15

ORIGINAL ARTICLE

Laila M Elbatal, Kariem Mostafa El Batouty, Maram Mohamed Obeid

Effect of Repeated Autoclaving on Cyclic Fatigue

of Three Nickel-Titanium Rotary Instruments
Laila M Elbatal, MD

Key words

autoclaving, cyclic fatigue, rotary nickel titanium instruments

Aim: The purpose of this study was to evaluate the effect of repeated autoclaving on the cyclic fatigue of NRT, Revo-S and ProTaper Universal rotary Ni-Ti instruments.
Methods: The instruments selected for this study (n = 120) were divided into three groups according
to the type of instrument: the NRT group, the Revo-S group and the ProTaper group. Each group
(n = 40) was further subdivided into four subgroups according to the number of sterilisation cycles:
subgroup A, in which the instruments were used without any exposure to sterilisation cycles; subgroup B (1 cycle); subgroup C (2 cycles); and subgroup D (3 cycles). All instruments were tested in
simulated canals with a 60 degree angle of curvature and a 5 mm radius. The number of cycles to
failure (NCF) was calculated and statistical analysis was carried out using the ANOVA test (P 0.05)
and Bonferronis post-hoc test. Scanning electron microscope analysis was used to analyse fractured
instruments.
Results: Statistically, signicant differences were found between the NCF mean values of the three
instruments when exposed to different sterilisation cycles (P < 0.001). There was no statistically signicant difference between the mean NCF amongst all subgroups in the ProTaper group. However,
in the Revo-S and NRT systems, a statistically signicant difference was found. The scanning electron
microscope analysis of the separated instruments of all groups did not show any signs of plastic deformation on the utes near the fracture site.
Conclusion: The number of sterilisation cycles has a variable effect on cyclic fatigue depending on
the type of Ni-Ti instrument.

Introduction
Despite the increased exibility of nickel-titanium
(Ni-Ti) rotary instruments, they still have an increased risk of fracture1,2. This is due to factors related to the instruments ability to resist fracture,
for example, the instruments taper, manufacturing
methods, degree of exibility, and the cross-section
and ute designs3,4, in addition to the anatomical
characteristics of the root canal anatomy, such as the
degree and the angle of curvature of the root canal5.

In the literature, there is a controversial debate

concerning the instruments mode of failure which is
due to either cyclic fatigue or torsional failure. Alapati et al6 assumed that instrument failure occurs due
to a single overload incident after a large number of
loading cycles that lead to a ductile fracture, rather
than cyclic fatigue. Cyclic fatigue occurs when the
instrument is rotating freely in a curved canal. The
maximum point of curvature is subjected to alternating compression/tension cycles, leading to metal
fatigue and subsequent fracture. Torsional failure

ENDO (Lond Engl) 2016;10(1):1522

Researcher, Department of
Endodontics, University of
Ain Shams, Cairo, Egypt

Kariem Mostafa
El Batouty, PhD
Assistant professor of
Endodontics, Department of
Endodontics, University of
Ain Shams, Cairo, Egypt

Maram Mohamed
Obeid, PhD
Lecturer of Endodontics,
Department of Endodontics,
University of Ain Shams,
Cairo, Egypt
Corresponding author:
Kariem Mostafa El Batouty
Assistant professor of
Endodontics,
Department of Endodontics,
University of Ain Shams,
7 Tayser tour,
Alanba Pola square,
Nasr city, Cairo, Egypt
Tel: +2/01000397491
Email:
kbatouty@hotmail.com

16

Elbatal et al

Effect of autoclaving on Nickel-Titanium files

occurs when part of the instrument is bounded in the

canal while the shaft keeps rotating, until fracture
occurs (taper lock)7. Whether rotary Ni-Ti instrument
fractures occur mainly due to the inuence of cyclic
fatigue, torsional failure, or their combined effect is
under debate. However, cyclic fatigue has proved
to be an important cause for instrument separation
during clinical use.
The sterilisation of root canal instruments before
use, as recommended by some manufacturers, or
after use is essential in endodontic practice8. Thermal
treatment is an important part of Ni-Ti manufacturing processes, therefore heat sterilisation procedures
and their possible effects on the mechanical properties of Ni-Ti instruments were of great interest in
several previous studies. A lot of controversy exists regarding the effect of autoclave sterilisation on
the cyclic fatigue of Ni-Ti instruments. Studies have
shown that heat sterilisation methods could increase
Ni-Ti cyclic fatigue resistance9,10. On the contrary,
a decrease in torsional fracture resistance after heat
sterilisation has also been reported for some Ni-Ti
instruments11,12. Bulem et al demonstrated that
Ni-Ti alloys exhibit higher corrosion rates at higher
temperatures13. According to Rapisarda et al, nickel
surface concentrations reduced during exposure to
autoclave sterilisation and titanium oxides increased,
conveying that oxidation may be taking place on
the surface, which was conrmed by auger electron
spectroscopy analysis14. However, some authors
found no relevant alterations in Ni-Ti torsional properties after heat sterilisation15,16.
Several clinical and laboratory studies investigated the cumulative effects of multiple sterilisation cycles on the incidence of cyclic fatigue and
instrument separation for Ni-Ti les. However, to
our knowledge, the effect of sterilisation on Revo-S,
ProTaper Universal and NRT Ni-Ti rotary instruments has not been investigated yet. Therefore, the
purpose of this study was to evaluate the effect of
autoclave sterilisation on the cyclic fatigue resistance
of Revo-S, ProTaper Universal and NRT. The null hypothesis was that repeated cycles of autoclave sterilisation had no effect on the Ni-Ti rotary instruments
used in this study.

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Materials and methods

All les under investigation (n = 120) were examined
under an operating microscope to detect manufacturing defects. All defective instruments were discarded and replaced. Instruments were divided into
three groups (n = 40) according to the type of instrument: the NRT group, which included NRT instruments, size 30 and 0.06 taper (Mani, Toshigi-Ken,
Japan); the Revo-S group, which included Revo-S
instruments, size AS30 and 0.06 taper (Micro Mga,
Besanon, France); and the ProTaper group, which included ProTaper Universal instruments size F3 (Dentsply Maillefer, Ballaigues, Switzerland). Each group
was further divided into four subgroups (n = 10/
subgroup) according to the number of sterilisation
cycles: subgroup A, the control group, in which the
instruments were used as received, without any exposure to sterilisation cycles; subgroup B, in which
the instruments were subjected to a single sterilisation cycle; subgroup C, in which the instruments
were subjected to two cycles; and subgroup D, in
which the instruments were subjected to three cycles. The instruments of subgroups B, C and D were
sterilised using a vacuum steam autoclave (MELAG,
Berlin, Germany) 23B+ at 134C, with a pressure of
2.1 bars, for a 3.5 min sterilisation cycle. The whole
cycle, including drying and cooling time, was 45 min
long. Instruments that underwent multiple autoclave
cycles were allowed to cool to room temperature
after sterilisation.
A cyclic-fatigue testing block (80 40 4 mm)
was custom-made with three simulated canals
grooved in with a 60 degree angle of curvature
and a 5 mm radius according to Pruetts method17.
The length of the curve was approximately 4.3 mm
from the apical end. Each groove was made as
an exact replica of the le to be tested, as recommended by Plotino et al18,19, with a 1 mm depth
increase and 0.1 mm relief, allowing the instrument
to rotate freely inside the articial canal. A positive
replica of each le was designed using Solidworks
3D design software and milled on a copper cube
using a computerised numerical-control machining
bench (Bridgeport VMC 760XP3, Hardinge Machine
Tools, Leicester, UK). The copper duplicates were
constructed according to the curvature parameters
selected for this study. Using these negative moulds,

Elbatal et al

the articial canals were made using a die-sinking

electrical-discharge machining process (Agietron Hyperspark 3, AGIE, Losone, Switzerland), in a stainless
steel block. The blocks were hardened through annealing then chrome-plated to ensure the reproducibility of the test. The articial canal was covered with
a tempered glass to prevent instrument slippage and
to allow for observation of the rotating instrument.
A custom-made cyclic fatigue-testing device was
designed and made specically for this study. The
steel base of the device was 23 cm long, 17 cm wide
and 2.5 cm thick. It weighed 8 kg ensuring maximum stability for functioning. Two main parts were
xed on top of the base. The rst part was a vice
(Fig 1) that holds the handpiece in position and can
be adjusted vertically via three screws. It was made
of galvanised steel except for half of the vice that was
made of stiff rubber to avoid scratching the handpiece. The second part was a plateau (Fig 1) designed
to receive the testing block on top of it. This plateau
was adjustable horizontally via two screws, which
were 10 cm long. Cyclic fatigue testing was done
using an endodontic motor (Endo-mate DT, NSK,
Tokyo, Japan) with a 20:1 reduction gear handpiece.
The motor was set according to the manufacturers
guidelines. Each groove was lubricated before testing
each le using synthetic handpiece lubricating oil.
All les were rotated at 250 rpm, and the torque
was as follows: ProTaper F3, 3.0 Ncm; NRT, 2.8 Ncm;
and Revo-S, 0.8 Ncm, according to each manufacturers recommendations. The les were monitored
visually, using 4 magnifying binocular loupes with a
light-emitting diode (LED), until fracture occurred. A
stopwatch was used to record the time each le took
to fracture. The number of cycles to failure (NCF)
was then calculated using the following equation:
NCF = T RPM / 60, where T is the time recorded
by the stopwatch in min and RPM (revolutions per
min) is the motor speed; the product of the two was
divided by 60 s.
Data were explored for normality using Kolmogorov-Smirnov and Shapiro-Wilk tests. All data showed
parametric distribution; therefore, two-way analysis
of variance (ANOVA) was used to test the signicance
of the effects of the system, the sterilisation cycles
and their interactions with NCF. Bonferronis post-hoc
test was used for pair-wise comparison between the
groups when ANOVA testing was signicant. The

Effect of autoclaving on Nickel-Titanium files

17

Fig 1 Longitudinal
SEM view after three
sterilisation cycles. a)
ProTaper le showing
machining grooves and
disruption of cutting
edges (the black arrow
shows the disruption
of the cutting edge).
b) Revo-S le showing
machining grooves, disruption of cutting edge
(black arrow), corrosion
of cutting edge and debris. c) NRT le showing
machining grooves.

signicance level was set at P 0.05. Statistical analysis was performed with the Statistical Package for
the Social Sciences (SPSS) Statistics software; Version
20 for Windows (IBM, New York, USA).

Scanning electron microscope

The separated instruments were further cleansed in
an ultrasonic bath of 70% alcohol for 2.5 min, in
order to remove the lubricating oil remnants and debris. Samples were examined under a scanning electron microscope (XL30, Philips, Eindhoven, Netherlands). Images captured with a scanning electron
microscope were categorised into three main groups:
1) longitudinal views of separated instruments;
2) fractographs with low magnication to reveal
the cross-section of the separated instruments; and
3) fractographs with high magnication to reveal

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Elbatal et al

Effect of autoclaving on Nickel-Titanium files

Table 1 Mean standard deviation (SD) of the NCF during cyclic fatigue testing for each Ni-Ti instrument, in terms of the
different subgroups.

P value

Groups

ProTaper

Revo-S

NRT

Subgroups

Mean SD

Mean SD

Mean SD

Control (subgroup A)

401.8

One cycle (subgroup B)

404.1

46.1

321.7

42.4

1188.0

Two cycles (subgroup C)

406.8

80.5

200.6

40.2

955.6

177.1

< 0.001*

Three cycles (subgroup D)

424.4

78.4

197.6

37.4

671.0

70.7

< 0.001*

P value

0.991

A
A
A

53.4

364.7

38.1

1280.0

151.8

< 0.001*

219.6

< 0.001*

0.039*

A
A

B
C

< 0.001*

*Signicant at P 0.05
Different small letters in the same row indicate signicant differences between systems. Different capital letters in the same column indicate
signicant differences between the different number of cycles.

the details and patterns of cyclic fatigue fractures.

The presence of plastic deformation adjacent to the
fractured surface was registered according to the
method described by Sattapan et al20.

Results
Mean values standard deviation, expressed as NCF,
are displayed in Table 1. The NRT system showed the
highest NCF mean value (1280.0 151.8) in subgroup A, followed by subgroup B (1188.0 219.6),
subgroup C (955.6 177.1) and subgroup D
(671.0 70.7). The Revo-S system showed the highest NCF mean value (364.7 38.1) in subgroup A,
followed by subgroup B (321.7 42.4), subgroup C
(200.6 40.2) and subgroup D (197.6 37.4). The
ProTaper system showed the lowest NCF mean value
(401.8 53.4) in subgroup A. The highest NCF mean
value was observed in subgroup D (424.4 78.4),
followed by subgroup C (406.8 80.5) and subgroup B (404.1 46.1).
A higher value of NCF was related to greater
resistance to cyclic fatigue fracture amongst the instruments tested. The one-way analysis of variance
indicated that there was a signicant difference between groups (P < 0.001). Comparing the results
between subgroups for the ProTaper system, the
Bonferronis post-hoc test showed no statistically signicant difference between the mean NCF amongst
all subgroups (P = 0.991). In the Revo-S (P = 0.039)
and NRT (P < 0.001) systems, statistically signicant
differences were found.

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The longitudinal views in the SEM analysis of

the separated instruments of each group showed no
signs of plastic deformation on the utes near the
fracture site (Fig 1). Manufacturing grooves were
evident in all samples. The corrosion site and the
disruption of the cutting edge were also visible. Fractographic analysis conrmed the cyclic fatigue pattern of the fracture with the presence of the three
characteristic zones: the crack initiation zone with
single or multiple crack origins, the slow crack propagation zone (striations) and the catastrophic (rough
dimpled) zone (Fig 2).

Discussion
The prevention of cross-contamination of infectious
diseases amongst clinicians, staff members, and patients is a major concern in dental practices. Aseptic techniques are especially important in root canal
treatments because microorganisms are the major
cause of endodontic failure21. To date, there is still
controversy regarding the effects of autoclave sterilisation on the mechanical properties of rotary Ni-Ti
instruments. Therefore, the aim of this study was to
evaluate the effects of autoclave sterilisation on the
cyclic fatigue of ProTaper Universal, Revo-S and NRT
instruments.
The chosen instruments indicated several differences, including the following: 1) the taper, as
the ProTaper instruments had a variable taper from
9% at D0 to 3% at D15; while Revo-S and NRT had
a constant taper of 6%; 2) the cross section that

Elbatal et al

varied from a modied convex triangle in ProTaper

to an asymmetrical shape in Revo-S and a modied rectangle in NRT; 3) the centre of rotation, as
only Revo-S instruments had an offset body centre;
4) the length of the working part for ProTaper was
15 mm, for Revo-S it was 18 mm, and for NRT it
was only 11 mm; 5) the fact that only NRT instruments had radial lands. However, this comparative
study was designed to provide clinically relevant
ndings for practitioners. In fact, it is difcult to
restrict the comparison according to standardisation, given the diversity of rotary instruments in the
market today.
Casper et al22 subjected instruments to seven
sterilisation cycles; however, the present study
followed the protocol of three sterilisation cycles,
as in several previous studies16,23,24. This sequence
was selected to incrementally examine the most
clinically relevant initial cycles, without proposing an
extreme number of cycles, as in a study conducted
by Seago et al, in which instruments were subjected
to 10 cycles25.
According to the results of this study, the null
hypothesis indicating that sterilisation has no effect
on Ni-Ti rotary les was rejected, except in the case
of the ProTaper system. The results of this study
showed that the type of Ni-Ti rotary system, the
sterilisation cycles and the interaction between these
two variables had statistically signicant effects on
the NCF. The difference in the effects of sterilisation
on each system is most probably due to different
manufacturing processes and different metallurgy.
This nding agrees with previous studies that reported different effects of sterilisation on different
systems7,23,26,27.
The NRT instruments showed the highest mean
NCF when compared to ProTaper and Revo-S instruments amongst all subgroups. This result may be attributed to the shorter working part and long shaft,
in addition to the rectangular-based cross-sectional
shape of the NRT instruments28. Pirani et al attributed the increased resistance to cyclic fatigue in NRT
instruments, to the fact that the inclusions (titanium
carbides and Ni-Ti oxides) in NRT instruments are
smaller and much more concentrated29. These inclusions are formed during the vacuum melting of
the Ni-Ti alloy in a graphite crucible. According to
Beremin, crack propagation is enhanced by large

Effect of autoclaving on Nickel-Titanium files

19

Fig 2 Fractograph of
the three systems under
investigation after 3
sterilisation cycles and
fatigue failure. a) ProTape le showing cracks
(black arrow) following
manufacturing grooves
(650x magnications).
b) Revo-S le showing the ductile pattern
of fracture with voids
(black arrow), dimples
and cracks (1000x
magnication). c) NRT
le showing two levels
of fracture planes (black
arrow) at 1500x magnication.

non-metallic inclusions that act as nucleation sites

of ductile fractures30. Small inclusions could slow
or stop propagation of a crack by acting as pinning
points. A pinning point in the material serves to halt
a dislocation movement, requiring the application
of a greater amount of force, in order to overcome
the barrier31.
The ProTaper system subgroups were signicantly more resistant to fracture after two or three
sterilisation cycles than the Revo-S counterparts.
This nding contradicts previous results indicating
that instruments with a larger size or larger taper

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Elbatal et al

Effect of autoclaving on Nickel-Titanium files

fracture more readily, as demonstrated by several

authors17,32,33. Hilfer et al23 described a similar nding, in which GTX size 20 taper 0.06 (Dentsply Tulsa,
Oklahoma, USA) instruments had a signicantly
higher NCF than the GTX size 20 taper 0.04 instruments. This nding may be attributed to the results
observed by Zhang et al34, who indicated that the
cross-sectional design of the instrument had a more
signicant impact on the development of stress than
its size or taper. Therefore, the modication in the
cross-section of ProTaper F3 rendered the instrument
more resistant to cyclic fatigue. However, this theory
contradicts the claims of the manufacturer of Revo-S
instruments, who insist that the asymmetrical crosssection and offset centre of rotation, where the three
cutting edges of the instrument are located on three
different radii, increase the exibility of these instruments. The increased cyclic fatigue resistance of ProTaper instruments, in comparison to Revo-S found in
the current study contradicts the results of da Silva
et al, who proposed that electrochemically polished
instruments keep their integrity for a signicantly
higher number of cycles35. Saghiri et al showed that
electrical treatment with 12-V DC was effective in
restoring the resistance of Ni-Ti instruments to cyclic fatigue. They found that the scanning electron
microscopy images and fractographs of samples exposed to 12-V electrical treatment showed a more
regular texture over the surface with less dimpling on
the fracture site36.
The effect of the number of sterilisation cycles on
the mean NCF of the tested Ni-Ti systems showed
no statistically signicant difference in the ProTaper
group. This may be due to the fact that the autoclaving process cannot provide enough energy to
enable any change in the crystalline structure of the
ProTaper instruments, rendering them less susceptible to fatigue crack nucleation and propagation. In
addition, the sterilisation temperature used in this
study was 134C; it is well documented in the literature that the Martinsite phase undergoes no crystalline changes at temperatures below 170C. From
170C to 640C, as the temperature increases, an
increasing amount of the crystal lattice reorders into
the Austenite phase. Above 640C, all crystals are
considered to have entered the Austenite phase37.
Thus, it is unlikely that heat treatment resulting from
autoclave sterilisation could have signicant effects

ENDO (Lond Engl) 2016;10(1):1522

on the reordering of the crystalline phases of Ni-Ti

instruments.
This result contradicts what was observed in the
fractographs, in which multiple cracks following the
manufacturing grooves were found to be deepened
after three sterilisation cycles, when compared to
when they were received and after two sterilisation
cycles.
The mean NCFs of the Revo-S and the NRT system were affected by the number of sterilisation cycles. This result differs from the ndings of many
reports in the sterilisation. For example, Silvaggio and
Hickes et al38 concluded that heat sterilisation of rotary Ni-Ti instruments (ProFile Series 29, 0.04 taper
[Dentsply Tulsa Dental]), which was up to 10 cycles
did not increase the likelihood of instrument fracture. In addition, Mize et al16 and Hilt et al15 showed
that heat treatment as a result of autoclave sterilisation did not extend the lifespan of Ni-Ti instruments.
Moreover, Melo et al39 demonstrated that the size
of the instrument, which determines the maximum
strain amplitude during cyclic deformation, was the
most important factor controlling fatigue resistance:
also the application of ve sterilisation procedures in
dry heat increased the average number of cycles to
failure of unused instruments by approximately 70%.
Sundaram et al24 found that Revo-S instruments
were not affected by autoclave sterilisation. This contradiction between the current study and the earlier
research may be due to the difference in sterilisation
temperature. Another possible cause is that Sundaram et al investigated a Revo-S instrument with a
smaller tip diameter (size AS25). Alexandrou et al26
also found that NRT was not affected by sterilisation
and that surface roughness was increased signicantly after 11 sterilisation cycles and suggested that
the NRT system is capable of superelastic behaviour
under clinical conditions.
With regard to the scanning electron microscope evaluation of all groups, an increase in surface
roughness was found in all sterilised instruments
when compared to the condition in which they
were received. This might be expected due to the
increased concentration of oxygen with repeated
sterilisation, leading to the formation of a passive
oxide layer (TiO2) on their surfaces40. Fractographic
examination of the samples conrmed that the
separation of Ni-Ti rotary instruments due to cyclic

Elbatal et al

fatigue occurs in a ductile manner that resembles a

brittle fracture at the end. Longitudinal examination
showed that all samples have machine grooves that
act as stress-concentration points and possible crack
origins.
In terms of the ProTaper system, striations characteristic of cyclic fatigue fracture were observed.
These striations indicate the slow progression rate of
the advancing crack, which could explain why the
NCF of the ProTaper system was not affected by the
number of sterilisation cycles. In the Revo-S group,
larger dimples were noticed in les sterilised for three
cycles, supporting the conclusion that the NCF decreased when the number of cycles increased. In
addition, the barely observable slow-overload zone
indicated the les short fatigue life. A previous
study investigating NRT and ProTaper instruments
showed that the inclusions in NRT instruments are
much more concentrated and smaller than those in
ProTaper instruments29. This would suggest higher
fatigue resistance in NRT than ProTaper instruments,
as the abundant small inclusions could slow or stop
the propagation of a crack. However, other studies
showed that crack propagation was enhanced by
large non-metallic inclusions that act as nucleation
sites of ductile fracture30.
A limitation of the current study is that the effect
of previous clinical usage of the instruments, in combination with subsequent heat sterilisation was not
examined. In addition, evaluation was carried out
in simulated canals lacking clinical effects. Clinical
procedures, especially in curved canals, exert complex stresses on Ni-Ti rotary instruments resulting in
reduced cyclic fatigue resistance41.

Conict-of interest
The authors conrm no nancial afliation (e.g. employment, direct payment, stock holdings, retainers,
consultantships, patent licensing arrangements or
honoraria) or involvement with any commercial organisation with a direct nancial interest in the subject or materials discussed in this manuscript, nor have
any such arrangements existed in the past 3 years.
Any other potential conict of interest is disclosed.

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Conclusions
Under the conditions of the present study it can
be concluded that the cyclic fatigue resistance of
both NRT and Revo-S Ni-Ti rotary instruments were
affected by an increase in the number of sterilisation cycles. However, the cyclic fatigue resistance of
ProTaper Universal instruments was not affected by
the same increase in cycles. Furthermore, an instruments increased surface roughness corresponded
with the number of autoclave cycles to which the
instrument was exposed.

Effect of autoclaving on Nickel-Titanium files

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