2013).
The treatment with sodium fluoride was
also in agreement with results reported in
the literature with no damage or particular
features observed. Besides a few reports
showing corrosion of titanium-based
implants after immersion in sodium fluo-
ride solutions, this is generally observed to
occur in acidic environments or with higher
concentrations of sodium fluoride (Huang
2002; Sartori et al. 2009; Noguti et al.
2012).
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
The rubbing treatment aimed to evaluate
the emergence of chemical and mechanical
conditions. The neutral or slightly acidic
treatments (sodium fluoride and hydrogen
peroxide 3%) had no effect on surface
morphology, which is due to the favorable
neutral electrochemical environment in
which the oxide layer is exposed (Suito
et al. 2013). The sodium fluoride treatments
are neutral to basic, and studies suggest
when titanium is present in this environ-
ment, corrosion is not likely to occur. (Ma-
thew et al. 2012). In addition, the fluoride
treatments used here had low concentra-
tions and therefore were considered unlikely
to cause surface damage as previously
reported (Noguti et al. 2012; Suito et al.
2013). A few studies showed corrosion of
titanium-based implants after immersion in
sodium fluoride solutions, but that generally
occurred in the presence of higher concen-
trations of sodium fluoride (K€ on€onen et al.
1995; Sartori et al. 2009; Noguti et al.
2012). Further investigation needs to be con-
ducted to conclude whether high concentra-
tions of fluoride cause oxidation or the
acidic medium in which the fluoride is sus-
pended is responsible for corrosive effects.
Similarly, the CO2 lasers mechanism of
action involves a reaction with water in liv-
ing tissues; this is irrelevant to the oxide
layer; therefore, no damage was observed
with laser treatment, which corroborate
with the results reported by Subramani &
Wismeijer (2012). The decrease in the pH
from hydrogen peroxide 3% to hydrogen
peroxide 15% was enough to cause remark-
able changes on the surface of both cpTi
and Ti6Al4V samples after rubbing as dem-
onstrated in Fig. 5. Comparing the surface
features of control (Fig. 5a,e) and treated
samples (Fig. 5b,f) for hydrogen peroxide
3%, no discoloration, pits, or change in
roughness were observed. On the other
hand, those features could be clearly distin-
guished when the concentration of hydrogen
peroxide increased to 15% as shown in
Fig. 5d,h. Previous experiments have indi-
cated that at low concentrations (~0.1%),
hydrogen peroxide can cause gel-like oxide
layer growth, while in higher concentra-
tions, several oxidation intermediates can be
generated and corrosion can take place,
which is in agreement with the present
results (Tengvall et al. 1989; Bearinger et al.
2003).
The effect of 0.01% chlorhexidine gluco-
nate (CHG) in normal saline (NS) on the
corrosion behavior of titanium has been
investigated by Bhola et al. (2011) with elec-
 Wheelis et al
Effects of decontamination on the surface of titanium

(a) (b) trochemical techniques (Bhola et al. 2013).

The results obtained in that study demon-
strated corrosion of titanium implant sur-
faces, which increased in severity with
exposure time (Bhola et al. 2013). Conse-
quently, the discoloration and increase in
roughness on the surface of samples treated
with CHG post-rubbing may have been
triggered by the synergistic effects of chlorh-
exidine and mechanical forces.
As hypothesized previously, the synergis-
tic effects of acidic pH and mechanical abra-
sion caused disruption of the oxide layer and
(c) (d)
increased surface damage (Rodrigues et al.
2009; Gilbert 2012). This can be confirmed
when the results obtained with the immer-
sion and rubbing treatments of the other
solutions with greater pHs are compared.
The effects of acidic rubbing treatments
(Figs 5–8) on the surface were more aggres-
sive than immersion (Figs 2–4) because there
is little to no opportunity for the oxide layer
to repassivate in the acids as mechanical
(e) (f) force is continuously applied (Mathew et al.
2012; Suito et al. 2013). Strong acids such as
peroxyacetic and citric acid were observed to
dissolute the oxide layer to a greater degree
under abrasion (Figs 6–8) in comparison with
immersion (Figs 2–4), which suggests that
stronger acids can have severe tribocorrosive
effects.
The influence of roughness in metal
implants has shown advantages but also dis-
advantages in terms of performance in vivo.
In general, an increase in surface roughness
(g) (h) of dental implants has been shown to
improve the biomechanical anchorage of the
implant with the bone (Novaes et al. 2010;
Garg et al. 2012). However, it has also been
observed as a critical factor for biofilm
growth, as reported in a review by Mellado-
Valero et al. (2013). The increase in surface
roughness (Fig. 9) observed after rubbing
treatments could be associated with surface
layers loss due to wear and corrosion. In
these regions, the localized dissociation can
(i) (j) result in metal particle or debris generation,
which has been discussed to potentially trig-
ger inflammation in vivo (Burbano et al.
2014). While differences in roughness from
control to treated surfaces with immersion
procedures were observed, comparatively
roughness change from the rubbing treat-
ments was often higher in magnitude, as
demonstrated in Fig. 9. Roughness changes
from immersion treatments can be attrib-
uted to the treatment properties, such as
Fig. 8. AFM images obtained for citric acid rubbing treatment: (a) CpTi control 2D image; (b) CpTi treated 2D; low pH, which can cause elution of surface
(c) CpTi control 3D; (d) CpTi treated 3D; (e) Ti6Al4V control 2D; (f) Ti6Al4V treated 2D; (g) Ti6Al4V control 3D; layers to a lesser extent in the absence of
and (h) Ti6Al4V treated 3D and OM images of (e) CpTi and (j) Ti6Al4V treated. abrasion.

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd 9 | Clin. Oral Impl. Res. 0, 2015 / 1–12
Wheelis et al
Effects of decontamination on the surface of titanium
Fig. 9. Roughness data of samples obtained by AFM.
Fig. 10. Mean and standard deviation of percent titanium on swab obtained by energy dispersive spectroscopy for
the rubbing treatment for the number of scans per sample (n).
Although removal of the oxide layer
does not automatically indicate corrosion of
titanium, when OM, AFM, roughness mea-
References
Alhag, M., Renvert, S., Polyzois, I. & Claffey, N.
(2008) Re-osseointegration on rough implant sur-
faces previously coated with bacterial biofilm: an
experimental study in the dog. Clinical Oral
Implants Research 19: 182–187.
Bearinger, J.P., Orme, C.A. & Gilbert, J.L. (2003)
Effect of hydrogen peroxide on titanium surfaces:
in situ imaging and step-polarization impedance
spectroscopy of commercially pure titanium and
titanium, 6-aluminum, 4-vanadium. Journal of
Biomedical Materials Research Part A 67: 702–
712.
Bhola, R., Bhola, S.M., Mishra, B. & Olson, D.L.
(2011) Corrosion in titanium dental implants/
prostheses - a review. Trends in Biomaterials and
Artificial Organs 25: 34–46.
Bhola, S.M., Kundu, S., Alabbas, F., Mishra, B. &
Olson, D.L. (2013) An electrochemical study on
chlorhexidine gluconate addition to normal saline
10 | Clin. Oral Impl. Res. 0, 2015 / 1–12
surements, and EDS scans are correlated,
there were important conclusions drawn from
the combination of the results. Future studies
for oral implant applications. International Jour-
nal of Electrochemical Science 8: 5172–5182.
Burbano, M., Russell, R., Huo, M., Welch, R., Roy,
D. & Rodrigues, D.C. (2014) Surface characteriza-
tion of retrieved metal-on-metal total hip
implants from patients with adverse reaction to
metal debris. Materials 7: 1866–1879.
De Waal, Y.C.M., Raghoebar, G.M., Meijer, H.J.,
Winkel, E.G. & van Winkelhoff, J. (2014)
Implant decontamination with 2% chlorhexidine
during surgical peri-implantitis treatment: a ran-
domized, double-blind, controlled trial. Clinical
Oral Implants Research 1–9. doi:10.1111/
clr.12419.
Duarte, A.R.C., Neto, J.P.S., Souza, J.C.M. & Bon-
achela, W.C. (2013) Detorque evaluation of dental
abutment screws after immersion in a fluoridated
artificial saliva solution. Journal of Prosthodon-
tics 22: 275–281.
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
will further investigate the electrochemical
properties of specimens subjected to the treat-
ments discussed. Additionally, the ability of
osteoblasts to regrow and reattach on tita-
nium surfaces that have been previously trea-
ted with the solutions and methods discussed
is under investigation and will be correlated
with the present observations.
Conclusion
In general, it was observed, when coupled
with rubbing, acidic environments inflicted
severe corrosion on the surface of the speci-
mens. The AFM scans showed pits of large
dimensions for both Ti grades investigated in
the peroxyacetic and citric acid treatment
groups, making them the most damaging
treatments to the surface. Neutral or basic
treatments such as sodium fluoride 0.12%,
0.20%, and 1.10%, showed no visibly evident
damage of the surfaces treated. These obser-
vations confirmed the hypothesis and showed
that surface damage of dental alloys may be
potentially induced after detoxification and
maintenance treatments with acidic solu-
tions. The results showed that there is poten-
tial damage induced by the use of the
treatments investigated in this study.
Acknowledgements: The authors
would like to thank the Texas A&M Baylor
College of Dentistry for the use of their
facilities and SEM and the Vascular
Mechanobiology laboratory at the University
of Texas at Dallas for the AFM facilities.
Ericsson, I., Persson, L.G., Berglundh, T., Edlund, T.,
Lindhe, J., Berglundh, T., Edlund, T. & Lindhe, J.
(1996) The effect of antimicrobial therapy on peri-
implantitis lesions. An experimental study in the
dog. Clinical Oral Implants Research 7: 320–328.
Finnegan, M., Linley, E., Stenyer, S.P., Mcdonnell,
G., Simons, C. & Maillard, J.Y. (2010) Mode of
action of hydrogen peroxide and other oxidizing
agents: differences between liquid and gas forms.
Journal of Antimicrobial Chemotherapy 65:
2108–2115.
Garg, H., Bedi, G. & Garg, A. (2012) Implant surface
modifications : a review. Journal of Clinical and
Diagnostic Research 6: 319–324.
Gilbert, J.L. (2012) Mechanically assisted corrosion
of metallic biomaterials. ASM Handbook: Materi-
als for Medical Devices 23: 79–89.
Gosau, M., Hahnel, S., Schwarz, F., Gerlach, T.,
Reichert, T.E. & B€ urger, R. (2010) Effect of six

different peri-implantitis disinfection methods on
in vivo human oral biofilm. Clinical Oral
Implants Research 21: 866–872.
Huang, H. (2002) Effects of fluoride concentration
and elastic tensile strain on the corrosion resis-
tance of commercially pure titanium. Biomaterials
23: 59–63.
Kamel, M.S., Khosa, A., Tawse-Smith, A. & Leichter,
J. (2013) The use of laser therapy for dental implant
surface decontamination: a narrative review of in
vitro studies. Lasers in Medical Science 29: 1977–
1985.
Kolonidis, S.G., Renvert, S., H€ammerle, C.H.F.,
Lang, N.P., Harris, D. & Claffey, N. (2003) Osseo-
integration on implant surfaces previously con-
taminated with plaque. An experimental study in
the dog. Clinical Oral Implants Research 14:
373–380.
K€
on€ onen, M.H., Lavonius, E.T. & Kivilahti, J.K.
(1995) SEM observations on stress corrosion crack-
ing of commercially pure titanium in a topical
fluoride solution. Dental Materials 11: 269–272.
Mathew, M.T., Abbey, S., Hallab, N.J., Hall, D.J.,
Sukotjo, K. & Wimmer, M.A. (2012) Influence of
pH on the tribocorrosion behavior of CpTi in the
oral environment: synergistic interactions of wear
and corrosion. Journal of Biomedical Materials
Research Part B 100: 1662–1671.
Mathew, M.T., Srinivasai Pai, P., Pourzal, R., Fi-
scher, A. & Wimmer, M.A. (2009) Significance of
tribocorrosion in biomedical applications: over-
view and current status. Advances in Tribology
2009: 1–12.
Mellado-Valero, A., Buitrago-Vera, P., Sola-Ruiz,
M.F. & Ferrer-Garcia, J.C. (2013) Decontamina-
tion of dental implant surface in peri-implantitis
treatment: A literature review. Medicina Oral
Patologia Oral y Cirugia Bucal 18: e869–e876.
Mouhyi, J., Dohan Ehrenfest, D.M. & Albrektsson,
T. (2012) The peri-implantitis: implant surfaces,
microstructure, and physicochemical aspects.
Clinical Implant Dental Related Research 14:
170–183.
Muguruma, T., Iijima, M., Brantley, W.A., Yuasa, T.,
Kyung, H.M. & Mizoguchi, I. (2011) Effects of
sodium fluoride mouth rinses on the torsional
properties of miniscrew implants. American Jour-
nal of Orthodontics Dentofacial Orthopedics 139:
588–593.
Noguti, J., de Oliveira, F., Peres, R.C., Renno,
A.C.M. & Ribeiro, D.A. (2012) The role of fluo-
ride on the process f titanium corrosion in oral
cavity. BioMetals 25: 859–862.
Novaes, A., de Souza, S.L., de Barros, R.R., Pereira,
K.K., Iezzi, G. & Piattelli, A. (2010) Influence of
implant surfaces on osseointegration. Brazilian
Dental Journal 21: 471–481.
Parlar, A., Bosshardt, D.D., Ҫetiner, D., Schaforth,
€
D., Unsal, B., Haytaҫ, C. & Lang, N.P. (2009)
Effects of decontamination and implant surface
characteristics on re-osseointegration following
treatment of peri-implantitis. Clinical Oral
Implants Research 20: 391–399.
Persson, L.G., Ara ujo, M.G., Berglundh, T.,
Gr€ ondahl, K. & Lindhe, J. (1999) Resolution
of peri-implantitis following treatment. An
experimental study in the dog. Clinical Oral
Implants Research 10: 195–203.
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
Wheelis et al
Effects of decontamination on the surface of titanium
Persson, L.G., Berglundh, T., Lindhe, J. & Sennerby,
L. (2001) Re-osseointegration after treatment of
peri-implantitis at different implant surfaces. An
experimental study in the dog. Clinical Oral
Implants Research 12: 595–603.
Persson, L.G., Mouhyi, J., Berglundh, T., Sennerby,
L. & Lindhe, J. (2004) Carbon dioxide laser and
hydrogen peroxide conditioning in the treatment
of periimplantitis: an experimental study in the
dog. Clinical Implant Dentistry and Related
Research 6: 230–238.
Renvert, S., Polyzois, I. & Maguire, R. (2009) Re-
osseointegration on previously contaminated sur-
faces: a systematic review. Clinical Oral
Implants Research 4: 216–227.
Rodrigues, D.B., Urban, R.M., Jacobs, J.J. & Gilbert,
J.L. (2009) In vivo severe corrosion and hydrogen
embrittlement of retrieved modular body tita-
nium alloy hip-implants. Journal of Biomedical
Materials Research Part B 88: 206–219.
Rodrigues, D.B., Valderrama, P., Wilson, T.G., Pal-
mer, K., Thomas, A., Sridhar, S., Adapalli, A.,
Burbano, M. & Wadhwani, C. (2013) Titanium
corrosion mechanisms in the oral environment: a
retrieval study. Materials (Basel) 6: 5258–5274.
Sartori, R., Correa, C.B., Marcantonio, E. & Vaz,
L.G. (2009) Influence of a fluoridated medium
with different pHs on commercially pure tita-
nium-based implants. Journal of Prosthodontics
18: 130–134.
Sato, N. (1989) Toward a more fundamental under-
standing of corrosion processes. Corrosion 45:
354–368.
Schou, S., Holmstrup, P., Jørgensen, T., Skovgaard,
L.T., Stoltze, K., Hjørting-Hansen, E. & Wenzel,
A. (2003a) Implant surface preparation in the sur-
gical treatment of experimental peri-implantitis
with autogenous bone graft and ePTFE membrane
in cynomolgus monkeys. Clinical Oral Implants
Research 14: 412–422.
Schou, S., Holmstrup, P., Jørgensen, T., Skovgaard,
L.T., Stoltze, K., Hjørting-Hansen, E. & Wenzel,
A. (2003b) Autogenous bone graft and ePTFE
membrane in the treatment of peri-implantitis. I.
Clinical and radiographic observations in cyno-
molgus monkeys. Clinical Oral Implants
Research 14: 391–403.
Schwarz, F., Hegewald, A., John, G., Sahm, N. &
Becker, J.L. (2013) Four-year follow-up of com-
bined surgical therapy of advanced peri-implanti-
tis evaluating two methods of surface
decontamination. Journal of Clinical Periodontol-
ogy 40: 962–967.
Schwarz, F., Sahm, N., Iglhaut, G. & Becker, J.
(2011) Impact of the method of surface debride-
ment and decontamination on the clinical out-
come following combined surgical therapy of
peri-implantitis: a randomized controlled clinical
study. Journal of Clinical Periodontology 38:
276–284.
Schwarz, F., Sculean, A., Berakdar, M., Georg, T.,
Reich, E. & Sculean, A. (2003a) Clinical evaluation
of an Er:YAG laser combined with scaling and root
planing for non-surgical periodontal treatment. A
controlled, prospective clinical study. Journal of
Clinical Periodontology 30: 26–34.
Schwarz, F., Sculean, A., Georg, T. & Becker, J.
(2003b) Clinical evaluation of the Er:YAG laser in
combination with an enamel matrix protein
derivative for the treatment of intrabony peri-
odontal defects: a pilot study. Journal of Clinical
Periodontology 30: 975–981.
Schwarz, F., Sculean, A., Romanos, G., Herten, M.,
Horn, N., Scherbaum, W. & Becker, J. (2005)
Influence of different treatment approaches on
the removal of early plaque biofilms and the via-
bility of SAOS2 osteoblasts grown on titanium
implants. Clinical Oral Investigations Journal 9:
111–117.
Shabalovskaya, S. (2002) Surface, corrosion and bio-
compatibility aspects of Nitinol as an implant
material. Biomedical Materials Engineering 12:
69–109.
Subramani, K. & Wismeijer, D. (2012) Decontami-
nation of titanium implant surface and re-osseo-
integration to treat peri-implantitis: a literature
review. International Journal of Oral Maxillofa-
cial Implants 27: 1043–1054.
Suito, H., Iwawaki, Y., Goto, T., Tomotake, Y. &
Ichikawa, T. (2013) Oral factors affecting
titanium elution and corrosion: an in vitro study
using simulated body fluid. Public Library of
Science One 8: e66052. doi:10.1371/jour-
nal.pone.0066052.
Swaminathan, V. & Gilbert, J.L. (2012) Biomaterials
fretting corrosion of CoCrMo and Ti6Al4V inter-
faces. Biomaterials 33: 5487–5503.
Swaminathan, V. & Gilbert, J.L. (2013) Potential
and frequency effects on fretting corrosion of
Ti6Al4V and CoCrMo surfaces. Journal of
Biomedical Materials Research Part A 101: 2602–
2612.
Takasaki, A.A., Aoki, A., Mizutani, K., Kikuchi, S.,
Oda, S. & Ishikawa, I. (2007) Er:YAG laser
therapy for peri-implant infection: a histological
study. Lasers in Medical Science 22: 143–157.
Tastepe, C.S., Liu, Y., Visscher, C.M. & Wismeijer,
D. (2012) Cleaning and modification of intraorally
contaminated titanium discs with calcium phos-
phate powder abrasive treatment. Clinical Oral
Implants Research 24: 1238–1246.
Tengvall, P., Elwing, H., Sj€oqvist, L., Lundstr€om, I.
& Bjursten, L.M. (1989) Interaction between
hydrogen peroxide and titanium: a possible role
in the biocompatibility of titanium. Biomaterials
10: 118–120.
Thierry, B., Tabrizian, M., Savadogo, O. & Yahia, L.
(2000) Effects of sterilization processes on NiTi
alloy: surface characterization. Journal of Bio-
medical Materials Research 49: 88–98.
Toniollo, M.B., Galo, R., Macedo, A.P., Rodrigues,
R.C.S., Ribiero, R.F. & Mattos, M. (2012) Effect of
fluoride sodium mouthwash solutions on cpTI:
evaluation of physicochemical properties. Brazil-
ian Dental Journal 23: 496–501.
Valderrama, P., Blansett, J., Gonzalez, M.G., Cantu,
M.G. & Wilson, T.G. (2014) Detoxification of
implant surfaces affected by peri-implant disease:
an overview of non-surgical methods. Open
Dental Journal 8: 77–84.
Velten, D., Biehl, V., Aubertin, F., Valeske, B., Poss-
art, W. & Breme, J. (2002) Preparation of TiO(2)
layers on cp-Ti and Ti6Al4V by thermal and ano-
dic oxidation and by sol-gel coating techniques
and their characterization. Journal of Biomedical
Materials Research 59: 18–28.
11 | Clin. Oral Impl. Res. 0, 2015 / 1–12
Sutton E. Wheelis Effects of decontamination solutions
Izabelle M. Gindri
Pilar Valderrama
on the surface of titanium:
Thomas G. Wilson Jr investigation of surface morphology,
Jessica Huang
Danieli C. Rodrigues
composition, and roughness

Authors’ affiliations: Key words: AFM, corrosion, detoxification, oxide layer, titanium
Sutton E. Wheelis, Izabelle M. Gindri, Danieli C.
Rodrigues, Department of Bioengineering,
University of Texas at Dallas, Richardson, TX, USA Abstract
Pilar Valderrama, Thomas G. Wilson Jr, Jessica Aim: To investigate the impact of treatments used to detoxify dental implants on the oxide layer
Huang, Private Practice of Periodontics, Dallas, TX,
morphology and to infer how changes in morphology created by these treatments may impact
USA
re-osseointegration of an implant.
Corresponding author: Materials and methods: Pure titanium (cpTi) and the alloy Ti6Al4V were subjected to a series of
Danieli C. Rodrigues
800 W. Campbell Road
chemical treatments and mechanical abrasion simulating surface decontamination of dental
Richardson, Texas 75080-3021, USA implants. The morphology and roughness of the surface layer before and after treatment with
Tel.: 972 883 4703 these solutions were investigated with optical and atomic force microscopy (OM, AFM). The
Fax: 972 883 4653
e-mail: Danieli@utdallas.edu solutions employed are typically used for detoxification of dental implants. These included citric
acid, 15% hydrogen peroxide, chlorhexidine gluconate, tetracycline, doxycycline, sodium fluoride,
peroxyacetic acid, and treatment with carbon dioxide laser. The treatments consisted of both
immersions of samples in solution and rubbing with cotton swabs soaked in solution for 1, 2, and
5 min. Cotton swabs used were analyzed with energy dispersive spectroscopy (EDS).
Results: The microscopy investigation showed that corrosion and pitting of the samples were
present in both metal grades with immersion and rubbing methods when employing more acidic
solutions, which had pH <3. Mildly acidic solutions caused surface discoloration when coupled with
rubbing but did not cause corrosion with immersion. Neutral or basic treatments resulted in no
signs of corrosion with both methods. EDS results revealed the presence of titanium particles on all
rubbing samples.
Conclusion: It was demonstrated in this study that acidic environments coupled with rubbing are
able to introduce noticeable morphological changes and corrosion on the surface of both titanium
grades.

Titanium has been placed at the forefront for
use in dental implants because of its mechan-
ical strength and protective oxide layer, which
ensures the bulk material of an implant is
protected against corrosion. This oxide layer
(e.g., TiO2, TiO) is naturally formed and
regenerated in the presence of air and/or aque-
ous media (Gilbert 2012; Swaminathan & Gil-
bert 2013). Besides providing protection
against corrosive environments, oxide layers
also aid in providing porosity to the surface.
Porosity and roughness are elements that
Date: allow bone surrounding an implant to osseo-
Accepted 4 December 2014 integrate, providing stability for the implant.
To cite this article: Titanium is unique in the fact that bone
Wheelis SE, Gindri IM, Valderrama P, Wilson TG Jr, Huang J, finds the oxide layer of the metal favorable
Rodrigues DC. Effects of decontamination solutions on the
surface of titanium: investigation of surface morphology, for integration. This is critical in dental
composition, and roughness.
implant applications because a well-osseoin-
Clin. Oral Impl. Res. 00, 2015, 1–12
doi: 10.1111/clr.12545 tegrated implant can survive the mechanical
requirements of the oral environment, mini-
mizing micromotion and the potential for
fretting-crevice corrosion associated with
occlusal forces (Mouhyi et al. 2012). To
obtain a favorable environment for osseointe-
gration to occur and continue, routine main-
tenance after placement has been suggested
(Wilson et al. 2014). These procedures are
designed to diagnose and treat inflammatory
responses known as peri-implant mucositis
and peri-implantitis (Mouhyi et al. 2012;
Wilson et al. 2014). Peri-implantitis is char-
acterized by inflammation and continued loss
of integrated bone around the implant. If
peri-implantitis does occur, the clinician has
the option to either remove the infected
implant or perform debridement and detoxifi-
cation of the implant surface to remove the
biofilm (Valderrama et al. 2014). The goal of

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd 1