Dental Materials Journal 2023; 42(4): 461–468

Improvement implant osseointegration through nonthermal Ar/O2 plasma

Chengzan WU1*, Min YANG2*, Kai MA1, Qian ZHANG1, Na BAI1 and Yanshan LIU3

1
Department of Prosthodontics, The Affiliated Hospital of Qingdao University, Qingdao 266003, China
2
Department of Oral and Maxillofacial Surgery, Shanxi Provincial People’s Hospital, Taiyuan 030001, China
3
Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266003, China
Corresponding author, Yanshan LIU; E-mail: yanshan_l@qdu.edu.cn

This study investigated the effects of nonthermal Ar/O2 plasma on the osseointegration of titanium implants. Through 8 weeks’ in
vivo evaluation of implants inserted into femoral bones of male Sprague-Dawley rats, the new bone mineralization apposition rate
(MAR) is increased by 1.87 and 2.14 times for implants of smooth machined (SM) and sand-blasted and acid-etched (SLA) after
plasma treatment. The bone volume fraction (bone volume/total volume, BV/TV) and bone-implant contact (BIC) ratios are improved
by 1.31, 1.26 times and 1.35, 1.15 times after 90 s plasma treatment. The improved hydrophilicity rather than implant surface
morphology is believed to play a critical role for the osseointegration improvement.

Keywords: Nonthermal plasma, Hydrophilicity, Osseointegration, Implants, New bone formation

Therefore, we are interested in the development of a

INTRODUCTION
The invention and use of dental implants have provided
a novel method of restoration for patients with tooth
loss. Moreover, dental implants have a success rate1).
Since implants were used clinically in the 1960s, the
clinical indications have become increasingly extensive,
due mainly to the research progress in the surface
modification of titanium implants.
Titanium and its alloys have been used widely in
dentistry for many years due to their high strength,
superior wear resistance, excellent corrosion resistance,
excellent fatigue properties, and good biocompatibility2).
Nevertheless, owing to the bioinertness of titanium,
implants with no hydrophilicity hardly combine with
osteoblasts, reducing osseointegration, while hydrophilic
implants can significantly enhance osseointegration,
shorten healing time, and increase the success rate of
implant operations3,4). Increasing the microroughness
on the implant surface and altering the surface
chemistry will increase the hydrophilicity of the implant
surface, improve the retention force, and enhance
osseointegration5,6). The method of sandblasting and
acid etching implants to modify their surface roughness
is commonly used to enhance cell migration, adhesion,
and osseointegration6).
However, studies have shown that the hydrophilicity
and biological properties of the titanium surface show
time-dependent degradation, that is, biological ageing7-9).
Over time, carbon molecules will accumulate on the
titanium surface in the form of hydrocarbons, causing
pollution and affecting the biological performance of
the implant surface10). Biologically ageing titanium
affects the adsorption of proteins such as albumin
and fibronectin and further affects the deposition,
proliferation, and mineralization of osteoblasts, which
in turn affects the integration of implants and bones8).
simple and easy method that can be directly operated
next to the dental chair to activate the surface of the
implant and improve its biological activity.
Representative examples of simple treatment
methods, such as ultraviolet (UV) irradiation11) or plasma
treatment12), do not change the surface topography of
the implant. Hydrophilic treatment in clinical practice,
however, may be difficult to accomplish due to the long
irradiation time of UV irradiation. In the contrast,
nonthermal plasma shows advantages in biomedicine
due to its low operating temperature (<40°C), and simple
and fast processing methods. A large high concentration
of bioactive substances produced by plasma can cause
changes in the surface energy and chemical properties
of the material, thereby promoting early wound recovery
and implant-bone interactions13-16).
Plasma is an electrically neutral ionized gas,
composed of reactive oxygen and reactive nitrogen
species (RONS), excited atoms and molecules, chemically
reactive and neutral particles, and plasma emits UV
irradiation15,17). The production of RONS has led to the
use of plasma treatment to clean implant surfaces and
enhance hydrophilicity, which in turn encourages protein
and cell adhesion15). In addition, according to Duske et al.,
titanium discs with human plaque biofilm were treated
with nonthermal plasma without imparing osteoblastic
cell development, and the results were comparable to
those of sterilized controls, demonstrating the role of
nonthermal plasma in sterilization18). Without affecting
the intricate surface geometry of the implant, the RONS
produced by plasma treatment could promote osteoblast
attachment and differentiation19,20). Nonthermal plasma
can introduce different functional groups to change the
physicochemical properties of the surface of the material
without affecting its microstructure, enhancing its
surface hydrophilicity21). Researchers found that oxygen

*These authors contributed equally to this work.

Received Jul 12, 2022: Accepted Jan 6, 2023
doi:10.4012/dmj.2022-158 JOI JST.JSTAGE/dmj/2022-158
462 Dent Mater J 2023; 42(4): 461–468
plasma introduced a mixture of predominantly -OH
functional groups and argon plasma introduced free
radicals22).
In the previous experiment, we used nonthermal
argon-oxygen plasma (NTAOP) to activate the surface of
the titanium sheets and carried out surface
characterization and in vitro experiments. The
experimental results showed that the NTAOP
can significantly increase the surface energy and
hydrophilicity of the pure titanium surface without
destroying the surface morphology of the titanium
sheets, accelerate the early adhesion and growth of
osteoblasts, and promote the expression of osteocalcin
and maturation of cytoskeletal actin10). The purpose of
this experiment was to investigate whether NTAOP
could activate implants in in vivo experiments and
improve the hydrophilicity of implant surfaces, thereby
promoting osseointegration. The null hypothesis of the
experiment was that pure titanium implants activated
by NTAOP do not promote osseointegration in vivo.
MATERIALS AND METHODS
Titanium materials
We used smooth machined titanium implants (SM Ti),
sandblasted, and acid-etched titanium implants (SLA
Ti) (grade 4, length 10 mm, diameter 2 mm). All implants
were made and sterilized by WEGO Company (Wei Hai,
China).
NTAOP activation
Implants in the experimental groups were treated
by NTAOP using an atmospheric pressure plasma
system (AS400+PFW10, PLASMA Treat, Steinhagen,
Germany), the same as the previous experiment10). The
schematic diagram is shown in Fig. 1. This plasma
system, powered by a low-frequency power supply (19
kHz) and matched network, is a glow-discharge plasma
Fig. 1 Schematic illustration of the atmospheric pressure
plasma system.
In this system, 95% Ar and 5% O2 were used as
inputting gases to generate plasma.
workspace with dimensions of 33 cm by 12.5 cm. The
parameters of the nonthermal plasma generating
equipment are the voltage of 320 V, a current of 0.3 A,
and the gas flow rate of 300 L/h. We placed the implants
in high-temperature resistant quartz glassware with a
diameter of 14 mm and a height of 30 mm and placed
them together at a distance of 4.5 cm from the nozzle of
the nonthermal plasma. Then the sample was exposed
to NTAOP generated with 95% argon+5% oxygen as the
input gas for 90 s.
Surface morphology observation
The SM Ti and SLA Ti surface morphologies were
examined before and after NTAOP treatment under
scanning electron microscopy (SEM; Phenom XL,
Phenom-World, Eindhoven, The Netherlands) at 10 kV.
Surface hydrophilicity
For wetting analysis, a contact angle analysis was used
to measure the contact angle of liquids on titanium
surfaces before and after NTAOP treatment. Disc-
shaped samples (n=4) were fabricated. And the surface
of the disc-shaped samples was given the same surface
treatment as the implants after they were fabricated. A
live video contact angle measurement system was used
to capture the image at room temperature after distilled
water (5 µL) was applied to the center of each sample
surface for ten s. Before and after NTAOP treatment,
changes in the hydrophilicity of SM Ti and SLA Ti were
evaluated by simulating implantation experiments, and
the changes in surface wetting property were observed
with the naked eye.
Experimental animals and surgical procedure
Animal selection, management, and surgical protocols
were in accordance with guidelines set forth by the
Institutional Animal Care and Use Committee of the
affiliated hospital of Qingdao University, Qingdao, China
(Approval No. AHQU-MAL20200619). The experimental
rats were kept in the animal room for at least one week to
adapt to the new environment. The rats were placed in a
constant temperature and humidity breeding room with
alternating light and dark every 12 h, and sufficient water
and food were provided. Twenty-four Sprague Dawley
rats, with a bodyweight of approximately 300 g, aged 3–6
months were selected as experimental objects. Before
the operation, the rats were prohibited from eating and
drinking for at least 12 h. After weighing, the rats were
injected intraperitoneally at a dose of 0.4–0.5 mL/100 g
body weight with chloral hydrate for anaesthetization.
When the rats were anaesthetized, the leg of the rats
was shaved, disinfected with iodophor disinfectant, and
positioned and clothed in sterile sheets on the table.
Then, 0.5 mL was injected into the rat femoral condyle
with articaine to reduce intraoperative bleeding and
pain. A curved incision measuring 10–15 mm was made
on the anterolateral knee surface; the joint capsule
was dissected, and incised, and then the patella was
subluxated, exposing the femur intercondylar fossa23).
At the center of the femoral intercondylar fossa as the
 Dent Mater J 2023; 42(4): 461–468
implant placement point, a split drill was used to prepare
an implant cavity with a depth of 10 mm and a diameter
of 1.8 mm at a low speed (800 rpm), and a large amount of
sterile normal saline was used to flush and cool the hole.
Then, one untreated SM Ti implant was inserted into
each left femur of the 12 rats, and one NTAOP-treated
SM Ti implant was inserted into each right femur.
The placement of 12 untreated and 12 NTAOP-treated
SLA implants was done in the same way. To avoid the
intrusion of surrounding soft tissues from affecting the
healing of the implant after implantation, it is necessary
to pay attention to the intrusion of surrounding soft
tissues during the implantation process. After the
implantation was completed, the joint capsule was reset,
the muscle, fascia, and subcutaneous tissue were sutured
layer by layer with 4-0 absorbable sutures, and the skin
was sutured with 4-0 silk suture. Antibiotics (penicillin
600,000 units/d) were injected intramuscularly for three
consecutive days after surgery to prevent postoperative
infection. To label bone metabolism, Alizarin Red was
injected at 30 mg/kg intraperitoneally on the 14th and
13th days before the rats were sacrificed, and Calcein
20 mg/kg was injected at 30 mg/kg intraperitoneally on
the 4th and 3rd days before the rats were sacrificed24).
After 8 weeks, the rats were sacrificed, and their femurs
along with implants were harvested for histology and
microcomputed tomography (micro-CT) examination.
Micro-CT
At eight weeks after implantation, the rat femur
specimens (n=3 specimens per group) with the implants
were obtained, the soft tissue surrounding the specimens
was removed, and then the specimens were subjected to
a micro-CT (Skyscan1076, Bruker, Antwerp, Belgium)
test. The specimens were scanned by micro-CT with a
voxel size of 18 μm and a voltage and electric current of
70 kV, 120 μA with an integration time of 700 ms. The
bone section around the implants from 1.8 mm below
the growth plate to 100 distal slices with a ring radius
of 0.5 mm from the implant surface was defined as the
Fig. 2 Schematic of the evaluation methodology.
(a) Position of inserted implants, (b) VOI of micro-
CT evaluation
463
volume of interest (VOI)25) (Fig. 2). The specimens were
measured by CTAN software to quantitatively analyse
the trabecular bone. The relevant parameters include
relative bone volume (BV/TV), trabecular number
(Tb. N), trabecular thickness (Tb. Th), and trabecular
separation (Tb. Sp).
Histologic analysis
After the micro-CT scan was completed, the specimens
were sliced for histological analysis. First, the rat
femurs containing implants were fixed in a 4% neutral
formalin buffer solution for 3 days. Then, the sample
was rinsed with running water to remove the residual
neutral formalin on the sample and then dehydrated
in ascending grades of ethanol of 70%, 80%, 90%, 100%
(two days are needed for each ethanol grade). After being
dehydrated, the samples were embedded in methyl-
methacrylate (Sigma–Aldrich, St. Louis, MO, USA).
After resin hardening, the specimens were cut parallel
to the longitudinal axis of the implants with a rotary
cutting saw (Exakt310CP, Exakt, Hamburg, Germany)
into 200-μm thick slices. Then, all histologic slices were
ground to approximately 50 μm with an Exakt400CS
(Exakt, Norderstedt, Germany). First, an evaluation
by fluorescence microscopy was performed (Zeiss
LSM 800, Carl Zeiss, Jena, Germany). The new bone
mineralization apposition rate (MAR) was determined
by measuring the distance between alizarin red (red) and
calcein blue labels. In all animals in each experimental
group, five measurements were taken at random points
between two labeling fronts along five different locations
of the implant surface. Then the MAR was calculated
by dividing the average by the interval between
administering each fluorochrome26). Subsequently, Van
Gieson staining, and microscopy (Zeiss Axio Imager M1,
Carl Zeiss, Oberkochen, Germany) were performed. For
histomorphometry analysis, the bone-implant contact
(BIC) was determined at various magnifications by
means of computer software (Image-Pro Plus; Media
Cybernetics, Silver Spring, MD, USA). The BIC was
defined as the length of BIC in relation to the implant
perimeter starting at the shoulder of the implant20). The
analyses were conducted by the same researcher, who
was blinded as to which group each sample belonged.
Statistical analysis
All data were collected and presented as the mean
and the standard error of the mean. The experimental
datasets were analysed using one-way analysis of
variance (ANOVA) and the Student-Newman-Keuls test
(SPSS statistics 26.0). The significance level was set to
a p value of 0.05.
RESULTS
Surface morphology
The application of NTAOP did not cause any changes in
the microtopography of the implant surface. As shown
in Fig. 3, under different magnifications, no changes
in the surface morphology of the implants were found.
464 Dent Mater J 2023; 42(4): 461–468

Fig. 3 Observation of implant surface morphology.

(a) SM Ti, (b) SM Ti+NTAOP, (c) SLA Ti, (d) SLA Ti+NTAOP

Fig. 4 Observation of different implant wetting property.

A: Water droplet images and water contact angle of Ti discs before and after
NTAOP treatment, B: Comparison of the wetting properties of untreated
and treated Ti implants. (a) SM Ti, (b) SM Ti+NTAOP, (c) SLA Ti, (d) SLA
Ti+NTAOP

Figures 3a and 3b show implants with a smooth surface.
The surface of the implants has uniformly aligned and
neatly arranged mechanical polishing traces. Figures 3c
and 3d show the SLA implants. The surface structure
of the implants can be observed to be a honeycomb-like
multilevel hole structure, but the microscopic morphology
of the implant surface before and after NTAOP treatment
did not change. The application of NTAOP indicates
that the geometry of the structure on the sample surface
was not destroyed by NTAOP treatment, preserving its
beneficial effects on osseointegration.
Surface wetting property
There was a statistically significant difference in the
contact angle between the untreated group and the
NTAOP treatment group (p<0.05). Before NTAOP
treatment, the contact angle of the SM Ti and SLA
Ti was 91.375±0.59° and 108.95±5.88°, respectively.
After NTAOP treatment, the contact angle of the SM
Ti+NTAOP and SLA Ti+NTAOP was 10.03±1.30° and
5.30±3.83°, respectively. As shown in Fig. 4B, the SM
Ti and SLA Ti before and after NTAOP treatment were
placed in the liquid to observe the changes in surface
wetting property with the naked eye. After placing the
NTAOP treated SM Ti and SLA Ti implants in liquid,
the liquid was found to spread rapidly along the surface
of the implant and quickly covered the implant surface,
while the untreated group had no obvious infiltration.
Based on the results, the implants showed increased
wetting property after NTAOP treatment, regardless of
their surface morphology (p<0.05) (Fig. 4B).
Clinical observation
Two rats died on the fourth and seventh days after the
 Dent Mater J 2023; 42(4): 461–468
operation. The cause of death might be a secondary
infection caused by wound dehiscence. One dead rat
belonged to the SLA Ti group, and the other belonged
to the SM Ti group. The remaining 22 rats were in
good condition after the operation, and there were no
infections or deaths before they were sacrificed.
Micro-CT evaluation
Micro-CT analysis was performed to evaluate new bone
formation around the implant and the osseointegration
of implants. In the image, the bone is characterized in
white, and the implants are characterized in yellow. An
obvious difference in the quantity of bone in the VOI
was observed in 3D micro-CT reconstructions (Fig. 5A).
Regarding the quantitative results, the micromorphology
of femurs in group SM Ti was typified by the lowest
values in BV/TV (11.98±0.51%), and Tb. N (1.31±0.01).
Compared to the SM Ti group, the BV/TV, and Tb. N was
remarkably higher in group SM Ti+NTAOP and SLA
Ti groups (p<0.001) (Fig. 5B). For the SLA Ti+NTAOP
group, there was a better recovery in bone apposition
around implants, with a significant increase in BV/TV
Fig. 5 A: Longitudinal and transverse micro-CT images
of proximal femurs. (a) SM Ti, (b) SM Ti+NTAOP,
(c) SLA Ti, (d) SLA Ti+NTAOP
B: Quantitative evaluation results. The asterisk
indicates significant differences by post-hoc
Student-Newman-Keuls test (***: p<0.001, ns: not
significant).
465
(29.08±0.61%) and Tb. N (2.54±0.09), but the lowest
value in Tb. sp (0.28±0.01 μm).
Histological analysis
The VG-stained tissue sections and the histomorphology
results presented as BIC are shown in Fig. 6. Figure 6A
shows a VG-stained tissue section of the bone-implant
interface, showing the newly formed bone around the
implant surface. After 8 weeks of bone healing around
the implants, a BIC of 31.54±0.89% was observed in
the SM Ti, while the BIC values in the SM Ti+NTAOP,
SLA Ti, and SLA Ti+NTAOP groups were 42.43±0.52%,
57.14±1.21%, and 65.71±1.13%, respectively. Compared
with the untreated group, the NTAOP group showed a
higher BIC value (Fig. 6B). The SLA Ti+NTAOP group
showed the highest influence on increasing BIC (p<0.05),
while the SM Ti+NTAOP group showed the next highest
influence (p<0.05).
The fluorescence microscopy results showed that red
and green fluorescence were enriched on the surface of
the implant, indicating bone growth at 6 and 8 weeks
after insertion (Fig. 7A). In the SM Ti group and SLA Ti
group, irregularly arranged fluorescence enrichment was
found scattered around the implant. The fluorescence
enrichment around the implants in the NTAOP group
was more obvious than the fluorescence enrichment
Fig. 6 Histological evaluation of the bone-to-implant
interface with Van Gieson staining.
A: (a) SM Ti, (b) SM Ti+NTAOP, (c) SLA Ti, and
(d) SLA Ti+NTAOP. B: Quantitative evaluation
results. The asterisk indicates significant
differences by post-hoc Student-Newman-Keuls
test (***: p<0.001).
466 Dent Mater J 2023; 42(4): 461–468
Fig. 7 Histological evaluation of the bone-to-implant
interface with fluorescence microscopy. Newly
formed bone is labelled by the enrichment of red
and green fluorescence.
A: (a) SM Ti, (b) SM Ti+NTAOP, (c) SLA Ti, (d)
SLA Ti+NTAOP. B: Fluorescently labelled bone
mineralization apposition rate. A varying number
of asterisks indicate significant differences by
post-hoc Student-Newman-Keuls test (*: p<0.05,
***: p<0.001, ns: not significant).
in the untreated group. The difference in the SLA
Ti group was more significant. As a result of NTAOP
treatment, there was an increase in the new bone MAR.
A statistically significant difference was found in the
SLA Ti group (p<0.001), and in the SM Ti group (p<0.05)
(Fig. 7B).
DISCUSSION
The success of implant surgery depends on
osseointegration after implant placement. Albrektsson
et al.27) defined the term osseointegration, that is, the
direct contact between the implanted implant surface
and the bone at the microscopic level. In the past few
decades, researchers have studied the modification
of the titanium surface to promote osseointegration.
However, studies have reported that carbon impurities
will inevitably accumulate on the titanium surface over
time, affecting osseointegration, that is, the biological
ageing phenomenon of titanium7-9). Therefore, how
to activate the surface of the implant immediately
before implantation has triggered people’s thinking.
Some scholars have improved the biological activity
of the surface of the titanium sheet by heating or UV
irradiation, but shortcomings such as time-consuming
and difficult operation methods limit its clinical
application28). NTAOP was used in the current study to
modify the implant surface, and the effect of promoting
osteogenic activity was thoroughly assessed.
In accordance with the microtopography observed
by SEM, the SLA Ti group surfaces showed the same
multilevel hole structure whether treated by NTAOP or
not, indicating that the geometry of the structure on the
sample surface was not destroyed by plasma treatment,
preserving its beneficial effects on osseointegration.
Implants with rough surfaces are reported to play a
vital role in simulating natural bone tissue to promote
bone healing after implantation29). This feature
might be very important, because the wettability of
nonbiological materials is not only determined by the
chemical composition, but also significantly determined
by the surface roughness and the micro-roughness of
the implant itself has been optimally adjusted through
extensive research as it is one of the important surface
properties for osseointegration that should not be easily
changed30).
In the hydrophilicity experiment, we observed
that the surface wetting property of the implant was
significantly improved after NTAOP treatment. This
might be due to the fact that the NTAOP treatment
increased the implants’ hydrophilicity by forming a
large number of polar oxygen groups (such as carboxyl,
carbonyl, and hydroxyl groups) on their surface31). Among
the properties of implants, surface hydrophilicity is the
first influencing factor that determines the formation
of osseointegration after implantation. Studies have
proven that hydrophilic implants can actively regulate
the pathway directly involved in the differentiation and
commitment of osteoblasts, thereby enhancing bone
formation at the implantation site4).
In this study, the postoperative scrutiny time was 8
weeks, which is a standard period of two-stage healing
of high-quality bone implants. In the eighth week, we
tested the BV/TV, Tb. n, Tb. Th and Tb. Sp by using
micro-CT. Measuring both trabecular microstructure and
bone density parameters will allow for a more accurate
evaluation of bone quality32,33). BV/TV is an important
index to describe the microstructure of bone, which
can reflect the content of bone tissue in the sample and
determine the main mechanical properties of trabecular
bone. Tb. N is the number of intersections between bone
tissue and nonbone tissue within a given measurement
range. Studies have shown that the increase in Tb.
N around the implant is beneficial to alleviate the
pressure of chewing. Tb. Sp is the average width of the
medullary cavity between trabecular bones. When there
is an increase in osteoporotic bone resorption, the Tb.
Sp value will increase accordingly. The experimental
results showed that the BV/TV and Tb. N values of
the SLA Ti group were higher than those of the SM
Ti+NTAOP group, which may indicate that although
the hydrophilicity of the implant surface increased
 Dent Mater J 2023; 42(4): 461–468
significantly in the SM Ti+NTAOP group, the geometry
structure of the implant surface is still the main factor
affecting osseointegration. and SLA Ti+NTAOP group
were significantly higher than those of the control
group. Moreover, there was a significant increase in
BV/TV and Tb. N in the group treated with NTAOP
compared to the untreated group, and the results were
statistically different. This might suggest that implants
with higher surface energy and hydrophilicity facilitate
osseointegration10). We also tested the BIC by making
hard tissue sections. The BIC reflects the osteogenic
ability of the implant. Therefore, a higher BIC is generally
considered a necessary condition for the stability of the
implant to achieve the functional reconstruction of the
implant denture34). The experimental results show that
the BIC was consistent with the micro-CT results and
that the implants treated with NTAOP had a higher BIC
compared to the untreated group.
These findings are consistent with other studies
describing the effects of nonthermal plasma treatment
in animal models. Teixeira et al.35) also conducted
experiments on implants activated by argon nonthermal
plasma in beagle dogs. In the experiment, they used
nonthermal plasma to activate each quadrant of the
implant for 20 s or 60 s. The experimental results show
that, compared with the control group, the surface
energy level of the implants in the experimental group
with different treatment times is higher, the adsorption
amount of carbon-containing impurities is lower,
and the torque level is higher after 2 and 4 weeks of
implantation.
From the histological level, to dynamically observe
the condition of the new bone around the implant, we
also adopted a double fluorescent labelling method. The
double fluorescent labelling method is often used to
observe the growth and mineralization of new bone. In
the experiment, we used two fluorescent dyes, Alizarin
Red and Calcein. The mechanism is to use the first step
of bone mineralization to combine with calcium ions in
the body to form a chelation reaction. The hard tissue
section was placed under a fluorescence microscope, and
the red and green fluorescence bands were displayed
under excitation at a wavelength of 605 nm (Alizarin
Red) and 522 nm wavelength (Calcein)36). Fluorescence
staining results show the new bone MAR after injection
of different fluorescent dyes. The results showed that the
rate of new bone mineralization apposition was highest
in the SLA Ti+NTAOP group, which was followed by the
SM Ti+NTAOP group and was higher than the SLA Ti
group, while the SM Ti group had the lowest rate. This
indicates that NTAOP treatment enhanced the new
bone MAR in the SLA Ti+NTAOP and SM Ti+NTAOP
group, possibly because hydrophilic implants promote
the adsorption of proteins after implantation. Studies
have shown that when the implant is infiltrated by blood
after implantation, hydrophilicity affects the quantity,
tridimensional conformation, and bonding strength of
the protein37).
However, implants treated with NTAOP are also time
sensitive. The higher surface energy and hydrophilicity
467
obtained by the implant is not lasting. Over time, the
hydrophilicity of the implant surface will gradually
weaken until it disappears. Through preliminary
experiments, after two days of NTAOP treatment, the
water contact angle on the titanium surface was found
to gradually increase from less than 1° to close to 10°
and to continue to increase over time. However, how long
the hydrophilic effect of plasma treatment can last have
not been elucidated. The limitation of this study is that
only the immediate implant insertion following NTAOP
treatment was considered; the impact of storage time on
the potential of the implants to stimulate osteogenesis
was not taken into account. Thus, the influence of the
change in surface hydrophilicity over time on in vivo and
in vitro experiments requires further exploration.
CONCLUSIONS
NTAOP treatment did not change the surface morphology
of the implant. However, within the limits of this study,
the results showed that the NTAOP treatment of pure
titanium implants enhance osseointegration. The
higher bone-to-implant contact is beneficial to improve
the stability of the implant to realize the functional
reconstruction of the implant denture. However, the
timeliness of implants treated with NTAOP still needs
to be studied. In addition, further studies should be
carried out according to the schedule to clarify the effect
of plasma treatment according to the healing stage.
ACKNOWLEDGMENTS
This work was supported by the National Natural
Science Foundation of China [Grant no. 81500882]
and Scientific Research Project of Affiliated Hospital of
Qingdao University [Grant no. QDFYQN202001015].
The authors would like to thank Dr. Peter P. Sun from
Intel Corporation for his thesis guidance. The authors
would like to thank Professor Dagang Miao’s lab at
Qingdao University for technical support and facility
utilization.
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