JOURNAL OF ENDODONTICS Printed in U.S.A.

Copyright © 2002 by The American Association of Endodontists VOL. 28, NO. 11, NOVEMBER 2002

An Assessment of Microbial Coronal Leakage of

Temporary Filling Materials in Endodontically
Treated Teeth

Hanan Balto, BDS, MSc

This in vitro study evaluated the microbial leakage restoration resulted in significantly more absence of periradicular
of Cavit, IRM, and Dyract when used as temporary inflammation than good root canal treatment (3).
filling materials after root canal treatment. The de- Studies of coronal leakage after completion of endodontic treat-
ment have shown that the canal obturating techniques and mate-
gree of coronal leakage was assessed by using a
rials do not provide a hermetic seal. Torabinejad et al. (4) showed
microbiological marker consisting of Streptococ-
that more than 50% of root canals were completely contaminated
cus faecalis and Candida albicans. For each of the when the coronal surfaces of their fillings were exposed to Staph-
two organisms, a set of 15 maxillary premolars ylococcus epidermidis. Similar observation was reported by
were prepared chemomechanically and obturated Khayat et al. (5). In addition to bacterial infection, recently there
with thermoplasticized gutta-percha. A 3.5-mm has been a growing concern about yeast infections of the root
thick layer of one of the three temporary filling canal. Microbiological investigations have shown that yeasts may
materials was inserted in the access cavities of the be present in the microbial flora of apical periodontitis (6, 7). They
teeth from each group (each group was compro- may enter the pulp through dentinal tubules, deep carious lesions,
mised of five teeth). The control teeth (four positive fractures, or as contaminants from the oral microflora during the
root-canal treatment (6, 8). Almost all isolated yeasts belong to
and four negative) lacked any filling material over
genus Candida, and Candida albicans is the most often isolated
the gutta-percha, whereas the orifice and the api- species. So far there has been no study that compares the coronal
cal foramen of the negative control were com- leakage of the temporary restorative filling materials when C.
pletely sealed with nail polish. Each tooth was albicans is used as a tracer.
placed in a well of a 24-well tissue culture plate and The aim of this in vitro study was to evaluate the microbial
embedded in trypticase soy broth and 0.5% Bacto- leakage of Cavit, IRM, and Dyract when used as temporary filling
agar. An organism suspension was inoculated in materials after root canal treatment in premolars. S. faecalis and C.
the access cavity, and microbial penetration was albicans were used as microbial tracers.
detected as an increase in turbidity of the broth. At
the end of 30 days, the results showed that all
positive control teeth leaked within 1 week, MATERIALS AND METHODS
whereas those that served as negative control re-
Thirty-eight extracted human maxillary premolars with intact
mained uncontaminated throughout the test pe- crowns were used in this study. These teeth were extracted for
riod. With both organisms, IRM started to leak after orthodontic reason. The teeth were stored in 0.9% physiological
10 days, whereas Cavit and Dyract leaked after 2 saline and were kept moist at all times throughout the experiment.
weeks. The periodontal ligament was removed from the root of the teeth
by using a curette. To ensure uniformity in the root canal length,
approximately 7 mm of the root length apical to CEJ was left
intact, and the apical part was sectioned and removed by using a
plain taper fissure bur in a high-speed handpiece under water spray.
A three-dimensional filling of the root canal system will prevent Access to the root canals was gained by using size #4 and #6 round
the penetration of microorganisms and toxins from the oral cavity burs in a high-speed handpiece under copious water spray. The
via the root canal into the periradicular tissues. Coronal leakage of access cavity was generally oval, 2.5 mm in width and 3.5 mm in
the root canal filling is considered to be an important cause of length. Working lengths were designated as 1-mm short of the
failure of root canal therapy (1). Weine (2) has indicated that point at which a #20 k-file exited the apical foramen. The canals
improper restoration leads to loss of more endodontically treated were instrumented to size 40k to obtain a standardized diameter of
teeth than actual failure of endodontic therapy. Good coronal the apical end of the canals. Approximately 2 ml of 1% NaOCl

762
Vol. 28, No. 11, November 2002 Coronal Leakage 763

were used to flush the canal between each file size. Coronal flaring TABLE 1. Means and standard deviations of starting day of
was accomplished with Gates Glidden burs, sizes 2 and 3. The microbial growth
specimens were steam autoclaved at 135°C for 20 min and after Materials Microorganisms Mean SD
that all the procedures were performed under a laminar airflow
hood using sterilized instruments. The canals were obturated with IRM S. Faecalis 10.00 2.24
IRM C. Albicans 12.00 2.74
thermoplasticized gutta-percha, Obtura II (Obtura Corporation,
Cavit S. Faecalis 16.40 1.34
Fenton, MO), using AH26 “silver free” as a sealer cement (De Cavit C. Albicans 15.20 1.64
Tray Dentsply, Milford, DE). Gutta-percha was cold burnished Dyract S. Faecalis 17.00 4.90
with a ball burnisher at the apical end and vertically condensed Dyract C. Albicans 15.20 1.64
coronally at the orifice opening of the canals. Excess root canal ⫹ ve control S. Faecalis & C. Albicans 6.00 0.00
sealer was removed coronally with a sterile cotton pellet moistened ⫺ ve control S. Faecalis & C. Albicans 30.00 0.00
in alcohol. The depth of the cavity was measured from the crest of
the marginal ridge with a periodontal probe and was approximately
5.5 mm. Teeth were placed in an incubator at 100% humidity at
37°C for 72 h to allow setting of the sealer. All teeth were bacteriological medium comprised of trypticase soy broth and
instrumented and obturated in the same manner by one operator 0.5% Bacto-agar (Detroit, MI) to cover the tooth up to CEJ (0.75
and then randomly divided into 3 groups of 10 teeth each. A ml).
3.5-mm thick layer of one of the three temporary filling materials The microorganism suspension was made in phosphate buffered
was inserted in each group. Cavit (ESPE America, INC., Norris- saline (PBS) containing 2% serum, and 50 ml of this suspension
town, PA) was used in group 1 and IRM (L.D., Caulk Division, was inoculated in the cavity on top of the filling material. Every
Milford, DE) in group 2. Both materials were placed incrementally 48 h, fresh culture was added and approximately 100 ml of PBS
in the access cavity with a plastic instrument, condensed with a was added to the bacteriological medium to keep it hydrated. The
plugger, and the excess material was removed with a sterile cotton plates were incubated at 37°C, and the samples were monitored
pellet lightly dampened with sterile saline. daily up to a maximum of 30 days. On a daily basis, the bottom of
For the teeth in group 3, a thin layer of bonding agent was the tissue culture wells was checked visually for turbidity as
placed with the applicator on the dentinal wall of the prepared compared with the negative control. If turbidity occurred, the day
cavity and over the gutta-percha, cured with a visible light for 10 s, of leakage was recorded for each sample. After microbial growth
and then a 3.5-mm thick layer of Dyract (Dentsply-De Trey, was noticed, a sample from the medium was plated onto Columbia
Konstanz, Germany) was injected directly into the prepared cavity agar. The plate was checked by macroscopic morphological ex-
from the dispensing syringe and cured with the visible light acti- amination and Gram staining to assure that it contained the same
vator for approximately 20 s. Group 4 consisted of eight teeth used type of organism as was placed in the prepared cavity.
as positive and negative controls (four teeth each). Parametric two-way analysis of variance (ANOVA) was uti-
The positive and the negative control teeth lacked any barrier lized to test the main factors (the organisms and the temporary
over the gutta-percha. All surfaces of the negative control teeth, filling materials) and the interaction between them. To check the
including the orifice and the apical foramen, were completely significant difference between the materials, Tukey’s multiple
sealed with three layers of nail polish (Clarins, France). The teeth range test was utilized.
in the three experimental groups and the positive controls received
three layers of nail polish leaving the area of canal’s orifice and the RESULTS
apical foramen exposed. The thickness of the temporary filling
materials was measured with a spring caliper (Hu-Friedy) with its The positive control teeth in groups A and B leaked within 1
one end placed at the proximal CEJ and the other end at the coronal week. Those that served as a negative control remained leakage-
level of the restoration. free at 30 days. A two-way ANOVA showed that there was no
The experimental and the control groups were divided into two significant difference in terms of coronal leakage between the two
equal subgroups, A and B. The teeth in subgroup A were inocu- organisms. Tukey’s test showed a significant difference in the
lated with S. faecalis, whereas teeth in subgroup B were inoculated coronal leakage between IRM and Cavit (p ⫽ 0.002) and between
with C. albicans. IRM and Dyract (p ⫽ 0.001). With both organisms, IRM started to
Before microbial inoculation, the filling materials in the access leak after 10 days, whereas Cavit and Dyract leaked after 2 weeks
cavity and the surrounding tooth structure were disinfected with (Table 1).
30% hydrogen peroxide for 1 to 2 min and swabbed with 5%
tincture iodine for approximately 2 min.
DISCUSSION

Preparation of the Microorganisms The importance of a well-sealed coronal restoration cannot be

overemphasized. Many in vitro methods have been used to eval-
The cultures of S. faecalis and C. albicans were obtained from uate the sealing quality of endodontic filling materials. These
the Microbiology Laboratory, King Faisal Specialist Hospital and methods are usually based on assessment of penetration of a tracer
Research Center, Riyadh, Kingdom of Saudi Arabia. The cultures along the obturated root canal. The tracers most often used are
were maintained on blood agar Petriplates (nutrient agar containing dyes, radioisotopes, bacteria, or bacterial by-products (9). Isotopes
10% defibrinated sheep blood) until needed. The experiments were and dye molecules, such as methylene blue, are much smaller than
performed in plastic tissue culture clusters (Heerbrugg, Switzer- bacteria and most of bacterial by-products. Although isotopes and
land) containing 24 wells each with an inner diameter of 15 mm. dyes may be good tools for comparing relative leakage, they do not
Each tooth was placed in the middle of a well and embedded in a simulate the types of microbial leakage that may occur clinically.
764 Balto Journal of Endodontics

S. faecalis is a facultative anaerobic Gram-positive coccus. This the interface, despite the antibacterial effect of the eugenol in the
bacterial species was chosen for use in this study because it is often cement.
involved in persistent endodontic infections (10) and is one of the Certainly this in vitro method of microleakage measurement
most resistant species found in the oral cavity, having the ability to cannot duplicate the environment that exists in vivo. However, the
survive under unusual environmental stresses (11). results from this study provide information that could aid the
The model used in this study was sensitive, simple, and prac- clinician in the selection of the best material to extend the leakage-
tical. Furthermore, the use of a culture on 0.5% Bacto-agar facil- free time period before the final restorative treatment is initiated.
itates the penetration of the organism through the bacteriological
This article (NF-1888) is registered with the College of Dentistry Research
medium. Center.
A total thickness of 3.5 mm for all restorations was used in this
The author expresses her appreciation to King Faisal Specialist Hospital
study to comply with the recommendation of Webber et al. (12), and Research Center for the opportunity to use the facility of Virology and
who found that a 3.5-mm thickness of Cavit was the minimum Infectious Disease Laboratory. Special thanks to Dr. Saad Al-Nazhan for his
advices and comments.
thickness necessary to prevent total leakage of the dye molecule.
The technique of placing these temporary filling materials into the Dr. Balto is affiliated with King Saud University, College of Dentistry,
access cavities might also have had some influence on the marginal Division of Endodontics, Riyadh, Kingdom of Saudi Arabia. Address requests
for reprints to Hanan Balto, BDS, MSc, King Saud University, College of
leakage. In this study, all the materials were introduced into the Dentistry, Division of Endodontics, P.O. Box 5967, Riyadh 11432, Kingdom of
access cavity by one operator to reduce the chances of a manipu- Saudi Arabia.
lative variable.
The results of this in vitro study indicate that Dyract and Cavit
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