Endodontology

Jesmy K. Antony, Liza George
1
, Josey Mathew
1
, Aleesha Joy
1
Department Of Conservative Dentistry and Endodontics, Kerala University of Health Sciences, 
1
Department of Conservative 
Dentistry and Endodontics, Annoor Dental College and Hospital, Kerala University of Health Science, Ernakulam, Kerala, India
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DOI:
10.4103/endo.endo_176_21
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How to cite this article: Antony JK, George L, Mathew J, Joy A. Sealing 
ability of mineral trioxide aggregate, Biodentine, and EndoSequence RRM 
putty used as retrograde restorative material: An 
in vitro bacterial leakage 
model study. Endodontology 2022;34:16-21.
Submitted: 22‑Sep‑2021 Revised: 15‑Dec‑2021
Accepted: 17‑Jan‑2022 Available Online: 25‑Mar‑2022
[Downloaded free from http://www.endodontologyonweb.org on Thursday, April 7, 2022, IP: 195.158.14.172]

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17
Antony, et al.: Bacterial microleakage of bioceramic root‑end filling materials
Endodontology / Volume 34 / Issue 1 / January‑March 2022
microorganism and their byproducts out of the periapical 
region.
[4]
Numerous materials have been proposed for root‑end fillings 
of which mineral trioxide aggregate (MTA) is the first calcium 
silicate‑based bioceramic material which is successfully 
used as a retrograde restorative material. Its excellent 
biocompatibility, sealing ability, cementogenesis, and 
several other advantages made it a landmark in the history 
of endodontics.
[5]
Biodentine is similar to MTA in terms of 
basic composition, and the manufacturers claim that the 
modification in its powder composition by addition of setting 
accelerators and softeners largely improved the physical 
properties of the material, making it more user‑friendly.
[6]
EndoSequence Root Repair Material (RRM) (ESRRM) putty is 
a premixed bioceramic material available as injectable paste 
or putty consistency. Endo Sequence RRM, with more of a 
putty consistency, is reported as being easier to physically 
manage and to place where it is necessary.
[7]
The aim of this study was to compare and evaluate the sealing 
ability of three retrograde filling materials ProRoot MTA, 
Biodentine, and EndoSequence RRM putty using bacterial 
leakage models.
MATERIALS AND METHODS
This in vitro study was completed in the Department of 
Conservative Dentistry and Endodontics (Ethical committee 
approval no: IHEC Ref No: 018‑A/04). Forty single‑rooted 
human premolar teeth were collected. The collected teeth 
were disinfected with 6% hydrogen peroxide,
[8]
followed by 
sample preparation.
The samples were decoronated and the lengths of all the 
roots were standardized to 16 mm. Cleaning and shaping 
was done along with intermittent irrigation using 2 ml of 
5% of sodium hypochlorite (Chemdent) alternating with 
17% ethylenediaminetetraacetic acid (Meta MD Cleanser, 
Biomed).
[9]
Irrigation was done using side‑vented needles. 
The apical preparation was done up to 40 #Kfile (Mani, INC.) 
and step back preparation was done up to size 55#K file 
(Mani, INC.).
The roots were sectioned 3 mm from the apex. The root end 
cavities were prepared using ultrasonic tips (Woodpecker 
EMS Endo Scaler Tip E3D) with a standard measurement 
of 3 mm depth and 1 mm diameter. The preparations were 
done using ultrasonic tips on which 3 mm markings were 
made.
Specimens were then randomly divided into four groups:
• Group 1 – ProRoot MTA (n = 10)
• (Dentsply Sirona Endodontics)
• Group 2 – Biodentine (n = 10)
• (Septodont, Saint Maur des Fosses, France)
• Group 3 – EndoSequence root repair putty (n = 10)
• (Brasseler USA, Savannah, GA)
• Group 4 – Control group– without any filling (n = 10).
A gutta percha point of 60 size and 2% taper was then placed 
in the root canal just above the root end cavity against which 
the samples condensed with the tested materials.
Retrograde cavities were filled with the three different materials 
according to the guidelines. The samples were then coated 
with two coats of three different colored nail varnish except 
on the apical 3 mm. The restorations were confirmed for the 
absence of voids with radiographs. Specimens were then stored 
for a period of 3 weeks in the presence of moisture in order 
to achieve the complete setting of the materials. Specimens 
were then mounted on the Enterococcus faecalis (ATCC strain 
29,212) bacterial leakage models in which 2–3 mm of the root 
tip was in contact with the brain heart infusion (BHI) broth. 
The whole models were placed in ultraviolet (UV) chamber 
for sterilization (Uniclave 5G UV chamber BioWarrior). Each 
root canal was then aseptically inoculated with 15 
µL freshly 
prepared culture and incubated in a humid environment at 37°C 
for 6 weeks. The turbidity (optical density [OD]) generated by 
the bacteria present in the BHI broth was measured using a UV 
spectrophotometer. The measurements were taken at every 
7
th
day for a 6‑week period. The sample obtained from the broth 
was then Gram stained for confirmation of cell morphology.
The data were subjected to statistical analysis using one‑way 
ANOVA to compare the difference in OD value between the 
materials in the 6‑week period. Repeated‑measure ANOVA 
was used to assess the changes in the OD value obtained 
for each material over 1–6 weeks. Post hoc was done using 
Bonferroni test. P < 0.05 was considered statistically 
significant.
IBM SPSS Version 25 – International Business Machines 
Corp. (IBM) 2017 Statistical package for Social Science (SPSS) 
statistics Version 25 (Armonk, NY: IBM Corp).
RESULTS
One‑way ANOVA was used to compare the OD value between 
the materials in each week (baseline, 1
st
, 2
nd
, 3
rd
, 4
th
, 5
th
, 
and 6
th
). There was a statistically significant difference 
between the materials in all the 6 weeks [Table 1].