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Review Article

Review of Microleakage Evaluation Tools

AlHanouf Abdullah AlHabdan
Department of Restorative Dental Sciences, King Saud University, Riyadh, KSA

Abstract
The advancement of restorative materials and techniques continues to enhance the clinical success of numerous restorative procedures. Despite
these new innovations, microleakage persists as one of the main causes of restoration failure. Microleakage tests provide useful information
on the performance of restorative materials, and different techniques for assessing microleakage have been developed and used. These tests
include the use of dyes, radioactive isotopes, air pressure, bacteria, neutron activation analysis, and artificial caries. However, little has been
done to determine the cause, mechanism, and nature of microleakage. Therefore, this review outlines and discusses the currently available
microleakage assessment tools.

Keywords: Dye penetration, in vitro, microleakage, tools

Introduction a material’s sealing ability and the quality of the hybrid layer,
which in turn affects the longevity of the restoration.[9,10]
The advancement of restorative materials and techniques
continues to enhance the clinical success of numerous Microleakage assays provide useful information on the
restorative procedures. Despite these new innovations, performance of restorative materials. Different techniques
microleakage persists as one of the main causes of restoration for assessing microleakage have been developed and used.
failure. Microleakage is the movement of bacteria, fluids, These tests which use dyes, radioactive isotopes, air pressure,
molecules, and/or ions between the tooth and restoration bacterial activity, neutron activation analysis, scanning
margins.[1] Microleakage is a result of the external environment electron microscope, dye penetration, and microcomputed
invasion through the margins of the restoration which also tomography (µCT) all come with both advantages and
can occur internally.[2,3] Microleakage can cause a variety of drawbacks.[11] Some of the older methods are no longer used
adverse effects, such as secondary caries, higher sensitivity because they do not represent the true nature of microleakage.[12]
of the restored tooth, and interfacial staining leading to pulp It is well known that microleakage affects the health of the
pathology.[4,5] Microleakage most commonly occurs when dental pulp and can cause pain, sensitivity, and discomfort.[2]
the gingival margin of any restoration is placed below the However, little is known about the cause, mechanism, and
cementoenamel junction because bonding to dentin is less nature of microleakage. Therefore, this review outlines and
predictable than enamel due to its complex pattern and lower discusses currently available microleakage assessment tools.
mineral content.[1,4,6]
Another type of leakage, known as nanoleakage, has also Discussion
been described. Nanoleakage is defined as when fluids move
Several factors are associated with microleakage, such
through the bonding between dentin and restorative resin.[7]
as polymerization shrinkage, which leads to dimensional
It is mainly caused by dentin acid etching that can provide
changes of the material, thermal contraction, water absorption,
a way for oral and dentinal fluids penetration into the hybrid
layer.[8] The amount of this penetration depends on several
Address for correspondence: Dr. AlHanouf Abdullah AlHabdan,
factors such as type and hydrophilicity of the bonding agent Department of Restorative Dental Sciences, King Saud University, P. O. Box:
and the application technique. Nanoleakage within the hybrid 54326, Riyadh 11415, KSA.
layer and adhesive–resin interface is an important indicator of E‑mail: [email protected]

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10.4103/jioh.jioh_160_17 How to cite this article: AlHabdan AA. Review of microleakage evaluation
tools. J Int Oral Health 2017;9:141‑5.

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AlHabdan: Microleakage evaluation tools

mechanical forces, as well as the changes in tooth structure resolution, which gives an overall view of the tooth structure
dimensions.[13] After polymerization, composite resin shrinks and microleakage at the interface.[25]
to a considerable amount which creates stresses at the
Mohapatra and Sivakumar reported that the acetate peel
restoration margins and thus gaps and microleakage occur.[14]
technique is a simple, inexpensive, and fast method for
Moreover, the adaptation of any bonded restorative depends
measuring microleakage. Moreover, peels are stable and
mainly on the shape of the cavity and number of walls bonded
can be preserved for further evaluation. However, the peel
(the C‑factor).[15] The coefficient of thermal expansion is
technique is delicate, which may produce artifacts that can be
another contributing factor to the occurrence of microleakage.
misinterpreted.[17]
Notably, the coefficient of thermal expansion for enamel and
dentin is far less than that of composite resin restorations.[13] Dye penetration
Leakage is typically evaluated with in vitro models rather The staining of microleakage and nanoleakage using colored
than in vivo methods, which can be qualitative as well as agents is the most commonly used technique. Dye penetration
quantitative.[16] The clinical performance of any new dental method involves the use of contrasting dyes as an immersion
restorative material can only be tested first using in vitro solution to stain the areas of microleakage, and then the
models. Theoretically, these findings can be transferred to the tooth–restoration interface is examined for evidence of
clinical environment.[17] staining. Notably, the most commonly used solutions are 0.5%
basic fuchsin, 2% methylene blue, and 50% silver nitrate.[26]
In the literature, in vitro studies use only 10–12 samples per
group, which may affect the power of the study statistically.[18] The dye penetration assay has many advantages over other
The International Organization for Standardization (ISO) has techniques. First, no reactive chemicals are used along
established guidelines for studying the physical properties with no radiation.[27] Second, different dye solutions are
of dental materials, such as flexural strength, water sorption, available; therefore, the technique is highly feasible and easily
and solubility (ISO/TS 11405: 2003, Dental Materials).[19] reproducible.[28]
For microleakage tests, the guidelines emphasize the need The current studies have failed to clearly establish which dyes
to standardize tooth quality, type of cavity preparation, and are suitable for use with microleakage test as some of the
method used to evaluate microleakage at the margin. [18] dyes can react with dentin such as basic fuchsin.[29] Another
New evaluation tools, such as µCT, can detect microleakage important issue with dye penetration methods is the particle
in vivo.[11] size of the used dye which could affect the reliability of the
Leakage tests can be subdivided into old and contemporary test.[26]
methods. Old methods were used to test the presence of gaps
Wu and Cobb [30] used silver nitrate because the strong
and the sealing ability of different restorative materials. These
optical contrast of silver particles is easily detected using
methods included air pressure, fluid filtration, electrochemistry,
microscopy. Silver nitrate staining is the most commonly used
neutron activation, bacteria, and artificial caries.[11] However,
material for nanoleakage evaluation as it easily penetrates the
these techniques were found to be nonrepresentative of leakage
interface zone due to its extremely small diameter (0.059 nm).
and thus have been replaced by more contemporary methods.
Following its penetration, silver nitrate molecules can become
Contemporary methods immobile, which prevents further penetration during specimen
Radioisotope method preparation. Silver nitrate was used to verify the discrepancy
A broad range of radioactive isotopes have been used in between the depth of the demineralized zone and monomer
microleakage studies, including the markers 45Ca, 131I, 35S, 22Na, diffusion, which is caused by the presence of water around
32
P, 86Rb, and 14C. Generally, autoradiography is used to detect collagen fibrils (nanoleakage).[31]
the leakage of isotopes at the restoration margins interface of Some authors reported possible problems arising from the use
a sectioned specimen.[20‑23] of 50% silver nitrate solutions or higher and recommended the
Isotopes can penetrate gaps equal to or larger than 40 nm, need to seal the specimens to ensure no other sources of silver
which is higher than the minimum detectable range of nitrate penetration. Moreover, a 24‑h immersion in silver nitrate
bacteria‑based studies. In addition, isotopes appear to be better decreased the pH to approximately 3.8.[31] In a study by Costa
at demonstrating microleakage than dye penetration tests.[24] On et al., a 24‑h immersion in an aqueous solution of 5% silver
the other hand, this technique requires the use of radiation, and nitrate was sufficient to detect a loss of the marginal seal in
the obtained autoradiograph does not represent microleakage composite restorations.[32]
as a three‑dimensional (3D) image.
Assessing a single section of the tooth is not representative
Acetate peel technique because dye penetration varies from one area to another. Thus,
Füsun et al. described the acetate peel technique as a rapid multiple‑surface scoring methods are regarded as superior
method for preparing many sequential replicas from an etched to single‑surface scoring methods because the results are
tooth surface to study dental hard tissue. Acetate films can more representative of the leakage pattern.[33] Raskin et  al.
be used to imprint the etched tooth surface at a micron scale recommended the utilization of three sections for each

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AlHabdan: Microleakage evaluation tools

restoration to accurately evaluate leakage.[34] Another technique primary molars prepared in vivo using µCT. These authors
introduced by Gale and Darvell[35] involves grinding of the concluded that µCT could be used to develop a standardized
specimen into sequential slices and then reconstruction of the method for measuring marginal leakage from in vivo samples.
images by the use of computer software. The authors stated In combination with a 4‑h immersion in 50% silver nitrate, the
that “3D techniques reveal markedly greater microleakage marginal leakage along the restoration–tooth interface was
than 2D assessments.”[35] Moreover, Majety and Pujar[36] accurately and reliably measured.[49] However, µCT alone is
recommended completely removing restorations to assess not a substitute for the histological examination of leakage in
the total amount of microleakage because leakage can vary in vitro samples.[50]
at different aspects of the cavity. The dye penetration method
requires an adequate evaluation tool to determine the true Confocal laser scanning microscopy
extent of microleakage. [11] Light microscopy, scanning CLSM is used to detect subsurface structures up to 100 µm in
electron microscopy, and transmission electron microscopy size without sample destruction.[51] Tangsgoolwatana et al.[52]
are widely used. However, recent methods to better evaluate compared the degree and pattern of microleakage in bonded
leakage include 3D methods, such as µCT 1072, confocal amalgam restorations treated with fluorescent dyes and a 45Ca
laser scanning microscopy (CLSM), and optical coherence radioisotope using CLSM and autoradiography, respectively.
tomography (OCT). The authors reported a high correlation between the results
of the fluorescent and radioisotope studies, indicating that
Three‑dimensional methods these two microleakage methods can be directly compared.[52]
The 3D analysis was first used by Gale et al.[37] The technique Another study by Grobler et  al.[53] stated that CLSM could
involves the production of sequential slices of the samples detect resin tag formation, penetration of the bonding agents
using a water‑cooled wire saw in 200‑μm thick and separated deep into the tubules, and hybrid layer formation for any
by 280 μm. Specialized computer software is then used to bonding material.
reconstruct the images and create 3D models. Then, the surface
area of dye leakage as well as volume of leakage is calculated
Optical coherence tomography
This evaluation tool was first introduced in 1998 by Colston
manually. 3D analysis was found to give more accurate
et al.[54,55] to evaluate dental hard and soft tissues in pigs. In
information than 2D analysis as the samples are examined
2008, Drexler and Fujimoto[56] used OCT to visualize dental
thoroughly.[38] However, the method is highly subjective and
tissue in vivo. Since then, OCT use in dentistry has become
destructive. The samples are destroyed during the sequential
increasingly more common.[57]
grinding and slicing procedure. Moreover, this slicing can
affect the restoration tested and thus alter the results.[39] OCT produces 3D images for the qualitative and quantitative
evaluation of dental hard and soft tissues in  vivo.[58] This
Microcomputed tomography
method enables clinicians to accurately and rapidly detect the
µCT is a modified version of medical computed tomography
early stages of dental caries, periodontal problems, and oral
that creates a 3D visualization of dental structures without
cancers.[59,60] Researchers have used OCT to clinically evaluate
destruction.[40] µCT starts from a set of 2D images taken along
dental restoration margins intraorally.[61] Moreover, OCT can be
the rotational axis, which is then transferred to a computer
used to quantify bubbles and voids within composites and most
program that produces a 3D image of the sample. These 3D
dental restorations by detecting the reflection of infrared light
reconstructions have a resolution of a few microns. One of
waves from internal microstructures, similar to the ultrasonic
the important features of µCT is that the 3D reconstruction
pulse echo method.[62] It has been also reported that OCT can
can be sectioned in any direction to gain accurate information
detect enamel cracks at the margins of composite restorations
of the sample’s internal geometric properties and structural
noninvasively.[63]
parameters. [41] Moreover, µCT results in a precise 3D
reconstruction of the sample.[42] Burghardt et al.[43] and Ibrahim Bakhsh et  al.[64] evaluated the tooth–restoration interface
et al.[44] recommended µCT as the gold standard for assessing using OCT and CLSM and reported that OCT can be used to
bone morphology and microarchitectures. quantify interfacial gaps at a micron scale. Another study by
Nazari et al.[65] used OCT to detect voids and gap formation
De Santis et al. was the first to use µCT in a microleakage
with flowable resin composite. The authors concluded that
evaluation study.[45] This technique was also been used to
OCT is a unique method to noninvasively and precisely assess
evaluate mechanically stressed dentin and adhesive–composite
restoration materials. The same conclusion was reported by
interfaces as well as detect marginal leakage at the
Shimada et al.[59] in 2015.
restoration–tooth interface in vivo. One of the main advantages
of µCT is that the 2D section information is maintained; thus,
all margins are available for visual inspection.[46,47] In addition, Conclusion
µCT was suggested by Özkan et al.[48] as an alternative method The microleakage assessment tools available to researchers
to histological examination for in vitro studies. Eden et al.[49] each have their own advantages and disadvantages. Notably,
evaluated the reliability of a marginal leakage assessment of the microleakage testing of different restorative materials
self‑etch adhesive Class II resin composite restorations in should use similar methodologies to reduce variability in the

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AlHabdan: Microleakage evaluation tools

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