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4 "MTT Assay"
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Research Article
A comparative evaluation of cytotoxicity of root canal sealers: an in vitro study
Gautam Pyarelal Badole, Manjusha Madhukar Warhadpande, Ganesh Kothiramji Meshram, Rakesh Namdeoraoji Bahadure, Shubha Gopal Tawani, Gopal Tawani, Shital Gautam Badole
Restor Dent Endod 2013;38(4):204-209.   Published online November 12, 2013
DOI: https://doi.org/10.5395/rde.2013.38.4.204
AbstractAbstract PDFPubReaderePub
Objectives

The objective of this in vitro study was to evaluate and compare the cytotoxicity of four different root canal sealers i.e. Apexit Plus (Ivoclar Vivadent), Endomethasone N (Septodont), AH-26 (Dentsply) and Pulpdent Root Canal Sealer (Pulpdent), on a mouse fibroblast cell line (L929).

Materials and Methods

Thirty two discs for each sealer (5 mm in diameter and 2 mm in height) were fabricated in Teflon mould. The sealer extraction was made in cell culture medium (Dulbecco's Modified Eagle's Medium, DMEM) using the ratio 1.25 cm2/mL between the surface of the sealer samples and the volume of medium in a shaker incubator. Extraction of each sealer was obtained at 24 hr, 7th day, 14th day, and one month of interval. These extracts were incubated with L929 cell line and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay was done. Two-way ANOVA for interaction effects between sealer and time and Post-hoc multiple comparison using Tukey's test across all the 16 different groups were used for statistical analysis.

Results

Apexit Plus root canal sealer was significantly less toxic than other sealers (p < 0.05) and showed higher cellular growth than control. Endomethasone N showed mild cytotoxicity. AH-26 showed severe toxicity which became mild after one month while Pulpdent Root Canal Sealer showed severe to moderate toxicity.

Conclusions

Apexit Plus was relatively biocompatible sealer as compared to other three sealers which were cytotoxic at their initial stages, however, they became biocompatible with time.

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Original Articles
Evaluation of the radiopacity and cytotoxicity of resinous root canal sealers
Chang-Kyu Kim, Hyun-Wook Ryu, Hoon-Sang Chang, Byung-Do Lee, Kyung-San Min, Chan-Ui Hong
J Korean Acad Conserv Dent 2007;32(5):419-425.   Published online September 30, 2007
DOI: https://doi.org/10.5395/JKACD.2007.32.5.419
AbstractAbstract PDFPubReaderePub

The aim of this study was to evaluate the radiopacity and cytotoxicity of three resin-based (AH 26, EZ fill and AD Seal), a zinc oxide-eugenol-based (ZOB Seal), and a calcium hydroxide-based (Sealapex) root canal sealers. Specimens, 10 mm in diameter and 1 mm in thickness, were radiographed simultaneously with an aluminum step wedge using occlusal films, according to ISO 6876/2001 standards. Radiographs were digitized, and the radiopacity of sealers was compared to the different thicknesses of the aluminum step wedge, using the Scion image software. Using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the cytotoxicity of each material was determined in immortalized human periodontal ligament (IPDL) cells.

The results demonstrated that EZ fill was the most radiopaque sealer, while Sealapex was the least radiopaque (p < 0.05). AH 26, AD Seal and ZOB Seal presented intermediate radiopacity values. All the materials evaluated, except for Sealapex, presented the minimum radiopacity required by ISO standards. The cell viabilities of resin-based root canal sealers were statistically higher than that of other type of root canal sealers through the all experimental time. Further, EZ fill showed statistically lower cell viability in 24 and 48 hours compared to AD Seal and in 72 hours compared to all other resin-based root canal sealers. However, there was no correlation between the radiopacity and cytotoxicity of three resin-based root canals sealers (p > 0.05).

These results indicate that resin-based root canal sealer is more biocompatible and has advantage in terms of radiopacity.

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Evaluation of periodontal ligament cell viability in rat teeth after frozen preservation using in-vivo MTT assay
Jae-Wook Kim, Eui-Sung Kim, Jin Kim, Seung-Jong Lee
J Korean Acad Conserv Dent 2006;31(3):192-202.   Published online May 31, 2006
DOI: https://doi.org/10.5395/JKACD.2006.31.3.192
AbstractAbstract PDFPubReaderePub

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen (-196℃) with 4℃ cold preservation group.

A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia.

Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium), Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in Viaspan®), Group 4 (10% DMSO in Viaspan®) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium), Group 6 (Viaspan®) at 4℃ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95% level of confidence. Another 2 teeth of each group were treated as the same manner and frozen sections 10 µm thick for microscopic observation.

The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P < 0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results.

In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

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The verification of the MTT assay on the viability of periodontal ligamental cells in rat molars through the histologic examination
Hyun-Ki Kim, Eui-Seoung Kim, In-Bok Choi, Jin Kim, Seung-Jong Lee
J Korean Acad Conserv Dent 2003;28(5):385-391.   Published online September 30, 2003
DOI: https://doi.org/10.5395/JKACD.2003.28.5.385
AbstractAbstract PDFPubReaderePub

The purpose of this study is to examine the viability of PDL cells in rat molars by using MTT assay and to verify the MTT assay through the histologic observation. Thirty of Sprague-Dawley white female rats of 4-weeks old with a body weight of about 100 grams were used. Groupings are as follows:

Immediate Group : Positive control group(n=10)-after extraction immediately.

Dried Group : Negative control group(n=10)-after drying for an hour under warm dry.

ViaSpan® Group : 1hour ViaSpan® group(n=10)-after storing in ViaSpan® at 4℃ for 1hour.

Ten teeth of each group were treated as same as above and replanted to the original socket of experimental animals. After two weeks of replantation, all the experimental animals were sacrificed. And after fixation, extracted maxillary jaw was dimineralized. After it was embedded in paraffin, serial section by 5µm was carried out and for construction of specimen, hematoxylin-eosin dye was used.

The mean MTT measurement of immediate group(positive control) is 2.81 and the mean measurement of dried group(negative control) is 0.98 which is significant differnt(P<0.05). The mean measurement of ViaSpan® group is 2.65 and there is significant difference between dried group and ViaSpan® group(P<0.05). However, there is no difference between immediate group and ViaSpan® group. The average resorption points of immediate group is 3.03 points. In the dried group, average 6.44 points resorption and 2.68 points showed resorption in the ViaSpan® group. Unlike with MTT assay, there was no significant difference between the immediate group and ViaSpan® group.

The usage of MTT assay as a viable cell marker may give us a better indication of the maintenance of periodontal ligament cell vitality.

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