The European Journal of Orthodontics Advance Access published January 18, 2016

European Journal of Orthodontics, 2016, 1–8

doi:10.1093/ejo/cjv097

Original article

Nanosilver coated orthodontic brackets: in vivo

antibacterial properties and ion release
Gamze Metin-Gürsoy*, Lale Taner* and Gülçin Akca**
Departments of *Orthodontics and **Medical Microbiology, Faculty of Dentistry, Gazi University, Ankara, Turkey

Correspondence to: Gamze Metin Gürsoy, Department of Orthodontics, Faculty of Dentistry, Gazi University, Bişkek cad.
1. Sok. No: 4 Emek, 06510 Ankara, Turkey. E-mail: gamgursoy@gmail.com

Summary
Introduction: Silver nanoparticles are currently utilized in the fields of dentistry. The aim of this
study was to evaluate the antibacterial properties and ion release of nanosilver coated orthodontic
brackets compared to conventional brackets.
Methods: Nanosilver coating process was applied to standard orthodontic brackets placed on the

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mandibular incisors of Wistar Albino rats in the study group and conventional brackets in the
control group. Dental plaque, mucosal vestibular smears, saliva, and blood samples were collected
from rats at various days. The amounts of nanosilver ions in blood and saliva were measured
and microbiological evaluation was made for Streptococcus mutans. For testing cariogenicity, all
rats were sacrificed at the end of 75 days under anaesthesia. Teeth were stained using a caries
indicator, then the caries ratio was assessed.
Results: Nanosilver coated orthodontic bracket favoured the inhibition of S.mutans on Day 30
and reduction of caries on the smooth surfaces. The nanosilver amounts in the saliva and serum
samples were significantly higher in the study group on Day 7.
Conclusion: It is suggested that nanosilver coated orthodontic brackets, as an antibacterial agent
without patient compliance, could be helpful for the prevention of white spot lesions during fixed
orthodontic treatment.

Introduction Fluoride is known as the most appropriate agent that prevents

the formation of WSL during orthodontic treatment. In order to
The purpose of orthodontic treatment with fixed appliances is
prevent WSL, fluoride is applied regularly in orthodontic patients
to improve function and aesthetics. After the placement of fixed
in many different ways including topical fluoride preparations, var-
orthodontic appliances, patients are at higher risk for caries
nish, gel, and solution implementation around brackets. In addi-
due to increased plaque and bacteria retention around brackets
tion, when orthodontic patients use a fluoride mouth rinse and
and bands. Orthodontic bands and brackets make it difficult for
tooth brushing with fluoride toothpaste, WSL has been shown
patients to achieve proper brushing and oral hygiene maintenance,
to reduce around fixed braces and bands; however, this applica-
which increases caries incidence (1–3). Streptococcus mutans and
tion is dependent on patient cooperation. Unfortunately, only
Lactobacilli are the most common bacteria that can produce sig-
less than 15% of orthodontic patients follow instructions (7–10).
nificant levels of lactic acid causing demineralization of teeth.
However, there is still a need to prevent WSL by acid-resistant anti-
Demineralization of the enamel results in a displeasing appear-
caries agents needing no patient cooperation during orthodontic
ance, which is called white spot lesions (WSL) (1, 4). WSL can
treatment.
become noticeable around the brackets within 1 month of bond-
The antimicrobial effects of silver ion or salts are known well
ing and the prevalence of WSL has been reported to be 38% in
since ancient times (11). Silver is currently used to control bacterial
6 months, 46% in 12 months after the initiation of the treatment,
growth in a variety of applications, including dental work, catheters,
and 50% at the end of the fixed orthodontic treatment (2, 5, 6).
and burn wound dressings (12–15). The term ‘nanotechnology’ was

© The Author 2016. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved.
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2 European Journal of Orthodontics, 2016

introduced by Professor Norio Taniguchi of Tokyo Science University cages with free access to food (soft biscuits) and water ad libitum.
in 1974 and the word ‘nano’ is used to indicate one billionth of a The rats were kept under an artificial 12-hour light/darkness cycle.
meter. Silver nanoparticles with their unique chemical and physical Lights were turned on at 7 AM and off at 7 PM. Room temperature
properties have been proved as an alternative for the development of varied between 22°C and 24°C, and appropriate room ventilation
new antibacterial agents (12, 16). was maintained.
The purpose of this study was to assess the antibacterial proper-
ties and ion release of nanosilver coated orthodontic brackets, to Collection of samples
compare the effects of silver regarding the amount of cariogenic All animals were anesthetized with 10 mg/kg xylazine hydrochlo-
streptococci adhesion, and to provide preliminary information for ride and 90 mg/kg ketamine hydrochloride by intraperitoneal injec-
the prevention of WSL in comparison with conventional orthodontic tion before all mentioned procedure. Saliva, blood, vestibular smear
brackets. The hypothesis tested was that silver nanoparticles coated from the lower lip epithelium, and the plaque sample of the incisor
to orthodontic bracket would decrease the adherence of S.mutans teeth were collected from all rats on Day 0 (before inoculation of
and caries during fixed orthodontic treatment. S.mutans cell suspension), and on Days 1, 3, 7, 14, 30, 45, and 75
(after inoculation).
Saliva was collected by placing a filter paper strip (Periopaper,
Materials and methods
Oraflow, Smithtown, New York, USA) under the tongue and the
All animal procedures were conducted with the approval of Gazi volume was measured by means of a precision balance (aeADAM,
University Animal Experiments Local Ethics Committee. All animal Kinston, UK). Samples were put into glass capped tubes containing
experiments were performed by one researcher who has a certificate 0.2 ml concentrated nitric acid. Blood samples were collected from
according to the guidelines for proper conduct of animal experi- the tail vein and serum samples were obtained from coagulated and
ments. Microbiological analyses were performed in Gazi University, centrifuged blood, which were then stored at −20°C.
Faculty of Dentistry, Department of Medical Microbiology. Silver analysis was performed by direct injection into the induc-
tively coupled plasma-mass spectrometer (ICP-MS) for saliva and
Coating procedure serum samples. The total silver concentration (ppb) was determined
Mandibular incisor orthodontic brackets (Gemini Roth; 3M Unitek, for each sample.
Monrovia, California, USA) were used in this study. All brackets Vestibular smear was collected by swabs on sterile coverslips

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were cleaned sonically with alcohol for 15 minutes. The coating and S.mutans was counted microscopically using the Gram-staining
of the brackets was made via physical vapour deposition (Midas method. Then, the number of S. mutans was counted at randomly
Thermal Evaporator, Vaksis, Ankara, Turkey). Brackets were fixated chosen 20 different microscopic areas, and it was calculated accord-
onto the substrate of the e-beam evaporator device using double- ing to arithmetic mean numbers.
sided bonding tapes. Electron beam evaporation method was per- The plaque sample of the incisor teeth was collected by dental
formed under <2 × 10−6 Torr vacuum pressure with oil-free pumping floss. Dental floss was put into a sterilized test tube containing 1 ml
for 8 hours and brackets were coated to 1 μm thickness by nanosil- Trypticase Soy Broth medium (TSB Merck, Germany), the test tubes
ver particles. were vortexed for 60 seconds and the suspensions were diluted
to 10–1, 10–2, and 10–3 serially. Then, 100 µl amounts of samples
Rat caries experiments were put onto tryptone-yeast extract-cysteine-sucrose-bacitracin
agar (TYCSB) for 48–72 hours in a 5% CO2 incubator to culture
Twelve male Wistar rats (4 months old) were used in this study. The
S.mutans. All plaque samples were investigated for bacterial coloni-
animals were divided randomly into two groups of six rats each. All
zation and the total numbers of viable S.mutans colonies were calcu-
rats were dosed by gavage with amoxicillin (25 mg/kg of rat) for two
lated as colony forming units (CFU). Therefore, the data analysis and
consecutive days (on Days −4 and −3) in order to enable the inocu-
statistics were performed according to the 1 ml amount of volume
lated organisms to establish themselves in the oral cavity to suppress
as CFU/ml.
the indigenous flora of the rats. After 2 days, all rats were started to
All animals were sacrificed on Day 75 by overdose anaesthetic
be fed with soft biscuit and water free of antibiotics for inoculation
solution. The left jaws were removed and the soft tissue was stripped
of S.mutans (on Days −2 and −1).
off. The left jaws were stored in plastic flasks containing 10% for-
On Day 0, nanosilver coated brackets in the study group and
maldehyde solution.
conventional brackets in the control group were bonded using a flu-
orine-free light-cured composite resin and adhesive (Transbond XT,
3M Unitek) in accordance with the manufacturer’s instructions on Caries assessment
the vestibular surface of the mandibular incisors as close as possible Caries evaluation was performed on smooth and occlusal sur-
to the gum. In addition to bonding the bracket, a ligature wire was faces of the maxillary molars and the sulcus of the mandibu-
used to ligate the bracket to the teeth in order to prevent the loss of lar molar teeth on the left side of each animal. To evaluate the
the bracket. sulcal caries, mandibular molar teeth were cut off through the
Immediately after the bonding process, the mouth of the rat sagittal plane. Before the pictures of the samples were taken
was infected with 0.1 ml of a cell suspension containing 6 × 108 cells using a digital camera, all sample teeth were stained using caries
of S.mutans ATCC#700610 serotype c (American Type Culture indicator-Sable Seek (Ultradent Products, Utah, USA) (Figure 1).
Collection) using a micropipette. Additionally, 1 ml of a cell suspen- The images were loaded in ArcGIS 10.2 for Desktop (ESRI,
sion was mixed into the drinking water of the rats in each cage and California, USA). The areas of caries were determined according
this mixed drinking water was accessible to the rats until the next to the extent of staining with Sable Seek (Ultradent Products)
morning. Then, this water mix was removed and replaced with regu- and the rates of caries (CR) were calculated for each surface
lar tap water. During the test period, the animals were kept in plastic (CR = Caries area/Tooth area).
G. Metin-Gürsoy et al. 3

Statistical analysis the study group (Table 2). Intra-group examination for the control
The statistical significance was determined using SPSS 20.0 software group showed that there were no differences between the serum con-
for Windows (SPSS Inc., Chicago, IL, USA). The Shapiro–Wilk test centrations of nanosilver on the other days.
was used to verify the normality of the data distribution. Differences
between the study and the control groups were tested by the Mann–
Whitney U-test and intra-group differences were tested by the Wilcoxon
test. Values were considered statistically significant at P < 0.05.

Results
In the present study, with a one-sided significance level of 0.05 and a
power of 75%, a minimum of six animals per group were included.
After the placement of the brackets, the animals did not dis-
play any behavioural or weight changes, and no mortality was
observed. During the experiment, brackets were checked every day
and, if dropped, a new bracket was bonded immediately, which
occurred 17 and 15 times for the study and the control groups,
respectively.

Saliva samples
There were not any significant differences between the study and the
control groups (Figure 2), whereas on Day 7, the salivary concentra-
tion of nanosilver was found significantly higher than the other days
in the study group (Table 1). Intra-group examination for the control
Figure 2. Silver saliva level in the study and the control groups.
group showed that there were no significant differences between sali-
vary concentrations of nanosilver on the other days. Table 1. Statistical comparison of saliva silver level in the study
group.

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Serum samples
There were no meaningful differences between the study and the Saliva silver level (ppb) Wilcoxon test
control groups (Figure 3). On Day 7, the serum concentration of
Days n Mean Median Min. Max. SD z P
nanosilver was found significantly higher than the other days in
0 6 0.03 0.02 0.00 0.08 0.03 −0.631 0.528
1 6 0.04 0.03 0.00 0.11 0.04
0 6 0.03 0.02 0.00 0.08 0.03 −0.946 0.344
3 6 0.05 0.04 0.00 0.13 0.05
0 6 0.03 0.02 0.00 0.08 0.03 −2.201 0.028*
7 6 0.35 0.08 0.03 1.75 0.68
0 6 0.03 0.02 0.00 0.08 0.03 −0.135 0.892
14 6 0.08 0.00 0.00 0.46 0.18
0 6 0.03 0.02 0.00 0.08 0.03 −0.105 0.917
30 6 0.03 0.01 0.00 0.09 0.04
0 6 0.03 0.02 0.00 0.08 0.03 −1.782 0.750
75 6 0.07 0.07 0.02 0.12 0.04

*P < 0.05.

Figure 1. Rats teeth decay. (a) occlusal, (b) smooth surface, and (c) sulcal. Figure 3. Silver serum level in the study and the control groups.
4 European Journal of Orthodontics, 2016

Plaque samples of incisor teeth group, the number of S.mutans was found to be greater on Day 3
The nanosilver coated bracket group exhibited significantly lower than on Day 0. In the control group, the number of S.mutans was
S.mutans counts compared to the control group on Day 30 (Table 3). greater on Day 3, 7, and 30 than on Day 0 (Figure 4).
In the study group, the number of S.mutans was found to be smaller
on Days 3, 7, 14, 30, and 45 than on Day 0 (Table 4). There were no Caries assessment
significant differences in the control group between the numbers of The rates of teeth with caries in the mandibular and maxillary
S.mutans counted on different days. molars are shown in Table 5. Significant differences were observed
between groups only on the smooth surface.
Vestibular smear
The number of S.mutans in the vestibular smear did not differ sig-
nificantly between the study and the control groups. In the study Discussion
Table 2. Statistical comparison of serum silver level in the study The sample size in animal studies of nanosilver cytotoxicity per
group. group varies considerably in the literature (17–21). In the present
study, the sample size (n = 6) per group was determined depending
Serum silver level (ppb) Wilcoxon test
on the decision of the ethics committee, and the sample size was
Days n Mean Median Min. Max. SD z P calculated by considering the mean differences of the serum concen-
tration of nanosilver between the study group and the control group.
0 6 0.04 0.04 0.03 0.05 0.01 −0.314 0.753 Metallic silver is a rare noble metal in the environment. In general
1 6 0.04 0.03 0.01 0.07 0.02 medicine, silver is used as medication in the form of silver sulfadia-
0 6 0.04 0.04 0.03 0.05 0.01 −1.753 0.080 zine in treatments due to its antibacterial activity, especially for burn
3 6 0.13 0.08 0.03 0.29 0.11 treatment. In addition, silver comes first among the topics of inter-
0 6 0.04 0.04 0.03 0.05 0.01 −2.201 0.028*
est in the field of orthopaedics (silver-coated megaendoprostheses),
7 6 0.16 0.07 0.05 0.39 0.15
for the prevention of catheter-related infection (silver-impregnated
0 6 0.04 0.04 0.03 0.05 0.01 −1.572 0.116
14 6 0.20 0.14 0.02 0.49 0.19 external ventricular drainage catheter, silver-impregnated haemodi-
0 6 0.04 0.04 0.03 0.05 0.01 0.314 0.753 alysis catheter), and for the inhibition of prosthetic valve endocardi-
30 6 0.05 0.03 0.02 0.14 0.05 tis (silver-coated prosthetic valve), and also metallic silver is used in

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0 6 0.04 0.04 0.03 0.05 0.01 −1.367 0.172 dental fillings materials since 1970s. Metallic silver is inert under in
75 6 0.02 0.01 0.00 0.08 0.03 vivo conditions, and when it contacts with moisture, like body fluids
or secretions, it turns to an active form which is the silver ion (Ag+).
*P < 0.05. Metallic silver shows very few antibacterial effects in its inert state

Table 3. Statistical comparison of groups for Streptococcus mutans counts on the plaque sample of the incisor.

S. mutans counts on the plaque sample of the incisor (CFU/ml) Mann–Whitney U-test

Days Groups n Mean Median Min. Max. SD Mean rank U P

0 Study 6 226.67 160.00 0.00 640.00 238.22 6.92 15.5 0.699

Control 6 246.67 80.00 0.00 880.00 351.83 6.08
Total 12 236.67 140.00 0.00 880.00 286.65
1 Study 6 53.33 0.00 0.00 320.00 130.64 7.00 15.0 0.699
Control 6 0.00 0.00 0.00 0.00 0.00 6.00
Total 12 26.67 0.00 0.00 320.00 92.38
3 Study 6 0.00 0.00 0.00 0.00 0.00 5.50 12.0 0.394
Control 6 86.67 0.00 0.00 480.00 193.36 7.50
Total 12 43.33 0.00 0.00 480.00 137.99
7 Study 6 0.00 0.00 0.00 0.00 0.00 6.50 18.0 1.000
Control 6 0.00 0.00 0.00 0.00 0.00 6.50
Total 12 0.00 0.00 0.00 0.00 0.00
14 Study 6 0.00 0.00 0.00 0.00 0.00 5.50 12.0 0.394
Control 6 30666.67 0.00 0.00 96000.00 47575.90 7.50
Total 12 15333.33 0.00 0.00 96000.00 35851.55
30 Study 6 0.00 0.00 0.00 0.00 0.00 4.00 3.00 0.015*
Control 6 553.33 580.00 0.00 920.00 381.51 9.00
Total 12 276.67 0.00 0.00 920.00 386.86
45 Study 6 0.00 0.00 0.00 0.00 0.00 4.50 6.00 0.065
Control 6 586.67 160.00 0.00 2600.00 1010.87 8.50
Total 12 293.33 0.00 0.00 2600.00 747.23
75 Study 6 1586.67 960.00 0.00 5360.00 2113.22 6.42 17.5 0.937
Control 6 3440.00 700.00 0.00 14240.00 5634.47 6.58
Total 12 2513.33 700.00 0.00 14240.00 4170.99

*P < 0.05.
G. Metin-Gürsoy et al. 5

Table 4. Statistical comparison of Streptococcus mutans counts in the study group.

S. mutans counts on the plaque sample of the incisor (CFU/ml) Wilcoxon test

Days n Mean Median Min. Max. SD z P

0 6 226.67 160.00 0.00 640.00 238.22 −1.367 0.172

1 6 53.33 0.00 0.00 320.00 130.64
0 6 226.67 160.00 0.00 640.00 238.22 −2.032 0.042*
3 6 0.00 0.00 0.00 0.00 0.00
0 6 226.67 160.00 0.00 640.00 238.22 −2.032 0.042*
7 6 0.00 0.00 0.00 0.00 0.00
0 6 226.67 160.00 0.00 640.00 238.22 −2.032 0.042*
14 6 0.00 0.00 0.00 0.00 0.00
0 6 226.67 160.00 0.00 640.00 238.22 −2.032 0.042*
30 6 0.00 0.00 0.00 0.00 0.00
0 6 226.67 160.00 0.00 640.00 238.22 −2.032 0.042*
45 6 0.00 0.00 0.00 0.00 0.00
0 6 226.67 160.00 0.00 640.00 238.22 −0.946 0.344
75 6 1586.67 960.00 0.00 5360.00 2113.22

*P < 0.05.

In the present in vivo study, the inhibited growth of S.mutans for

the prevention of WSL via infection of nanosilver coated bracket was
shown. Our results showed that the number of S.mutans around the
nanosilver coated brackets was significantly less, especially on Day
30, compared to the control group. This is a very important finding
as WSL can become noticeable around the brackets within 1 month

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after bonding (2). In our study, one bracket was used on each rat due
to the narrow width of the mandibular teeth. Nanosilver concentra-
tion of the saliva was found significantly higher on Day 7 than the
other days in the study group, however, there were no meaningful
differences in comparison to the control group.
Previous studies suggested that the antibacterial effectiveness of
nanosilver is dependent on the silver ion release and the direct con-
tact to the bacteria (30, 31, 35). Studies showed that materials with
higher nanosilver loading release more nanosilver ions and have a
higher inhibitory effect (29, 36).
In our study, an analysis of the caries showed that maxillary
Figure 4. Counts of vestibular Streptococcus mutans in the study and the smooth caries in the study group was significantly less compared to
control groups. the control group. However, occlusal and sulcal caries scores in the
study and the control group did not differ. This suggests that nanosil-
while silver ions have stronger features against a wide spectrum of ver ions may be more effective for superficial bacteria compared to
bacteria (22–27). the bacteria located in occlusal fissures and deep cavities. This is
Nanosilver has already been in use for the treatment of burn beneficial with regard to the occurrence of WSL on the vestibular
wounds in clinical practice (15, 28). Currently, nanosilver is a lead- smooth surfaces of teeth in contact with orthodontic attachments.
ing subject at the field of dentistry and orthodontics (14, 29–31). Antibacterial effectiveness of nanosilver coated materials has
The silver nanoparticles show efficient antimicrobial properties been shown with (36) and without (37) the release of nanosilver ions
compared to other salts due to their extremely large surface area, in in vitro studies. In addition, long-term inhibitory effects against
which provides better contact with microorganisms. When nanosil- S.mutans (38) have been found while no nanosilver ions were
ver is evaluated for its antimicrobial activity, it has been observed released. Li et al. (31) showed that bonding agents with nanosil-
that the nanosilver particles get attached to the cell membrane and ver were effective against bacterial growth inhibition, not only for
can penetrate inside the bacteria. Cell death occurs because it dis- S.mutans present on surface, but also for the S.mutans away from
turbs the respiratory chain and leaks through the holes in the cell the surface in the culture medium.
wall (12, 32). We found that S.mutans counts were significantly less on Day
Therefore, we decided to apply nanosilver for the prevention 30 in the study group than the control group. However, in the
of WSL, which is a side-effect of the fixed appliance therapy and a intra-group comparisons, the concentration of nanosilver in the
major issue that is needed to be solved in orthodontics. Orthodontic saliva was found significantly higher only on Day 7, whereas
brackets coated with nanosilver may lead to a new approach and be S.mutans counts were significantly less on Day 3, 7, 14, 30, and
a novel solution. Most of the in vitro studies showed that nanosilver 45 in the study group. This indicates that the antibacterial activity
is in interaction with the inhibition of S.mutans when supplemented of nanosilver has contact-inhibition characteristics more than the
with composite or coated wire, but there is no in vivo study on its release of nanosilver ions, which leads us to prefer this recently
effectiveness on S.mutans (14, 29, 33, 34). invented bracket.
6 European Journal of Orthodontics, 2016

Table 5. Statistical comparison of groups for caries ratio.

Caries ratio Mann–Whitney U-test

Caries
surface Groups n Mean Median Min. Max. SD Mean rank U P

Occlusal Study 6 4.31 4.39 3.41 5.12 0.60 5.17 10.00 0.240
Control 6 6.05 7.06 1.68 8.83 2.66 7.83
Total 12 5.18 4.60 1.68 8.83 2.05
Smooth Study 6 1.46 1.40 0.29 3.37 1.11 4.42 5.50 0.041*
Control 6 3.83 3.25 1.23 9.12 2.82 8.58
Total 12 2.65 1.80 0.29 9.12 2.39
Sulcal Study 6 1.82 1.47 0.05 5.34 1.92 6.83 16.00 0.818
Control 6 1.28 1.42 0.79 1.59 0.35 6.17
Total 12 1.55 1.42 0.05 5.34 1.35

*P < 0.05.

Even if nanosilver ions released from the nanosilver coated Animal studies for nanosilver cytotoxicity were conducted
orthodontic brackets decrease during the prolonged orthodontic via respiratory and gastrointestinal tracts because these are the
treatment, the antibacterial activity will continue via contact-inhibi- main entry portals of nanosilver into the human body. Nanosilver
tion as shown above. The smooth surface caries was reduced at the (5000 mg/kg) was given at once via gavage to mice (46) and, for rats
end of the experiment, in spite of the concentration of nanosilver in (17), daily intake oral doses of up to 9 mg/kg were found to be safe
the saliva, which did not show significant differences between the according to chemical parameters and histopathological evaluations
study and the control group. One of the most important reasons of at the end of 14 days. The lowest adverse effects were observed in
this condition may be S.mutans counts were insignificantly smaller the rats (47) given high doses of nanosilver particles in the long-term
in the study group than the control group from Day 3 to Day 75 (90 days).
and significantly smaller on Day 30. Also we suppose that contact- We assume that even if the patient swallows the bracket during

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inhibition feature of nanosilver is efficient especially for the preven- orthodontic treatment, it is not possible to reach the above-men-
tion of WSL around the orthodontic bracket. tioned daily dose.
Bacteria have a highly developed recognition system capa- Argyria, a permanent blue grey discoloration, is the most com-
ble of recognizing and interacting with specific macromolecules monly side-effect of silver, which is stored in skin, nails, and mucous
on tissue surfaces. Adhesions of the oral pathogens to the oral membranes (48, 49). Especially, burn patients are treated by nanosil-
mucosa take place via the glycoproteins and glycolipids, which ver coated wound dressings (15, 28, 50). Vlachou (15) and Moiemen
are receptors on the surface of the epithelial cells. There are com- (50) noted that there were no signs of argyria while nanosilver
plementary structures having molecular architecture on the sur- serum concentration was 226 and 436 µg/L, respectively. However,
face of the bacterial cell for adhesion. Bacteria can hold easily, Trop (48) showed that the blood nanosilver level was 107 µg/L in
strongly, fast, and persistently to epithelium using these surface a patient who suffered from acute silver toxicity. So, the lethal dose
molecules (39). or the blood nanosilver level causing the argyria symptom is not
In the present study, counts of S.mutans in the vestibular smear yet known.
were proliferated quickly after inoculation, and vice versa, in the In the present study, the serum concentration of nanosilver was
plaque sample on the teeth. It was found significantly higher on Day found as 0.00175 µg/L in rats. This value could increase during
3 in the study group and on Day 3, 7, 30 in the control group. These orthodontic treatment due to the use of 20 nano-coated brackets.
findings may be an indication of S.mutans being accumulated easily However, it is clear that the serum nanosilver count still remains far
on the oral epithelium. However, the number of S.mutans on the oral below the levels, as shown in other clinical trials (15, 50).
epithelium was found to be normal after Day 3 in the study group, The antibacterial effects in inhibition of S.mutans, low levels of
whereas it was normalized after Day 30 in the control group. We can nanosilver ion release, and reduction of smooth surface caries have
predict that the growth of S.mutans was inhibited by the nanosilver been shown by the nano-coated orthodontic bracket as applied to
coated bracket. the mandibular incisors of rats in this study. This initial study will be
Nanosilver particles have been reported to be cytotoxic in in a guide for further advanced work.
vitro studies for several types of cells, however, the toxicity mecha-
nism of nanosilver is not clear yet. The common view is that it is
similar to bacterial death, which changes the permeability of the cell Conclusion
membrane by blocking the ion channels, causing a mitochondrial Our results show that nano-coated orthodontic bracket is effective
dysfunction, and producing oxidative stress (40–43). On the con- in the inhibition of S.mutans and reduction in smooth surface caries.
trary, it has been reported that nanosilver does not lead to signifi- This newly introduced nano-coated orthodontic bracket may be an
cant cytotoxicity in mouse fibroblasts, human osteoblasts (44), and essential non-compliant appliance in avoidance of WSLs for patients
human gingival fibroblasts cell lines (14). The most important factor especially with poor oral hygiene during fixed orthodontic treat-
in the formation of different test results is size and concentration of ment and may provide solution to patients with immune deficiency,
nanosilver particles and also testing with nanosilver particles alone diabetics or in the need of protection from infections such as with
or with a medical device. It should be noted that in vitro tests cannot subacute bacterial endocarditis. We need more detailed research to
entirely predict the overall biocompatibility of a material and in vivo predict the blood silver level during orthodontic treatment to esti-
use of the material must be questioned (45). mate contingent side effects of nanosilver.
G. Metin-Gürsoy et al. 7

Funding inhaled ultrafine silver particles in rats. Environmental Health Perspective,

109, 547–551.
Gazi University Scientific Research Committee (03/2013-01). 19. Tian, J., Wong, K.K., Ho, C.M., Lok, C.N., Yu, W.Y., Che, C.M., Chiu,
J.F. and Tam, P.K. (2007) Topical delivery of silver nanoparticles promotes
wound healing. ChemMedChem, 2, 129–136.
Acknowledgements 20. Ji, J.H., et al.et al. (2007) Twenty-eight-day inhalation toxicity study of
We thank National Nanotechnology Research Center (UNAM) for kind silver nanoparticles in Sprague-Dawley rats. Inhalation Toxicology, 19,
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