Smear layer removal by EGTA

The purpose of this study was to compare the effects of ethylene glycol-bis (beta-aminoethyl ether)-N,N,N', N'-tetraacetic acid (EGTA) and EDTA on removal of the smear layer by using scanning electron microscopy. Fifteen single-rooted teeth were instrumented to size 60. Three teeth served as the control group, and the remaining teeth were assigned into two groups. After instrumentation, root canals of the first group were irrigated with 10 ml of 17% EDTA, and the second group was irrigated with 10 ml of 17% EGTA for 2 min. Both were then irrigated with 10 ml of 5% NaOCl. The teeth in the control group were irrigated with only 10 ml of 5% NaOCl. Then the specimens were sectioned longitudinally and processed for scanning electron microscopy. The results showed that the smear layer was completely removed by EDTA, but it caused erosion of the tubuli. EGTA was somewhat effective in removing the smear layer without inducing erosion. Our results suggest that EGTA is an alternative chelator for removing the smear layer.     

5种冲洗剂组合对前牙直根管清洁效果的比较

目的：比较5种冲洗剂组合对前牙直根管的清洁效果。方法：25颗离体直根管前牙,随机分为5组,不锈钢K锉常规法预备根管．应用5种冲洗剂组合进行冲洗。第1组：根管器械预备期间和预备结束后依次用1％NaOCl和3％H2O2冲洗;第2组：根管器械预备期间用1％NaOCl冲洗,预备结束后用17％EDTA冲洗;第3组：根管器械预备期间和预备结束后依次用1％NaOCl和17％EDTA冲洗：第4组：根管器械预备期间依次用17％EDTA和1％NaOCl冲洗,器械预备结束后用17％EDTA冲洗;第5组：根管器械预备期间依次用17％EDTA、1％Triton X-100(表面活性剂)和1％NaOCl冲洗．器械预备结束后用17％EDTA冲洗。每组冲洗剂剂量和冲洗时间均为22ml和7min。将牙纵劈后进行扫描电镜观察。结果：第1组,根管壁上见典型玷污层结构和大量杂质和残屑。第2组在根管冠、中1／3能部分去除玷污层．根尖1／3残留大量玷污层。第3组虽然能有效去除玷污层．但会引起牙本质小管中度腐蚀。第5组在根管冠、中1／3能部分去除玷污层,但根管壁上黏着大量杂质和残屑．此外还存在重度腐蚀现象。第4组根管清洁效果最好,且对牙本质小管无腐蚀性。结论：在严格控制冲洗时间和顺序的情况下,联合应用17％EDTA和1％NaOCl能有效去除玷污层．且不会腐蚀牙本质小管。     

Smear layer influence on the apical seal of four different obturation techniques

The purpose of this study was the in vitro evaluation of four techniques for the obturation of the root canal system in the presence or absence of a smear layer. Ninety-six human upper central incisors were instrumented using the pressureless crown-down technique and irrigated with 5.25% NaOCl. The teeth that had the smear layer removed were irrigated with this solution in combination with 17% EDTA. The teeth were obturated with lateral condensation with an accessory or standardized cone as the main cone, with vertical condensation of warm gutta-percha or with thermoplasticized injectable gutta-percha. Apical leakage was assessed by measuring the linear penetration of methylene blue dye with a stereomicroscope. The results showed no significant differences in the degree of leakage with and without the smear layer when the samples were considered as a whole. However, when the groups were assessed separately, teeth in the lateral condensation with an accessory main cone group and teeth in the thermoplasticized group leaked less with a smear layer present. In contrast teeth with lateral condensation and a standardized main cone leaked more with a smear layer present. In the vertical condensation groups there was no difference attributable to the smear layer.     

Effect of removal of the smear layer on apical microleakage

It has been shown previously that the smear layer created during instrumentation of the root canal promotes apical microleakage. The purpose of the present study was to investigate the effect of removal of the smear layer on apical microleakage, using the fluid filtration method. Thirty-six single-rooted human premolar teeth were used. Six of the teeth were selected randomly as controls, and the remaining 30 teeth were randomly divided into two groups of 15 teeth each. For the first group (group 1) teeth were irrigated with NaOCl only (smear layer not removed). In the second group (group 2) the teeth were irrigated with 15% EDTA, followed by NaOCl to remove the smear layer. Both groups were filled with thermoplasticized gutta-percha (Obtura II) using glass-ionomer cement as sealer. All teeth were stored at 37 degrees C and 100% humidity for 2 days. Apical microleakage of the filled root canal was measured by the fluid filtration technique. Hydraulic conductance was measured by the movement of an air bubble in a capillary tube after a pressure of 200 mm Hg was applied to the system. The mean microleakage rate of group 1 and group 2 were 13.0 (SD +/- 5.1) and 16.9 (SD +/- 15.3) nl/s, respectively. Removal of the smear layer caused significantly more apical microleakage (p < 0.05, Student's t test) than when the smear layer was left intact.     

次氯酸钠配合SmearClear清除根管玷污层的研究

目的:比较4种根管冲洗方法清除根管玷污层的效果,研究出有效去除根管玷污层的方法。方法:20个无龋单根前牙随机分为4组,A组:60℃30 g/L次氯酸钠液加170 g/L EDTA,#15K锉提拉冲洗;B组:60℃30 g/L次氯酸钠液加SmearClear,#15K锉提拉冲洗;C组:60℃30 g/L次氯酸钠液加170 g/LEDTA液超声荡洗;D组:60℃30 g/L次氯酸钠液加SmearClear超声荡洗。扫描电镜下放大2 500倍和4 000倍观察根尖1/3、根中1/3和根上1/3玷污层清除情况。按照根管壁玷污层清除情况和根管壁腐蚀程度的分级标准进行统计学分析。结果:①根管壁玷污层清除情况:4组在根上2/3区玷污层均有效去除,各组间差异无统计学意义(P>0.05),B组根尖1/3基本无玷污层,A、C、D组仅见少部分玷污层被去除,B组与A、C、D组差异有统计学意义(P<0.05);②根管壁腐蚀性比较:C、D组根上2/3区腐蚀度较大,而A、B组未见明显腐蚀。C、D组与A、B组相比差异有统计学意义(P<0.05),各组根尖1/3根管壁腐蚀度差异无统计学意义(P>0.05)。结论:SmearClear与60℃30 g/L次氯酸钠液联合应用能更有效的去除根管玷污层,且对根管壁表面结构无明显破坏。     

The effect of application time of EDTA and NaOCl on intracanal smear layer removal: an SEM analysis

AIM: To verify, under the scanning electron microscope (SEM), the influence of irrigation time with ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl) on intracanal smear layer removal. METHODOLOGY: Twenty-one extracted human permanent teeth with single straight root canals were included. The root canals of the teeth were instrumented and, at the end of preparation, were irrigated with 3 mL of 15% EDTA, followed by 3 mL of 1% NaOCl for 1 min (group 1), for 3 min (group 2), and for 5 min (group 3). The canals of teeth in group 4 (control) did not receive the final irrigation. The teeth were sectioned longitudinally and prepared for an SEM. The dentinal wall of cervical, middle and apical thirds was graded according to the amount of debris and smear layer remaining on the walls. The results were analysed using the Kruskal-Wallis and Conover-Inman tests. RESULTS: In all the canals of experimental groups irrigation with EDTA and NaOCl completely removed the smear layer from the cervical and middle thirds. In the apical third, the dentine surface were partially covered, particularly in the teeth of group 1, where there was significantly more smear layer when compared with the other thirds in the same group (P<0.007). However, the Kruskal-Wallis test showed overall that there were no significant differences between groups 1, 2 and 3 (P>0.05). CONCLUSION: In this limited laboratory study, canal irrigation with EDTA and NaOCl for 1, 3 and 5 min were equally effective in removing the smear layer from the canal walls of straight roots.     

Post-operative symptoms and healing after endodontic treatment of infected teeth using pulsed Nd: YAG laser

Abstract— Forty-four teeth in 38 patients, diagnosed with chronic apical periodontitis, were endodontically treated. Root canals were shaped using a step-back technique with 5% NaOCl and 3% H₂O₂ as irrigants. In half of the teeth the canal terminus was irradiated with pulsed Nd:YAG laser (1 W, 15 pps, 1 s). All canals were then obturated with laterally condensed gutta-percha points and sealer, and final radiographs were obtained. Occurrence of spontaneous pain was recorded 1 day after treatment. Percussion pain was recorded after 1 week, and then at 3 and 6 months after treatment. Radiographic follow-up was performed at 3 and 6 months. Percussion pain was significantly less ( P >0.05) in the laser-treated group than in the control group, both 1 week and 3 months after treatment. Other differences between the groups were not significant. These results suggested that the clinical application of pulsed Nd: YAG laser might be advantageous for the treatment of infected root canals.     

Effectiveness of 1 mol L-1 citric acid and 15% EDTA irrigation on smear layer removal

AIM: The aim of this study was to evaluate in vitro the cleansing and smear layer removal capability of alternate canal irrigation with citric acid and NaOCl. METHODOLOGY: Eighty-one teeth were divided into three groups on the basis of the type of instrumentation, namely, manual stainless steel, Ni-Ti mechanized ProFile .04 taper or MACXim. The groups were further divided on the basis of irrigation protocol: 5% NaOCl alone, NaOCl alternated with 1 mol L-1 citric acid solution or a combination of 15% EDTA and Cetrimide solution. After longitudinal sectioning, dentinal walls were microphotographed with scanning electron microscopy at x300 and x1000 magnifications. Qualitative and quantitative cleansing level evaluations were performed using computerized image analysis software. Data were statistically evaluated using Kruskal-Wallis analysis and t-test. RESULTS: Qualitative evaluation at x300 and x1000 showed no statistically significant differences in cleansing ability between citric acid, EDTA and NaOCl groups. Quantitative evaluation of smear layer removal, measured as open tubules/total dentinal surface ratio, showed that 1 mol L-1 citric acid solution was comparable to EDTA (11.97% vs. 10.36%) (NS); in samples treated with ProFile .04 taper instruments citric acid was most effective (16.17%), whilst in the group treated with manual instrumentation EDTA and Cetrimide were the most effective (11.94%). Specimens irrigated with 5% NaOCl demonstrated significantly more cleansing than those obtained in the other two groups (P < 0.001). CONCLUSIONS: 1 mol L-1 citric acid solution was as effective in removing smear layer as EDTA, but was superior in specimens treated with ProFile .04 taper instruments.     

The effectiveness of different endodontic irrigation procedures on the removal of the smear layer: a scanning electron microscopic study

To compare the effectiveness of different irrigation procedures on the removal of the smear layer, 40 curved canals were manually prepared in vitro under copious irrigation with 3 per cent NaOCl. Ten canals served as controls. The remaining 30 were equally distributed in three experimental groups and subsequently irrigated with the following: ultrasound with NaOCl; EDTA; ultrasound with EDTA. In the scanning electron microscope, the appearance of the smear layer was rated over the distribution of dentinal tubule openings for the coronal, middle and apical segments of the canals. Irrigation with NaOCl produced consistently smeared surfaces. Ultrasonic stirring of NaOCl removed the smear layer moderately, while EDTA produced almost smear-free surfaces. Ultrasound in association with EDTA did not enhance the dissolving capability of this chelating agent. A definite decline in the efficiency of the irrigation procedures was also observed along the apical part of the canals.     

The effect of German chamomile (Marticaria recutita L.) extract and tea tree (Melaleuca alternifolia L.) oil used as irrigants on removal of smear layer: a scanning electron microscopy study

AIM: To compare the cleaning effectiveness of chamomile hydroalcoholic extract and tea tree oil to 2.5% sodium hypochlorite (NaOCl) solution as an intracanal irrigant for the removal of the smear layer. METHODOLOGY: Forty extracted, single-rooted, mature, permanent, human teeth were allocated at random into one of three experimental groups of ten teeth and two control groups of five teeth. For each tooth, the pulp chamber was accessed and the canal prepared using K-type files and Gates-Glidden burs, using a step-back technique; the apical stop was prepared to a size 30. Each canal was subsequently irrigated with one of the following solutions: distilled water (as a negative control), 2.5% NaOCl + 17% ethylenediamine tetraacetic acid (EDTA) (as a positive control), chamomile or tea tree oil or 2.5% NaOCl. Each tooth was split longitudinally and prepared for examination by scanning electron microscopy (SEM). The quantity of smear layer remaining on the three levels of each canal (coronal, middle and apical) was examined using magnifications of 2000 and 5000x. The data were analysed using nonparametric Kruskal-Wallis and Mann-Whitney U-tests. RESULTS: The most effective removal of smear layer occurred with the use of NaOCl with a final rinse of 17% EDTA (negative control) followed by the use of a chamomile extract. Chamomile extract was found to be significantly more effective than distilled water and tea tree oil (P < 0.008).The use of a 2.5% NaOCl solution alone, without EDTA and that of tea tree oil, was found to have only minor effects. There was no statistical difference between distilled water, 2.5% NaOCl and tea tree oil. CONCLUSIONS: The efficacy of chamomile to remove smear layer was superior to NaOCl alone but less than NaOCl combined with EDTA.     

An apical leakage study in the presence and absence of the smear layer

The apical seal produced by low-temperature injection of gutta-percha, with or without root canal sealer, in the presence or absence of the smear layer was evaluated quantitatively using an electrochemical technique. The root canals of 44 teeth were prepared with the smear layer intact following a final flush of NaOCl (group 1), while the root canals of a further 44 teeth were prepared and the smear layer removed with a final flush with EDTA followed by NaOCl (group 2). Subgroups were obturated using Ultrafil System alone or accompanied by Calciobiotic Root Canal Sealer. Microleakage data was expressed as a percentage of apical leakage evident in each group. Statistical analysis of the results revealed that the groups obturated with Ultrafil alone showed no significant differences in leakage compared with the groups where sealer was used, either when the smear layer had been left intact or removed. (P>0.05). However, comparison of the combined two groups with smear layer present versus the combined two groups with smear layer removed showed a highly significant difference (P<0.01), with the incidence of leakage reduced in the absence of the smear layer.     

Comparison of the cytotoxic effects and smear layer removing capacity of oxidative potential water, NaOCl and EDTA

This study compared the smear layer removing capability and cytotoxicity of NaOCl, EDTA and Oxidative Potential Water (OPW). Fifteen extracted single-rooted human upper incisors were examined in three groups. The root canals were enlarged to the apical foramen with K files to size #60 and irrigated with: (a) NaOCl followed by OPW, (b) OPW during and after instrumentation and (c) NaOCl followed by EDTA and NaOCl. The effect of these irrigants on the smear layer was evaluated using a scanning electron microscope. In vitro cytotoxicity of these irrigants was examined by MTT colorimetric assay. We found that the combination of NaOCl and OPW as well as the application of OPW alone, failed to remove the smear layer from the apical third, whereas the EDTA and NaOCl combination achieved complete removal. OPW, when used during and after instrumentation, removed the smear layer in the middle third more effectively than NaOCl followed by OPW. EDTA exerted more cytotoxic effects at all concentrations tested when compared with OPW and NaOCl. In conclusion: (a) OPW was less cytotoxic than other irrigants but did not effectively remove the smear layer, (b) treatment with EDTA followed by NaOCl efficiently removed of the smear layer, but their cytotoxicity should be considered during endodontic therapy.     

Effects on smear layer and debris removal with varying volumes of 17% REDTA after rotary instrumentation

The purpose of this study was to quantify the volume of 17% ethylene diamine tetra-acetic acid (EDTA) needed to efficiently remove the smear layer after rotary instrumentation, and to determine if additional irrigation has any effect on debris removal. Forty single canal teeth were instrumented with ProFile GT rotary instruments. Experimental groups were irrigated with 1, 3, or 10 ml of 17% EDTA for 1 min, followed by a final rinse with 3 ml of 5.25% sodium hypochlorite (NaOCl). Samples were scored for debris remaining and examined under SEM to determine quality of smear layer removal. There were no significant differences among groups when comparing either debris remaining or quality of smear layer removal. EDTA irrigation volume greater than 1 ml did not improve debris removal. Efficient removal of the smear layer was accomplished with a final rinse of 1 ml of 17% EDTA for 1 min, followed by 3 ml of 5.25% NaOCl.     

Effect of calcium hydroxide and four irrigation regimens on instrumented and uninstrumented canal wall topography

The topography of instrumented and uninstrumented canal walls exposed to calcium hydroxide and four different irrigation regimens was observed by scanning electron microscopy. After chemomechanical debridement, one tooth in each matched pair was medicated with calcium hydroxide. One week later, the teeth were irrigated and split longitudinally for evaluation. When no calcium hydroxide was used, predentin and pulpal debris covered the dentinal tubules of the uninstrumented surfaces in specimens irrigated with water or EDTA, but was absent on uninstrumented surfaces in specimens irrigated with NaOCl or NaOCl and EDTA. A typical smear layer was absent in instrumented specimens irrigated with NaOCl and EDTA, but covered the dentinal tubules of the instrumented surfaces of the EDTA irrigated specimens (partially) and the water or NaOCl irrigated specimens (completely). Calcospherites or their remnants were seen on the uninstrumented canal walls of specimens irrigated with NaOCl or NaOCl and EDTA, respectively. Calcium hydroxide use did not alter the surface topography in specimens irrigated with water, EDTA, or NaOCl, but seemed to erode the intertubular dentin in specimens irrigated with NaOCl and EDTA. All irrigants seemed to effectively remove most of the calcium hydroxide.     

Evaluation of the smear layer removal effectiveness of EDTA using two techniques: an SEM study

Obtaining the cleanest canal possible before obturation is one of the goals of endodontic treatment. The purpose of this study was to compare the smear layer removal capability of ethylenediamine tetra-acetic acid (EDTA) application with passive ultrasonic and cotton wrapped on reamer activation. Twelve extracted human teeth with single root canals were used for the study. They were conventionally hand instrumented using reamers and K files. The apical aspect of the canals was enlarged to a #40 file. The teeth were divided into 4 four groups, instrumented and irrigated as follows: Group-A EDTA agitated at the end of preparation with cotton wrapped on a reamer for 1 min; Group-B EDTA applied with ultrasonic agitation for 1 min; Group-C irrigated with EDTA+sodium hypochlorite (NaOCl) (negative control group); and Group-D irrigated with distilled water (positive control group). After scanning electron microscopic study at three different levels, smear layer and dentinal tubules were scored. Means were tested for significance using the Z test. When the techniques were compared, the cotton wrapped on reamer agitation method was as successful as the ultrasonic activation of the files. Although all groups had significantly higher smear layer scores at apical compared to coronal sections, no significant differences were recorded.     

Comparison of the Efficacy of Maleic Acid and Ethylenediaminetetraacetic Acid in Smear Layer Removal from Instrumented Human Root Canal: A Scanning Electron Microscopic Study

IntroductionThe aim of this study was to assess, by scanning electron microscopic (SEM) analysis, the ability of 17% EDTA and 7% maleic acid in the removal of the smear layer from the human root canal system.MethodsEighty single-rooted human anterior teeth were subjected to standardized root canal instrumentation (step-back technique) and were irrigated with 2.5% NaOCl after each instrument. Based on the final irrigating solution used, samples were divided randomly into three groups: (1) the EDTA group: 17% EDTA+ 2.5% NaOCl (n = 30), (2) the maleic acid group: 7% maleic acid + 2.5% NaOCl (n = 30), and (3) the control group: 0.9% saline (n = 20). After final irrigation, teeth were prepared for SEM analysis to evaluate the cleaning of the coronal, middle, and apical thirds of radicular dentin by determining the presence or absence of smear layer. The data was statistically analyzed using the Kruskall-Wallis test.ResultsAt the coronal and middle thirds, there was no significant difference between EDTA and maleic acid. Both were equally efficient in the removal of smear layer. In the apical third, maleic acid showed significantly better smear layer removing ability than EDTA.ConclusionFinal irrigation with 7% maleic acid is more efficient than 17%EDTA in the removal of smear layer from the apical third of the root canal system, which is a crucial area for disinfection.     

A Comparative Study of Smear Layer Removal and Erosion in Apical Intraradicular Dentine With Three Irrigating Solutions: A Scanning Electron Microscopy Evaluation

IntroductionThis study compared the efficacy of BioPure MTAD (Dentsply Tulsa, Tulsa, OK), 17% EDTA, and 42% citric acid in endodontic smear layer removal and degree of erosion in the apical third of endodontic canals.MethodsNinety-six extracted single-rooted human teeth were randomized into four groups (n = 24) and instrumented using System GT nickel-titanium rotary instruments (Dentsply Tulsa, Tulsa, OK). Each canal was irrigated with one of the following solutions: BioPure MTAD, 17% EDTA, 42% citric acid, or 5.25% NaOCl (control). Next, all specimens were irrigated with 5.25% NaOCl.ResultsEvaluation by scanning electron microscopy showed no significant differences among test irrigants in removing the smear layer. However, the efficacy of BioPure MTAD and 17% EDTA in removing the smear layer was significantly greater than 5.25% NaOCl (control). The erosive effects of irrigating solutions could not be evaluated.ConclusionsIn conclusion, the protocols used in this study were not sufficient to completely remove the smear layer in the apical third of prepared root canals.     

Push‐out bond strength of gutta‐percha with a new bioceramic sealer in the presence or absence of smear layer

The purpose of this study was to compare the bond strength of a new bioceramic sealer (EndoSequence BC Sealer) and AH Plus in the presence or absence of smear layer. Extracted single‐rooted human teeth were prepared and randomly divided into four groups. In groups 1 and 3, the root canals were finally irrigated with 5.25% NaOCl and smear layer was not removed, but in groups 2 and 4, the root canals were finally irrigated with 17% EDTA followed by 5.25% NaOCl in order to remove the smear layer. In groups 1 and 2, the root canals were obturated with gutta‐percha/AH Plus, but in groups 3 and 4, obturation was performed with gutta‐percha/EndoSequence BC Sealer. Push‐out bond strength and failure modes were evaluated. The bond strength of gutta‐percha/AH Plus and gutta‐percha/EndoSequence BC Sealer was not significantly different (P = 0.89). The presence or absence of smear layer did not significantly affect the bond strength of filling materials (P = 0.69). The mode of bond failure was mainly cohesive for all groups. In conclusion, the bond strength of the new bioceramic sealer was equal to that of AH Plus with or without the smear layer.     

Comparison of the efficacy of three chelating agents in smear layer removal

The purpose of this study was to compare the efficacy of SmearClear (Sybron Endo, Orange, CA), 17% EDTA, and 10% citric acid in smear layer removal. Forty-eight extracted single-rooted human teeth were randomly divided into 4 groups (n = 12) and instrumented using Mtwo nickel-titanium rotary instruments. Each canal was subsequently irrigated with one of the following solutions: 5.25% NaOCl (control), SmearClear, 17% EDTA, or 10% citric acid. After that, all the specimens were subjected to irrigation with 5.25% NaOCl. The teeth were then processed for scanning electron microscopy (SEM), and the removal of the smear layer was examined in the coronal, middle, and apical thirds. The results showed that there were no significant differences in the efficacy of three chelating agents at all levels of the root canals. The comparison of three one thirds in each group showed no significant difference in the SmearClear and EDTA groups. However, the efficacy of citric acid was significantly less in the apical third compared with the coronal and middle thirds of the canals. In conclusion, the protocol used in this study was not efficient to completely remove the smear layer especially in the apical third of the canal.     

Trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on cultured odontoblast cell lines after consecutive applications

Aim  To evaluate the trans-enamel and trans-dentinal cytotoxic effects of a 35% H₂O₂ bleaching gel on an odontoblast-like cell lines (MDPC-23) after consecutive applications.
Methodology  Fifteen enamel/dentine discs were obtained from bovine central incisor teeth and placed individually in artificial pulp chambers. Three groups ( n  = 5 discs) were formed according to the following enamel treatments: G1: 35% H₂O₂ bleaching gel (15 min); G2: 35% H₂O₂ bleaching gel (15 min) + halogen light (20 s); G3: control (no treatment). After repeating the treatments three consecutive times, the extracts (culture medium + gel components that had diffused through enamel/dentine discs) in contact with the dentine were collected and applied to previously cultured MDPC-23 cells (50 000 cells cm⁻²) for 24 h. Cell metabolism was evaluated by the MTT assay and data were analysed statistically (α = 5%; Kruskal–Wallis and Mann–Whitney U -test). Cell morphology was analysed by scanning electron microscopy.
Results  Cell metabolism decreased by 92.03% and 82.47% in G1 and G2 respectively. G1 and G2 differed significantly ( P  < 0.05) from G3. Regardless of halogen light activation, the application of the bleaching gel on the cultured odontoblast-like cells caused significantly more severe cytotoxic effects than those observed in the nontreated control group. In addition, significant morphological cell alterations were observed in G1 and G2.
Conclusion  After three consecutive applications of a 35% H₂O₂ bleaching agent, the diffusion of the gel components through enamel and dentine caused severe toxic effects to cultured pulp cells.