According to the results of this study, the incidence of cracks was observed in all studied preparation systems and in apical, median and coronal sections from the apex, with no relationship with the anatomical location of the apex or the system. Overall, in the groups of Reciproc, ProTaper and Mtwo files, dentinal crack was observed in seven (9/3%), twelve (7/6%) and seven (9/3%) roots, respectively. Despite the larger number of dentinal crack incidence in ProTaper preparation system, there was no significant difference. On the other hand, dentinal cracks were observed in 3, 5 and 9 mm apical, middle and coronal sections from the anatomic apex, and in 6.5%, 9.3% and 5% of the specimens. In the study of Milani et al. (2012) the number of damages in 3 and 6 mm sections and the frequency of surface cracks in these sections had no obvious difference, thus this report is consistent with the results of our study, despite the fact that in the current study, sections were created at 6 mm from apex (
8). In addition, Burklein et al. (2013) investigated the incidence of dentin cracks after root canal preparation by rotary files and Reciproc, and observed no significant difference in the incidence of damage in various sections (3, 6 and 9 mm ) (
21). Previous studies have shown that dentinal crack can be associated with system of preparation and canals filling method, and currently there is no complete prevention of these types of damages (
4,
5). The difference between the various root canal preparation instruments in terms of dentinal cracks can be associated with preparation techniques and the cross sectional design of files. Mtwo and Reciproc preparation systems have S-shaped cross-sectional design and their cutting tips are extremely sharp, while the ProTaper system has a triangular cross-section and lower cutting performance and also it has limited filings space (
14). High cutting ability is usually associated with increased cleaning efficiency (
22,
23). Reciprocating motions can also increase the movement of debris towards the apex (
24), and there is a possibility of increasing torsional forces following these motions. It seems that the effects of increased torsional forces along the sharp tips, as with reciproc files, can be effective in the formation of dentinal cracks. Bier et al. (2009) investigated the ability of Ni-Ti rotary instruments in cracks formation in the root canal preparation process (
4). In the study of Yoldas et al. (2012), as well as the current study, no significant differences were observed regarding the number of dentinal cracks following preparation with four rotary files, including Twisted File, Revo-S, HERO shaper and ProTaper (60%, 25%, 40% and 30%) (
7). In this study, molar teeth were used, as like our study PDL was simulated but performed with a different method.
In one study, Burklein and colleagues (2013) showed that preparation with the Reciproc files led to more complete cracks compared to Mtwo and Reciproc files, and at apical surfaces (3mm), reciprocating instruments led to more incomplete cracks compared to Mtwo and ProTaper rotary files; thus cracks were not considered as complete or incomplete (
21). In the study of Burklein, unlike the present study, central mandibular teeth were used and master apical file was also different from the present study, so that in the Reciproc file, the master file was R40, which had 0.06 convergence, in ProTaper f4 file convergence was 06/0, and in Mtwo 40.04 was used as a master apical file, which obviously produces more stress on the root canal and this may explain the difference between these results and those of the current study. On the other hand, Liu and colleagues (2013), compared the frequency of cracks incidence following the use of three single-file OneShape, Reciproc and SAF systems with the ProTaper system and the reported incidence of cracks in the ProTaper was 50%, OneShape 35% and Reciproc 5% (
25). In a recent study, the experimental group difference was significant in this regard thus the findings were not consistent with the current study. The only similarity between the present study and this recent study was finding more dentin damage frequency in preparation with the ProTaper system. Liu and colleagues (2013) reported that 25% of cracks forming in the roots were prepared with the ProTaper system in apical regions (
26). Jalali et al. (2015) in a similar study concluded that the frequency of crack incidence following preparation with Mtwo and ProTaper files is significantly higher than the prepared tooth with Reciproc files, however this study was conducted on mandibular premolars, which can be a reason for the differences in results between this study and the present study (
27). In the study of Ustun and colleagues (2015), the incidence of dentinal crack was compared between handmade K file, ProTaper universal, ProTaper next, and Reciproc systems. In this study ProTaper universal and ProTaper next systems caused more cracks than Reciproc systems, which corresponds with the data of this study (
28).
Reviewing of various studies indicates that the ProTaper system causes more dentinal damage. In the ProTaper system, finishing files have been used to prepare the apical areas and due to larger taper of apical finishing files in this system (up to about 0.09 mm), more stress is generated on the canal dentinal walls compared to other system (
4) that may cause a crack. Although today many rotary systems are used with the crown-down method, ProTaper files have been used with single length techniques (
29) and at the same time, some research results have shown increased crack incidence following preparation with the crown-down technique in the rotary system (
5,
11,
30). However, the effects of single length preparation techniques in the development of dentinal cracks still remain unknown.
There are no definitive conclusions about the clinical outcomes of dentine damages during long-term periods (
31,
32). Also, it is not clear if incomplete cracks or craze lines can lead to complete cracks and vertical fractures of the root following root canal treatments. In addition, the role of therapeutic processes such as canal preparation or second treatments (
3), and applying masticatory forces, as well as occlusal loading (
33), as factors involved in dentinal cracks or fracture forming, still have not been investigated. Obviously, fractures can develop in teeth even without having root canal treatment (
1). Also, no reasonable relationship was observed between the obtained results in experimental studies and clinical conditions for the incidence of cracks. Despite the efforts made under laboratory conditions to make the situation similar to clinical conditions, yet elimination of the differences and discrepancies in the two groups seem to be impossible as cracks may be out of section areas. Hence, external conditions still affect the findings of various researches. Specimens maintenance and the number of samples before, during and after preparation may also be effective on the results, especially when the mechanical properties of the specimens are examined. Sectioning techniques used in present study allowed us to investigate the effects of root canal treatment on dentin by direct observation. These processes have been adopted in previous studies (
4,
21). However, other methods such as computation of distributed stresses, direct observation of damages in teeth sections and resistance of teeth with root therapy have been used for this purpose (
34-
37). Regarding the method used in this study, no external force was applied and the effects of the preparation of root canal at canal walls and adjacent dentin were observed directly. In the present study, the roots were sectioned 3 mm, 5 mm and 9 mm from the anatomic apex and dentin crack incidence was assessed in them. The reason for 5-mm sections in this study was that the canal curve usually starts at 5 mm from apex (
1). On the other hand, in present study molars were used, which was because of more linear fractures of roots in these teeth (
1,
38). In previous research, mostly single-rooted teeth were used for evaluation of dentin damages. In the present study, multi-root teeth, especially the mesial root of lower first molar and maxillary mesiobuccal root were used, which are at a higher risk of strip perforation due to the anatomical issues. Also it has been suggested that most teeth with fracture development, are mandible and maxilla first molars (
38). Periodontal ligament because of its viscoelastic properties plays a major role in distributing stress in the teeth. Therefore, PDL simulation has always been considered in studies that examined the effects of applied forces in crack forming and fracture strength, including the present study. In the present study, the polyether impression material was used to simulate the periodontal ligament (
8,
39). Despite previous research, further studies are still needed to evaluate the results of reciprocating motions during root canal preparation and evaluation of factors involved in this field of treatment outcomes.
5.1. Conclusion
According to the limitation of this in vitro study and its confiding factors, dentin cracks have been found in preparations with different systems and at different distances from the anatomic apex and despite their higher frequency with the ProTaper system, in this regard there was no significant differences between these systems or different sections of the anatomic apex.