Journal of Pioneering Medical Sciences, 12, 1: 48–53

https://www.jpmsonline.com/
Received 7 August 2023, Accepted 9 November 2023, Published 21 December 2023

Research Article
Microleakage assessment of Hydrophilic pit and fissure sealants fortified with green
synthesized silver nanoparticles-An in vitro study
Mahalakshmi Kumaraguru , Jayashri Prabakar , Meignana Arumugham Indiran , Ganesh J , Rajesh
1 1, * 1 2

Kumar. S and Jishnu Krishna Kumar

3 1

1
Department of Public Health Dentistry Saveetha Dental College and Hospitals, Saveetha Institute of Medical
and Technical Sciences, Saveetha University, Tamil Nadu 600077, India.
2
Department of Pedodontic and Preventive Dentistry Saveetha Dental College and Hospitals, Saveetha
Institute of Medica and Technical Sciences, Saveetha University, Tamil Nadu 600077, India.
3
Department of Pharmacology Saveetha Dental College and Hospitals, Saveetha Institute of Medical and
Technical Sciences, Saveetha University, Tamil Nadu 600077, India.

* Correspondence: Jayashri Prabakar: jayashri.sdc@saveetha.com

Abstract: Background and Aim: Pit and fissure sealants play a significant role in preventive dentistry. The
sealant’s physical properties lend a notable contribution to its long-term success. The invention of nanoscience
has led to breakthrough discoveries in improving the mechanical properties of sealants. The current work aimed to
evaluate the microleakage of Hydrophilic sealant fortified using silver nanoparticles compared to the Hydrophilic
sealant and conventional sealant on permanent molars. Materials and methods: Silver nanoparticles were green
synthesized using beetroot extract and incorporated into hydrophilic Ultraseal XT Hydro sealant. A sample of
extracted third molars, thirty in number, were allocated randomly to three different groups as follows: Group
I: conventional Clinpro sealant, Group II: hydrophilic Ultraseal XT hydro, and Group III: Ultraseal XT hy-
dro fortified with silver nanoparticles. Microleakage assessment was performed using a stereomicroscope. The
Kruskal-Wallis test was employed to compare the microleakage values of the three groups. Results: Group III
had the lowest values about mean microleakage score (0.60±0.548), and the highest scores were noted for Group
I (1.40±0.548). A statistically significant difference was observed in comparing the three groups employing the
Kruskal-Wallis test. Conclusion: The results of the current study have established that the microleakage was the
lowest in fortified UltraSeal XT Hydro (Group III) when compared to Ultraseal XT Hydro (Group II) and Clinpro
sealant (Group I).
Keywords: Fluoride releasing sealants; Minimal Invasive dentistry; Primary prevention

1. Introduction timal treatment of choice by protecting tooth regions

that are more vulnerable to plaque and bacterial ac-
cumulation by transforming them into smoother self-
Most dental caries tend to originate from occlusal cleansing areas [7]. Despite advancements in restora-
pits and fissures. This is because these are anatomi- tive dentistry, contemporary sealant materials are re-
cally inconspicuous regions that lack exposure to sali- stricted by their bioinert nature and negligible interac-
vary flow, its remineralizing properties, and the effects tion with the oral environment [8]. This calls for for-
of thorough brushing [1]. Regarding the total area con- mulating innovative and bioactive materials with added
stituted by tooth surfaces, occlusal surfaces contribute valuable physical and functional properties to manage
only 12.5% and yet succumb to a more significant per- and treat dental caries efficiently [8, 9]. This could be
centage of the total caries prevalence observed in chil- done by harnessing the potential of nanotechnology.
dren [2]. For this reason, dental caries should be inter- Nanotechnology is a scientific field that handles mat-
vened at the very beginning to prevent serious compli- ter on a nanoscale (diameter less than 100nm) that dis-
cations [3–5]. In light of this fact, various approaches plays pioneering physical, optical, chemical, and bio-
are sought to reduce the probability of dental caries for- logical characteristics [10]. The nanoparticle synthesis
mation, including the habit of flossing, fluoride prophy- process can be carried out by various chemical, physi-
lactic agents such as rinses and varnish, and the reg- cal, and biological techniques. As the chemical method
ular use of fluoride toothpaste. However, more than is limited by the production of by-products that are haz-
routine brushing may be required concerning deep pits ardous to the environment, there exists a demand for
and fissures, given that brush bristles cannot reach the more eco-friendly methods [11]. This has led to the
inaccessible depths of these areas [6]. To overcome invention of ”green nanotechnology,” which uses a bi-
this limitation, dental sealants could serve as an op-
Journal of Pioneering Medical Sciences 49
ological approach wherein microorganisms and plants der was incorporated and brought to a boil, after which
are involved in the process of extracellular and intracel- it was filtered to form the beetroot extract. 50 ml of
lular nanoparticle synthesis [12, 14]. In this study, the the beetroot extract was added to a beaker containing
potential of beetroot extract as a reducing agent was ex- a mixture of 0.0169 grams of 1 millimole silver nitrate
plored and used to formulate silver nanoparticles. Beet- and 50 ml of distilled water. This mixture was then kept
root is a readily available vegetable rich in folate con- for 32 hours in an orbital shaker, which was then dis-
tent. Moreover, it is a powerful antioxidant that con- tributed and poured into six centrifugation tubes. The
tains Vitamins A, B6, and C [15]. The science behind tubes were placed in the centrifugation machine and
sealant synthesis has widened its branches to incorpo- centrifuged at 10,000 rpm for 20 minutes. The super-
rate the attribute of moisture resistance. Hydrophilic natant liquid was disposed of from each tube, and the
sealants with moisture-tolerant capabilities are now at a remaining material present at the bottom of each tube
drawing level with conventional hydrophobic sealants. (pellet solution) was transferred to a single tube and
Being readily miscible with water, they flow into etched then refrigerated for future use. The pellet solution was
enamel coated with moisture, thus forming a retention- laid out on a petri dish, placed in the hot air oven, and
enhancing strong bond [16]. This is highly relevant as subjected to a temperature of 100 degrees Celsius for 8
sealing pits and fissures are technique-sensitive and rely hours. The petri dish, now containing dry solution, was
on effective moisture control for their long-term suc- scraped out to yield silver nanoparticles.
cess. Sealants that tolerate moisture, such as Ultraseal
XT Hydro, have been introduced to combat this. This
light-cured sealant exhibits excellent thixotropic prop- Addition of silver nanoparticles into the sealant
erties apart from being fluoride-releasing. Moreover, Following a proportion of 1:10 by weight, the silver
an amalgamation of benefits of both hydrophobic and nanoparticles were added to Ultraseal XT hydro. This
hydrophobic sealants is noted in this material. The hy- mixture was then subjected to mixing in an amalgama-
drophilicity of the sealant ensures its uninterrupted flow tor for a period of 10 minutes.
into the depths of the fissures, even in the presence of
saliva [17]. Though many studies [17, 18] have been
done to assess the properties of hydrophilic sealants, Placement of sealant
very few have been done concerning sealants modi-
fied with silver nanoparticles. In the present study, we The teeth sample were first etched for 30 seconds us-
aimed to assess the microleakage of Ultraseal XT hydro ing 37% orthophosphoric acid. Next, the teeth were
sealant fortified with silver nanoparticles compared to rinsed with water and dried thoroughly with the help of
the Ultraseal XT hydro sealant and conventional Clin- a three-way syringe to make the enamel surface appear
pro sealant on permanent molars. white and frosty for Clinpro sealant. For Groups II and
III, the sample teeth were dried gently and left in mod-
2. Materials and methods erate wetness to yield a shiny appearance. The sealants
allocated for each group were placed on the teeth, fol-
This in vitro study was conducted to compare three lowed by light-curing for 30 seconds.
different sealant materials. The sample for the study
was constituted of permanent molars. A study by Microleakage assessment
Prabakar J et al. was used to arrive at the sample size
[18] employing G*Power 3.1.2 software with a power A solution of 5% sodium hypochlorite was used to
of 0.95 and (p ≤ 0.05). A sample size of 10 teeth soak the teeth with sealant placed on it. Any resid-
per group was derived, resulting in a total of 30 teeth. ual periodontal tissue or calculus particles were cleaned
Only teeth that were extracted for surgical or orthodon- and removed thoroughly. Following this, all the teeth
tic reasons, with intact occlusal surfaces, were included were scrutinized under the microscope to rule out the
in the study. Teeth exhibiting caries or developmental presence of caries, cracks, or defects. The specimens
anomalies were excluded. The scientific review board not fulfilling the inclusion criteria were rejected, while
of Saveetha University granted its approval before ini- those fulfilling were stored in 10% formalin solution
tiating the study. Following computer-generated ran- until further use. After the placement of sealants, mo-
domization, ten molars were allotted to each of the three lars were immersed inverted in 0.1% silver nitrate so-
groups, which were Clinpro sealant (Group I), Ultraseal lution at 37 C for 8 hours, followed by placement in
◦

XT hydro (Group II), and Ultraseal XT hydro fortified developer solution for 4 hours. All the teeth were then
with silver nanoparticles (Group III). exposed to a thermocycling procedure for 30 seconds,
with temperature ranges between 5 C and 55 C. Mo-
◦ ◦

Green synthesis of silver nanoparticles using beetroot lars were sliced buccolingually [Figure 1], and the tooth
extract sections were evaluated for microleakage using a stere-
omicroscope (LEICA M205C) [Figure 2,3] and scored
After washing the beetroots with clean drinking wa- by an examiner who was blinded to the study. Ovrebo
ter to remove impurities, they were diced and left to dry and Raadal [19] guidelines were used to assess the mi-
at room temperature. The dry pieces were added into croleakage and the interpretation of the scores.
a grinder mixer and powdered. In a beaker containing
100mL of distilled water, 20 grams of the beetroot pow-

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Journal of Pioneering Medical Sciences 50
Descriptive Statistics Group I Group II Group III
Number of samples 10 10 10
Mean microleakage scores 1.40 1.00 0.60
Standard deviation 0.548 0.707 0.548
Median 1.00 1.00 1.00
Minimum 1 0 0
Maximum 2 2 1
Table 1. Mean microleakage scores of Groups
I, II, and III.
Groups N Mean Rank Kruskal Wallis Score Significance
I 10 10.40
II 10 8.00 3.733 0.04*
III 10 5.60
Table 2. Mean difference in microleakage
score between Group I, II, and III
Figure 1. Buccolingual sectioning of the tooth
specimen.
3. Results
Table 1 shows the mean microleakage scores for
Group I, II, and III, ranging from 1-2, 0-2, and 0-1,
respectively. The median value of all three groups is
1.0. The mean microleakage scores were found to be
the lowest for Group III (0.60 ± 0.548) and highest for
Group I (1.40 ± 0.548). Kruskal Wallis test exhibited a
significant difference between the three groups regard-
ing microleakage. Hence, group 3 was found to be su-
perior to the other groups (Table 2).
4. Discussion

Nanotechnology aids in the treatment of dental caries

in two ways. The first approach is remineralization,
which employs nano-materials capable of releasing flu-
oride and calcium. The method includes using antibac-
terial nanoparticles such as silver, quaternary ammo-
Figure 2. Microleakage assessment using nium polyethylene amine, and zinc oxide [20]. Because
stereomicroscope. of their increased surface-to-volume ratio and bioavail-
ability toward cells and tissues, nanosized systems have
unique characteristics [21]. Increased surface area leads
to improved mechanical interlocking of nanoparticles to
the resin matrix [22]. Because of the minimization in
stress concentration areas, there is superior crack prop-
agation resistance and increased fatigue strength [23].
Silver nanoparticles are broad-spectrum antimicro-
bial agents that can be used to prevent dental caries [24].
Because of the particles’ high surface area, they adhere
to the outer cell membrane of bacteria, influencing the
bacteria’s permeability and cell structure [25]. Recent
investigations have shown that a dental sealant contain-
ing Ag NPs can be more effective than a traditional
sealant in preventing enamel caries in first permanent
molars [26–31]. In the current study, silver nanoparti-
cles were green synthesized from beetroot extract, in-
corporated into Ultraseal XT hydro, and subjected to
microhardness and microleakage testing. In the cur-
rent study, the mean microleakage scores for Clinpro,
Figure 3. Silver nitrate dye penetration as Ultraseal XT Hydro, and reinforced Ultraseal XT Hy-
seen through the stereomicroscope dro were 1.40 ± 0.548, 1.00 ± 0.707, 0.60 ± 0.548 re-
spectively. Though there was no significant difference
in the microleakage (p > 0.05) of the three sealants,
Clinpro exhibited the highest microleakage scores, and

Journal of Pioneering Medical Sciences Volume 12,Issue 1, 48–53

Journal of Pioneering Medical Sciences 51
reinforced Ultraseal XT Hydro exhibited the lowest. Journal of Contemporary Dental Practice, 20(7),
Borsatto et al. reported similar findings [32] Clinpro 812-817.
sealant had a higher level of microleakage than its glass 3. Jayachandar, D., Gurunathan, D., & Jeevanandan,
ionomer counterpart. Under saliva contamination, the G. (2019). Prevalence of early loss of primary mo-
article has demonstrated that the glass ionomer offered lars among children aged 5-10 years in Chennai: A
better marginal sealing than the resin-based Clinpro cross-sectional study. Journal of Indian Society of
sealant. This is in contrast to a study by Kusgoz A et al., Pedodontics and Preventive Dentistry, 37(2), 115-
[33] in which Fuji Triage had significantly higher mi- 119.
croleakage scores than Clinpro and Grandio Seal. This
result is by Ganesh and Shobha’s results [34], and Ri- 4. Jayakumar, A., Gurunathan, D., & Mathew, M. G.
ratt Tanapong et al. [35], which revealed that the resin- (2020). Correlation of Protein Level with the Sever-
based sealant outperformed the Fuji VII GIC Sealant ity of Early Childhood Caries - An Observational
in terms of sealing ability (in dry conditions). How- Study. Journal of Research in Medical and Dental
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who found no difference in microleakage score between 5. Gandhi, J. M., Gurunathan, D., Doraikannan, S.,
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age than conventional Clinpro sealant, though the dif- tistry, 39(4), 369-372.
ference was not statistically significant. The author at- 6. Subramaniam, P., Konde, S., & Mandanna, D. K.
tributes this difference to the enhanced thermal stability (2008). Retention of a resin-based sealant and a
of the resultant nano-polymer caused by AgNPs. This glass ionomer used as a fissure sealant: a compara-
may reduce marginal defects between the material and tive clinical study. Journal of Indian Society of Pe-
the tooth substrate, especially when subjected to ther- dodontics and Preventive Dentistry, 26(3), 114-120.
mal cycling. The results agreed with Morales-Quiroga 7. Beauchamp, J., Caufield, P. W., Crall, J. J., Donly,
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Journal of Pioneering Medical Sciences Volume 12,Issue 1, 48–53