Arılı Öztürk et al.
BMC Oral Health (2025) 25:352 BMC Oral Health
[Link]
RESEARCH Open Access
Effect of various final irrigation solutions
at different temperatures on dentin tubule
penetration of bioceramic-based root canal
sealer
Esra Arılı Öztürk1, Burhan Can Çanakçi1* and Ceren Turan Gökduman1
Abstract
Background To evaluate the effect of 4 final irrigation solutions at different temperatures on dentin tubule
penetration of bioceramic-based sealer using the warm vertical compaction method with a laser scanning confocal
microscope (CLSM).
Methods In total, 180 human maxillary incisors were prepared. Different final irrigation solutions [distilled water,
ethylenediaminetetraacetic acid (EDTA), peracetic acid, or 1-hydroxyethylidene-1,1-bisphosphonate (HEBP)] at
various temperatures (25, 36, or 60˚C) were used. Obturation was performed via warm vertical compaction using a
bioceramic-based sealer. Three horizontal sections were obtained from each specimen and observed using CLSM.
The maximum penetration depth, penetration area, and penetration percentage were measured. Comprasions were
performed using three-way robust ANOVA, and Bonferroni correction (P < 0.05).
Results The effects of different solutions and temperatures on the maximum sealer penetration depth were similar
(P > 0.05). Different temperatures significantly affected the penetration area in the apical section at EDTA (60 °C > 25 °C,
P < 0.05), and in middle section at peracetic acid (36 °C > 60 °C, P < 0.05). In addition, different temperatures
significantly affected penetration percentage in the middle section of the HEBP (60 °C > 25 °C, P < 0.05). The effect of
cross-sectional area was significant in all parameters (Coronal > Middle > Apical, P < 0,05).
Conclusions According to the results of this study, heating of the final irrigation solution may affect sealer
penetration.
Keywords Dentin tubule penetration, Heating, Temperature increase, Bioceramic sealer, HEBP, Peracetic acid
*Correspondence:
Burhan Can Çanakçi
burhancanakci@[Link]
1
Faculty of Dentistry, Department of Endodontics, Trakya University,
Edirne, Turkey
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Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 2 of 9
Background been widely studied [2, 10, 11], but the impact of final
The main purpose of root canal treatment is to reduce irrigation solutions at different temperatures on dentin
microbial load in the root canal system. Chemomechani- tubule penetration of the sealer have not been investi-
cal preparation of the root canal may be inadequate in gated. This in-vitro study aimed to examine the effect
complex anatomical areas, such as the lateral canals, isth- of different irrigation solutions (EDTA, peracetic acid,
muses, and dentin tubule depths [1]. The root canal obtu- HEBP) at different temperatures (25, 36, and 60˚C) on
ration materials should hermetically seal the root canal in the dentin tubule penetration of bioceramic-based sealer
both the lateral and apical directions [1]. The penetration after root canal obturation using the warm vertical com-
of the root canal sealers into the dentin tubules is essen- paction method with a laser scanning confocal micro-
tial to ensure a tight connection between the root canal scope. The null hypothesis is that different irrigation
filling materials and dentin, ensure sealing, and prevent solutions at different temperatures have similar effects on
the colonization of residual microorganisms and reinfec- dentin tubule penetration of the sealer.
tion of the root canal system [1].
The smear layer is a layer containing organic (pulp and Methods
necrotic tissue residues, microorganisms, and residues) This in-vitro study was approved by the Ethics Commit-
and inorganic (dentin chips) tissues that form on the den- tee of Trakya University, Medical Sciences (Decision no:
tin surface during preperation of the root canal and cause 2021/003). The sample size calculation based on data
blockage of the dentin tubules [2]. The smear layer may obtained from a previous study [12] indicated that 14
prevent the penetration of irrigation solutions, medica- samples were sufficient per group [alpha (α) level = 5%,
ments, and sealers into dentin tubules and reduce their beta (b) level = 5%, power = 90%]. To account for any sam-
effectiveness [2]. Removal of the smear layer may result ple loss, a sample size of n = 15 was selected.
in greater penetration of the canal sealer and improve the In total, 180 freshly extracted human maxillar incisor
sealing properties of the filling materials [2]. teeth with a fully formed apex, single (Vertucci I), and
Various irrigation solutions have been suggested to straight canals, without internal or external resorption,
remove the smear layer [2]. Alternating the use of sodium were used. All teeth were recently extracted for periodon-
hypochlorite (NaOCl) and ethylenediaminetetraacetic tal reasons. Teeth were examined using bucco-lingual
acid (EDTA) is recommended to remove both the organic and mesio-distal radiographs and an operating micro-
and inorganic portions of the smear layer, soft tissue, scope at ×10 magnification (Zumax OMS2380; Suzhou,
and debris. However, EDTA reduces the effectiveness of China). Organic and inorganic residues on the tooth sur-
NaOCl, and its repeated use triggers root canal dentin faces were gently cleaned with a sharp-edged periodon-
erosion, decalcification of peritubular dentin, and disso- tal curette, and the teeth were washed in an ultrasonic
lution of the exposed organic matrix [3]. Peracetic acid is bath. The teeth were immersed in 0.1% thymol solution
a chelating agent that has an antimicrobial effect similar for 48 h for disinfection and stored in distilled water until
to that of the combination of 17% EDTA + 2.5% NaOCl use. A single operator performed all the test procedures.
[4], even at low concentrations, as well as smear removal The teeth were decoronated, and the roots were stan-
efficiency similar to that of 17% EDTA [5]. Etidronate, dardized to a length of 16 mm. After coronal access
an aqueous solution containing 1-hydroxyethylidene- cavity preparation, a #10 K file (Dentsply Maillefer, Bal-
1,1-bisphosphonate (HEBP), is another chelating agent laigues, Switzerland) was inserted into the root canal
with similar smear removal efficacy as EDTA [5]. HEBP is until it became apparent under ×10 magnification at the
a less destructive\abrasive to root dentin than EDTA [5] apical foramen. The working length (WL) was deter-
and does not negatively affect the organic tissue dissolv- mined by subtracting 1 mm from the total length of the
ing capacity or antibacterial efficacy of NaOCl [3]. root canal. Root canals were shaped to ProTaper Univer-
Various factors, such as the concentration, pH, appli- sal F4 NiTi files (Dentsply Maillefer) using an endodontic
cation time, amount, activation, and temperature of the motor (Motopex, Guilin Woodpecker Medical Instru-
irrigation solution, can affect the effectiveness of the ment, China) in accordance with the manufacturer’s rec-
solution [6]. Higher temperatures enhance the tissue dis- ommendations. 2 ml of 3% NaOCl solution (Cerkamed,
solving and antibacterial efficacy of NaOCl [7, 11], and its Stalowa Wola, Poland) was applied between each file. A
dentin tubule penetration [8]. Organic tissue dissolution total of 14 ml of NaOCl was used for each root canal.
and smear layer removal capacity increase when a mix- The root canals were then irrigated using 2 ml of distilled
ture of HEDP and NaCl is heated [9]. In addition, heating water and dried with sterile paper points (Meta Biomed,
EDTA reduces surface tension and increases wettability Korea).
[6]. The teeth were randomly distributed into 4 main
The effects of different irrigation solutions and devices groups as follows, according to the final irrigation solu-
on the penetration of different root canal sealers have tion to be applied;
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 3 of 9
• Group 1: 17% EDTA (Cerkamed). samples were polished with composite polishing discs
• Group 2: 9% HEBP (Sigma Aldrich, St Louis, MO, (Sof-Lex, 3 M ESPE, USA) under water cooling. Sam-
USA). ples were mounted on glass slides and examined with a
• Group 3: 1% peracetic acid (Sigma Aldrich). 532/580 nm helium laser using CLSM (Leica, TCS SP8
• Group 4 (Control): Distilled water. MP, Mannheim, Germany) at ×5 magnification. If the
entire canal could not be examined in one image, several
The groups were then randomly distributed into 3 sub- partial images were taken and assembled using Adobe
groups according to the temperature of the final irriga- Photoshop CS10 software (Adobe Systems, San Jose,
tion solution to be used (25, 36, or 60 ˚C; n = 15). The California, USA). Measurements were performed using
glass tube containing the irrigation solution was placed Image J software (ImageJ software, National Institute of
in a container filled with hot water. The temperature of Health, Bethesda, MD, USA) after placing a measure-
the irrigation solution in the glass tube was measured ment ruler on the images in Leica LAS X Core 3.7.4 soft-
using a digital thermometer. When the irrigation solution ware (Leica). Three parameters in the sample sections
reached the test temperature, root canal irrigation was were examined; Maximum penetration depth (mm), pen-
immediately performed. The final irrigation of the sam- etration area (mm²) [11], and penetration percentage (%)
ples was performed with different irrigation solutions at [10].
different temperatures in a volume of 2 ml for 1 min. All The data were analyzed using R software (Lucent Tech-
irrigation procedures were performed using a 30-gauge nologies, New Providence, NJ, United States of America).
endodontic irrigation needle (NaviTip FX; Ultradent The normality of the data distribution was examined
Products Inc, South Jordan, UT, USA) with the tip of the using the Shapiro-Wilk test. The non-normally distrib-
needle 1 mm behind the WL. The root canals were then uted penetration values were compared using a three-
irrigated with 2 ml of distilled water and dried using ster- way robust ANOVA with the WRS2 package. Multiple
ile paper points. comparisons were performed using Bonferroni correc-
For root canal obturation, a #40.04 α phase gutta‑per- tion. The significance level was set at P < 0.05.
cha (VDW GmbH, Munich, Germany) was fitted to the
WL to check tugback. For observation using a confocal Results
laser scanning microscope (CLSM), bioceramic-based Maximum sealer penetration depth
root canal sealers (Bioserra, Dentac, Türkiye) were mixed Regardless of the different temperatures and cross-sec-
with 0.1% fluorescent rhodamine B isothiocyanate dye tional areas, the values of the experimental groups were
(Sigma Aldrich) and placed into the root canal to 1 mm similar (P > 0.05) and higher than those of the control
short of the working length using a size 30-lentulo spiral. groups (P < 0.001, Fig. 1; Table 1). The effects of different
The 0.5‑mm tip of the gutta‑percha was trimmed, coated temperatures were also similar, regardless of the differ-
with a sealer, and placed into the root canal 0.5 mm short ent cross-sectional areas and solutions (P > 0.05, Fig. 2).
of the WL. BeeFill™ (VDW) downpack device was used Regardless of the different temperatures and solutions,
for obturation of the apical part, as recommended by the the effect of the cross-sectional area was significant (Cor-
manufacturer. A hot plugger (Dentsply Maillefer) was onal > Middle > Apical, P < 0,001).
applied approximately 3–4 mm from the apex. BeeFill™
(VDW) backfill device was used for the remainder of the Sealer penetration area
root canal system, as recommended by the manufacturer. Regardless of the different temperatures and cross-sec-
Vertical condensation was then performed using a plug- tional areas, the values of the experimental groups were
ger (Dentsply Maillefer). The quality of the root canal fill- similar, and higher than those of the control groups
ings was assessed using radiography. If canal filling was (P > 0.05), except in the middle section at 25 °C (Per-
not radiologically sufficient/appropriate, the sample was acetic acid > HEBP, P < 0.05), and in the apical section at
renewed. The access cavities were sealed with a light- 60 °C (EDTA > HEBP, P < 0.05, Fig. 1; Table 2). The effect
cured flowable adhesive (EsFlow, Spident, Korea). The of the cross-sectional area was significant regardless of
samples were stored in an incubator at 37 °C and 95% the temperature and solution (Coronal > Middle > Api-
humidity for 21 days. cal, P < 0,05). The effects of different temperatures
After the root canal sealer had set, the roots were were significant in the apical section at EDTA groups
embedded in a block of acrylic resin. Each specimen was (60 °C > 25 °C, P < 0.05), and in the middle section at per-
sectioned perpendicular to its long axis at depths of 2 acetic acid groups (36 °C > 60 °C, P < 0.05, Fig. 3).
(apical section), 5 (middle section), and 8 mm (coronal
section) from the root apex, respectively, using a low- Sealer penetration percentage
speed saw (Isomed, Buehler Ltd, Lake Bluff, IL, USA) Regardless of the different temperatures and cross-
under water cooling to a thickness of 1 ± 0.1 mm. Then, sectional areas, the values of
the experimental groups
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 4 of 9
Fig. 1 Representative confocal laser scanning microscopy images of the calculation of the maximum sealer penetration depth (a) and the sealer penetra-
tion area (b)
Table 1 Mean and standard deviation values of penetration depths of sealer (mm)
Distilled water EDTA Peracetic acid HEBP P
25˚C Apical 0,228 ± 0,047A 0,371 ± 0,064B 0,338 ± 0,025B 0,356 ± 0,063B < 0.001
Middle 0,668 ± 0,097A 0,812 ± 0,103B 0,948 ± 0,093B 0,847 ± 0,045B
Coronal 0,97 ± 0,1A 1,343 ± 0,104B 1,467 ± 0,088B 1,151 ± 0,073B
36˚C Apical 0,216 ± 0,027A 0,462 ± 0,112B 0,46 ± 0,065B 0,294 ± 0,025B
Middle 0,426 ± 0,075A 1,102 ± 0,146B 1,034 ± 0,073B 0,833 ± 0,093B
Coronal 0,74 ± 0,057A 1,517 ± 0,085B 1,332 ± 0,101B 1,148 ± 0,050B
60˚C Apical 0,212 ± 0,057A 0,453 ± 0,05B 0,519 ± 0,071B 0,246 ± 0,028B
Middle 0,688 ± 0,084A 0,97 ± 0,052B 1,061 ± 0,133B 0,773 ± 0,137B
Coronal 1,037 ± 0,094A 1,252 ± 0,057B 1,498 ± 0,119B 1,253 ± 0,107B
The different uppercase letters indicate significant differences between groups in the same line (P < 0.001)
were similar, and higher than those of the control advantages, such as providing a detailed view of inter-
groups (P > 0.05), except in the middle section at 25 °C facial adaptation and the distribution of sealers, clear
(EDTA > HEBP, P < 0.05), at 60 °C (HEBP > EDTA, visualization of sealer tags using fluorescence, and visu-
P < 0.05) and in apical section at 60 °C (EDTA > HEBP, alization of the infiltration of sealer tags into dentin
P < 0.05, Table 3). The effect of the cross-sectional area tubules with few artifacts [10]. CLSM is a nondestructive
was significant regardless of the temperature and solu- technique since it allows the use of the same sample for
tion (Coronal > Middle > Apical, P < 0,05). The effects of additional analysis [14]. Furthermore, CLSM does not
different temperatures were significant in the middle sec- require sample processing, such as dehydration, which
tion of the HEBP groups (60 °C > 25 °C, P < 0.05, Fig. 4). causes shrinkage of the material [10]. CLSM requires the
addition of a small amount of dye to label the sealer. The
Discussion addition of 0.1% Rhodamine B provides adequate quality
Many factors can influence the depth of penetration, during fluorescence imaging while not affecting the flow
including the structure of the dentin, the irrigation solu- properties of root canal filling sealers [10]. Therefore, in
tion used, the removal of the smear layer, and the physical this study, the bioceramic sealer was labeled with 0.1%
and chemical properties of the sealer [2, 13]. The current Rhodamine B before CLSM evaluation, similar to previ-
study is the first to examine the dentin tubule penetration ous studies [15].
of a bioceramic-based root canal sealer used with warm In this study, three parameters were measured to
vertical compaction obturation method after the use of evaluate dentin tubule penetration in CLSM; maximum
EDTA, Peracetic acid, HEBP, and distilled water at differ- penetration depth, penetration area [10], and sealer
ent temperatures (25, 36, and 60˚C) as the final irrigant. penetration percentage [11]. The assessment of maxi-
According to this study, the temperature increase of the mum penetration depth may not be clinically relevant if
final irrigation solution may affect the dentin tubule pen- the sealer penetration is not homogeneous throughout
etration of the bioceramic-based root canal sealer. the canal, deep sealer penetration is at one point, or the
There are several methods for investigating sealer sealer is not well dispersed in the canal circumferentially
penetration, including scanning electron microscopy, [13]. In contrast, the assessment of the sealer penetration
light microscopy, and CLSM [2, 10]. CLSM has several percentage allows the measurement of the penetration of
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 5 of 9
Fig. 2 Bar chart presenting mean and standard deviation values of maximum penetration depths (mm) of sealer according to the temperature, root
canal third, and irrigation solution
Table 2 Mean and standard deviation values of penetration area of sealer (mm2)
Distilled water EDTA Peracetic acid HEBP P
25˚C Apical 0,124 ± 0,021A 0,303 ± 0,051B* 0,403 ± 0,098B 0,376 ± 0,059B < 0.05
Middle 1,215 ± 0,319A 2,329 ± 0,288BC 2,719 ± 0,323B¥ 1,554 ± 0,183C
Coronal 3,136 ± 0,47A 5,609 ± 0,614B 5,495 ± 0,379B 3,964 ± 0,532B
36˚C Apical 0,117 ± 0,024A 0,567 ± 0,152B 0,34 ± 0,083B 0,202 ± 0,038B
Middle 0,856 ± 0,222A 1,81 ± 0,456B 2,418 ± 0,224B 2,161 ± 0,267B
Coronal 1,883 ± 0,3A 6,186 ± 1,178B 4,786 ± 0,575B 5,061 ± 0,456B
60˚C Apical 0,151 ± 0,038A 0,441 ± 0,029B* 0,37 ± 0,057BC 0,175 ± 0,015C
Middle 1,374 ± 0,266A 1,925 ± 0,385B 1,803 ± 0,347B¥ 2,011 ± 0,672B
Coronal 3,063 ± 0,486A 4,159 ± 0,353B 4,679 ± 0,835B 4,888 ± 0,584B
P < 0.05
The different uppercase letters indicate significant differences between groups in the same line (P < 0.05). The same symbols (* or ¥) in the same column represent
statistical differences between different temperature groups (25, 36 or 60˚C) in the same sections (apical, middle or coronal)
material into the entire walls of the root canal. The pen- [19]. The use of calcium silicate-based sealers in clinical
etration percentage and maximum penetration depth practice is increasing because of their biocompatibility,
results of the sealer have been reported to be inconsistent antimicrobial substantivity, and bioactivity [20]. Calcium
[16]. Therefore, the evaluation of sealer penetration per- silicate-based sealers are recommended using the single-
centage may be more meaningful, reliable, and clinically cone technique because heat can affect their physical
relevant than the measurement of the maximum depth of properties by decreasing bond strength, shortening set-
sealer penetration [13]. ting times, and reducing flow rates [18]. However, it has
The sealer’s physical and chemical characteristics and been reported that silicate-based sealers are not actually
dentin permeability may affect sealer penetration [17, influenced by heat [21], and lateral canals are more easily
18]. Bioceramic sealers are characterized by their high filled with warm vertical compaction technique [22]. In
flowability, small particle size, and hydrophilic properties this study, premixed, injectable tricalcium silicate sealer
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 6 of 9
Fig. 3 Bar chart presenting mean and standard deviation values of the penetration area (mm2) of sealer according to the temperature, root canal third,
and irrigation solution The statistical difference in the solution at different temperatures is shown with the same symbol (* or ¥, P < 0.05)
Table 3 Mean and standard deviation values of sealer penetration percentage (%)
Distilled water EDTA Peracetic acid HEBP P
25˚C Apical 21,9 ± 2,7A 30,7 ± 4,1B 40,1 ± 7,5B 27,4 ± 2,6B < 0.05
Middle 37,6 ± 3,4A 59,5 ± 3,9B 50,3 ± 3,6BC 43,6 ± 2,5C*
Coronal 43,3 ± 3,7A 69,9 ± 3,7B 52,2 ± 4,4B 59,5 ± 3,5B
36˚C Apical 28,8 ± 2,3A 45 ± 5,3B 37 ± 2,7B 36,2 ± 3,5B
Middle 41,9 ± 3,9A 53,2 ± 2,6B 44,3 ± 5,4B 52,2 ± 6,9B
Coronal 50,5 ± 4,9A 69,7 ± 1,9B 55 ± 4,1B 64,2 ± 3,8FGH
60˚C Apical 23,6 ± 1,2A 41,3 ± 4B 34,8 ± 5,7B 30,8 ± 2,2B
Middle 48,5 ± 3,2A 54,4 ± 2,5B 51,8 ± 6,4BC 58,5 ± 2,9C*
Coronal 58,2 ± 3,5A 64,8 ± 2,4B 68,4 ± 3,6B 70,9 ± 3,9B
P < 0.05
The different uppercase letters indicate significant differences between groups in the same line (P < 0.05). The same symbol (*) in the same column represent
statistical differences between different temperature groups (25, 36 or 60˚C) in the same sections (apical, middle or coronal)
was used with gutta-percha via warm vertical compac- apical region are fewer in number and smaller in diam-
tion technique [17]. eter, and the dentin is more atubular and sclerotic [2, 24].
According to the results of this study, all experimental Second, since irrigation efficiency decreases from coronal
groups achieved better penetration values than the con- to apical, the smear layer removal efficiency of irrigation
trol groups, similar to the results of several studies [2, protocols decreases in the apical region [25]. In addi-
12, 15, 23]. Sealer penetration is directly related to smear tion, greater compressive forces during obturation at the
removal capacity [2]. The lack of use of a chelator as the coronal and middle thirds may be another factor. Finally,
final irrigation solution in the control groups may have when warm obturation techniques are used, devices such
caused this result in the study. as heat carriers and pluggers may find it more difficult to
Sealer penetration values are
found highest in the cor- access the apical third [17], which may also affect the api-
onal section, then middle and apical sections, similar to cal obturation efficiency.
the results of previous studies [2, 10, 13, 15]. The possi- According to the results of this study, when the max-
ble reason for this may be that the dentin tubules in the imum penetration depth was evaluated, the use of
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 7 of 9
Fig. 4 Bar chart presenting mean and standard deviation values of sealer penetration percentage (%) according to the temperature, root canal third, and
irrigation solution
The statistical difference in the solution at different temperatures is shown with the same symbol (*, P < 0.05)
different solutions gave similar [Link] results are study of Ulusoy et al. [30]. Additionally, similar smear
consistent with those of previous studies [2, 26]. Alim et removal potentials of EDTA with 9% and 18% HEBP [31],
al. reported that a mixture of 2.5% NaOCl + 9% HEBP and 18% HEBP [5], 1% peracetic acid [27], and 0.5% and 2.5%
EDTA provided similar sealer penetration in their study peracetic acid have been reported [32]. Different irrigants
using Endosequence BC (Brasseler USA, Savannah, GA) may lead to different demineralization patterns for dentin
and a single-cone technique [23]. Conversely, Gawdat et [32] and may change the composition of the dentin sur-
al. reported that a mixture of 2.6% NaOCl + 9% HEBP face, affecting the bond and penetration between sealer
provided better sealer penetration than EDTA in their and root dentin [33].
study using Well-Root ST sealer (Vericom, GangwonDo, Based on the findings of this research, the effects of
Korea) and a single-cone technique [12]. Additionally, different temperatures were similar, except for the pen-
Keine et al. reported similar penetration results for 1% etration area values in the apical section of the EDTA
peracetic acid and EDTA using AH-Plus (Dentsply) and groups (60 °C > 25 °C, P < 0.05), and in the middle section
the single-cone technique [27]. The strong chelating effect at peracetic acid groups (36 °C > 60 °C, P < 0.05), and the
of EDTA, which can produce a deep zone of demineral- penetration percentage valuesin the middle section of
ized dentin compared with HEBP, may explain the better the HEBP groups (60 °C > 25 °C, P < 0.05).Because there
penetration observed with EDTA [15]. In addition, De- are no similar studies, these results cannot be directly
Deus et al. reported that the demineralization kinetics of compared. With increasing temperature, a decrease in
9% HEDP were slower than those of 17% EDTA over time the surface tension of the solutions and an increase in
[28]. Additionally, the highest penetration area values their wettability may occur, resulting in an increase in
without significance at 25˚C were found in the peracetic the smear removal capacity and speed [6, 9]. Since the
acid groups, followed by the HEBP and EDTA groups, temperature of the irrigation solution heated outside
similar to the results of several studies [12, 29]. Although decreases rapidly inside the root canal [7], the effect of
it has been reported that the smear removal potential is the temperature increase on penetration may have been
proportional to sealer penetration [27], contrary to the limited.
results of the present study, 9% HEBP removed smear This study has several limitations. First, in this study,
better than 1% peracetic acid and EDTA according to the single-rooted teeth were used. Different results may be
Arılı Öztürk et al. BMC Oral Health (2025) 25:352 Page 8 of 9
Competing interests
observed for multi-rooted teeth and teeth with complex The authors declare no competing interests.
anatomy. The reason for extraction and the physiological
age of teeth can affect the structure, diameter, and irregu- Received: 21 November 2024 / Accepted: 26 February 2025
larity of dentin tubules [34]. In addition, due to the higher
density of dentin tubules in the bucco-lingual direction
compared with the mesio-distal, and positioning of the
tubules along the root canal, an optical phenomenon References
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