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Dental Materials Journal 2013; 32(1): 96–100

Effect of sodium ascorbate on degree of conversion and bond strength of
RealSeal SE to sodium hypochlorite treated root dentin
Deepti SHRESTHA1, Wan-Cui WU2, Qing-Yin HE1, Xi WEI1 and Jun-Qi LING1

1
Operative Dentistry and Endodontics, Guanghua School of Stomatology, Sun Yat-Sen University, Guangzhou, Guangdong, China
2
The 2nd Affiliated Hospital of Guangzhou Medical College, Guangzhou, Guangdong, China
Corresponding author, Xi WEI; E-mail: weixi@mail.sysu.edu.cn

The aim of this study was to investigate the effect of sodium ascorbate (Sa) on degree of conversion (DC) and bond strength (BS) of
RealSeal SE to sodium hypochlorite (NaOCl) treated root dentin. Two hundreds simulated canals were prepared and irrigated with
Distilled water(DW), 1.3% NaOCl (1.3% N), 5.2% NaOCl (5.2% N), MTAD, 17% EDTA (EDTA), 10% Sa, 1.3% NaOCl/MTAD (N-M),
1.3% NaOCl/Sa/MTAD(N-Sa-M), 5.2% NaOCl/EDTA(N-E), and 5.2% NaOCl/Sa/EDTA (N-Sa-E) respectively. They were subsequently
bulk filled with RealSeal SE and analyzed with micro-Raman spectroscopy and universal testing machine for DC and BS respectively.
One-way ANOVA and post hoc Tukey’s test showed DC of 1.3% N, 5.2% N, N-M and N-E were significantly lower (p<0.01) than other
six groups. BS of DW, Sa, N-M were significantly lower than 1.3% N, 5.2% N, MTAD, EDTA, N-Sa-M and N-E (p<0.01), and group
N-Sa-E achieved the highest BS among all groups (p<0.01). NaOCl negatively affected DC and BS of RealSeal SE, which could be
reversed with 10% Sa.

Keywords: Degree of conversion, Micro-Raman spectroscopy, Push-out bond strength, RealSeal SE, Sodium ascorbate

layer7,8). BioPure MTAD (a mixture of 3% doxycycline
INTRODUCTION
hyclate, 4.25% citric acid, and 0.5% polysorbate-80
Dental adhesive technologies have been borrowed from detergent; DENTSPLY Tulsa, Tulsa, US), has been
restorative dentistry in an attempt to seal the root canal introduced to remove the smear layer along with NaOCl
system more effectively1). Among various methacrylate- with less destruction to the tooth structure. Moreover,
based sealers, RealSeal SE (a mixture of EBPADMA, previous study by Torabinejad et al.9) has demonstrated
HEMA, BIS-GMA, acidic methacrylate resins, silane- that there were no significant differences between the
treated bariumborosilicate glasses, silica, hydroxylaptite, ability of 1.3% NaOCl and 5.25% NaOCl as root canal
Ca-Al-F-silicate, bismuth oxychloride with amines, irrigations with MTAD to remove the smear layer.
peroxide, photo initiator, stabilizers and pigment, Therefore, it seems prudent to use 1.3% NaOCl with
SybronEndo, Orange, CA) is the new generation of self- MTAD to remove the smear layer to avoid toxic effect
etching adhesive sealer which may reduce the technique of high concentrations of NaOCl9). Chemical substances
sensitivity and application time of bonding to root canal2). used as irrigation during instrumentation might affect
RealSeal SE is claimed to bond to the radicular dentin the dentin substrate10). NaOCl, an oxidizing agent, has
to form monoblock structure aiming in achieving a fluid- been shown to retard polymerization of resin based
imperious seal3,4). The self-etching bonding mechanism materials, thus, affecting the degree of conversion (DC)
is complex, including resin monomer infiltration and bond strength (BS)11). Therefore, the manufacturer
into the conditioned dentine surface and subsequent recommends the use of EDTA as the final rinse to
polymerization to create micromechanical interlocking overcome the adverse effect of NaOCl while achieving
between the dentin collagen and resin3,4). It was reported adequate removal of smear layer before obturation
that imperfect polymerization of the adhesive resin with RealSeal SE. Other studies on coronal restorations
could create water-rich and stress concentration zones have demonstrated the adverse effect of NaOCl on BS
that might reduce the resin-dentin bonds and leave and marginal seal with its reversal with ascorbic acid
the interface susceptible to water degradation over and sodium ascorbate (Sa)12-15). Bonding of the root
time5). It would be important to investigate the degree filling materials to intraradicular dentine is similar
of polymerization and adaptation of RealSeal SE to root to contemporary adhesive strategies used for coronal
canal dentin. restorations16). However, no study has been done to
Successful endodontic therapy depends on investigate the effect of an antioxidant like sodium
three-dimensional obturation as well as sufficient ascorbate on the DC and BS of RealSeal SE sealer to
chemomechanical preparation of root canal system. NaOCl/EDTA and NaOCl/MTAD treated root dentin.
Mechanical instrumentation of root canals leaves a Thus, this study was performed to observe the effect
smear layer on the canal dentinal walls6). Alternating of EDTA, MTAD or sodium ascorbate as final rinse on
use of ethylenediaminetetraacetic acid (EDTA) and 5.2% DC and BS of RealSeal SE to NaOCl treated root dentin
NaOCl has been proved to be efficient in removing smear by micro-Raman spectroscopic analysis and universal

The first two authors contributed equally to this study.
Received May 8, 2012: Accepted Oct 31, 2012
doi:10.4012/dmj.2012-125 JOI JST.JSTAGE/dmj/2012-125

in order to figure out an Group DW: Distilled water (control group) appropriate irrigation regimen when obturating with Group 1. All spectra (1. Three spots on sealer (2 µm next to resin-dentin interface) were chosen randomly for Raman analysis and their mean was calculated. and the canals were bulk filled with the irrigated with distilled water. The DC was calculated according to the following formula: Fig. Each simulated were dried with multiple paper points until the last canal was instrumented with 6% tapered Hero-Shaper paper point appeared dry. tooth with a low speed diamond saw (Isomet. Tooth slices were stored in 100% humidity and absolute dark for 1 week to allow self curing of the material.3% NaOCl+MTAD collected and stored in 0. The coronal portion of each tooth Group N-E: 5. Dent Mater J 2013. RealSeal SE. UK) following Wu’s Protocol20). The slice was distilled water.. according to the irrigation solution used. They were covered with glass cover slips they were randomly divided into 10 groups (N=20) to avoid oxygen interruption during polymerization19).2% NaOCl+EDTA was embedded in epoxy-resin. Gloucestershire.500–1.08 with auto-mix syringe according to the manufacturer’s mm in diameter).609 cm−1 (aromatic C=C) in cured of the tooth. Vertically oriented truncated canals and uncured material respectively. During irrigation of the simulated canals.2% NaOCl+sodium ascorbate+EDTA thick longitudinal slice was prepared by making bucco. Buehler Ltd. sodium ascorbate (pH 7) and/or sectioned perpendicular to the long axis of the tooth at the 17% EDTA (pH 7. When cemento-enamel junction to obtain the radicular portion MTAD was used. Raman-spectra of uncured RealSeal SE (N=2) were obtained immediately after mixing with an automix syringe. after which samples were subjected to micro-push out bond test. DC=(1−Rcured/Runcured)×100 1. For each tooth. micro-push out bond . 1 Representative experimental specimen. samples from all 10 groups (N=20) were analyzed with a micro-Raman spectrometer (Renishaw in Via. Five mL of distilled water was used as irrigation lingual sections parallel to the longitudinal axis of the subsequently after each solution to avoid interactions18). After micro-Raman spectroscopy.3% NaOCl RealSeal SE.0 mm Group N-Sa-E: 5. R referred to the ratio of peak heights at 1. canals were instructions. The simulated canals following Huffman’s protocol (Fig.800 cm−1) were taken with one accumulation of 60 s exposure time.02% sodium azide at room Group N-Sa-M: 1. 5 mL of Lake Bluff. All 100 teeth slices (200 simulated canal spaces) were subjected to micro-Raman spectroscopic analysis. IL.2% N: 5. NaOCl. USA) under water cooling.3% N: 1. RealSeal SE was prepared Nickel-Titanium rotary file number 30 up to D13 (1.5) were used for 1 min each.94 mW laser power. Group 5. a 1. After instrumentation. The Raman spectra were excited by a 785 nm laser line at a resolution of 1 cm−1. Origin 8 (OriginLab Corp. were prepared on either side of the radicular dentin in the middle third with equal distance Micro-push out bond test from cementum and the canal wall.0 mm thick longitudinal slice below the cemento- enamel junction was prepared by making bucco. The laser beam was focused with an optical microscope at ×50 magnification with approximately 4.2% NaOCl Group MTAD: MTAD MATERIALS AND METHODS Group EDTA: 17% EDTA Group Sa: Sodium ascorbate One hundred single rooted extracted human teeth were Group N-M: 1. 1 mL of the solution was slowly only.639 cm−1 lingual sections parallel to the longitudinal axis (aliphatic C=C) and 1.3% NaOCl+sodium ascorbate+MTAD temperature until use. Then the canal was rinsed from cementum and the canal wall) were prepared on with the remaining 4 mL of MTAD according to the either side of the radicular dentin in the middle third manufacturer’s instructions15). Micro-Raman spectroscopy One week after bulk filling of simulated canals with RealSeal SE. Vertically oriented truncated canals (equidistant placed in the canal for 5 min. Northampton. 1)17). During instrumentation. 32(1): 96–100 97 testing machine respectively. MA) was used as the software for spectral analysis.

75) and N-E (72.59±3.08±3. differences among the groups (p<0.00±2.48e Groups represented by different superscript letters are significantly different (p<0.60±2.84).3% NaOCl 71.58±2.05±1.88±1.35±2.71±1.22b 9.75b 4.44±2. IL).88) C=C) was listed in descending order.59±3.59) (Table 1).74±1. 32(1): 96–100 test was carried out using a universal-testing machine was significantly lower (p<0.3% N (71.34±1.35±2.2% NaOCl 70.03±1. 5.3% NaOCl/MTAD 73.22). introducing shear stresses along the interfaces. Table 1 Degree of conversion (in percentage) and micro-push out bond strengths (in MPa) of different groups (mean±standard deviation) Treatment Degree of conversion (%) Bond strength (MPa) Distilled water 80. peak heights at 1. groups. Post hoc Tukey’s test Using 1.51±1.96).01). A 0. (9. rejecting the null hypothesis.0.37a 4. .35).88±1. MTAD h.37).02c 1. 5.04c 1. one- way Analysis of variance (ANOVA) was used to test the difference among groups at 95% confidence interval with SPSS software package (version 16.844c 1.08±3.88b 9. The load at One-way ANOVA showed statistically significant failure indicated by the sharp drop was recorded. Post hoc Tukey’s test was performed to compare the difference between the groups.15±1. N-Sa-M (80.56) and The push-out bond strength was calculated according N-E (9.04) were significantly lower (p<0.2% N standard. (78.71±1. One-way ANOVA showed Fig. SPSS Inc.59a 12.43d 10% Sodium ascorbate 80.05±1. N-M a vernier calliper.56d 5. 2 Representative micro-Raman spectra of all the statistical differences among the groups (p<0. Post hoc Tukey’s test showed height (h) of the simulated canals were measured using that the BS of DW (3.98).5(π×cd+π×ad) those of 1.83d 5.70±0. A compressive force at a speed of 1 mm/min was exerted to Micro-push out bond test the inverted truncated cavity until failure. EDTA (8. The bonded area of the root filling (4.01). MTAD (79.76d 5. The null hypothesis was that there was no significant difference among the different groups.76).2% NaOCl /10% sodium ascorbate/17% EDTA 78.02).48) achieved to the following formula: the highest BS among various groups (p<0.62d 17% EDTA 79.98 Dent Mater J 2013.43a 9. N-Sa-M (7.34±1.62). N-M (73.35b 8.28±1.45).2% NaOCl/17%EDTA 72.639 cm−1 (aliphatic (70.88±3.98a 8.55±3.74±1.03±1. apical diameter (ad) and hypothesis was rejected. Group N-Sa-E (12.52±2.70±0.01) than material was calculated as Area (mm2)=0.58±2.52±2.90d MTAD 79.52a 7.43).00±1.60±2.51±1. cylindrical stainless steel plunger tip was positioned Sa (80.3% N (8.3% NaOCl/sodium ascorbate/MTAD 80.01) and the null The coronal diameter (cd). EDTA (79.8 mm diameter (80.609 cm−1 (aromatic C=C) as an internal showed that the DC of 1.2% N (9. Chicago.28±1.96 a 3. Sa (4.88±3. 2.83). Micro-push out bond strength (MPa)=Maximum Load (N)/Area (mm2) Statistical analysis The data were normally distributed.55±3. RESULTS Micro-Raman spectroscopy Representative micro-Raman spectra of different groups were presented in Fig.01) than those of DW (Zwick/Roell Z010.43).15±1.44±2.52) and N-Sa-E so that it contacted only the filling material on loading. Germany).01) (Table 1). Therefore.

Further study is needed to bond strength11).88.25% NaOCl/EDTA recommend 5.25). be attributed to the smear layer which could not be it was possible that the prevention was not thorough. 32(1): 96–100 99 EDTA or 1.28) has demonstrated the photo- Dentin surface treatment with different irrigation redox reaction between NaOCl and MTAD involved regimens may cause alteration in the chemical and oxidation of doxycycline from MTAD triggered by structural composition of human dentin. Although combined use of NaOCl and MTAD ACKNOWLEDGMENT was able to remove smear layer. significantly different after irrigation with 5. the application of sodium As vitamin C and its salts (e. the Ministry of Health of China (no. adhesive materials31).3% NaOCl/MTAD irrigation. and they have some potential (AODTC) by photo-acoustic spectroscopy29). Dent Mater J 2013. Similarly. Some studies have among NaOCl. Shokouhinejad et al. while sodium ascorbate with strength of resin-based obturation system30) and other a neutral pH had no effect on dentin demineralization23). removed by combination irrigation with NaOCl and In summary. between irrigation of 1. Since DC is an important ascorbate between 5.28) revealed that such water and sodium ascorbate treated dentin. with the manufacturer’s recommendation. and performed irrigation with light exposure.2% NaOCl and EDTA achieved aspect of methacrylate-based material affecting the highest push-out BS and optimal DC. layer. Our study demonstrated that tooth surface after 1. However. had previously demonstrated that product from 5. which would inhibit the the compromised DC of RealSeal SE to NaOCl/EDTA or polymerization of monomers in RealSeal SE11. DC and BS of RealSeal SE are adversely EDTA.22). present study. combination of 1. which was inconsistent DISCUSSION with our study. photo-redox interaction between NaOCl and doxycycline The present study showed lower bond strength in from MTAD. NaOCl/EDTA obtained similar bond affected by NaOCl root canal irrigation. Therefore. containing both organic and inorganic component. When NaOCl is used for reaction might be confined partially. There was no significant difference which would affect the bonding. Use of 10% It was possible that EDTA was not able to inactivate sodium ascorbate after NaOCl irrigation can reverse the oxidizing effect of NaOCl.27) revealed the funding from Department of Science and Technology in push-out bond strength of Resilon/Epiphany SE was not Guangdong Province(NO:00272400127680054). Therefore it will be interesting to observe the effect of could not be effectively removed by irrigation with NaOCl on other methacrylate-based resin sealers and NaOCl. Orange resulting in premature chain termination and incomplete precipitates were observed on simulated root canal polymerization11. MTAD or EDTA alone9. resulting NaOCl/MTAD treated root dentin. there might be some reactive used simulated canal spaces produced on thin root slice free radicals which inhibit the polymerized reaction.44±2. their use on resulted in a significant increase in bond strength in the dentine might create no adverse biological effect23). acid and sodium ascorbate (salt of ascorbic acid) both 12 alpha-anhydro-4-oxo-4-dedimethylaminotetracycline have potent anti-oxidants. we bond strength26).27) performed irrigation in affecting the quality and adhesion of root canal filling root canal where light was absent. thereby NaOCl. Such result was in disagreement when RealSeal SE is used in root canal obturation. Such orange precipitate may be the not dependent on the concentration of NaOCl. It was because smear treated dentin is dependent on adhesive systems32). sodium ascorbate) are ascorbate.g. a reducing agent. EDTA or MTAD could not reverse this effect. Although Tay et al.3% NaOCl with MTAD as irrigation in the present study This study was supported by Key Clinical Program of obtained similar bond strength to groups DW and Sa. This may interaction could be prevented by sodium ascorbate. bond strength of RealSeal SE According to the results. used instead of ascorbic acid because ascorbic acid was Use of 10% sodium ascorbate. Tay et al. MTAD and EDTA treated groups. sodium ascorbate was determine the exact nature of such oxidation product. However.25% NaOCl/ . bonding in group 5. Accordingly. It could be effectively its reversal with sodium ascorbate.3% NaOCl/MTAD. to alter the oxidizing agents via redox reaction on the similar reaction product was inferred from the oxidation NaOCl-treated substrate and reverse the compromised reaction of doxycycline. It might be attributed to the no effect on the bond strength of RealSeal SE sealer.2% NaOCl/Sa/EDTA as irrigation regimen was adversely affected. non-toxic and are used in food industry. etched with RealSeal SE sealer leading to reduction of leaving some orange precipitates in the dentinal tubules the bond strength24). Final rinse with strength to NaOCl. we rejected the null in group NaOCl/Sa/MTAD was statistically lower than hypothesis that different irrigation regimens will have group NaOCl/Sa/EDTA. since oxidation product of doxycycline by NaOCl28). [2010]439) and the Previous study by Shokouhinejad et al. MTAD. and such photo-redox material to root dentin21). Whereas our study root canal irrigation.22). Thus it slices irrigated with NaOCl achieved comparatively would be possible that the formation of orange precipitate lower DC and could be reversed after 10% sodium on root dentin adversely affected the bonding between ascorbate treatment. or NaOCl/EDTA treated shown that the compromised bond strength to NaOCl groups in terms of bond strength. acidic in nature and may affect the bond strength by after NaOCl irrigation has demonstrated increased bond demineralizing the dentin. EDTA. Such compromised effect was sealer and dentin.2% NaOCl were statistically insignificant. Application of sodium in lower DC of 72.3% NaOCl and et al. Davies differences in DC between Groups 1.3% NaOCl and MTAD. Ascorbic photo-oxidation reaction of tetracycline was 4 alpha. In our study.

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