Original Article

A comparative evaluation of the effect of 5.25%

sodium hypochlorite and 2% chlorhexidine on the
surface texture of Gutta‑percha and resilon cones
using atomic force microscope
Mahima Tilakchand, Balaram Naik, Abhijith S Shetty
Department of Conservative Dentistry and Endodontics, S.D.M. College of Dental Sciences and Hospital, Sattur, Dharwad, Karnataka, India

Abstract
Aims & Objectives: The purpose of this study was to investigate the effects of 5.25% sodium hypochlorite (NaOCl) and 2%
chlorhexidine (CHX) on Gutta‑percha and Resilon cones using an atomic force microscope (AFM).
Materials and Methods: Gutta‑percha cones (n = 15) and Resilon cones (n = 15) were cut 3 mm from their tip, attached to a glass
slide with cyanoacrylate glue and immersed in 5.25% NaOCl and CHX for 1, 5, 10, 20 and 30 min. Five each of Gutta‑percha and
Resilon cones not treated with any disinfectant were used as control. The analysis of the surface topography was performed on the region
between 1 and 2 mm from the tip using the AFM. The root mean square (RMS) parameters for contact mode imaging were measured.
The differences between RMS values were tested by SPSS‑16.0 version statistical software [IBM SPSS (Statistical Product and Service
Solutions) Data Software, Chicago, US] using Kruskal‑Wallis ANOVA, Mann‑Whitney U‑test and Wilcoxon matched pairs test.
Results: There was no deterioration in the surfac e topography of Gutta‑percha and Resilon when treated with 2% CHX in comparison
to baseline (P < 0.05). Resilon exhibited no deterioration in topography when immersed in 5.25% NaOCl. There was a significant
decrease in the mean RMS values of Gutta‑percha treated with NaOCl from the control at time intervals of 1, 5, 10, 20 and 30 min.
Keywords: Chlorhexidine; Gutta‑percha; Resilon; sodium hypochlorite and atomic force microscope

INTRODUCTION provided sterile by the manufacturer. Even then, they should

Obturation is an important procedure in endodontic
treatment. Gutta‑percha, is at present, the most commonly
used material for the obturation of the root canal system.
It is biocompatible, dimensionally stable, radiopaque and
thermoplastic. Recently, a new thermoplastic, filled polymer
that has the potential to challenge Gutta‑percha as a root
filling material has been introduced. The thermoplasticity
of Resilon is provided by polycaprolactone which is a
biodegradable polyester with a moderately low melting
point. Its bond ability is derived from the inclusion of resin
with methacryloxy groups.[1]
be chemically disinfected chair side before use in patients.[2]
A large variety of chemical disinfectants have been
used to sterilize Gutta‑percha cones before root
canal filling. These include sodium hypochlorite
(NaOCl), glutaraldehyde, alcohol, chlorhexidine (CHX),
hydrogen peroxide, polyvinylpyrrolidone iodine and
MTAD, Mixture of doxycycline, 10% citric acid and
tween‑80. A 5.25% NaOCl solution is recognized as an
effective decontaminant. It was found to be effective
against a variety of Gram‑positive, Gram‑negative and
spore‑forming microorganisms.[3‑5]

According to concepts of infection control, every instrument Studies have shown, that as a strong oxidizing agent, 5.25%
and material placed within the root canal should be sterile. NaOCl causes extreme topographic alterations in the cones,
Core filling materials, such as Gutta‑percha and Resilon which might be indicative of aggressive deterioration.
cones should be sterilized before placing them within Moreover, crystal formation on the surface of Gutta‑percha
the root canal. These materials are thermolabile and are
cones has been identified after rapid sterilization with 2.5%
Address for correspondence:
Dr. Abhijith S Shetty, Department of Conservative Dentistry and Access this article online
Quick Response Code:
Endodontics, S.D.M. College of Dental Sciences and Hospital,
Website:
Sattur, Dharwad ‑ 580 009, Karnataka, India. www.jcd.org.in
E‑mail: mahima702002@yahoo.co.in
Date of submission: 10.09.2013 DOI:
Review completed: 24.09.2013 10.4103/0972-0707.124102
Date of acceptance: 21.10.2013

18 Journal of Conservative Dentistry | Jan-Feb 2014 | Vol 17 | Issue 1

 Tilakchand, et al.: Evaluation of irrigants on surface texture of obturation materials using AFM

and 5.25% NaOCl. Therefore an effective, but safer chemical recorded in the contact mode operation of the AFM (M/S
is desirable for disinfecting Gutta‑percha cones. It has been Digital Instruments, USA). Typical AFM probes (length
shown that a 30 min exposure to 2% CHX does not change 115 µ and curvature radius 10 nm) mounted on silicon
the properties of Gutta‑percha. This suggests that CHX may nitride cantilever with spring constant of 0.58 N/m were
be less prejudicial to the structure of Gutta‑percha. However, used. Resonance frequency nominal of 57 kHz was used.
the effect of 2% CHX on the integrity of Gutta‑percha points Scanned areas (5 Hz) were a perfect square.
has not been fully investigated.
AFM images were processed with the Scanning Probe
The advent of atomic force microscope (AFM), a member of Microscope (SPM) lab 4.0 Software [BME (BioMedical
the scanning probe microscopes family; has opened several Engineering) 499/Biostat 642] and analyzed with WSxM
applications in surface studies of different materials.
The key principle of the AFM is the probing of a sample
surface with a small tip attached to a flexible cantilever.
The detection of the several parameters between the
tip and the sample interactions provides qualitative and
quantitative information about the sample.[6,7]

The proposed study evaluates the effect of 2% CHX and

5.25% NaOCl on surface topography of Gutta‑percha and
resilon cones, using an AFM.

MATERIALS AND METHODS

This study was conducted in the Department of

Conservative Dentistry, SDM college of Dental sciences
and Hospital, Dharwad. The purpose of this study was to
evaluate the effect of 5.25% NaOCl and 2% CHX on surface
texture of Gutta‑percha and Resilon cones.

15 40 size Gutta‑percha (Dentsply) and Resilon (Epiphany)

cones each were cut 3 mm from their tip and attached to a
glass slide measuring 5 mm × 5 mm using the rapid setting Figure 1: Atomic force microscope image of Gutta-percha
cyanoacrylate glue. These glass slides were prepared with the
help of a diamond glass cutter. Following these procedures
the samples were divided into four groups as following:

• Group I: Five Gutta‑percha cones were immersed in

5.25% NaOCl (Rankem) for 1, 5, 10, 20 and 30 min
respectively.
• Group II: Five Gutta‑percha cones were immersed
in 2% CHX (V‑Consept) for 1, 5, 10, 20 and 30 min
respectively.
• Group III: Five Resilon cones were immersed in 5.25%
NaOCl for 1, 5, 10, 20 and 30 min respectively.
• Group IV: Five Resilon cones were immersed in 2% CHX
for 1, 5, 10, 20 and 30 min respectively.

Five each of Gutta‑percha and Resilon cones, which were

not treated with any disinfectant, were used as control.
After immersion, the samples were thoroughly rinsed with
5 ml of distilled water and the specimen was dried with filter
paper. Each slide was viewed under an AFM using contact
mode and the analysis was performed at a region located
between 1 and 2 mm from the tip of the cones [Figure 1 and,
Figure 2]. AFM images of the Gutta‑percha samples were Figure 2: Atomic force microscope image of Resilon

Journal of Conservative Dentistry | Jan-Feb 2014 | Vol 17 | Issue 1 19

 Tilakchand, et al.: Evaluation of irrigants on surface texture of obturation materials using AFM

SPM software 2.0. For the purpose of comparison, root
mean square (RMS) roughness was chosen to investigate
the topographic alteration of Gutta‑percha and resilon
cones.
Statistical analysis
Statistical analysis was performed using statistical
software‑SPSS‑16.0 version using Kruskal‑Wallis ANOVA,
Mann‑Whitney U‑test and Wilcoxon matched pairs test.
RESULTS
There was a significant decrease in the mean RMS values
of group I from the baseline (P < 0.05) at time intervals of
1 min, 5 min, 10 min, 20 min and 30 min [Table 4].
There was a significant difference in the RMS values
between the groups.
There was a significant difference (P < 0.05) between group
I and group II at time intervals of 5, 10, 20 and 30 min.
There was no significant difference between group III and
IV at all-time intervals [Table 5].

The following tables show the mean values of RMS for CMI Comparison of time intervals in each group by
profiles: Wilcoxon matched pairs test
Showed that there was a significant difference (P < 0.05) in
For group I RMS values were significantly lower when the RMS values for group I at all the different time intervals
compared to the control with respect to 1 min, 5 min, 10 min, when compared to the baseline. There was a significant
20 min and 30 min of exposure to NaOCl [Tables 1 and 2]. difference in the RMS value for group II at time period of
Group II, III and IV showed no significant difference in the 1 min, 5 min and 20 min.
RMS values with different time exposures when compared
with the control [Tables 2 and 3]. DISCUSSION
Table 1: Baseline RMS values for CMI profile The prevention of contamination of the root canal system is
Baseline RMS values an important aspect of endodontic therapy. Materials such
Gutta‑percha Resilon as Gutta‑percha and resilon are manufactured under aseptic
95 95 conditions, but they can be contaminated by handling, by
86 96 aerosols and by physical sources during the storage process. The
91 98 sterilization by conventional processes using moist or dry heat
87 95
88 99 is not possible because of their thermoplastic characteristics.
Mean 89.40 96.60 Therefore, a rapid chair‑side chemical disinfection is needed.[8]
SD 3.65 1.82
RMS: Root mean square, CMI: Contact mode imaging Gutta‑percha cones exposed in clinics can be
contaminated (19.4%) and the contaminating bacteria are all
Table 2: RMS values for CMI profiles of group I, II at vegetative rather than resistant bacterial spores. Gutta‑percha
different time intervals cones might also play a role in initiating biofilm infections in
Group I (min) Group II (min) cases of over obturation showing periapical lesions.[3]
1 5 10 20 30 1 5 10 20 30
82.9 73.6 54.1 40.6 29.8 78.9 89.3 81.9 86.7 77.9 Table 4: Comparison of four groups by Kruskal Wallis
78.4 74.6 58.4 38.1 31.2 82.3 83.7 76.5 84.6 92.2 ANOVA
79.3 79.1 51.7 45.4 36.6 79.1 88.2 84.9 83.8 89.5
75.8 68.9 61.5 44.3 34.8 84.9 86.7 86.7 86.5 88.4 Group I Group II Group III Group IV
83.2 66.1 63.6 43.7 33.9 87.4 75.5 89.2 79.9 83.4 Control 89.4±3.647 89.4±3.647 96.6±1.816 96.6±1.816
Mean 79.92 72.46 57.86 42.42 33.26 82.52 84.68 83.84 84.30 86.28 1 minute 79.92±3.135 82.52±3.685 93.6±4.980 93.6±4.037
SD 3.13 5.08 4.96 3.00 2.75 3.69 5.55 4.89 2.75 5.67 5 minutes 72.46±5.077 84.68±5.546 95±5.657 95.2±4.919
RMS: Root mean square, CMI: Contact mode imaging 10 minutes 57.86±4.964 83.84±4.891 95.6±4.929 94.2±5.933
20 minutes 42.42±3.001 84.3±2.752 94±4.123 96.6±5.272
30 min 33.26±2.745 86.28±5.666 97.2±2.863 96.6±4.159
Table 3: RMS values for CMI profiles of group III, IV at
different time intervals ANOVA: Analysis of Variance

Group III (min) Group IV (min)

Table 5: Pair wise comparison of four groups by
1 5 10 20 30 1 5 10 20 30
Mann‑Whitney U‑test
87 97 98 97 99 98 95 102 99 99
98 89 89 94 97 96 101 98 88 97 Groups BL 1 min 5 min 10 min 20 min 30 min
99 101 102 87 95 88 89 89 102 101 I vs II 1.0000 0.3472 0.0163 0.0090 0.0090 0.0090
93 89 96 97 94 95 99 95 98 96 I vs III 0.0163 0.0090 0.0090 0.0090 0.0090 0.0090
91 99 93 95 101 91 92 88 96 90 I vs IV 0.0163 0.0090 0.0090 0.0090 0.0090 0.0090
Mean 93.60 95.00 95.60 94.00 97.20 93.60 95.20 94.20 96.60 96.60 II vs III 0.0163 0.0163 0.0283 0.0163 0.0090 0.0090
SD 4.98 5.66 4.93 4.12 2.86 4.04 4.92 5.93 5.27 4.16 II vs IV 0.0163 0.0090 0.0163 0.0283 0.0090 0.0163
RMS: Root mean square, CMI: Contact mode imaging III vs IV 1.0000 0.9168 0.9168 0.6761 0.1745 1.0000

20 Journal of Conservative Dentistry | Jan-Feb 2014 | Vol 17 | Issue 1

 Tilakchand, et al.: Evaluation of irrigants on surface texture of obturation materials using AFM
NaOCl solutions of 0.5‑5.25% have a recognized tradition of
use as an irrigating solution in endodontic practice.[9] 5.25%
NaOCl solution is recognized as an effective decontaminant.[5]
It has antibacterial and sporocidal activity related to the
liberation of active chlorine.[3]
A 5.25% NaOCl solution is routinely used as an effective
chemical for rapid decontamination of Gutta‑percha cones
before use, killing microorganisms in 1 min of exposure
time. Studies have shown, that as a strong oxidizing agent,
5.25% NaOCl causes extreme topographic alterations in the
cones, which might be indicative of aggressive deterioration.
Moreover, crystal formation on the surface of Gutta‑percha
cones has been identified after rapid sterilization with 2.5%
and 5.25% NaOCl. Valois et al. reported that 5.25% NaOCl
resulted in surface deterioration of Gutta‑percha cone and that
it was due to the loss of Gutta‑percha cone components by
the oxidizing agent (NaOCl). Therefore an effective, but safer
chemical is desirable for disinfecting Gutta‑percha cones.[1,3,4]
Currently, there is an increasing interest in the antibacterial
activity of CHX in endodontic practice. It has antibacterial
properties with broad spectrum and relatively low
toxicity. Nevertheless, it has been reported that it is
ineffective against spores except at high temperatures.
CHX has additional properties such as substantivity
and biocompatibility compared with NaOCl. CHX is less
prejudicial to the structure of Gutta‑percha. However, the
effect of 2% CHX on the integrity of Gutta‑percha points has
not been fully investigated.[2‑4,8,9]
The scanning electron microscope (SEM) has long been the
standard means of investigation of surface characteristics
of Gutta‑percha cones. They needs to be operated under
vacuum and uses an electron beam to give a 2‑Dimensional
“photographic” image of the sample covered with a layer
of a conductive material such as gold; but cannot provide
quantitative data regarding the topography. SEM and AFM
resolve structures down to the nanometer scale. However,
the image formation mechanism is quite different,
resulting in different types of information. The AFM is now
a well‑established technique to study quantitatively the
topography of a wide variety of materials.[6]
The present study shows decrease in the RMS values
for Gutta‑percha when exposed to 5.25% NaOCl for 1,
5, 10, 20 and 30 min and no significant alteration in the
surface topography of Gutta‑percha and resilon when
exposed to CHX. Resilon showed no changes in the
surface topography when exposed to NaOCl as well. The
result of the study is consistent with the study conducted
by Valois et al. indicating a deteriorative tendency of the
intermediate (2.5%) and high (5.25%) NaOCl concentrations
on Gutta‑percha cone surface topography.[9]
Pang et al. detected that Gutta‑percha cones soaked in 5.25%
Journal of Conservative Dentistry | Jan-Feb 2014 | Vol 17 | Issue 1
NaOCl showed a precipitate with a cuboidal crystal structure
over the entire surface with particular concentration in the
defective or folded area of cones. A large variety of rounded
structures were detected in the deeper strata of the defective
area of the Gutta‑percha cones, which are lysis products of
some components of Gutta‑percha cones. They also showed
significant effects on elongation rate.[3]
Isci et al. found that CHX had no effect on the surface
topography, whereas NaOCl caused alteration of the surface
topography of the Gutta‑percha. Similarly, Seabra Pereira et al.
observed that when 2% CHX was used, no topographic
changes were observed with or without rinse.[2,6]
CONCLUSION
Based on the results of this study, it can be inferred that 5.25%
NaOCl causes severe surface topographic changes of the
Gutta‑percha points. There is no effect on surface topography
of Gutta‑percha when treated with 2% CHX. Resilon showed
no alteration in surface topography when treated with either
5.25% NaOCl or 2% CHX. Thus, the study shows that 2% CHX
could be a better alternative to 5.25% NaOCl for chair‑side
disinfection of Gutta‑percha and resilon cones.
REFERENCES
1. Gomes BP, Berber VB, Montagner F, Sena NT, Zaia AA, Ferraz CC, et al.
Residual effects and surface alterations in disinfected gutta‑percha and
Resilon cones. J Endod 2007;33:948‑51.
2. Seabra Pereira OL, Siqueira JF Jr. Contamination of gutta‑percha and
Resilon cones taken directly from the manufacturer. Clin Oral Investig
2010;14:327‑30.
3. Pang NS, Jung IY, Bae KS, Baek SH, Lee WC, Kum KY. Effects of
short‑term chemical disinfection of gutta‑percha cones: Identification
of affected microbes and alterations in surface texture and physical
properties. J Endod 2007;33:594‑8.
4. Valois CR, Silva LP, Azevedo RB. Effects of 2% chlorhexidine and 5.25%
sodium hypochlorite on gutta‑percha cones studied by atomic force
microscopy. Int Endod J 2005;38:425‑9.
5. Short RD, Dorn SO, Kuttler S. The crystallization of sodium hypochlorite
on gutta‑percha cones after the rapid‑sterilization technique: An SEM
study. J Endod 2003;29:670‑3.
6. Isci S, Yoldas O, Dumani A. Effects of sodium hypochlorite and
chlorhexidine solutions on Resilon (synthetic polymer based root canal
filling material) cones: An atomic force microscopy study. J  Endod
2006;32:967‑9.
7. Valois CR, Silva LP, Azevedo RB, Costa ED Jr. Atomic force microscopy
study of gutta‑percha cone topography. Oral Surg Oral Med Oral Pathol
Oral Radiol Endod 2004;98:250‑5.
8. Prado M, Gusman H, Gomes BP, Simão RA. The importance of final rinse
after disinfection of gutta‑percha and Resilon cones. Oral Surg Oral Med
Oral Pathol Oral Radiol Endod 2011;111:e21‑4.
9. Valois CR, Silva LP, Azevedo RB. Structural effects of sodium
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How to cite this article: Tilakchand M, Naik B, Shetty AS. A
comparative evaluation of the effect of 5.25% sodium hypochlorite
and 2% chlorhexidine on the surface texture of Gutta-percha and
resilon cones using atomic force microscope. J Conserv Dent
2014;17:18-21.
Source of Support: Nil, Conflict of Interest: None declared.
21
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