Microbiological analysis of endodontically treated teeth with apical periodontitis before and after endodontic retreatment.

OBJECTIVE: To characterize the microbiota of teeth with endodontic treatment failure by 16S ribosomal RNA genetic sequencing (GS) and PCR at the different phases of the endodontic retreatment and to associate the presence of specific bacteria with clinical and radiographic features in teeth with apical periodontitis. MATERIALS AND METHODS: Twenty infected root canals of single-rooted teeth were selected. Samples were collected with sterile paper points before chemo-mechanical preparation (CMP) (S1), after CMP (S2) and after 30 days of intracanal medication (ICM) (S3). Microbial identification was performed using GS and PCR. Tukey-Kramer post hoc test and post hoc ANOVA were used for intergroup analysis. Paired t test and repeated-measures ANOVA were applied for intragroup analysis, at a significance level of 5%. RESULTS: A total of 89 strains were identified using GS. Sixty-five strains were recovered in S1 and 15 strains in S2, and 9 strains remained in S3. Enterococcus faecalis was the most predominant bacteria. Gram-positive cocci bacteria predominated. Gram-negative species were also detected. Using species-specific PCR primers to detect seven species, the most prevalent ones at all the phases of the endodontic retreatment were E. faecalis and Porphyromonas gingivalis. However, Parvimonas micra and P. gingivalis were associated with previous pain, P. gingivalis was associated with tenderness to percussion and E. faecalis, Fusobacterium nucleatum and P. gingivalis were associated with periapical lesion > 3 mm. CONCLUSIONS: In conclusion, the microbiota of persistent infection is polymicrobial with predominance of E. faecalis and P. gingivalis in all phases of the endodontic retreatment, regardless of the method used for microbial identification. Associations were found between specific bacteria and clinical/radiographic features. CLINICAL RELEVANCE: The characterization of the bacteria present at all phases of the endodontic retreatment is important for the monitoring of the effectiveness of the techniques used and to better understand the susceptibility of these species to the disinfection agent used during the procedures.

Evaluation of the presence of microorganisms from root canal of teeth submitted to retreatment due to prosthetic reasons and without evidence of apical periodontitis.

AIM: The objective of this study was to evaluate the presence of microorganisms in the root canals (RC) of teeth submitted to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. MATERIAL AND METHODS: Twenty teeth referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis were included in this study. Gutta-percha (GP) was collected from each third of the RC. After GP removal, microbial samples were also collected from the full extension of RC using paper points and files. The samples were cultivated in Fastidious Anaerobe Agar and the colony-forming units (CFU/mL) were counted. They were also semi-quantitatively analyzed by checkerboard DNA-DNA hybridization using 40 DNA bacterial probes. RESULTS: Microorganisms were found in all samples. The coronal third of GP was more contaminated than the apical third (p ≤ 0.05). There was prevalence of Enterococcus hirae and Enterococcus faecalis in all RC thirds and also in the samples collected from the full extension of RCs. CONCLUSION: Microorganisms were found in all cases referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. Enterococcus was the genus most frequently detected. CLINICAL SIGNIFICANCE: An endodontic retreatment should be considered before replacing a prosthesis.

Effectiveness of calcium hydroxide-based intracanal medication on infectious/inflammatory contents in teeth with post-treatment apical periodontitis.

OBJECTIVE: The objective of this work was to investigate in vivo the effects of calcium hydroxide-based intracanal medication (ICM) on the levels of bacteria, pro-inflammatory cytokines (PICs), and matrix metalloproteinases (MMPs) in root canals and periradicular tissues of teeth with failure of the root canal treatment and apical periodontitis. MATERIALS AND METHODS: Twenty infected root canals of single-rooted teeth were randomly assigned into two groups according to the irrigant used for chemomechanical preparation (CMP) (n = 10 per group): G1 - 2% chlorhexidine (CHX) gel and G2 - 6% sodium hypochlorite (NaOCl). Root canal contents were taken by using paper points before CMP (S1) and after 30 days of calcium hydroxide-based ICM (S2). Microbial reduction was calculated by means of colony-forming unit count (CFU/mL), with PICs and MMPs (pg/mL) being measured by using enzyme-linked immunosorbent assay (ELISA). RESULTS: Culturable bacteria (101.2 ± 79.2), PICs (IL-1ß 1.2 ± 0.4 and TNF-α 8.8 ± 4.7), MMP-2 (803.7 ± 96.4), MMP-3 (453.9 ± 229.3), MMP-8 (245.9 ± 122.4), MMP-9 (129.4 ± 29.6), and MMP-13 (70.8 ± 12.8) were present in all S1 samples. After 30 days of ICM (S2), a 99.5% microbial reduction was observed, together with a significant reduction of PICs in all groups. Overall, it was observed a decrease in the levels of MMPs (S2), except MMP-13, which was found in increased levels after ICM (P < .05), independently of the groups. CONCLUSIONS: Calcium hydroxide-based intracanal medications have had a positive effect on the microbial reduction by decreasing the levels of PICs and MMPs. Both auxiliary chemical substances (i.e., 2% CHX and 6% NaOCl) presented similar effects when calcium hydroxide was used as intracanal medication. CLINICAL RELEVANCE: Teeth with failure of the root canal treatment and apical periodontitis, and consequently with high levels of bacteria, PIC, and MMP, may present a better prognosis after a 30 days of a calcium hydroxide-based ICM.

Antibiotic Resistance Profile of Clinical Strains of Enterococci from Secondary/Persistent Endodontic Infections: What do We Know? A Systematic Review of Clinical Studies.

INTRODUCTION: Enterococcus faecalis is the most common enterococcal species associated with infective endocarditis and 1 of the most commonly detected bacteria in cases of secondary/persistent endodontic infection (SPEI). Antimicrobial resistance is a global public health concern. This review aimed to answer the following research question: "Is there a change in the antibiotic resistance profile in clinical strains of E. faecalis over the years?". P (population) - patients with SPEI, I (intervention) -endodontic retreatment, C (comparison) -not included, O (outcome) - profile of Enterococci resistance and susceptibility to systemic antibiotics used. METHODS: Two authors independently performed study selection, data extraction, and risk of bias assessment. The literature search was conducted using the following electronic databases: PubMed, Scopus, EMBASE, Web of Science, and Medline. Clinical studies in which Enterococci strains were isolated to assess their antimicrobial resistance were included. RESULTS: Eleven clinical trials were included. Overall, E. faecalis isolated from teeth with SPEI presented an intermediate resistance to 16 antibiotics. In recent years, E. faecalis showed a little resistance to amoxicillin (without clavulanate) and benzylpenicillin. Erythromycin and rifampicin presented an increase in the intermediate-resistance status between the first and the last studies. E. faecium presented intermediate-resistance results. CONCLUSION: The most effective drugs remain the combination of amoxicillin and clavulanate, followed by amoxicillin and benzylpenicillin. In patients allergic to penicillin derivatives, moxifloxacin and azithromycin may be indicated with caution. The antibiotics with the highest pattern of resistance against E. faecalis are clindamycin, gentamicin, metronidazole, and rifampicin and are therefore, contraindicated in cases of SPEI. Very few clinical studies using a microbiological approach in teeth with endodontic failure have been carried out to improve the efficacy of prophylactic regimens. However, as bacteria periodically develop resistance to the main drugs used, regular studies should be carried out on the action of these drugs in infection control.

Efficacy of 6% Sodium Hypochlorite on Infectious Content of Teeth with Symptomatic Irreversible Pulpitis.

INTRODUCTION: The objective of this study was to monitor the effects of chemomechanical preparation (CMP) performed with 6% sodium hypochlorite and calcium hydroxide-based intracanal medication (ICM) on the levels and diversity of bacteria, endotoxins (lipopolysaccharides [LPS]), and lipoteichoic acid (LTA) in root canals of teeth with symptomatic irreversible pulpitis. METHODS: Samples were collected from 10 teeth with symptomatic irreversible pulpitis before CMP (S1), after CMP (S2), and after ICM (S3). The levels of bacteria, LPS, and LTA were assessed by using checkerboard DNA-DNA hybridization, LAL Pyrogent 5000, and enzyme-linked immunosorbent assay, respectively. Wilcoxon test, repeated-measures analysis of variance, and Tukey post hoc test were used for statistical analysis at a significance level of 5%. RESULTS: Forty species were detected at S1. Two species were eliminated after CMP and 5 after ICM. Resistant and pain-related species were detected in the root canals. Higher levels of culturable bacteria were detected at S1. However, CMP and ICM effectively reduced the microbial load in the root canals. Higher levels of LPS and LTA were detected at S1. CMP was effective in reducing both LPS and LTA (P < .05). ICM produced additional reduction in the levels of LPS (P > .05) and LTA (P < .05). CONCLUSIONS: Chemomechanical preparation using 6% sodium hypochlorite and calcium hydroxide-based intracanal medication were effective in reducing the levels of bacteria, LPS, and LTA in teeth with vital pulp and irreversibly inflamed pulp.

Identification of Culturable and Nonculturable Microorganisms, Lipopolysaccharides, and Lipoteichoic Acids From Root Canals of Teeth With Endodontic Failure.

AIM: To elucidate the presence of apical periodontitis in the root canal of teeth with secondary/persistent infection, including composition of microbiota, levels of endotoxins and lipoteichoic acid (LTA), and clinical implications of these findings. METHOD: Samples were collected from root canals of 50 patients who needed endodontic retreatment and had radiographic evidence of apical periodontitis. Microorganisms were identified by using the culture technique and biochemical tests. Nested-polymerase chain reaction (nested-PCR) was used to identify 17 species of specific bacteria. Lipopolysaccharides (LPS) and LTAs were quantified by using, respectively, limulus amebocyte lysate and enzyme-linked immunosorbent assay tests. RESULTS: Bacteria were detected in all samples by culture and molecular methods. A total of 154 gram-positive strains, of 188 strains isolated, were found in the root canals by culture. Enterococcus faecalis and Gemella morbillorum were the most prevalent species identified by the biochemical tests, whereas molecular analyses (nested-PCR) showed a high frequency of P. gingivalis, E. faecalis, and Fusobacterium nucleatum. LPS and LTA were detected in all samples, with mean values being 3.52 EU/mL and 597.83 pg/mL, respectively. Significant statistical correlations were found between levels of LTA and clinical features. CONCLUSION: Despite the prevalence of gram-positives, the microbiota present in secondary/persistent infections showed a large variety of species. Within this diversity, associations were found between specific bacteria and clinical features. In addition, higher levels of LTA were statistically associated with larger periapical radiolucent areas, but no correlation between this feature and LPS was found.

Microbiological Investigation in Teeth with Persistent/Secondary Endodontic Infection in Different Stages of Root Canal Retreatment.

OBJECTIVE: The present clinical study investigated the microbiota of teeth with persistent secondary endodontic infection in the different phases of root canal retreatment. METHODS: Twenty filled single-rooted teeth with apical periodontitis were included. Samples were collected with sterile paper points before chemo-mechanical preparation (CMP) (S1), after CMP (S2) and after 30 days of calcium hydroxide-based intracanal medication (ICM) (S3). Cultivable bacteria were assessed by colony forming units count (CFU/mL). DNA was extracted and assessed by using nested PCR. Paired t-test and repeated measures ANOVA were applied for intragroup analysis in the stages of endodontic therapy at a significance level of 5%. RESULTS: Cultivable bacteria were detected in all initial samples. CMP reduced bacteria by 99.4% and ICM by 99.5%. The most prevalent species found in the initial samples were E. faecalis (20/20), P. gingivalis (20/20), F. nucleatum (17/20) and A. actinomycetemcomitans (10/20), whereas D. pneumosintes, F. alocis, P. nigrescens and T. socranskii were not detected. After CMP, A. israelii, A. naeslundii, G. morbillorum, T. forsythia and T. denticola were not detected (P<0.05) either. E. faecalis and P. gingivalis had a low reduction (P>0.05) and F. nucleatum had its DNA significantly reduced after CMP (P<0.05). ICM had no additional effect on microbial reduction. CONCLUSION: The microbiota of teeth with persistent/secondary endodontic infection consists of a polymicrobial community with Gram-positive and Gram-negative species, bacillus and cocci, facultative and strict anaerobes. E. faecalis and P. gingivalis were frequently detected in all stages of root canal retreatment, evidencing their great resistance to endodontic procedures. The endodontic procedures were effective in reducing the levels of bacteria from teeth presenting with persistent/secondary endodontic infection.

Apically Extruded Debris Using Passive Ultrasonic Irrigation Associated with Different Root Canal Irrigants.

The present study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) associated with passive ultrasonic irrigation (PUI) using four different root canal irrigants, namely, 6% sodium hypochlorite (NaOCl), 2% chlorhexidine gel + saline solution (2% CHXg+SS), 2% chlorhexidine solution (2% CHXs) and SS alone. Sixty mandibular premolars with single straight root canals were selected and randomly assigned into 4 groups (n=15) according to the root canal irrigant used as follows: G1 (PUI + NaOCl), G2 (PUI + CHXg+SS), G3 (PUI+CHXs) and G4 (PUI+SS). Reciproc® R25 files (25/.08) were used during CMP and the debris extruded from each tooth were collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed by using an analytical microbalance. Data were statistically analysed by using ANOVA and post-hoc Tukey's test (a=0.05). Debris extrusion was observed in all groups, irrespective of the root canal irrigating, with 2% CHXg + SS being associated with lower debris extrusion compared to other irrigants (p<0.05). No significant differences were observed between 6% NaOCl, 2% CHXs and SS. In conclusion, passive ultrasonic irrigation did not completely prevent apically extrusion of debris. PUI performed with 2% chlorhexidine gel + saline solution significantly minimized debris extrusion compared to 6% sodium hypochlorite, chlorhexidine solution and saline solution.

Evaluation of Apically Extruded Debris Using Positive and Negative Pressure Irrigation Systems in Association with Different Irrigants.

This study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) using positive and negative pressure irrigation systems [Conventional irrigation (CI) and EndoVac (EV)] in association with different irrigants [6% Sodium Hypochlorite (NaOCl), 2% Chlorhexidine gel + saline solution (CHXg + SS), 2% Chlorhexidine solution (CHXs) or Saline solution (SS)]. Eighty mandibular premolars with single root canals were selected and randomly assigned into 8 groups (n = 10) according to the irrigation system and the irrigant used during CMP: G1 (EV + NaOCl), G2 (EV + CHXg + SS), G3 (EV + CHXs), G4 (EV + SS), G5 (CI + NaOCl), G6 (CI + CHXg + SS), G7 (CI + CHXs) and G8 (CI + SS). Reciproc® R25 files (25/.08) were used during the CMP and the extruded debris from each tooth was collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed using a microbalance, and the data were statistically analyzed using ANOVA and the post hoc Tukey's test (a = 0.05). All groups were associated with debris extrusion. EV was the irrigation system with less extruded debris (p < 0.05). No differences were observed regarding the irrigant when EV was used. When CI was used, CHXg + SS were associated with lower debris extrusion (p < 0.05). It was concluded that no irrigation protocol succeeded in preventing debris extrusion. EV resulted in lower levels of debris extrusion than CI. The use of CHXg + SS resulted in lower debris extrusion.

Effectiveness of final decontamination protocols against Enterococcus faecalis and its influence on bond strength of filling material to root canal dentin.

AIM: The aim of this study was to evaluate the effectiveness of final decontamination protocols against Enterococcus faecalis and their influence on bond strength of filling material to root canal dentin. MATERIAL AND METHODS: Ninety root canals were enlarged with ProTaper system and inoculated with E. faecalis for 15days. Sixty samples were randomly divided into six groups (n=10) and subjected to following protocols: G1-distilled water(control), G2-2% chlorhexidine, G3-QMix, G4-6.5% grape seed extract, G5-photodynamic therapy with optical fiber and G6-photodynamic therapy without optical fiber. The percentage of bacterial reduction was checked by counting of CFUs. The remaining 30 samples were subjected to the same decontamination protocols (n=5) and filled with gutta-percha and AH Plus sealer in order to perform the push-out test. Data from both tests were subjected to one-way ANOVA followed by Tukey's post hoc procedure (α=0.05). RESULTS: The greatest bacterial reduction was observed for 2% chlorhexidine, QMix and 6.5% grape seed extract, with no statistically significant difference between them. Photodynamic therapy, with and without optical fiber, demonstrated a significantly higher reduction than distilled water, with no statistically significant difference between them (p<0.05). For the push-out test, final decontamination protocols showed similar bond strength values (p<0.05), with the highest incidence of cohesive failure in all groups. CONCLUSIONS: The tested final decontamination protocols showed effectiveness against E. faecalis and did not interfere with the bond strength of filling material to root canal dentin.

Quantification of Lipoteichoic Acid Contents and Cultivable Bacteria at the Different Phases of the Endodontic Retreatment.

INTRODUCTION: The infectious content of root canals, including bacteria and lipoteichoic acid (LTA), cause injuries to the periapical tissues. The purpose of this clinical study was to quantify the levels of both LTA and cultivable bacteria at the different phases of endodontic retreatment (ER) of teeth with post-treatment apical periodontitis. It also aimed to investigate the presence of gram-positive microorganisms before and after chemomechanical preparation (CMP) and intracanal medication (ICM). METHODS: Twenty infected root canals of single-rooted teeth were randomly assigned into 2 groups according to the chemical substance used for CMP (n = 10 per group): chlorhexidine (CHX) group, 2% CHX gel, and the sodium hypochlorite (NaOCl) group, 6% NaOCl. Root canal samples were taken using paper points before (S1) and after CMP (S2) and after 30 days of ICM with calcium hydroxide + 2% CHX gel (S3). Microorganisms were identified by the culture technique using biochemical tests. Cultivable bacteria were determined by counting the colony-forming unit. LTA levels were measured using the enzyme-linked immunosorbent assay (pg/mL). RESULTS: A total of 70 gram-positive species, out of 102 species isolated, were found in the root canals (54 in S1, 4 in S2, and 12 in S3). Enterococcus faecalis was the most frequent isolated taxon in all phases of the ER. LTA (574.0 ± 94.7) and cultivable bacteria (101.2 ± 79.2) were present in all S1 samples. CMP decreased the overall levels of cultivable bacteria by 99.4% and LTA by 24.8% (P < .05), whereas the total overall reduction level of ICM on viable bacteria was 99.5% and on LTA it was 38.6% (P < .05). CMP with 2% CHX gel (CHX group, 99.3%) was more effective (P < .05) than 6% NaOCl (NaOCl group, 92.1%) on bacterial reduction. Likewise, ICM showed a 100% reduction in the CHX group and 98.5% in the NaOCl group. Regarding the reduction of LTA, CMP with 2% CHX gel (CHX group, 26.9%) was more effective (P < .05) than 6% NaOCl (NaOCl group, 22.6%). In addition, ICM showed a 43.2% reduction in the CHX group and 36.2% in the NaOCl group (P > .05). CONCLUSIONS: The reduction rates of bacteria were higher than the LTA. Moreover, gram-positive microorganisms were present in all phases of the endodontic retreatment.

Antimicrobial Susceptibility and Characterization of Virulence Genes of Enterococcus faecalis Isolates from Teeth with Failure of the Endodontic Treatment.

INTRODUCTION: The aim of this study was to investigate the prevalence of virulence factors and the antimicrobial resistance of Enterococcus faecalis isolates of teeth with failure of the endodontic treatment. METHODS: Twenty root canal samples were collected from teeth with apical periodontitis. E. faecalis was firstly identified based on phenotypic features and then by 16S ribosomal RNA gene sequencing. The antimicrobial susceptibility was determined by the minimum inhibitory concentration (MIC) of amoxicillin, amoxicillin + clavulanate, azithromycin, benzylpenicillin, ciprofloxacin, clindamycin, chloramphenicol, doxycycline, erythromycin, gentamicin, metronidazole, moxifloxacin, rifampicin, tetracycline, and vancomycin using the E test method. Virulence factors (ace, asa, asa373, cylA, efaA, esp, and gelE) were detected by polymerase chain reaction assay. RESULTS: Amoxicillin + clavulanate was effective against all strains. Intermediate and total resistance was found against the majority of the tested antimicrobials. The susceptibility of some microorganisms to some antimicrobial agents changed according to the evaluation time. MIC50 and MIC90 also varied according to the evaluation time. In relation to the virulence factors of the E faecalis isolates, ace was detected in 100% of the strains, asa (60%), asa373 (15%), efaA (95%), esp (70%), and gelE (75%), whereas cylA was not detected. CONCLUSIONS: It was concluded that E. faecalis isolates from persistent endodontic infections showed varied degrees of intermediate/total resistance to several antimicrobial agents, with amoxicillin + clavulanate being the most effective agent. Moreover, the strains showed different patterns for virulence gene detection.

Evaluation of apically extruded debris using positive and negative pressure irrigation systems in association with different irrigants

Abstract This study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) using positive and negative pressure irrigation systems [Conventional irrigation (CI) and EndoVac (EV)] in association with different irrigants [6% Sodium Hypochlorite (NaOCl), 2% Chlorhexidine gel + saline solution (CHXg + SS), 2% Chlorhexidine solution (CHXs) or Saline solution (SS)]. Eighty mandibular premolars with single root canals were selected and randomly assigned into 8 groups (n = 10) according to the irrigation system and the irrigant used during CMP: G1 (EV + NaOCl), G2 (EV + CHXg + SS), G3 (EV + CHXs), G4 (EV + SS), G5 (CI + NaOCl), G6 (CI + CHXg + SS), G7 (CI + CHXs) and G8 (CI + SS). Reciproc® R25 files (25/.08) were used during the CMP and the extruded debris from each tooth was collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed using a microbalance, and the data were statistically analyzed using ANOVA and the post hoc Tukey's test (a = 0.05). All groups were associated with debris extrusion. EV was the irrigation system with less extruded debris (p < 0.05). No differences were observed regarding the irrigant when EV was used. When CI was used, CHXg + SS were associated with lower debris extrusion (p < 0.05). It was concluded that no irrigation protocol succeeded in preventing debris extrusion. EV resulted in lower levels of debris extrusion than CI. The use of CHXg + SS resulted in lower debris extrusion.

Resumo Este estudo avaliou a quantidade de debris extruídos apicalmente após o preparo químico-mecânico (PQM) utilizando sistemas de irrigação com pressão positiva e negativa [irrigação convencional (IC) e EndoVac (EV)] em associação com diferentes irrigantes [hipoclorito de sódio 6% (NaOCl), clorexidina gel + solução salina (CLXg + SS), solução de clorexidina 2% (CLXs) ou solução salina (SS)]. Oitenta pré-molares inferiores com único canal radicular foram selecionados e aleatoriamente alocados em 8 grupos (n=10) de acordo com o sistema de irrigação e irrigante utilizado durante o PQM: G1 (EV + NaOCl), G2 (EV + CLXg + SS), G3 (EV + CLXs), G4 (EV + SS), G5 (IC + NaOCl), G6 (IC + CLXg + SS), G7 (IC + CLXs) e G8 (IC + SS). Limas Reciproc® R25 foram utilizadas durante o PQM e os debris extruídos de cada dente foi coletado em tubos pré-pesados e secos. O peso médio de debris foi avaliado por meio de microbalança, e os dados foram analisados estatisticamente utilizando ANOVA e teste de Tukey (a = 0.05). Todos os grupos foram associados com extrusão de debris. EV foi o sistema de irrigação com menos debris extruídos (p<0.05). Não foram observadas diferenças entre os irrigantes quando o EV foi utilizado. Quando foi utilizada IC, CLXg + SS foram associados a menor extrusão de debris (p<0.05). Concluiu-se que nenhum protocolo de irrigação conseguiu prevenir extrusão de debris. EV resultou em menores níveis de extrusão de debris que a IC. A utilização da CLXg + SS resultou em menor extrusão de debris.

Canal transportation and centering ability of curved root canals prepared using rotary and reciprocating systems

Abstract Aim: To evaluate canal transportation and centering ability of Protaper and WaveOne systems in curved root canals by cone-beam computed tomography. Methods: Twenty human molars were randomly divided into two groups according to the system used: Group GP (ProTaper Universal(r)) and group GW (WaveOne(tm)). CT scans were performed before and after the chemo-mechanical preparation on three points: 2 mm, 3 mm and 4 mm from the tooth apex. The cuttings in dentin were measured and results were statistically analyzed. Results: In GP, there was higher percentage in transport at 2 mm and 4 mm toward the mesial wall of the tooth root compared to GW (p<0.05). There was no difference between the systems regarding their centering ability. Conclusions: None of the systems was able to touch all the dentin walls and stay centered during the chemical-mechanical preparation.