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Evaluation of Clinical and Antimicrobial Efficacy of Silver Nanoparticles and Tetracycline Films in the Treatment of Periodontal Pockets

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Periodontitis is a multifactorial infection associated with a variable bacterial pattern. The treatment focuses mainly on the reduction of the total bacterial count. Local delivery of antimicrobials has been investigated as an adjunct to conventional therapy. Tetracycline was proved to inhibit collagenases and was thus proposed to be useful in treating diseases. In recent years, silver nanoparticles have attracted considerable attention for medical applications due to their antibacterial activity. This study aims to evaluate the clinical and the microbiological findings following intrasulcular applications of tetracycline films and silver nanoparticles in periodontal pockets. A total of 48 periodontal pockets were studied. Group (A) received scaling and root planing with tetracycline film application, Group (B): scaling and root planing with silver nanoparticles application and Group (C): scaling and root planing only. The drugs were applied once weekly for three weeks. Clinical parameters were taken at baseline, after one and three months. Samples of gingival crevicular fluid were obtained at baseline and after one month for microbiological analysis. Groups A and B showed a significant decrease in probing depth and clinical attachment level as well as the reduction in the bacterial count compared to Group C. Thus, local application of tetracycline films and silver nanoparticles were effective in improving the clinical outcome and elimination of bacterial infection in periodontal pockets.
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IOSR Journal of Dental and Medical Sciences (IOSR-JDMS)
e-ISSN: 2279-0853, p-ISSN: 2279-0861.Volume 14, Issue 7 Ver. I (July. 2015), PP 113-123
www.iosrjournals.org
DOI: 10.9790/0853-1471113123 www.iosrjournals.org 113 | Page
Evaluation of Clinical and Antimicrobial Efficacy of Silver
Nanoparticles and Tetracycline Films in the Treatment of
Periodontal Pockets
Heba A. Shawky1, Soha M. Basha2, Gihan A. EL Batouti3, Abeer A. Kassem4
1 Department of Periodontology and Oral Medicine, Faculty of Dentistry, Pharos University in
Alexandria, Egypt.
2Department of Basic Dental Science, Faculty of Dentistry, Princess Nora BintAbdulrahman University,
Saudia Arabia.
3Department of Microbiology and Immunology, Faculty of Pharmacy, Pharos University in
Alexandria, Egypt.
4Department of Pharmaceutics, Faculty of Pharmacy, Princess Nora BintAbdulrahman University,
Saudia Arabia.
Abstract: Periodontitis is a multifactorial infection associated with a variable bacterial pattern. The treatment
focuses mainly on the reduction of the total bacterial count. Local delivery of antimicrobials has been
investigated as an adjunct to conventional therapy. Tetracycline was proved to inhibit collagenases and was
thus proposed to be useful in treating diseases. In recent years, silver nanoparticles have attracted considerable
attention for medical applications due to their antibacterial activity. This study aims to evaluate the clinical and
the microbiological findings following intrasulcular applications of tetracycline films and silver nanoparticles
in periodontal pockets. A total of 48 periodontal pockets were studied. Group (A) received scaling and root
planing with tetracycline film application, Group (B): scaling and root planing with silver nanoparticles
application and Group (C): scaling and root planing only. The drugs were applied once weekly for three weeks.
Clinical parameters were taken at baseline, after one and three months. Samples of gingival crevicular fluid
were obtained at baseline and after one month for microbiological analysis. Groups A and B showed a
significant decrease in probing depth and clinical attachment level as well as the reduction in the bacterial
count compared to Group C. Thus, local application of tetracycline films and silver nanoparticles were
effective in improving the clinical outcome and elimination of bacterial infection in periodontal pockets.
Key words: chronic periodontitis, silver nanoparticles, tetracycline, antibacterial activity.
I. Introduction
Periodontitis is an inflammatory disease of the gingiva and the adjacent attachment apparatus and is
characterized by the loss of both, connective tissue attachment and alveolar bone. Chronic periodontitis, the
most common form, is characterized by pocket formation and gingival recession.1 The disease is initiated by
polymicrobial infection which is composed of more than 300 different species of bacteria. The bacteria of
supragingival plaque are predominantly Gram-positive cocci, whereas periodontal pathogens in subgingival
plaque are dominated by Gram-negative anaerobic organisms.2,3
Predisposing factors include heredity, systemic disorders as well as environmental factors as smoking.
These risk factors play a crucial role in determining the progression and severity of the disease.4 Although strict
anaerobic periodontal pathogenic microorganisms are directly involved in the onset and progression of chronic
periodontitis, it is also important to mention that for several years, facultative anaerobe Gram-negative
Enterobacteriaceae have also been found in the gingival sulcus of patients with chronic periodontitis.58
The normal immune response to bacterial invasion is the activation of inflammatory cells. The host
inflammatory response is responsible for the majority of the hard- and soft-tissue breakdown that takes place in
periodontitis.9, 10 Thus periodontal destruction is caused either directly by the action of bacteria on the invaded
tissues, or indirectly through the release of biologic mediators in the form of enzymes and chemical mediators
from the host tissue cells that lead to host tissue destruction.11 Interleukin I and tumor necrosis factor-alpha
induce the release of other mediators that aggravate the inflammatory response, such as prostaglandins.12-14
These host-derived mediators have the potential to stimulate bone resorption and activate or inhibit other host
immune cells.15
The great challenge for successful periodontal therapy is to eliminate pathogenic organisms present in
the dental plaque. Different treatment modalities were attempted including surgical intervention, non-surgical
procedures, mechanical therapy and the use of pharmacological agents.1,16 Mechanical therapy that comprises
scaling and root planing (SRP) has become the “gold standard” nonsurgical treatment for periodontitis.17 Several
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studies have indicated that the reduction of the microbial level can effectively improve clinical parameters like
Papillary Bleeding Index (PBI), Gingival Index (GI), Probing Depth (PD) and Clinical Attachment Loss
(CAL).18,19 However, in case of deeper pocket sites, manual SRP alone, is not sufficient to completely eradicate
pathogenic bacteria. Subsequently, power-driven ultrasonic mechanical instruments were developed to enhance
the capability of the operator enabling it to reach through the depth of the pocket more conveniently.20,21 On the
other hand, various studies have reported that the microbiological and clinical effects attained by ultrasonic
debridement are similar to those of manual SRP.22,23
Therefore, the adjunct use of antimicrobial agents with mechanical debridement could be more
effective.24 Nevertheless, the concentration of systemic antibiotics is usually less in the gingival crevicular fluid
(GCF) compared to its concentration in the blood stream. The systemic use of antibiotics causes different side
effects as hypersensitivity and gastrointestinal intolerance, in addition to the bacterial resistance that may
arise.25,26 Thus, currently systemic antibiotics are prescribed only for the treatment of aggressive or refractory
periodontitis.27 The shortcomings of systemic antimicrobial treatment led to the development of local drug
delivery systems.28
The intra-pocket application of antimicrobial therapy has evoked great interest as it overcomes the
limitations of systemic antimicrobial therapy and is also considered as a local site-specific modality. The
periodontal pocket acts as a natural reservoir for the application of a local delivery device. It is bathed by
gingival crevicular fluid which provides a leaching medium for the release of a local delivery drug and
facilitates its distribution throughout the pocket.29,30 Different local drug delivery devices were approved for the
treatment of periodontal pockets; such as those constituted from chlorhexidine gluconate,31 doxycycline
hyclate,32 minocycline hydrochloride33 and tetracycline fibers.34
Tetracyclines have been widely used in the field of periodontal therapy since the 1940s. They are semi-
synthetic chemotherapeutic agents with a bacteriostatic action and hence are effective against rapidly
multiplying bacteria.35 Tetracycline and its derivatives have been used as local delivery drugs in the treatment of
periodontal pockets in order to limit the drug to its target site with little or no systemic uptake, which in turn will
avoid most of the side effects associated with systemic therapy.34 They have been incorporated into various non-
resorbable or bio-resorbable delivery systems for their insertion into periodontal pockets.35,36 These systems
include hollow fibers,37ethyl cellulose fibers,38 acrylic strips,39ethylene vinyl acetate copolymer fibers,40 collagen
preparations,41,42 and poly (D, L-lactic-co-glycolic acid) microspheres.43 Freisen et al. (2002) demonstrated that
local delivery of tetracycline with multiple polymer strips was effective in reducing papillary bleeding index,
and that it was superior to root planing alone in reducing probing depth.44
As a result of development of microbial resistance to multiple antibiotics, the antibiotic-free delivery
systems for treatment of periodontal infections have been tried. Recent advances in nanotechnology introduce
new therapeutic materials for periodontal regeneration. Nanoparticles are clusters of atoms in the size range of
1-100 nm.45 Inorganic nanoparticles and their nano-composites are good antibacterial agents. Focus has been
made upon the medical and chemical applications of silver nanoparticles (Ag-NPs) due to their unique
properties; including antibacterial activity, high resistance to oxidation, and high thermal conductivity.46
The exact mechanism by which Ag-NPs employ an antimicrobial effect is not clearly known and is a
debatable topic. There are however different theories concerning their antibacterial activity. Ag-NPs have the
ability to anchor to the bacterial cell wall and subsequently penetrate it, thereby causing structural changes in the
cell membrane affecting its permeability and hence death of the cell.47Amro et al. (2000) reported that metal
depletion may lead to formation of irregularly shaped pits in the outer membrane due to the progressive release
of lipopolysaccharide molecules and membrane proteins, which in turn, increases the membrane’s
permeability.48 Hence, ‘pits’ are formed and nanoparticles are accumulated on the cell surface.49 The formation
of free radicals by the Ag-NPs may be considered as another mechanism by which the cells die. Some studies
suggested that free radicals are formed when the Ag-NPs come in contact with bacteria, and these free radicals
have the ability to damage the cell membrane and rendering it porous, thus can ultimately lead to cell death .50,51
It has also been proposed that there can be release of silver ions by the nanoparticles,52 and these ions can
interact with the thiol groups of many vital enzymes and inactivate them.53
Nowadays, Ag-NPs display different applications in the field of dentistry owing to their antimicrobial
effect. They are used in restorative dentistry, where Ag-NPs were incorporated into nano-composites of
quaternary ammonium dimethacrylate and calcium phosphate.54,55 Results showed that these nano-composites
are promising as they possess the double benefits of remineralization and antibacterial capabilities to inhibit
dental caries. Ag-NPs were also incorporated into tissue conditioners for patients using dental prosthesis. It was
demonstrated that a dose of 0.1% Ag-NPs combined to tissue conditioners displayed a minimal bactericidal
effect against Staphylococcus aureus and Streptococcus mutans strains, whereas 0.5% was lethal for fungal
strains.56 In addition, Ag-NPS have been used with endodontic retrofill cements; where the combination of
Ag-NPs to Angelus white mineral trioxide aggregate enhanced its antimicrobial activity against Enterococcus
faecalis, Candida albicans and Pseudomonas aeruginosa.57 In the field of implantology, Ag-NPs were
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incorporated in the coating of titanium implants. Results revealed that titania nanotubes incorporated with
Ag-NPs possess acceptable osteoconductivity, a relatively long-term antibacterial effect and good tissue
integration.58,59
The present study was carried out to evaluate and compare the clinical improvement and antimicrobial
efficacy of tetracycline films and Ag-NPs as an adjunct to SRP in persistent periodontal pockets.
II. Materials And Methods
A randomized, controlled, clinical study was conducted, and the inclusion criteria for patient selection
were females ranging from 30 to 40 years of age. Patients diagnosed with mild to moderate chronic periodontitis
with at least three non-adjacent periodontal pockets with 4 to 6 mm pocket depth were included in the study. All
selected females were ascertained to be in good general health with no history of systemic disorders (diabetes,
osteoporosis, cardiovascular diseases, hyperthyroidism, glucose-6-phosphate-dehydrogenase deficiency or
severe myasthenia) and with no history of antibiotic therapy, oral prophylaxis, or periodontal surgery during the
last six months.60,61 Pregnant, lactating females and smokers were excluded from the study.62
A total of 48 pockets from 16 patients were selected for the study. Three periodontal pockets in each
patient were identified for the study in which SRP were performed followed by impressions for the fabrication
of acrylic stents required for the standardized measurement of pocket depths and clinical attachment loss in the
test and control groups throughout the study period63 (Fig. 1). During the second visit baseline data were
recorded. The selected pockets were assigned randomly to three groups. The three treatment groups were Test
Group (A): SRP with tetracycline film application, Test Group (B): SRP with Ag-NPs application and Control
Group (C): SRP alone.
2.1 Preparation and application of Tetracycline Film
A concentration of 1:10 tetracycline hydrochloride (TcHCl) drug to Carboxymethyl cellulose sodium
(Na CMC)
was prepared. Freshly prepared drug solution (100 mg TcHCl dissolved in 5 mL distilled water) was
added to 15 ml of polymer solution, dropwise with continuous stirring using a glass rod. The mixture was left to
stand until all air bubbles disappeared and was then casted in a horizontally leveled Teflon plate of 5 cm internal
diameter. The polymeric drug solution was dried under ambient conditions with the aid of a fan in a dark place
to avoid photo-decomposition of the drug. The dried films were carefully removed from the Teflon cups,
checked for any imperfections or air bubbles64 (Fig. 2). Tetracycline films were aseptically cut into portions that
were smaller than the pocket dimensions. These portions were inserted into the pocket once weekly for three
successive weeks (Fig.3).
2.2 Application of Silver Nanoparticles
100 nm Cytodiagnostic spherical Ag-NPs
(20ml) of concentration 0.02 mg/ml were used in this study
(Fig.4). 2 ml of the Ag-NPs solution were injected directly inside the periodontal pocket using an insulin syringe
and were allowed to fill the pocket (Fig. 5). The intrasulcular application of Ag-NPs was repeated once weekly
for three successive weeks. Periodontal dressing
§
was applied to achieve retention of the drug to the pocket for
the required period (Fig. 6).
2.3 Post-procedure Instruction and Clinical Parameters
Patients were instructed to carry out normal oral hygiene procedures, without using dental floss, any
mouth washes or oral irrigation devices. They were asked to report immediately if pain, swelling or any other
problem occurred. Clinical measurements included, PBI,65 GI,66 PD and CAL.67 All the clinical parameters were
recorded at baseline, after one and three months.
2.4 Microbiological Analysis
Samples of GCF were obtained using sterile paper points, inserted into the pocket for 30 seconds until
resistance was felt or the paper points had bent (Fig. 7).68 They were immediately transferred to 5 ml screw
capped test tubes containing brain heart infusion broth
**
that served as a transport and enrichment medium (Fig.
8). The GCF samples were taken at baseline and after one month. The test tubes were incubated under anaerobic
conditions for 4 hours at 370C; after which they were shaken by vortexing to ensure homogeneous mixture of
the broth. Immediately 50μl were aseptically inoculated onto blood and MacConkey agar
††
plates and incubated
Alexandria Pharmaceuticals Co., Alexandria, Egypt.
Prolaboproducts Pour Laboratoires, Rhône-Poulenc, France.
Bio-Synthesis, Inc. Lewisville; Texas, Usa.
§
Coe-Paktm (Gc America Inc., Alsip, Il, Usa).
**
Bd Bbl™ , Becton, Dickinson And Company, Usa.
††
Oxoid Ltd, Waderoad,Basingstoke, Hampshire, Uk.
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aerobically at 370C for 48 hours. The resulting isolated colonies were counted to determine the bacterial load
and were subjected to further identification by Gram stain and biochemical tests.69, 70
2.5 Statistical analysis of the data71
Data were fed to the computer and analyzed using IBM SPSS software package version 20.0.72
Quantitative data were described using range (minimum and maximum) mean, standard deviation and median.
The distributions of quantitative variables were tested for normality using Kolmogorov-Smirnov test, Shapiro-
Wilk test and D'Agstino test. If it reveals normal data distribution, parametric tests was applied. If the data were
abnormally distributed, non-parametric tests were used. For abnormally distributed data, comparison between
the different studied groups were done using Kruskal Wallis test and pair wise comparison was assessed using
Mann-Whitney test. To compare between the different periods Wilcoxon signed ranks test was used.
Significance of the obtained results was judged at the 5% level.
Fig. 1: PD and CAL measurements using Williams
probe
Fig. 2: Tetracycline films
Fig 3: Insertion of tetracycline film portions into the
periodontal pocket
Fig. 4: Silver nanoparticles solution
Fig. 5: Intra-pocket injection of silver nanoparticles
Fig. 6: Application of Coe-pack at the site of
injection of silver nanoparticles
Fig. 7: Insertion of paper points into the pocket for
GCF sampling
Fig. 8:Paper points in screw capped tubes
containing brain heart infusion broth
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III. Results
The clinical findings at baseline, one month and three months postoperative are displayed in Tables
(1 4) for the three treatment modalities; Group A: SRP with tetracycline film application, Group B: SRP with
Ag-NPs application and Group C: SRP only (control group). At first, there was no significant difference
between all treatment groups regarding the PBI, GI, PD, and CAL.
Table (1) shows the gradual reduction of PBI that occurred in all studied groups. This decrease was
greater in Group A than in Group B. Group C showed the least decrease. The difference in PBI of all groups was
insignificant. Similarly, a significant decrease in GI was observed in Group A after three months (Zp = 0.046)
followed by Group B, while the least amount of decrease was denoted in Group C; Table (2).
The PD of all groups showed a significant decrease after one and three months (Zp < 0.001); Table (3). This
decrease was most prevalent in Group B followed by Group A. The least amount of reduction was reported in
Group C. Accordingly; Table (4) revealed more gain of attachment in Group B compared to Groups A and C.
Table (1): Comparison between the Papillary Bleeding Index in the 48 periodontal pockets
before and after the application of three treatment modalities
Papillary bleeding index
Group A
(n = 16)
Group B
(n = 16)
Group C
(n = 16)
p
Baseline
Min. Max.
0.0 2.0
0.0 2.0
0.0 2.0
0.950
Mean ± SD
0.94 ± 0.68
0.94 ± 0.68
1.0 ± 0.63
Median
1.0
1.0
1.0
Sig. bet. groups
p1 = 1.000 , p2 = 0.781 , p3 = 0.781
After 1 month
Min. Max.
0.0 2.0
0.0 2.0
0.0 2.0
0.715
Mean ± SD
0.75 ± 0.68
0.81 ± 0.66
0.94 ± 0.68
Median
1.0
1.0
1.0
Sig. bet. groups
p1 = 0.769 , p2 = 0.428, p3 = 0.598
Zp
0.405
0.564
0.317
After 3 months
Min. Max.
0.0 1.0
0.0 1.0
0.0 2.0
0.407
Mean ± SD
0.63 ± 0.50
0.63 ± 0.50
0.88 ± 0.62
Median
1.0
1.0
1.0
Sig. bet. groups
p1 = 1.000, p2 = 0.250, p3 = 0.250
Zp
0.132
0.166
0.157
KW2: Chi square for Kruskal Wallis test
p1: p value for Mann Whitney test for comparing between Group A and B
p2: p value for Mann Whitney test for comparing between Group A and C
p3: p value for Mann Whitney test for comparing between Group B and C
Zp: p value for Wilcoxon signed ranks test for comparing between baseline with after 1 month and 3 months
Table (2): Comparison between the Gingival Index in the 48 periodontal pockets before and after the
application of three treatment modalities
Gingival index
Group A
(n = 16)
Group B
(n = 16)
Group C
(n = 16)
KW
p
Baseline
Min. Max.
0.0 1.0
0.0 1.0
0.0 1.0
0.164
0.921
Mean ± SD
0.44 ± 0.51
0.44 ± 0.51
0.50 ± 0.52
Median
0.0
0.0
0.50
Sig. bet. groups
p1 = 1.000, p2 = 0.727 , p3 = 0.727
After 1 month
Min. Max.
0.0 1.0
0.0 1.0
0.0 1.0
1.285
0.526
Mean ± SD
0.25 ± 0.45
0.31 ± 0.48
0.44 ± 0.51
Median
0.0
0.0
0.0
Sig. bet. groups
p1 = 0.699 , p2 = 0.272 , p3 = 0.472
Zp
0.083
0.157
0.317
After 3 months
Min. Max.
0.0 1.0
0.0 1.0
0.0 1.0
1.446
0.485
Mean ± SD
0.19 ± 0.40
0.25 ± 0.45
0.38 ± 0.50
Median
0.0
0.0
0.0
Sig. bet. groups
p1 = 0.674, p2 = 0.246, p3 = 0.453
Zp
0.046*
0.083
0.157
KW2: Chi square for Kruskal Wallis test
p1: p value for Mann Whitney test for comparing between Group A and B
p2: p value for Mann Whitney test for comparing between Group A and C
p3: p value for Mann Whitney test for comparing between Group B and C
Zp: p value for Wilcoxon signed ranks test for comparing between baseline with after 1 month and 3 months
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Table (3): Comparison between the Probing Depth in the 48 periodontal pockets before and after the
application of three treatment modalities
Probing depth
Group A
(n = 16)
Group B
(n = 16)
Group C
(n = 16)
KW
p
Baseline
Min. Max.
4.0 6.0
4.0 5.0
4.0 5.0
1.390
0.499
Mean ± SD
5.0 ± 0.73
4.81 ± 0.40
4.75 ± 0.45
Median
5.0
5.0
5.0
Sig. bet. groups
p1 = 0.419 , p2 = 0.293, p3 = 0.674
After 1 month
Min. Max.
3.0 5.0
3.0 4.0
3.0 4.0
0.553
0.759
Mean ± SD
3.56 ± 0.73
3.50 ± 0.52
3.63 ± 0.50
Median
3.0
3.50
4.0
Sig. bet. groups
p1 = 1.000, p2 = 0.554, p3 = 0.483
Zp
<0.001*
<0.001*
<0.001*
After 3 months
Min. Max.
2.0 3.0
1.0 3.0
2.0 4.0
18.573*
<0.001
*
Mean ± SD
2.25 ± 0.45
2.19 ± 0.66
3.25 ± 0.68
Median
2.0
2.0
3.0
Sig. bet. groups
p1 = 0.857, p2<0.001*, p3<0.001*
Zp
<0.001*
<0.001*
<0.001*
KW2: Chi square for Kruskal Wallis test
p1: p value for Mann Whitney test for comparing between Group A and B
p2: p value for Mann Whitney test for comparing between Group A and C
p3: p value for Mann Whitney test for comparing between Group B and C
Zp: p value for Wilcoxon signed ranks test for comparing between baseline with after 1 month and 3 months
*: Statistically significant at p ≤ 0.05
Table (4): Comparison between the Clinical Attachment Loss in the 48 periodontal pockets before and
after the application of three treatment modalities
Clinical attachment loss
Group A
(n = 16)
Group B
(n = 16)
Group C
(n = 16)
KW
p
Baseline
Min. Max.
2.0 4.0
2.0 3.0
2.0 4.0
0.742
0.690
Mean ± SD
2.88 ± 0.72
2.75 ± 0.45
2.94 ± 0.57
Median
3.0
3.0
3.0
Sig. bet. groups
p1 = 0.660 , p2 = 0.748, p3 = 0.338
After 1 month
Min. Max.
2.0 - 3.0
2.0 3.0
2.0 4.0
3.776
0.151
Mean ± SD
2.56 ± 0.51
2.44 ± 0.51
2.81 ± 0.54
Median
3.0
2.0
3.0
Sig. bet. groups
p1 = 0.486, p2 = 0.204, p3 = 0.059
Zp
0.025*
0.025*
0.157
After 3 months
Min. Max.
1.0 3.0
1.0 3.0
2.0 3.0
14.021*
0.001*
Mean ± SD
2.0 ± 0.63
1.88 ± 0.62
2.69 ± 0.48
Median
2.0
2.0
3.0
Sig. bet. groups
p1 = 0.570, p2 = 0.003*, p3 = 0.001*
Zp
0.001*
0.001*
0.046*
KW2: Chi square for Kruskal Wallis test
p1: p value for Mann Whitney test for comparing between Group A and B
p2: p value for Mann Whitney test for comparing between Group A and C
p3: p value for Mann Whitney test for comparing between Group B and C
Zp: p value for Wilcoxon signed ranks test for comparing between baseline with after 1 month and 3 months
*: Statistically significant at p ≤ 0.05
Table (5) displays the type of bacteria in the total 48 periodontal pockets before and after the
application of tetracycline films in Group A, Ag-NPs in Group B and in control Group C. It is clear from this
table that the application of tetracycline films reduced the percentage of the total bacterial count in periodontal
pockets revealing Gram positive bacteria from 43.8% to 6.3%, and pockets revealing Gram negative bacteria
from 56.3% to 31.3%; while the pockets that revealed complete elimination of bacterial infection increased from
0.0% to 62.5% (highly significant, p = 0.006). Nearly similar results were recorded in the pockets that were
treated by Ag-NPs, where the percentage of Gram positive pockets decreased from 43.8% to 12.5%, and Gram
negative pockets from 50.0% to 31.3%; while the completely free pockets increased from 6.3% to 56.3%
(significant, p = 0.041). On the other hand, the least percentage of reduction in periodontal pockets was
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observed in Group C; where the percentage of Gram positive bacteria decreased only from 18.8% to 12.5% and
Gram negative bacteria from 37.5% to 31.3% (insignificant, p = 0.631). Thus, it is evident that elimination of
bacterial infection in the control pockets increased only from 43.8% to 56.3%.
Table 5: Comparison between the type of bacteria in the 48 periodontal pockets before and after the
application of three treatment modalities
Type of bacteria
Group A
(n = 16)
Group B
(n = 16)
Group C
(n = 16)
2
MCp
No.
%
No.
%
No.
%
Before
Gram positive
7
43.8
7
43.8
3
18.8
11.476*
0.017*
Gram negative
9
56.3
8
50.0
6
37.5
Free
0
0.0
1
6.3
7
43.8
After
Gram positive
1
6.3
2
12.5
2
12.5
0.761
1.000
Gram negative
5
31.3
5
31.3
5
31.3
Free
10
62.5
9
56.3
9
56.3
p1
0.006*
0.041*
0.631
2: Value for Chi square
MC: Monte Carlo test
p1: p value for Marginal Homogeneity test for comparing between before and after in each group
*: Statistically significant at p ≤ 0.05
IV. Discussion
Periodontitis is a chronic inflammation of the periodontium that results in periodontal tissue destruction
and alveolar bone loss. Tissue destruction occurs as a consequence of the host’s attempt to eliminate bacteria
from the gingival sulcus by evoking an immuno-inflammatory response.3,9 The main objective of periodontal
therapy is to reduce the pathogenic bacterial count to the level at which the periodontal destruction is arrested.19
The nonsurgical periodontal treatment remains the gold standard for managing the patients with
periodontitis. Matthews (2005)73, Cobb (2008)74 and Apatzidou et al. (2010)75 postulated that the nonsurgical
treatment can result in reduction of inflammation, decrease in pocket depth and gain of attachment. However,
mechanical therapy may fail to eliminate the pathogenic bacteria because of their location within gingival
tissues or in other areas inaccessible to periodontal instrumentation.76 Although an additional clinical benefit of
adjuvant systemic antibiotics has been described, it is only recommended in cases of refractory or aggressive
periodontitis to prevent the development of antimicrobial resistance.77 Previous studies reported that local
delivery of adjunctive antimicrobial therapy is considered a safe and effective alternative to systemic
administration.78,30,28
A nonsurgical approach using the repeated intrasulcular application of tetracycline films and Ag-NPs
was used in this study. The main advantage of tetracycline films is its’ ease of insertion inside the pocket and
that the dimension of the films could be easily adjusted according to the size of periodontal pocket, causing no
or only minimal discomfort to the patient.79 However, the application of Ag-NPs may be an alternative modality
for patients hypersensitive to tetracycline.
The PBI and GI were used to monitor the changes in gingival inflammation throughout the study. The
PBI of all groups decreased throughout the study period. This decrease was greater in Group A [tetracycline
film] by one month (mean of PBI = 0.75 ± 0.68), followed by Group B [Ag-NPs] (0.81 ± 0.66). After three
months, both groups A and B showed similar results (0.63 ± 0.50) while in Group C [control] showed the least
decrease (mean = 0.88 ± 0.62). Similar results were observed concerning the GI as it showed a decrease in all
groups after one and three months. This decrease was greater in Group A by one month (mean= 0.25 ± 0.45),
followed by Group B (0.31 ± 0.48). The least amount of decrease was denoted in Group C (0.44 ± 0.51). This
decrease was insignificant in all groups. By three months, the decrease in Group A showed significant
difference compared to baseline (mean = 0.19 ± 0.40, Zp = 0.046). The decrease in Group B and C was
insignificant 0.25 ± 0.45 and 0.38 ± 0.50 respectively.
Thus, the least amount of inflammation was denoted in Group A followed by Group B. The
tetracycline films used in the present study provided a sustained release for the TcHCl which showed positive
effects on reducing the inflammation. These results are in agreement with those of Sachdeva et al. (2011) who
demonstrated a significant decrease in GI from baseline to one month (difference was 0.95 ± 0.33) and from
baseline to three month (difference was 1.79 ± 0.35) in the test group treated by tetracycline films.80 This could
be attributed to the better substantivety and good binding and/or penetration into the root surfaces offered by
tetracycline as a bacteriostatic antibiotic.81 It interferes with bacterial protein synthesis and has a broad spectrum
of activity inhibiting both Gram negative and Gram positive organisms.82
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On the other hand, the reduction of inflammation in Group B could be attributed to the antibacterial
activity of Ag-NPs which plays an important role in subsiding inflammation. This suggestion is in accordance
with that reported by Nadworny et al. (2008) when they found that Ag-NPs had direct anti-inflammatory
effects.83
Nanoparticles, owing to their small size, show high penetration capability to deep periodontal pockets
which may be inaccessible to other delivery systems.84 In addition, nanoparticles provide a uniform distribution
of the active agent over an extended period of time, and thus the frequency of administration of these systems is
reduced.79
Regarding the reduction in pocket depth and attachment gain, both PD and CAL showed significant
decrease in all groups after one month (Zp < 0.001). This could be attributed to a great extent to the resolution of
inflammation. The degree of probe penetration is greatly influenced by the inflammation of the gingival tissue.80
After three month significant difference existed between Group A and C and between Group B and C.
The high levels in attachment gain in the tetracycline group could be attributed to its bacteriostatic
effect. Tetracyclines have long been considered useful adjuncts in periodontal therapy based on their
antimicrobial efficacy against putative periodonto-pathogens.42,43 Moreover, these drugs were found to inhibit
collagenases and several other matrix metalloproteinases (MMPs) from cells such as neutrophils, macrophages,
osteoblasts and chondrocytes in gingival tissue.85 Tetracyclines inhibit MMPs directly by an interaction between
the tetracycline molecule and metal ions within the MMP and also indirectly by inhibiting the expression of
MMPs.86
Although no significant difference in PD or CAL existed between Group A and B; Group B revealed
the lowest results (mean = 2.19 ± 0.66, 1.88 ± 0.62 for PD and CAL respectively), showing better improvement
in attachment gain. The positive role of Ag-NPs in fibroblast maturation and proliferation was supported by
previous studies.87,88 Liu et al. (2010) stated that the presence of Ag-NPs may have a key role in the
enhancement of fibroblast maturation and proliferation by providing an inflammatory free environment. 89
In the present work, the bacteria revealed from the periodontal pockets were all facultative anaerobes
that included mainly Enterobacter cloacae, Escherichia coli, followed by Enterococcus faecalis, Streptococcus
mutans and Streptococcus viridans. In the human oral environment, Enterobacteriaceae have been isolated
from mucosa and teeth in addition to the gingival sulcus.90,91 Its presence in the oral cavity can be attributed to
orofecal transmission, deficient oral hygiene, or contamination from food or drink.92,93Enterobacteriaceae are
considered as key pathogens in some cases of refractory periodontitis.94,95 This is due to the inadequate use of
antibiotics leading to suppression of normal oral microbiota which may result in persistent colonization of the
oral cavity by the opportunistic microorganisms.96
It is well known that Ag-NPs possess an antibacterial activity against both Gram positive and negative
species.97 According to Okafor et al. (2013), Ag-NPs at a concentration of 2 parts/million (2 ppm) are not
cytotoxic for human healthy cells but inhibit bacterial growth.98 In the current study the utilized concentration,
0.2 ppm of Ag-NPs was found to be lethal against most of the pathogenic bacteria revealed from the periodontal
pockets. Lansdown (2002)99 and Castellano et al. (2007)100 attributed the antibacterial activity of silver ions to
its high reactivity. The bacterial cells in contact with silver take in silver ions, which thereby inhibit several
functions in the cell and lead to its damage. The antimicrobial activity of silver depends upon its ions, which
bind strongly to electron donor groups in biological molecules containing sulfur, oxygen or nitrogen.101 It has
also been postulated that the generation of reactive oxygen species, which are possibly produced through the
inhibition of a respiratory enzyme by silver ions, may attack the cell itself.102
Another mode of action for silver ions is the interaction with the bacterial DNA which will prevent the
cell reproduction.103 DNA has sulfur and phosphorus as its major components. Nanoparticles can act on these
soft bases and destroy DNA.104 The interaction of the Ag-NPs with sulfur and phosphorus of DNA, can inhibit
DNA replication of the bacteria and result in cell death. It has also been found that the nanoparticles can
modulate the signal transduction in bacteria. It is a well-established fact that phosphorylation of protein
substrates in bacteria influences bacterial signal transduction. Dephosphorylation is noted only in the tyrosine
residues of Gram-negative bacteria. The phosphotyrosine profile of bacterial peptides is altered by the
nanoparticles. It was found that the nanoparticles dephosphorylate the peptide substrates on tyrosine residues,
which leads to signal transduction inhibition and thus the inhibition of growth.105
V. Conclusion
Based on the results of the present study, it can be concluded that, intrasulcular injection of Ag-NPs
resulted in a pronounced improvement in clinical parameters and reduction of microbial infection. Ag-NPs were
as effective as local application of tetracycline films in treatment of periodontal pockets. However, more
comprehensive and long-term studies that monitor clinical and microbiological activity together are also
necessary.
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... Although an additional clinical benefit was reported when systemic antibiotics have been described, it is only preferred in cases of aggressive or refractory types of periodontitis to prevent the development of antibiotic resistant bacteria (11) . ...
... Our findings coincided with those previously reported by Heba A. Shawky et al. who found marked improvement in clinical manifestations and decrease in microbial infection after intrasulcular injection of silver nanoparticles asits effect was similar to tetracycline films application in treatment of periodontal pockets (11) . ...
... Nanoparticles can act on soft bases in DNA as sulfur and phosphorus as its main components and destroy it. This can inhibit DNA replication of the bacteria leading to cell death and can modify bacterial transduction signals (11) . ...
... Although an additional clinical benefit was reported when systemic antibiotics have been described, it is only preferred in cases of aggressive or refractory types of periodontitis to prevent the development of antibiotic resistant bacteria (11) . ...
... Our findings coincided with those previously reported by Heba A. Shawky et al. who found marked improvement in clinical manifestations and decrease in microbial infection after intrasulcular injection of silver nanoparticles asits effect was similar to tetracycline films application in treatment of periodontal pockets (11) . ...
... Nanoparticles can act on soft bases in DNA as sulfur and phosphorus as its main components and destroy it. This can inhibit DNA replication of the bacteria leading to cell death and can modify bacterial transduction signals (11) . ...
... [31] Shawky et al. (2015) introduced silver NP solution into the periodontally involved deep pockets and found a significant reduction in the number of microorganisms 1 month post procedure. [32] Gram staining showed that Gram-positive microorganisms predominated the outer surface of gingival mucosa in both the groups at presurgery. The genus included Streptococcus mutans, Enterococcus faecalis, and Staphylococcus aureus. ...
... [34,35] These microaerophiles have also been recently discussed as potential pathogenic microorganisms in periodontal diseases. [32] Hence, our study suggests that the shift of Gram-positive bacteria into Gram-negative is also seen in facultative anaerobes/ aerobes on the gingival surface, but it is happening when the periodontal condition is reverting back to health. The presence of Gram-negative bacteria in health has been also supported by Kumar et al., wherein they concluded that Gram-negative bacteria were associated with periodontal health. ...
Article
Background: Periodontal dressings are used for wound protection and patient comfort. Nano-silver particles have the ability to promote wound healing through anti-inflammatory properties. Hence, the present study aims to evaluate early wound healing parameters following periodontal surgery using nano-crystalline silver membrane as periodontal dressing. Materials and methods: Forty-two systemically healthy patients diagnosed with chronic periodontitis indicated for periodontal flap surgery were enrolled for the present study. Post surgery, the patients were randomly allocated to either a nano-crystalline silver dressing (Acticoat™) group (test group) or only the noneugenol dressing group (control group). Plaque index (PI) and wound healing index were recorded at the 7th- and 14th-day postsurgery. The microbiological analysis and vascular endothelial growth factor (VEGF) levels were evaluated at baseline and 7th-day postsurgery. Results: The healing index was significantly higher in the test group as compared to the control group at days 7 and 14 (P < 0.001; P < 0.001). The colony-forming units/ml count of bacteria were significantly reduced postsurgery in the test group (P = 0.019). VEGF levels increased significantly 7th-day postsurgery in the test group (P = 0.001). There was no statistically significant difference in the PI on the 7th-day postsurgery between the two groups (P = 0.173). Conclusion: The results of the study revealed that silver can be used as a potent periodontal dressing ingredient that can decrease the microbial colonization beneath the pack and promote faster healing postsurgery due to its antimicrobial activity.
... This raises the need for the use of local antiseptics or Chapter One Introduction 3 systematic antibiotics (Montevecchi et al., 2013). Unfortunately, prolong use of such antimicrobials may lead to disagreeable side effects and development of microbial resistance which is considered a major threat in medicine and public health (Shawky et al., 2015). In addition, differences in the effect of antimicrobial agents against oral bacteria and between biofilm and planktonic cells highlight the importance of screening for new oral antimicrobial formulations with more predictive clinical activity (Masadeh et al., 2013;Ali, 2018 (Jain et al., 2019). ...
... For the effective use of antimicrobial agents in the treatment of periodontal diseases, delivering adequate concentrations of drugs at the site of action for a sufficient duration are required (Mehta et al., 2011). In spite of that, systemic antibiotics can reach via blood stream to the deeper pockets; their concentration in GCF is usually less than in blood stream (Shawky et al., 2015). Topical antimicrobials, like mouthwashes, are in much higher concentrations in GCF rather than a systemic one for successful control of microbial plaque (Bogdanovska et al., 2012); however their action is higher at time of application when they are in their initial salivary concentration of 20 to 50 times bactericidal levels; but they will rapidly fall to approximately one tenth level following expectoration (Kumar et al., 2010). ...
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The current study was designed as the first one in Mosul City to isolate and characterize the three species of red complex pathogens from the mixed population of periodontal anaerobes and identify them by a new molecular method as thirty samples of gingival fluid were obtained from periodontal pockets with ≥ 4mm depth suffering from chronic periodontitis in patients attending the Teaching Hospital- College of Dentistry at the University of Mosul. Three types of media were prepared for culturing the samples on, Schaedler Anaerobic Blood Agar; Tannerella forsythia (TF) Agar and Trypton Yeast extracts Gelatin Volatile fatty acids and Serum (TYGVS) Agar. After 4-7 days of anaerobic incubation, colonies with different morphologies were picked and are subcultured for routine purification and molecular diagnosis by the new Loop Mediated Isothermal Amplification (LAMP) technique. Confirmed isolates by LAMP were more characterized by phenotypic features and their mono- and polymicrobial biofilms were formed in a microtitter plate. The results showed that the three pathogens of red complex were identified in the same specimen and great variations in the phenotypic characters of the same isolate were noticeable. The three pathogens were also able to form mono- and polybacterial biofilms in a synergistic mode. The current study also searched more rapid and easier methods for testing the ability of two natural materials, olibanum and alum and two standard antibacterial agents, Ciprofloxacin (CIP) and Chlorhexidine (CHX) to inhibit many aspects in the pathogenicity of these periodontopathogens. The Minimal Inhibitory Concentration (MIC) of these four antibacterial agents was determined and the manner of interaction between these four agents was studied. The results showed that the easiest method was resazurin- based microdilution. The four agents recorded different MIC values on the three types of bacteria and the MICs of the aqueous crude extract of the two natural materials were higher than that of the standard antibacterial agents. The four agents showed synergistic inhibitory effect against the three types of red complex pathogens. The study also investigated the efficacy of the natural materials to prevent coaggregation between red complex pathogens and it is proved that 2- 12 mg/ ml concentrations prevented this phenomenon. The effect of the four antibacterial agents on the mono- and polymicrobial growth of red complex pathogens and biofilm formation was also estimated. They inhibited the polymicrobial growth but with less significant than that on the monomicrobial growth and interfered with the formation of homotypic biofilms but their activity, except that of olibanum, reduced on polymicrobial biofilms. The study also utilized the fluorescent dyes of LIVE/ DEAD BackLight Bacterial Viability kit to differentiate between live and dead bacteria after treating the polymicrobial plankton and mature biofilm with the four antibacterial agents. The results showed that exposure for one hour to all agents, except CIP, was effective to loss cell viability but not effective against the mature polymicrobial biofilm. Another goal of this study was to design an experiment to make a polymer of antibacterial agents using disk- like film of olibanum and investigate its ability to liberate the antibacterial agents by agar diffusion method. The study successfully made this film which maintained its ability to liberate the antibacterial agents and form an inhibition zone. For the first time locally, this study was successfully able to manufacture a Medicated Chewing Gum (MCG) from olibanum and test its therapeutic activity in several participants infected with chronic periodontitis. Bacterial inhibition was followed by tracking the hydrolytic enzymes' level of periodontal pathogens in the gingival crevicular fluid (GCF) using APIZYM system. The manufactured olibanum- MCG proved its efficacy in reducing the bioburden of periodontal pathogens in term of diminishing the levels of hydrolytic enzymes in the GCF in participants chewing the medicated gum compared to the control group that were only treated with mechanical cleaning of periodontal pocket.
... Scaling and root planing coupled with silver nanoparticles, with or without tetracycline films as local drug delivery systems have shown reduced pocket depths, better gain in attachment levels, and a decrease in bacterial count than mere scaling and root planning [24]. ...
Article
Nanotechnology is the study of materials, devices, and systems that display features that are distinct from those found in larger systems. It is no surprise that medical and dental researchers have taken notice of this rapid advancement in technology, which has piqued their curiosity about how it may be used to cure and prevent disease. It is a relatively new science that deals with the manipulation of matter at the molecular level, including the manipulation of individual molecules and their relationships. High-level control of position and chemical characteristics are the primary goals of this method. Nanotechnology has become one of the most promising and significant areas of scientific research as a result, thanks to an increased interest in interpreting matter's property at this dimension. Nanotechnology has been used in dentistry to produce new materials and procedures for diagnosing, preventing, treating, and regenerating periodontal disease, which is the subject of this article. Cargoes and materials such polymeric nanoparticles, non-porous nanogels, nanotube scaffold matrices, and nanofibers have shown promise effectiveness, and their functions in disease treatment are of significant interest. The purpose of this review paper is to offer thorough recent updates on the numerous nanotechnology-based methods to periodontal disease treatment.
... Inclusion of nanoparticles into new materials has opened up doors for broad span application in multiple fields, including dentistry. Silver nanoparticles in particular have been of great interest due to their excellent antimicrobial action [18], use in caries prevention [19], and high activity against periodontal disease [20]. Although previous generations of silver nanoparticles suffered from toxicity concerns, it is becoming clear that highly stable and purified plant compounds can now be utilized for improving these materials downsides. ...
... Unfortunately, prolong use of such antimicrobials may lead to disagreeable side effects and development of microbial resistance which is considered a major threat in medicine and public health [6]. In addition to, differences in the effect of antimicrobials among oral bacteria and between biofilm and plankton cells, less activity of the common antimicrobials in the treatment of anaerobic bacterial infections and multidrug resistant species along with the development of several resistance mechanisms were recorded [7,8]. ...
Article
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The current study aimed to investigate the effectiveness of four regents; two naturals, olibanum and alum, and two standards, ciprofloxacin (CIP) and chlorhexidine (CHX) to affect the growth and biofilm of three types of periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, "the red complex group". Clinical isolates of the red complex pathogens were isolated from chronic periodontitis. They were identified by phenotypic properties and molecular method. The inhibitory activity of the four reagents was tested by microdilution method. Minimal inhibitory concentration (MIC) on the bacterial plankton and minimal biofilm inhibitory concentration (MBIC) on biofilm of the four reagents in a single and combinational use was determined on mono-and polymicrobial populations. Simple linear regression modeling was used to explore the effect of each reagent and determine MICs and MBICs. All reagents showed inhibition activity against the growth of mono-and polymicrobial planktonic population. MIC values on polymicrobial growth were higher than on monomicrobial growth and MBICs were much higher. All reagents had antibacterial activity on a monomicrobial biofilm with greater significant effect on T. denticola then T. forsythia and P. gingivalis. On polymicrobial biofilm, just olibanum continued showing its effect whilst CHX was less effect and both alum and CIP had no effect. Combinational use with Olibanum encouraged the effect of other regents on polymicrobial biofilm. This combination is a promising medicated preparation to combat the subgingival plaque of red complex pathogens.
... The antibacterial effects of NS in the current study is in agreement with a study conducted by Halkai et al. 21 who found that NS revealed effective antibacterial ability against P. gingivalis, also with numerous studies that demonstrated; reduction of microbial infections after use of NS [22][23][24][25] . In the current study; also the bacterial counts was lower in the NS group than in CHX group, but there was no significant difference between them at the three ISSN: 2456-8058 4 CODEN (USA): UJPRA3 experimental time points (p>0.05). ...
Article
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The necessitate for frequent application of Chlorhexidine (CHX), and other side effects has encouraged the search for option that are more suitable for patients as nanosilver mouthwash (NS). So the aim of this study was to determine the effects of a mouthwash made with nanosilver on dental plaque microbial counts and compare it with commercially available Chlorhexidine. Sixty-two plaque-induced gingivitis patients were allocated into two groups and asked to rinse with 10 ml of NS and CHX, immediately after brushing, for 1 min, in the morning and evening. Sub gingival plaque microbial counts were taken at baseline, two weeks, and finally at four weeks for each patient. Subsequently, the samples were collected, transferred and cultured in blood agar in anaerobic media. The colonies were counted and expressed as CFUs. The statistical analysis between CFUs variables within groups was calculated and the variation significance was calculated by performed t-test. It is very obvious that the values of CFU decreased significantly (p<0.001) as the time of use nanosilver until reaching the highest value when the time of use was 4 weeks [70.3±47 to 32.4±24.6 (2 weeks), and 14.2±9.9 (4 weeks) with inhibition of growth rate after 2 weeks was 46% and after 4 weeks was 79.7%. The effect of commercially available CHX mouthwash was approximately similar to the effect of NS mouthwash used. In conclusion, both Group I and Group II showed similar effect on inhibition anaerobic periodontal pathogens counts and gingival health. There was significant inhibitory effect on microbial counts where NS mouthwash had shown better results than CHX, but there was no significant difference between the nanosilver mouth wash and the Chlorhexidine mouthwash.
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Over the past few decades, nanoparticles of metals, and in particular silver, with a diameter of less than 100 nm have significantly expanded their field of application for various biomedical purposes. So, silver nanoparticles have great potential in a wide range of applications as antimicrobial agents, coatings for biomedical products, carriers for drug delivery, bioengineering, since they have discrete physical properties and wide biochemical functionality. Studies have shown that the size, morphology, stability and properties (chemical and physical) of metal nanoparticles are strongly influenced by the conditions of the experiment, the kinetics of the interaction of metal ions with reducing agents and the adsorption processes of the stabilizer with metal nanoparticles. This review aims to analyze the use of silver nanoparticles in modern medicine based on data from domestic and foreign literature over the past five years. The study confirmed the high biological activity of drugs with nanoserebrum as anti-inflammatory, antimicrobial agents, antifungals, the presence of an inhibitory effect on protozoa, antioxidant and anticancer effects, and substantiated the relevance of use in bioengineering and dentistry. However, rapid advances and advances in technology have led to concerns about the potential risk associated with the use and application of silver nanoparticles to human health and the environment. Therefore, this review attempts to characterize and quantify the potential harmful effects of silver nanoparticles on the health of laboratory animals and humans, and focuses on ways to neutralize or reduce the toxic effects of silver nanoparticles on the human body.
Article
Background New materials are currently designed for efficient treatment of oral tissue lesions by guided tissue regeneration. The aim of this study was to develop a multifunctional 3D hybrid biomaterial consisting of extracellular matrix components, collagen, chondroitin 4-sulfate and fibronectin, functionalised with silver nanoparticles, intended to improve periodontitis treatment protocols. Methods Structural observations were performed by autometallography, scanning and transmission electron microscopy. In vitro tests of 3D constructs of embedded gingival fibroblasts within hybrid biomaterial were performed by MTS and Live/Dead assays. Genotoxicity was assessed by comet assay. In vivo experiments using chick embryo chorioallantoic membrane (CAM) assay analysed the degradation and nanoparticles release, but also angiogenesis, new tissue formation in 3D constructs and the regenerative potential of the hybrid material. Biological activity was investigated in experimental models of inflamed THP-1 macrophages and oral specific bacterial cultures. Results Light micrographs showed distribution of silver nanoparticles on collagen fibrils. Scanning electron micrographs revealed a microstructure with interconnected pores, which favoured cell adhesion and infiltration. Cell viability and proliferation were significantly higher within the 3D hybrid biomaterial than in 2D culture conditions, while absence of the hybrid material's genotoxic effect was found. In vivo experiments showed that the hybrid material was colonised by cells and blood vessels, initiating synthesis of new extracellular matrix. Besides the known effect of chondroitin sulfate, incorporated silver nanoparticles increased the anti-inflammatory activity of the hybrid biomaterial. The silver nanoparticles maintained their antibacterial activity even after embedding in the polymeric scaffold and inhibited the growth of F. nucleatum and P. gingivalis. Conclusion The novel biomimetic scaffold functionalised with silver nanoparticles presented regenerative, anti-inflammatory and antimicrobial potential for oral cavity lesions repair.
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Periodontitis, a disease involving supportive structures of the teeth prevails in all groups, ethnicities, races and both genders. The relationship between bacterial plaque and the development of periodontal disease and caries is well established. Antibacterial agents have been used effectively in the management of periodontal infection. The effectiveness of mechanical debridement of plaque and repeated topical and systemic administration of antibacterial agents are limited due to the lack of accessibility to periodontopathic organisms in the periodontal pocket. Systemic administration of drugs leads to therapeutic concentrations at the site of infection, but for short periods of time, forcing repeated dosing for longer periods. Local delivery of antimicrobials has been investigated for the possibility of overcoming the limitations of conventional therapy. The use of sustained release formulations to deliver antibacterials to the site of infection (periodontal pocket) has recently gained interest. These products provide a long-term,effective treatment at the site of infection at much smaller doses. Biodegradable polymers are extensively employed in periodontal drug delivery devices because of their abundant source, lack of toxicity, and high tissue compatibility. A major advantage of natural polymers is that they do not affect periodontal tissue regeneration. Amongst various natural polymers, chitosan, a deacetylated product of chitin is widelyused in drug delivery devices. Since it exhibits favourable biological properties such as non-toxicity, bioco patibility, biodegradability and wound healing traits, it has attracted great attention in the pharmaceutical and biomedical fields. The conventional treatment consists of tooth surface mechanical cleaning and root planning, associated or not to the systemic use of high concentrations of antibiotics, but with reduced effectiveness, and adverse effects. The patient compliance to the therapeutic is committed too. In the last decades, the treatment has been optimized for the use of drug delivery systems to the periodontal pocket, with the advantage of delivering the drug in the specific site, sustaining and/or controlling the drug concentration. Recently, the use of new drug delivery systems has been receiving great interest. This review approaches the main delivery systems for the administration of drugs to the periodontal pocket, their usefulness, as well as the advancement of these systems effectiveness in the periodontal therapy.
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The aim of the study was to compare the efficacy of chlorhexidine gluconate 2.5 mg (Periochip) and Minocycline hydrochloride 1 mg (Arestin) as local drug delivery agents in the management of chronic periodontitis. Twenty patients in the age group of 30-50 years suffering from chronic periodontitis (12 males and 8 females), with almost identical probing depth bilaterally (5-8 mm), and exhibiting bleeding on probing were selected and divided into two groups: Group I consisted of periodontal pockets on the left side and received Periochip and group II consisted of periodontal pockets on the right side and received Arestin. Patients were recalled after 6 weeks and 3 months intervals from the baseline visit to record plaque index, gingival index, and probing depth. There was reduction in all the parameters in both the groups at 6 weeks and 3 months as compared to baseline. From the results of the present study, it was concluded that both the drugs were equally effective in reduction of plaque scores as well as gingival scores. It was further observed that Arestin resulted in better results at 6 weeks while Periochip showed better results at 3 months with respect to probing depth reduction.
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The bactericidal efficiency of various positively and negatively charged silver nanoparticles has been extensively evaluated in literature, but there is no report on efficacy of neutrally charged silver nanoparticles. The goal of this study is to evaluate the role of electrical charge at the surface of silver nanoparticles on antibacterial activity against a panel of microorganisms. Three different silver nanoparticles were synthesized by different methods, providing three different electrical surface charges (positive, neutral, and negative). The antibacterial activity of these nanoparticles was tested against gram-positive (i.e., Staphylococcus aureus, Streptococcus mutans, and Streptococcus pyogenes) and gram-negative (i.e., Escherichia coli and Proteus vulgaris) bacteria. Well diffusion and micro-dilution tests were used to evaluate the bactericidal activity of the nanoparticles. According to the obtained results, the positively-charged silver nanoparticles showed the highest bactericidal activity against all microorganisms tested. The negatively charged silver nanoparticles had the least and the neutral nanoparticles had intermediate antibacterial activity. The most resistant bacteria were Proteus vulgaris. We found that the surface charge of the silver nanoparticles was a significant factor affecting bactericidal activity on these surfaces. Although the positively charged nanoparticles showed the highest level of effectiveness against the organisms tested, the neutrally charged particles were also potent against most bacterial species.
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To investigate the mechanism of inhibition of silver ions on microorganisms, two strains of bacteria, namely Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus), were treated with AgNO3 and studied using combined electron microscopy and X-ray microanalysis. Similar morphological changes occurred in both E. coli and S. aureus cells after Ag⁺ treatment. The cytoplasm membrane detached from the cell wall. A remarkable electron-light region appeared in the center of the cells, which contained condensed deoxyribonucleic acid (DNA) molecules. There are many small electron-dense granules either surrounding the cell wall or depositing inside the cells. The existence of elements of silver and sulfur in the electron-dense granules and cytoplasm detected by X-ray microanalysis suggested the antibacterial mechanism of silver: DNA lost its replication ability and the protein became inactivated after Ag⁺ treatment. The slighter morphological changes of S. aureus compared with E. coli recommended a defense system of S. aureus against the inhibitory effects of Ag⁺ ions. © 2000 John Wiley & Sons, Inc. J Biomed Mater Res, 52, 662–668, 2000.
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The antimicrobial effect of the nanosilver on Escherichia coli and the antimicrobial mechanism was studied elementarily. Experimental results indicated that the silver nanoparticles of 20μg/mL could inhibit completely the growth of 106 cfu/mL cells in liquid LB medium. The growth curves showed that silver nanoparticles prolonged the lag phase of E. coli, and the higher of the concentration of silver nanoparticles, the longer of the lag phase of E. coli. Transmission electron microscopy (TEM) was used to evaluate the cell morphology of both the normal and the treated E. coli. The observation with TEM suggested that silver nanoparticles lead to the formation of "pits" in cell wall of the bacteria, and silver nanoparticles could enter into periplasm through the pits and destroyed the cell membrane. Then the silver nanoparticles could enter into the bacterial cell, which not only condensed DNA, but also combined and coagulated with the cytoplasm of damaged bacteria. Finally, silver nanoparticles resulted in the leakage of cytoplasmic component. Moreover, the analysis of agar gel electrophoresis demonstrated that silver nanoparticles could increase the decomposability of genome DNA.
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Background: The selective removal or inhibition of pathogenic microbes with locally delivered antimicrobials when combined with scaling and root planing is often an effective approach for the managment of chronic periodontitis. Aim: To compare the clinical efficacy of tetracycline fibers and a xanthan based chlorhexidine gel in the treatment of chronic periodontitis. Methods and materials: Thirty systemically healthy patients in the age group of 30-50 years suffering from generalized chronic moderate periodontitis were selected. For each subject, two experimental sites were chosen that had probing depth >5mm and were located in symmetric quadrants and the sites were randomized at split mouth level with one receiving tetracycline fibers and the other chlorhexidine gel. Plaque score, bleeding score, probing pocket depth and relative attachment level gain was recorded on day 0 and at the end of 3 months.Results and conclusion: In both groups, there was statistically highly significant reduction in all the clinical parameters i.e. plaque score, bleeding score and probing pocket depth and relative attachment level gain was seen at different time intervals. Local delivery of tetracycline and chlorhexidine is a safe, easy and efficacious method along with scaling and root planing in the treatment of chronic periodontitis. Inter-comparison of both local drug delivery agents with respect to clinical changes shows that tetracycline fibers are better than chlorhexidine gel for treatment of chronic periodontitis. Nevertheless, long term studies with more samples are suggested to further evaluate and compare the efficacy of both materials.
Article
A critical summary of Matesanz-Perez P, Garcia-Gargallo M, Figuero E, Bascones-Martinez A, Sanz M, Herrera D. A systematic review on the effects of local antimicrobials as adjuncts to subgingival debridement, compared with subgingival debridement alone, in the treatment of chronic periodontitis.
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Background: Scaling and root planing is the basic treatment modality for periodontal disease. Mechanical treatment is limited by physical impediments and biochemical considerations. Antimicrobial agents may be used as an adjunct to overcome limitations of mechanical therapy. Methods: A case-control study was carried out on 30 patients suffering from chronic periodontitis. In Group A only scaling and root planing was carried out whereas in Group B tetracycline fibers were used along with scaling and root planing. Result: Tetracycline fibers as an adjunct to scaling and root planing was found to be more effective in reducing inflammation. The number of sites with bleeding on probing were 12 in Group A as compared to four in Group B after 30 days. The mean decrease in probing depth was more in Group B than Group A after 30 and 90 days. General linear model showed that decrease in probing depth was statistically significant with both mechanical therapy as well as adjunctive use of tetracycline fibers. Conclusion: Local drug delivery with tetracycline fiber is an effective and simple non surgical method to improve periodontal health which can be practiced even by a general dental practitioner.