ORIGINAL RESEARCH

https://doi.org/10.5005/jp-journals-10015-2229
World Journal of Dentistry
Volume 14 | Issue 5 | Year 2023

Microleakage Assessment of Calcium Silicate-based Root-end Filling Materials Using Dye Penetration: An In Vitro Study

Bassem M Eid¹, Bedour A Alarfaj², Heba M Abdelaal³, Tarek M Elsewify⁴

^1,2Department of Restorative Dental Sciences, Gulf Medical University, College of Dentistry, Ajman, United Arab Emirates

³Department of Restorative Dental Sciences, Gulf Medical University, College of Dentistry, Ajman, United Arab Emirates; Department of Operative Dentistry, Faculty of Dentistry, Ain Shams University, Cairo, Egypt

⁴Department of Restorative Dental Sciences, Gulf Medical University, College of Dentistry, Ajman, UAE; Department of Endodontic, Faculty of Dentistry, Ain Shams University, Cairo, Egypt

Corresponding Author: Bedour A Alarfaj, Department of Restorative Dental Sciences, Gulf Medical University, College of Dentistry, Ajman, United Arab Emirates, Phone: +971 503004485, e-mail: bedour.abdullah.a@gmail.com

Received on: 03 April 2023; Accepted on: 05 May 2023; Published on: 02 August 2023

ABSTRACT

Aim: To evaluate the sealing ability of different types of calcium silicate-based cement (CSC) when used as retrograde filling materials.

Materials and methods: A total of 85 extracted single-rooted teeth were decoronated, instrumented, and obturated. A 3 mm root end resection was performed, followed by a 3 mm retrograde cavity preparation. Teeth were randomly divided into five equal groups (n = 15) according to the retro-filling material applied; group I, ProRoot mineral trioxide aggregate; group II, Biodentine; group III, NeoPutty MTA; group IV, Produits Dentaires (PD) MTA White; group V, TheraCal LC, in addition to positive and negative controls. After 24 hours, the roots were covered with nail polish and immersed in methylene blue dye for 24 hours. After longitudinal sectioning using a diamond disk, roots were examined using a stereomicroscope at 8× magnification for dye penetration, and maximum linear dye penetration was measured in millimeters using ZEN 3.4 software. Statistical analysis was performed using a one-way analysis of variance (ANOVA) test followed by Tukey’s post hoc test.

Results: All tested retro-filling materials showed dye penetration and failed to totally seal the root end. ProRoot MTA and Biodentine showed the least dye penetration, followed by NeoPutty and PD MTA White with a statistically significant difference. TheraCal LC showed maximum dye penetration with a statistically significant difference.

Conclusion: Altering the consistency and manipulation of different CSC significantly affects their sealing ability.

Clinical significance: Proper sealing of the retrograde cavities is mandatory. ProRoot MTA and Biodentine provided a better sealing ability for retrograde cavities.

How to cite this article: Eid BM, Alarfaj BA, Abdelaal HM, et al. Microleakage Assessment of Calcium Silicate-based Root-end Filling Materials Using Dye Penetration: An In Vitro Study. World J Dent 2023;14(5):435–439.

Source of support: Nil

Conflict of interest: None

Keywords: NeoPutty, Retrograde filling, Sealing ability, TheraCal LC

INTRODUCTION

Proper cleaning and shaping, in addition to three-dimensional fluid-tight sealing of the root canal system, are required for the success of the root canal treatment. Surgical endodontics is indicated to address some mishaps and pathologies that could not be managed using conventional orthograde nonsurgical root canal treatment.¹

Following root-end resection and retrograde cavity preparation during periapical surgery, the cavity should be properly sealed in order to provide a favorable environment for the healing of the periradicular tissues. The retrograde filling material should therefore prevent bacterial leakage and provide a three-dimensional apical seal.²^,³

Retrograde filling materials should be biocompatible, insoluble, easily manipulated, and insensitive to moisture during application and setting. Amalgam, gutta-percha, intermediate restorative, super ethoxy benzoic acid, glass ionomer, and composites were previously used as retrograde filling materials, but they failed to demonstrate all of the desired properties.⁴

Mineral trioxide aggregate cement (MTA) is a calcium silicate-based cement (CSC) developed at Loma Linda University in the mid-1990s.⁵ ProRoot MTA (Dentsply Sirona, York, USA) was the first commercial MTA launched in the market and was extensively studied and tested.⁶ Hence, it became the gold standard for retrograde filling materials owing to its superior biocompatibility, bioactivity, good sealing ability, and antimicrobial properties. Yet, ProRoot MTA has shown some limitations, such as the long setting time, discoloration, and poor handling properties.⁷ Recently, other brands/formulations of MTA have been developed in the market. These formulations share the same original formula, with some minor variations in an attempt to improve the original MTA’s limitations. Produits Dentaires (PD) MTA White (Produits Dentaires SA, Vevey, Switzerland) is one of these formulations, which has been produced to be used for pulp capping, pulpotomy, apical plugs, and retrograde filling.⁸

Biodentine (Septodont, Saint-Maur-des-Fossés, France) is another formulation that solved the problem of the prolonged setting time by the addition of calcium chloride as an accelerator to the liquid. Bismuth oxide was replaced by titanium oxide, yielding better radiopacity of Biodentine.⁹

More recently, CSC has been manufactured in a putty consistency to improve the handling properties without changing the physical property of the material upon setting.¹⁰ NeoPutty (Avalon BioMed, Houston, TX) is a novel premixed CSC version of the Neo MTA composed of fine tricalcium silicate material. Tantalum oxide was used as a radiopacifying agent.¹¹

Another form of CSC is TheraCal LC (Bisco Inc, Schaumburg, IL). The addition of the resin to the calcium silicate helped control the material consistency in addition to the controlled setting time through light curing of the resin. It has been introduced to the market as a direct and indirect pulp capping material or as a liner under restorative materials. It shows physiochemical bonding to dentin, good sealing abilities, and is well tolerated by immortalized odontoblast cells. It is comprised of Portland cement, polyethylene glycol, dimethacrylate polymers, barium zirconates as a radiopacifier, and hydrophilic fumed silica as a thickening agent.¹²

These recently introduced CSC products have not been sufficiently evaluated, and we lack research-based conclusions. The sealing ability/microleakage of TheraCal LC and NeoPutty has not been previously tested as retrograde filling materials. Therefore, the objective of the current study is to evaluate the sealing ability/microleakage of different types of CSC used as retrograde filling materials. The null hypothesis tested is that there is no statistically significant difference between the tested CSC materials’ sealing ability/microleakage when used as retrograde fillings.

MATERIALS AND METHODS

This in vitro study was approved by the local ethical committee.

Samples Selection

A power analysis was performed based on the results of Benz et al.,³ the predicted sample size (n) was found to be a total of 75 samples (i.e., 15 samples per experimental group). Sample size calculation was performed using G*Power version 3.1.9.4.

The selected teeth had been extracted due to orthodontic or periodontal causes and were not related to this study. The selected teeth were free from fractures, resorption, or evidence of more than one canal.

Sample Preparation

All the teeth were cleaned from soft tissues, debris, or calculus using an ultrasonic scaler and then placed in 0.1% thymol solution for 24 hours. All teeth were decoronated at the cementoenamel junction level using a diamond disc mounted on IsoMet saw, leaving the roots approximately 16 mm long to ensure straight-line access and providing a reference plane. All the samples were then stored in distilled water until they were used in the study.

Following working length determination and glide path preparation using K-file #15, the canals were prepared using WaveOne Gold large file (40/0.8) (Dentsply Maillefer, Ballaigues, Switzerland) according to the manufacturer’s recommendation. First, irrigation was performed using 3% sodium hypochlorite followed by 5 mL of ethylenediaminetetraacetic acid (EDTA) 17%. Subsequently, the root canals were dried using paper points and obturated with matching gutta-percha points and BioRoot sealer (Septodont, Saint-Maurdes-Fosses, France) using a single cone technique. Obturated canals were radiographed to determine the quality of the obturation. Then, a 4 mm resin composite filling (3M-ESPE, St. Paul, MN) was applied coronally to provide a corona seal. The roots were wrapped in moist gauze to ensure 100% humidity and stored in an incubator at 37°C for 7 days.

A 3 mm root end resection was performed perpendicular to the long axis using a diamond stone (Mani Inc., Tochigi, Japan) mounted on a high-speed handpiece with water coolant. Root end cavity preparation was performed to a depth of 3 mm using ultrasonic tip AS3D (Satelec, Acteon, France). The cavity’s depth was checked using a calibrated periodontal probe. Cavities were irrigated using saline and then conditioned using EDTA 17% for 3 minutes.

Sample Grouping

The samples were randomly divided into five experimental groups using www.random.org according to the type of root-end filling material used (n = 15), in addition to two control groups (n = 5).

Group I—ProRoot MTA (Dentsply, Tulsa, Oklahoma).
Group II—Biodentine (Septodont, Saint-Maur-des-Fossés, France).
Group III—NeoPutty (Avalon BioMed, Houston, Texas).
Group IV—PD MTA White (Produits Dentaires SA, Vevey, Switzerland).
Group V—TheraCal LC (Bisco Inc., Schaumburg, Illinois).
Group VI—negative control (roots totally sealed using nail polish).
Group VII—positive control (roots left without any root end filling or nail polish).

The retrograde filling materials were mixed and applied to the retrograde cavities according to the manufacturer’s recommendations for all the experimental groups. A double layer of nail polish was then applied to all roots, excluding the apical-most 1 mm. Roots were incubated for 24 hours at 37°C and 100% humidity. All the sample preparation was done by the first author.

Five roots did not receive any filling or nail polish on the external surface of the root and the apical foramen, which served as a positive control. Five roots were totally covered by nail polish on the apical and lateral surfaces, which served as a negative control.

Microleakage Evaluation

All samples were immersed in 2% methylene blue dye for 24 hours. Samples were then washed under running water for 30 minutes. Samples were air-dried, and the nail polish was removed from the root surface using a scalpel. Finally, the samples were sectioned in a buccolingual direction using a diamond disc. All the samples were examined under a stereomicroscope with a magnification of 8× (Carl Zeiss, Jena, Germany) to detect dye penetration. The software ZEN 3.4 was used to measure the maximum linear dye penetration distance along the dentinal walls in millimeters. The evaluation was done by the second and the third authors, who were totally blind to the sample group.

Statistical Analysis

As shown in Flowchart 1, the acquired data were recorded and subjected to statistical analysis. Numerical data were represented as mean and standard deviation (SD) values. Shapiro–Wilk’s test was used to test for normality, while Levene’s test tested the homogeneity of variances. The significance level was set at p < 0.05 within all tests. The data were found to be parametric and showed variance homogeneity, so the analysis was performed using one-way analysis of variance (ANOVA) test followed by Tukey’s post hoc test. Statistical analysis was performed using Jeffreys’s Amazing Statistics Program statistical analysis software version 0.16.0.0 for Windows.

Flowchart 1: Flowchart of workflow

RESULTS

The mean and SD, and intergroup comparison of the dye penetration distance for all the tested groups are displayed in Table 1. There was a statistically significant difference amongst the tested groups p < 00001. The TheraCal group showed the maximum dye penetration followed by the PD MTA White, NeoPutty, Biodentine, then MTA ProRoot, with values of (1.976 ± 0.5442), (1.3224 ± 0.3397), (1.1706 ± 0.36), (0.6667 ± 0.1769), and (0.5724 ± 0.1875), respectively.

**Table 1:** Comparison of means of dye penetration distance among experimental groups
Dye penetration (mm) (mean ± standard deviation)					f-value	p-value
MTA ProRoot	Biodentine	NeoPutty	PD MTA White	TheraCal LC	f-value	p-value
0.5724 ± 0.1875^C	0.6667 ± 0.1769^C	1.1706 ± 0.36^B	1.3224 ± 0.3397^B	1.976 ± 0.5442^A	39.4764	<00001*

*Significant (p < 0.05); means with different superscript letters within the same horizontal row are significantly different ; MTA ProRoot and Biodentine, (p = 0.94609) ns; MTA ProRoot and NeoPutty, (p = 0.00002) s; MTA ProRoot and PD MTA White, (p = 0.00163) s; MTA ProRoot and TheraCal LC, (p = 0.00000) s; Biodentine and NeoPutty, (p = 0.00000) s; Biodentine and PD MTA White, (p = 0.00012) s; Biodentine and TheraCal LC, (p = 0.00000) s; NeoPutty and PD MTA White, (p = 0.75560) ns; NeoPutty and TheraCal LC, (p = 0.00002) s; PD MTA White and TheraCal LC, (p = 0.00000) s

There was no statistically significant difference between the PD MTA White and Neoputty (p = 0.75560). There was no statistically significant difference between the MTA ProRoot and the Biodentine groups (p = 0.94609). Representative stereomicroscopic photographs of the root-end fillings showing the dye penetration of different groups are shown in Figure 1. No dye penetration was shown in the negative control group. Diffuse dye penetration was shown throughout the whole length of the dentinal walls of the positive control group.

Figs 1A to E: Stereomicroscopic photograph of the root-end filling showing the dye penetration in different groups: (A) MTA ProRoot; (B) Biodentine; (C) NeoPutty; (D) PD MTA White; (E) TheraCal LCw

DISCUSSION

The creation of a proper three-dimensional apical seal is deemed mandatory for the success of the periapical surgery. Proper selection and proper manipulation of the root-end filling material are fundamentals for the long-term success of periradicular surgery.²^,³ Leakage of bacteria and bacterial byproducts from coronal or apical pathways leads to failure of the endodontic treatment.¹³^,¹⁴

CSCs are considered the material of choice for sealing retrograde cavities in the micro-endodontics era.¹⁵^-¹⁷ Different brands/formulations of CSC are available nowadays in the market showing slight variations in their chemical compositions.¹⁸^,¹⁹ The main characteristic of tricalcium silicate-based cement is the precipitation of hydroxyapatite crystals in the presence of tissue fluids, followed by the formation of an interfacial layer that consolidates binding between the CSC and the dentinal wall.²⁰

Different methodologies can be used for the evaluation of the sealing ability/microleakage, such as radioactive isotopes, bacterial leakage, fluid filtration, dye penetration, dye extraction, and scanning electron microscopy. Until now, we do not have a standardized method of evaluation of the sealing ability/microleakage. Even within each methodology, we lack a standardized protocol to follow. For fluid filtration, the diameter of the capillary tube, the air bubble length, the evaluation time, in addition, to the used pressure can dramatically affect the results.²¹ No correlation was shown between the dye penetration method and the fluid filtration method.²² Bacterial leakage studies mimic the clinical scenario more than other methodologies, yet, they tend to be more qualitative.²³ Scanning electron microscopic studies might be misleading due to the cracks formed as a result of dehydration and heating during sample preparation.²⁴

The dye penetration method was used in the current study to compare and evaluate the sealing ability of ProRoot MTA, Biodentine, NeoPutty, TheraCal, and PD MTA White using methylene blue dye owing to its bright staining and low molecular weight.²⁵^-³¹ Methylene blue dye was preferred to rhodamine dye and black India ink, which possess an extremely small molecular size and weight, leading to false positive results.³² Linear measurement of the deepest point of dye penetration was used as it provides reliable data about apical sealing ability/microleakage.³³^,³⁴

In the current study, all the positive control samples showed microleakage, confirming the necessity of retrograde filling material. Failure of the dye to penetrate the dentinal walls in all the teeth of the negative control group confirmed the reliability of the methodology adopted. The nail polish totally prevented microleakage, with dye only penetrating the apical portion of the roots. This validates the methodology adopted in the current study.

The results of the present study showed that all the tested materials showed a variable amount of leakage. The null hypothesis is rejected as Biodentine, and ProRoot MTA showed statistically significant better sealing ability and less microleakage compared to the other tested CSC.

The superior sealing ability results of Biodentine and ProRoot MTA in the current study come in full agreement with previous studies.²⁹^,³⁵^-³⁷ This may be attributed to the hydrophilic properties and formations of an interfacial layer between the CSC material and dentin. This interfacial layer is composed of tag-like structures of calcium or phosphate-rich crystalline deposits, which decrease the gap.³⁸ Although there was no statistically significant difference between Biodentine and ProRoot MTA, the less microleakage noted in the Biodentine group could be attributed to the shorter setting time of the Biodentine as testing was performed after 24 hours. This also comes in agreement with Nabeel et al., who showed mean leakage values of 0.80 ± 0.63, 1.40 ± 0.52, and 0.60 ± 0.52 after one day, 1 week, and 1 month for ProRoot MTA and 0.20 ± 0.42, 1.40 ± 0.52, and 2.10 ± 0.57 for Biodentine.³⁹

The more dye penetration values recorded for the Neo MTA Putty compared to Biodentine and ProRoot MTA could be attributed to the viscous or heavy consistency, which might affect the reaction rate and formation of the interfacial layer and hydroxyapatite structure. These results could not be compared to other studies as none could be found in the literature testing the NeoPutty.

Mineral trioxide aggregate cement (MTA) PD showed dye penetration values statistically significantly more than those of ProRoot MTA, which might be related to differences in minor ingredients between different brands affecting the manipulation of the material and its setting time. The calcium tungstate and calcium oxide added to the MTA PD, together with the resultant relatively short final setting time of 15 minutes, might have affected the quality of the interfacial layer and hydroxyapatite structure. These results also could not be directly compared to other studies, as none could be found in the literature addressing MTA PD.

The more dye penetration values recorded for the TheraCal LC come in agreement with Camilleri et al., findings of incomplete hydration and low calcium ion leaching.⁴⁰ This could be easily explained by the slow reaction rate of TheraCal LC. Another explanation could be due to the application of the material on dried dentin. Whereas the manufacturer instructs placing this material on moist dentin in order to achieve the best properties and the best bonding to the dentin subtract.⁴¹

Limitations of this study include the in vitro nature of the study, which does not actually mimic the clinical situation, and performing the study at a certain point in time rather than at different intervals. Another limitation is the longitudinal sectioning, where the cut section might not show the section of the deepest dye penetration.

CONCLUSION

Within the limitations of this in vitro study, it can be concluded that altering the consistency and manipulation of different CSC significantly affects their sealing ability. Further investigation of different newly launched CSC at different intervals is needed. Standardization of microleakage/sealing ability assessment is deemed mandatory to properly evaluate and compare different retrograde filling materials.

REFERENCES

1. Estrela C, Holland R, Estrela CR, et al. Characterization of successful root canal treatment. Braz Dent J 2014;25(1):3–11. DOI: 10.1590/0103-6440201302356

2. Shahriari S, Faramarzi F, Alikhani MY, et al. Apical sealing ability of mineral trioxide aggregate, intermediate restorative material and calcium enriched mixture cement: a bacterial leakage study. Iran Endod J 2016;11(4):336–340. DOI: 10.22037/iej.2016.16

3. Benz K, Markovic L, Jordan RA, et al. In vitro evaluation of the apical seal of two different retrograde filling materials. Aust Endod J 2017;43(1):29–33. DOI: 10.1111/aej.12161

4. Kadam A, Gaikwad A, Misra H. Comparative evaluation of the sealing ability of two different retrograde filling materials using dye penetration method: an in-vitro study. IP Indian J Conserv Endod 2020;4(4):121–125. DOI: 0.18231/j.ijce.2019.030

5. Torabinejad M, Hong CU, McDonald F, et al. Physical and chemical properties of a new root-end filling material. J Endod 1995;21(7):349–353. DOI: 10.1016/S0099-2399(06)80967-2

6. Abusrewil SM, McLean W, Scott JA. The use of Bioceramics as root-end filling materials in periradicular surgery: a literature review. Saudi Dent J 2018;30(4)273–282. DOI: 10.1016/j.sdentj.2018.07.004

7. Dawood AE, Parashos P, Wong RHK, et al. Calcium silicate-based cements: composition, properties, and clinical applications. J Investig Clin Dent 2017;8(2):1–15. DOI: 10.1111/jicd.12195

8. Kassab P, El Hachem C, Habib M, et al. The pushout bond strength of three calcium silicate-based materials in furcal perforation repair and the effect of a novel irrigation solution: a comparative in vitro study. J Contemp Dent Pract 2022;23(3):289–294. DOI: 10.5005/jp-journals-10024-3309

9. Nowicka A, Lipski M, Parafiniuk M, et al. Response of human dental pulp capped with biodentine and mineral trioxide aggregate. J Endod 2013;39(6):743–747. DOI: 10.1016/j.joen.2013.01.005

10. Walsh RM, Woodmansey KF, Glickman GN, et al. Evaluation of compressive strength of hydraulic silicate-based root-end filling materials. J Endod 2014;40(7):969–972. DOI: 10.1016/j.joen.2013.11.018

11. Siboni F, Taddei P, Prati C, et al. Properties of NeoMTA plus and MTA plus cements for endodontics. Int Endod J 2017;50(Suppl 2):e83–e94. DOI: 10.1111/iej.12787

12. Poggio C, Arciola CR, Beltrami R, et al. Cytocompatibility and antibacterial properties of capping materials. Sci World J 2014;2014:181945. DOI: 10.1155/2014/181945

13. Saunders WP, Saunders EM. Coronal leakage as a cause of failure in root- canal therapy: a review. Endod Dent Traumatol 1994;10(3):105–108. DOI: 10.1111/j.1600-9657.1994.tb00533.x

14. Tabassum S, Khan FR. Failure of endodontic treatment: the usual suspects. Eur J Dent 2016;10(1):144–147. DOI: 10.4103/1305-7456.175682

15. Abou ElReash A, Hamama H, Comisi JC, et al. The effect of retrograde material type and surgical techniques on the success rate of surgical endodontic retreatment: systematic review of prospective randomized clinical trials. BMC Oral Health 2021;21(1):375. DOI:10.1186/s12903-021-01731-9

16. von Arx T, Peñarrocha M, Jensen S. Prognostic factors in apical surgery with root-end filling: a meta-analysis. J Endod 2010;36(6):957–973. DOI: 10.1016/j.joen.2010.02.026

17. Caron G, Azérad J, Faure MO, et al. Use of a new retrograde filling material (Biodentine) for endodontic surgery: two case reports. Int J Oral Sci 2014;6(4):250–253. DOI: 10.1038/ijos.2014.25

18. Ferreira Guimarães Xavier V, Felipe Moreira L, Guimarães Xavier D, et al. Bioceramic cements in endodontics. Oral Dis 1–14 (2020) DOI: 10.5772/intechopen.89015

19. Wang Z. Bioceramic materials in endodontics. Endod Top 2015;32(1):3–30. DOI: https://doi.org/10.1111/etp.12075

20. Shalabi M, Saber S, Elsewify T. Influence of blood contamination on the bond strength and biointeractivity of Biodentine used as root-end filling. Saudi Dent J 2020;32(8):373–381. DOI: 10.1016/j.sdentj.2019.11.005

21. Pommel L, Camps J. Effects of pressure and measurement time on the fluid filtration method in endodontics. J Endod 2001;27(4):256–258. DOI: 10.1097/00004770-200104000-00003

22. Youngson CC, Jones JC, Fox K, et al. A fluid filtration and clearing technique to assess microleakage associated with three dentine bonding systems. J Dent 1999;27(3):223–233. DOI: 10.1016/s0300-5712(98)00048-7

23. Barthel CR, Moshonov J, Shuping G, et al. Bacterial leakage versus dye leakage in obturated root canals. Int Endod J 1999;32(5):370–375. DOI: 10.1046/j.1365-2591.1999.00235.x

24. Fitzpatrick EL, Steiman HR. Scanning electron microscopic evaluation of finishing techniques on IRM and EBA retrofillings. J Endod 1997;23(7):423–427. DOI: 10.1016/S0099-2399(97)80295-6

25. Nepal M, Shubham S, Tripathi R, et al. Spectrophotometric analysis evaluating apical microleakage in retrograde filling using GIC, MTA and biodentine: an in-vitro study. BMC Oral Health 2020;20(1):37. DOI:10.1186/s12903-020-1025-9

26. Ahirwar A, John J, Paul M, et al. Assessment of sealing ability of two root-end filling materials at different depths: a comparative study. Int J Oral Care Res 2019;7(3):61. DOI: 10.4103/INJO.INJO_31_19

27. Karobari MI, Basheer SN, Sayed FR, et al. An in vitro stereomicroscopic evaluation of bioactivity between Neo MTA plus, Pro Root MTA, biodentine & glass ionomer cement using dye penetration method. Materials (Basel) 2021;14(12):3159. DOI: 10.3390/ma14123159

28. Bhavsar B, Ramchandran R, Kharat N, et al. A comparative stereomicroscopic evaluation of bioactivity of different biomimetic root end filling materials-an in vitro study. Int J Med Sci Diagnosis Res Orig Res Artic 2020;4:6–11.

29. Gelda A, Prahlad A, Agrawal M, et al. A comparative evaluation of sealing ability and marginal adaptation of four root-end filling materials using stereomicroscope and scanning electron microscope – an in vitro study. J Res Adv Dent 2021;12:213–217. DOI: 10.53064/jrad.2021.12.5.51

30. Jain A, Parihar A, Saxena A, et al. An in vitro dye penetration study : evaluating apical sealing ability of mineral trioxide aggregate compared to intermediate restorative material. 2018;7:143–148.

31. Camps J, Pashley D. Reliability of the dye penetration studies. J Endod 2003;29(9):592–594. DOI: 10.1097/00004770-200309000-00012

32. Veríssimo DM, do Vale MS. Methodologies for assessment of apical and coronal leakage of endodontic filling materials: a critical review. J Oral Sci 2006;48(3):93–98. DOI: 10.2334/josnusd.48.93

33. Schäfer E, Olthoff G. Effect of three different sealers on the sealing ability of both thermafil obturators and cold laterally compacted gutta-percha. J Endod 2002;28(9):638–642. DOI: 10.1097/00004770-200209000-00003

34. Dandakis C, Kaliva M, Lambrianidis T, et al. An in vitro comparison of the sealing ability of three endodontic sealers used in canals with latrogenic enlargement of the apical constriction. J Endod 2005;31(3):190–193. DOI: 10.1097/01.don.0000137644.48860.d8

35. Saraswathi DD, Tejavath SK, Babu MR, et al. A comparative evaluation of sealing ability of three recent root-end filling materials: an in vitro study. J Adv Oral Res 2015;6(2):33–39. DOI: 10.1177/2229411220150206

36. Kokate S, Pawar AM. An in vitro comparative stereomicroscopic evaluation of marginal seal between MTA, glass inomer cement & Biodentine as root end filling materials using 1% methylene blue as tracer. Endodontology 2012;2(2):36–42. DOI: 10.4103/0970-7212.352091

37. Khandelwal A, Karthik J, Nadig RR, et al. Sealing ability of mineral trioxide aggregate and Biodentine as the root end filling material, using two different retro preparation techniques - an in vitro study. Int J Contemp Dent Med Rev 2015;2015:1–6. DOI: 10.15713/ins.ijcdmr.48

38. Singh FJ, Ahuja L, Kakkar G, et al. An in vitro comparative evaluation of the sealing ability of five different root end filling materials using high copper amalgam, resin modified glass ionomer cement, cermet cement, mineral trioxide aggregate and biodentine: an analysis by confocal laser microscopy. Nat Res Dent 2020;9(1):36–43. DOI: 10.4103/ccd.ccd_662_18

39. Nabeel M, Tawfik HM, Abu-Seida AMA, et al. Sealing ability of Biodentine versus ProRoot mineral trioxide aggregate as root-end filling materials. Saudi Dent J 2019;31(1):16–22. DOI: 10.1016/j.sdentj.2018.08.001

40. Camilleri J, Laurent P, About I. Hydration of Biodentine, Theracal LC, and a prototype tricalcium silicate-based dentin replacement material after pulp capping in entire tooth cultures. J Endod 2014;40(11):1846–1854. DOI: 10.1016/j.joen.2014.06.018

41. Makkar S, Kaur H, Aggarwal A, et al. A confocal laser scanning microscopic study evaluating the sealing ability of mineral trioxide aggregate, Biodentine and anew pulp capping agent-theracal. Dent J Adv Stud 2015;3(1):020–025. DOI: 10.1055/s-0038-1672009

________________________
© The Author(s). 2023 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and non-commercial reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.