Tooth discoloration induced by endodontic materials: a laboratory study
Tooth discoloration induced by endodonticmaterials: a laboratory study
P. Lenherr1*, N. Allgayer2*, R. Weiger2,3, A. Filippi3,4, T. Attin5 & G. Krastl2,3,61Clinic for Reconstructive Dentistry and Temporomandibular Disorders, University of Basel, Basel; 2Department of Periodontology,Endodontology and Cariology, University of Basel, Basel; 3Center of Dental Traumatology, University of Basel, Basel; 4Departmentof Oral Surgery, Oral Radiology and Oral Medicine, University of Basel, Basel; 5Clinic for Preventive Dentistry, Periodontology andCariology, Centre for Dental and Oral Medicine and Craniomaxillofacial Surgery, University of Zurich, Zurich, Switzerland; and6Department of Oral Surgery, School of Dentistry, University of Birmingham, Birmingham, UK
1 month (T3), 3 months (T4), 6 months (T5) andafter 1 year (T6). Colour change (DE) values were
Lenherr P, Allgayer N, Weiger R, Filippi A, Attin T, Krastl
calculated. A two-way analysis of variance was used
G. Tooth discoloration induced by endodontic materials: a
to assess significant differences between the endodon-
laboratory study. International Endodontic Journal.
tic materials. The mean values of all groups were
Aim To investigate the discoloration potential of
compared using the Tukey multiple comparison test
endodontic materials using a bovine tooth model.
Methodology Two hundred and 10 dentine-en-
Results Significant differences were detected amongst
amel cuboid blocks (10 · 10 · 3.5 mm) were pre-
the experimental groups after 12 months (P < 0.0001).
pared out of the middle thirds of bovine tooth
The lowest colour change values were observed in the
crowns. Standardized cavities were prepared in the
groups N (AH Plus, 3.2 ± 1.5), A (empty, 3.8 ± 1.4), L
walls of the pulp chamber leaving 2 mm of enamel
(PC, 4.1 ± 1.7), C (calcium hydroxide, 4.7 ± 1.5), E
and dentine on the labial wall of the crown. The
(Ultracal XS, 5.1 ± 1.9) and J (WMTA, 7.9 ± 6.7). The
specimens were randomly assigned to 14 groups
most discoloration was measured in groups G (3Mix,
(n = 15). Endodontic materials were placed into the
66.2 ± 9.9) and F (Ledermix, 46.2 ± 11.6). PC showed
cavities as follows: group A: empty, group B: blood,
the best colour stability amongst the Portland cement-
group C: calcium hydroxide, group D: ApexCal,
based materials; however, when contaminated with
group E: Ultracal XS, group F: Ledermix, group G:
blood (group M), a significantly higher DE value
(13.6 ± 4.2) was detected (P = 0.032).
MTA(GMTA), group I: GMTA + blood, group J: white
Conclusion Materials used in endodontics may stain
MTA (WMTA), group K: WMTA + blood, group L:
teeth. Therefore, the choice of material should not rely
Portland cement (PC), group M: PC + blood and
solely on biological and functional criteria, but also
group N: AH Plus. The cavities were sealed with
take aesthetic considerations into account.
composite and stored in water. Standardized colour
Keywords: bovine tooth model, discoloration, end-
measurement (VITA Easyshade compact) was per-
odontic materials, sealer, staining.
formed at the following intervals: prior to (T0) andafter placement of the filling (T1), 1 week (T2),
Received 5 February 2012; accepted 15 March 2012
Correspondence: Gabriel Krastl, Center of Dental Traumatol-ogy, Department of Periodontology, Endodontology and Cari-
Tooth discoloration induced by endodontic materials is
ology, University of Basel, Hebelstrasse 3, CH-4056 Basel,
a common finding and may impair the aesthetic
Switzerland (Tel.: 0041 61 2672622; e-mail: gabriel.krastl@
outcome of endodontically treated teeth (van der Burgt
et al. 1986a,b). A progressive discoloration is suggested
*Both authors contributed equally to this study.
to be primarily a result of materials ingressing into
Discoloration from endodontic materials Lenherr et al.
dentinal tubules (van der Burgt et al. 1986a,b). How-
materials, which are used in endodontics. This study
ever, it has been shown that a visible crown discolor-
was undertaken to develop a new model for the
ation may not necessarily be associated with tubule
assessment of tooth discoloration and to investigate
penetration and may be caused by material remnants
the staining ability of endodontic materials.
in the pulp chamber, which get darker over time and
The tested hypotheses were (i) there is no difference
transmit through the hard tissues (Davis et al. 2002).
in discoloration amongst the tested materials after
The staining ability of several endodontic materials
12 months and (ii) all materials show a similar
including Walkhoff’s paste, Grossman’s cement, zinc
progression of discoloration over time.
oxide eugenol, endomethasone and N2 has beendemonstrated (Gutierrez & Guzman 1968, van der
Burgt et al. 1986a,b). However, most of these materialsare no longer used.
Limited data are available on the staining ability of
recent endodontic materials. Most studies that focused
Two hundred and 10 bovine incisors were extracted,
on root canal sealers concluded they stained teeth. This
cleaned and stored in water at room temperature.
was shown in laboratory studies for AH26, Kerr Pulp
Following the removal of the roots, the labial surface of
Canal Sealer, Roth 801, Sealapex, Endofill, Tubliseal,
each tooth was cleaned meticulously with scalers. A
zinc oxide eugenol, Apatite Root Canal Sealer, Cavizol,
cuboid enamel-dentine block (10 · 10 mm) was pre-
AH Plus and EndoREZ (Parsons et al. 2001, Davis et al.
pared from the middle third of each crown using a
2002, Partovi et al. 2006, Elkhazin 2011). The discol-
diamond-coated disc (Intensiv SA, Grancia, Switzer-
oration potential of Ledermix has been evaluated ex
land). The height of each block was standardized at
vivo (Kim et al. 2000a,b) and in a recent clinical study
3.5 ± 0.1 mm, measured with a calliper (Iwanson,
(Day et al. 2011). In the same three studies, calcium
Ustomed, Tuttlingen, Germany). A cylindrical-shaped
hydroxide pastes were used as controls. It was
hole with the diameter of 2.5 mm was drilled with a
concluded that, compared with Ledermix, calcium
form-congruent bur (Dentsply Maillefer, Ballaigues,
hydroxide may cause only minor but measurable
Switzerland) in the middle of each specimen to leave
staining. However, there are various formulations of
2 mm of the labial tooth structure (Fig. 1). The
calcium hydroxide available with different constitu-
specimens were placed in 1% sodium hypochlorite for
ents, which have been added to the powder toimprove properties such as antibacterial action, radio-pacity, flow and consistency (Fava & Saunders 1999). These additives may affect the staining ability of thepastes.
Several case reports reveal discoloration produced by
MTA. However, there is only one study documentingsevere discoloration caused by MTA when the materialwas used as a pulpotomy agent in primary molars(Naik & Hegde 2005).
Although manufacturers such as Medcem GmbH
(Weinfelden, Switzerland) claim that a better colourstability is achieved when using Portland cement (PC)instead of MTA, there are no studies available to provethat statement.
Discoloration after canal medication with triple
antibiotic pastes (3Mix), as used in the field of regen-erative endodontics, has only been scarcely mentionedin the literature (Trope 2010). Only one laboratoryexperiment proved clear evidence of tooth discolorationafter use of the paste (Kim et al. 2010).
Figure 1 Schematic showing the tooth piece cut out of the
In summary, no systematic approach has been used
middle third of the crown of a bovine incisor. The spectro-
to evaluate and compare the discoloration induced by
photometer is held on to the tooth piece.
Lenherr et al. Discoloration from endodontic materials
30 min and, after drying with air, placed in 20% EDTA
Polymerization was initiated with a LED curing light
(lege artis, Detthausen, Germany) for two additional
(Smart Lite PS Series, Dentsply International, York, PA,
minutes to remove the smear layer. After a final 3 min
in sodium hypochlorite, the specimens were stored in
Every specimen was placed into a single tube with
tap water (Standard Micro Test Tube 3810; Eppendorf
The specimens were randomly assigned to 14 groups
AG, Hamburg, Germany). The tubes were stored at
(n = 15), and different endodontic materials (Table 1)
room temperature and kept in the dark during the first
were placed into the cavities. The materials were
3 months. During the following period and up to
prepared as indicated by the manufacturer’s guidelines/
12 months, the specimens were exposed to indirect
recommendations. In group G, a triple antibiotic paste
sunlight until the end of the experiment.
including metronidazole, minocycline and ciprofloxacinwas produced according to the composition and mixing
instructions described by Trope (Trope 2008). Ingroups I, K and M, 1.5 lL of bovine blood was placed
Colour measurements were taken in a dark room with
on top of the material with a pipette (0.5–10 lL;
a spectrophotometer (VITA EasyshadeÒ compact; VITA
Eppendorf AG, Hamburg, Germany) to simulate a
Zahnfabrik, Bad Sa¨ckingen, Germany) under standard-
typical clinical situation in which calcium silicate-
ized conditions in a custom-fabricated measuring
based materials are in contact with vital and vascular-
station (Fig. 2). The station consisted of a wooden
ized tissue. This situation was not simulated for the root
board with a fixed lamp, a carrier for the specimens
canal dressings and sealers because their application
and the VITA EasyshadeÒ Compact unit. The instru-
implies that vital tissues have been removed from the
ment was calibrated before the measurement in each
The cavities were sealed with a self-adhesive resin
Seven sessions of colour measurements were ob-
material (RelyX Unicem; 3M ESPE, Seefeld, Germany).
tained at the following intervals: prior (T0 = baseline)
Table 1 Materials used in the different experimental groups and their compositions
Produits Dentaires, SA, Vevey, Switzerland
Injectable calcium hydroxide paste containing
Injectable calcium hydroxide paste containing
Glucocorticosteroid-antibiotic root canal
Riemser Arzneimittel, Greifswald, Germany
Triple antibiotic mixture (ciprofloxacin,
Grey mineral trioxide aggregate, containing
White mineral trioxide aggregate (similar to
GMTA, but lower amounts of iron, aluminium
Portland cement (similar to MTA but absence
of bismuth ions and presence of potassium
Epoxy–amine resin-based root canal sealer,
Discoloration from endodontic materials Lenherr et al.
The DE values at several time intervals are presented inFig. 3. The most severe discoloration after 12 monthswas caused by Ledermix and 3Mix. However, duringthe period of dark storage (first 3 months), Ledermixdid not reveal a significant difference compared withbaseline (P = 0.3057), whilst a highly significantdifference was observed for 3Mix (P < 0.0001). Pro-gressive discoloration in terms of a significant differencebetween baseline and 12 months was also observed forApexCal (P < 0.0001), WMTA + blood (P = 0.0146)and PC + blood (P = 0.0206).
Figure 2 Custom-built measuring station with VITA Easy-
GMTA showed severe discoloration from the time of
shadeÒ Compact instrument in the measuring position.
placement (T1). After a slight increase during the firstweek, the mean DE value seemed to remain stable at
and after placement of the filling (T1), after 1 week
(T2), 1 month (T3), 3 months (T4), 6 months (T5) and
DE mean values and standard deviations after
12 months (T6). To prevent optical changes caused by
12 months are given in Table 2. At this time interval,
dehydration, the excess water was removed briefly by
PC caused the least discoloration amongst the Portland
air-drying for 1 s. The entire measurement was com-
cement-based materials when no blood was added, the
pleted within 5 s for every specimen, and each speci-
difference being statistically significant compared with
men was measured once. The CIE L*a*b* data were
GMTA (P < 0.0001) but not compared with WMTA
For each specimen, colour change (DE) values were
calculated with the following formula.
This study investigated the potential of endodontic
DEÃ ¼ ððDLÃÞ2 þ ðDaÃÞ2 þ ðDbÃÞ2Þ1=2; DLÃ
materials to induce discoloration in a new experimental
¼ Là À Là ; Daà ¼ aà À aà ; Dbà ¼ bà À bà :
set-up using a bovine tooth model. The majority of
For each group, the means of the DE values were
previous studies have used human teeth to assess the
calculated at the given time intervals.
degree of staining. However, the significant variability
A two-way analysis of variance was used to assess
in tooth morphology combined with small sample sizes
significant differences between the tested endodontic
may influence the results considerably. To overcome
materials. The mean value of all groups were compared
these limitations, the present laboratory model used
using the Tukey multiple comparison test (a = 0.05).
standardized bovine tooth pieces with a similar shape
Figure 3 DE values of the experimental groups at the different measurement points, directly after placement of the filling (T1),
1 week (T2), 1 month (T3), 3 months (T4), 6 months (T5) and after 1 year (T6). Each error bar is constructed using a 95%
confidence interval of the mean. A time dependency is obvious for groups D, F and G.
Lenherr et al. Discoloration from endodontic materials
Table 2 Mean colour change after 12 months (mean ± stan-
and to provide optimal penetration of the endodontic
materials into the dentinal tubules, the smear layer was
Despite the standardized experimental set-up, the
present model has limitations in fully imitating the
clinical situation. Interaction of the endodontic mate-
rial with salivary components and bacteria may occur
if there is leakage at the restoration margins. This may
lead to different staining mechanisms in vivo. Further-
more, placing medicaments or sealers intentionally for
up to 12 months inside the tooth crown may exagger-
ate the clinical situation in which materials in the
access cavity are removed before the final restoration is
The present survey revealed significant differences
between the endodontic materials with regard to their
ability to discolour teeth. Furthermore, the materials
Levels not connected by same superscript are significantlydifferent.
showed a different progression of discoloration overtime. Thus, both hypotheses had to be rejected.
and thickness. Furthermore, the colour analysis wasconducted under standardized conditions in a custom-
Discoloration by Portland cement-based materials
fabricated measuring station to ensure that the con-secutive measurements were taken at the same region
Portland cement-based materials have gained much
popularity in endodontics because of their biocompat-
For colour determination, the Vita Easyshade Com-
ibility and good sealing properties (Parirokh & Tora-
pact Device was chosen because of its high data
binejad 2010, Torabinejad & Parirokh 2010).
stability and excellent repeatability (Lehmann et al.
Amongst these materials, in the present study, the
2010). Even though deviations from the spectrophoto-
best colour stability could be achieved with PC even
metric reference may occur with most commercially
though the difference was not significant compared
available devices, this discrepancy may not be relevant
with WMTA after 12 months. PC differs from the MTA
for this study, because the assessment of colour
by the absence of bismuth ions and presence of
changes and not the determination of the exact tooth
potassium ions (Song et al. 2006). According to Steffen
and van Waes (2009), bismuth oxide, which has been
Although bovine root dentine has a significantly
added to PC to increase radiopacity, is a possible factor
higher tubule density than human samples, coronal
responsible for the discoloration of teeth treated with
dentinal layers do not differ significantly in terms of
MTA. After contamination of the specimens with blood,
density or diameter of the tubules (Schilke et al. 2000).
all Portland cement-based materials showed an in-
This suggests that bovine incisor crowns may represent
creased discoloration. Namazikhah et al. (2008) dem-
suitable substitutes for human teeth in laboratory
onstrated that the microstructure of the materials
studies dealing with tooth discoloration.
shows pH-dependent porosities. These porosities may
The severity of tooth discoloration depends on
uptake blood components and may be responsible for
whether the smear layer is removed or not (Davis et al.
the observed discoloration. This is of clinical relevance
2002). It has been reported that the smear layer can
because Portland cement-based materials are usually
markedly reduce the permeability of dentine (White
placed in direct contact to vital, vascularized tissue.
et al. 1987). In studies performed without the removal
Thus, the development of biocompatible materials with
of smear layer, tooth discoloration was less evident or
a reduced porosity level may be beneficial.
took longer (Parsons et al. 2001, Davis et al. 2002). Recent protocols for root canal irrigation recommend
Discoloration by triple antibiotic pastes
the removal of the smear layer to facilitate thedisinfection of the root canal system (van der Sluis
The most severe discoloration was seen when the triple
et al. 2007). To reproduce a realistic clinical situation
antibiotic paste containing ciprofloxacin, metronidazole
Discoloration from endodontic materials Lenherr et al.
and minocycline was used. The specimens in this group
corticosteroid dressing may be a reasonable alternative
became almost black after 12 months. Discoloration
after canal medication with the triple antibiotic pastehas scarcely been mentioned in the literature (Trope
2010). Kim et al. (2010) identified minocycline as thecause for discoloration in vitro. Furthermore, the latter
AH Plus, the only sealer tested in this study, seemed to
study demonstrated that sealing the dentine wall with a
exhibit satisfactory colour stability, presumably owing
bonding agent prior to application of the paste could
to the stable radiopacifier. Zirconium oxide has a high
reduce the overall colour change, without being able to
radiopacity but is not known to be involved in
prevent it. Even though the triple antibiotic paste
discoloration. However, the present results are in
originally introduced by Hoshino et al. (1996) reliably
contrast to the findings of Elkhazin (2011) who showed
eradicates bacteria from infected root canals setting the
distinct discoloration induced by AH Plus after
conditions for subsequent re-vascularization in the field
6 weeks, which tended to decline after 8 weeks. The
of regenerative endodontic procedures (Trope 2008), it
present study also revealed a greater colour change
may cause severe aesthetic problems. Thus, future
during the first 3 months and declining DE values
research should focus on alternative mixtures with a
substitute for minocycline. Trope (2010) showed that
with Arestin as a substitute discoloration could bemarkedly reduced but not prevented.
As a comparable alternative a prefabricated trianti-
biotic mixture with cefuroxim as a substitute for
The calcium hydroxide dressings varied in their discol-
minocycline is available (TreVitaMix; Medcem GmbH).
oration ability. Pure calcium hydroxide and Ultracal XS
However, there are no studies available neither regard-
did not show any discoloration or difference to the
ing the disinfection potential of the mixture in the root
negative controls at any time. Interestingly, the Apex-
canal, nor its discoloration ability.
Cal specimens showed an increase in their DE values inthe second half of the observation period. The differenceto all the other calcium hydroxide materials was
statistically significant after 12 months. As bismuth
Ledermix has regained attention in the field of dental
carbonate is part of the chemical composition of
traumatology. Placed as an intracanal medication
ApexCal (22 weight per cent, according to the manu-
after severe luxation injuries, it has the potential to
facturers specification), a discoloration produced by
inhibit inflammatory root resorption (Pierce et al.
bismuth ions may explain this finding.
1988, Bryson et al. 2002, Wong & Sae-Lim 2002).
A recently published randomized controlled clinical
However, in the present study, severe and increasing
trial on replanted teeth demonstrated that even Ultr-
discoloration occurred in the Ledermix specimens
acal XS produced measurable colour changes (DE
especially after the specimens had been exposed to
change in colour = 3.0) in a clinical situation. How-
indirect sunlight. This is in accordance with other
ever, only 1 of 12 patients was concerned about the
laboratory (Kim et al. 2000a,b) and clinical studies
colour of the tooth compared with 7 of 10 patients in
(Day et al. 2011). From a clinical aspect, it may not be
the Ledermix group (Day et al. 2011). In the laboratory
possible to completely avoid contamination of the
tests for this study, the mean DE change in colour for
coronal dentine with the medicament or to meticu-
the Ultracal XS group was 5.1. In a clinical situation, a
lously clean the surfaces after placement. Thus,
minor colour mismatch between DE 2.6 and 3.7 is
alternative nonstaining alternatives are needed. This
sufficient to be identified by a dentist. However, a
may be Odontopaste (Australian Dental Manufactur-
difference of DE 5.5–6.8 is needed to classify the
ing, Kenmore Hills, Australia), a recently released
discoloration as unacceptable and recommend further
alternative with substitution of the tetracycline com-
ponent by clindamycin (Athanassiadis et al. 2011). Furthermore, as the inhibiting effect of Ledermix on
external root resorption after severe dislocation inju-ries is primarily attributed to the corticosteroid com-
Materials used in endodontics may stain teeth. There-
ponent (Chen et al. 2008, Kirakozova et al. 2009), a
fore, the choice of material should not rely solely on
Lenherr et al. Discoloration from endodontic materials
biological and functional aspects, but also take aes-
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The authors are very grateful to VITA Zahnfabrik (Bad
color-measuring devices compared with a spectrophotomet-
Sa¨ckingen, Germany) for providing the study with the
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SECTION 15121 PIPING EXPANSION COMPENSATION PART 1 - GENERAL MIL-E-17814E - Expansion Joints, Pipe, Slip-Type, Packed. Provide piping offsets and equipment required to control expansion and contraction of piping in accordance with the Drawings. Heating Hot Water: 200 degrees F. maximum. Submit under provisions of Section 01330 as applicable. Flexible Pipe Connectors: Indicate maxim