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 Table of Contents  
ORIGINAL ARTICLE
Year : 2019  |  Volume : 16  |  Issue : 3  |  Page : 136-141

Enhanced bioactivity and interfacial marginal adaptation of mineral trioxide aggregate and Biodentine materials modified by eggshell powder at root dentine


1 Department of Endodontics, Faculty of Dentistry, Beni Suef University, Beni Suef, Egypt
2 Department of Dental Bio-Materials, Faculty of Dentistry, Al-Azhar University, Cairo, Egypt

Date of Submission11-Mar-2019
Date of Acceptance22-May-2019
Date of Web Publication14-Jan-2020

Correspondence Address:
Assistant Professor Ramy A. Abdelrahim
Department of Dental Biomaterials, Faculty of Dentistry, Al-Azhar University, Mokattam, Elhadaba Elwasta, 1082
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/tdj.tdj_16_19

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  Abstract 

Objectives: Mineral trioxide aggregate (MTA) as silicate-based cement material may provide better bioactivity than Biodentine material which used as a replacement to MTA in dental practice in different endodontic uses. Also the modifying MTA and Biodentine by adding eggshell powder (ESP) may improve their bioactivity and promote the formation of a reliable sealing through formation a thick and continuous interfacial layer at root dentine/material interface.
Aim: This study was aimed to investigate marginal adaptability, the presence or absence of interfacial layer, and enhanced bioactivity's of MTA and Biodentine modified by addition of ESP, at the root dentine/material interface.
Materials and methods: Thirteen root dentine segments of about 2 mm thickness were obtained from extracted human teeth and mechanically prepared by endodontic files to enlarge their canals. The specimens were then divided into six groups (n=5), each group were filled with unmodified MTA and Biodentine and modified with 0.5 and 1% ESP. The specimens were immersed in Hank's Balanced Salt Solution simulating body fluid for 30 days. The specimens were then, examined by scanning electron microscopy and energy disperse x-ray spectroscopy (EDX) to examine the presence or absence of interfacial layer and Ca/P ratio.
Results: The interfacial layer can be seen in MTA and Biodentine groups that modified with 1% ESP. Also, MTA showed better marginal adaptability than Biodentine groups. There was a statistically significant difference in the Ca/P ratio among the all investigated groups.
Conclusion: Addition of ESP to MTA and Biodentine can improve the mineral deposition at root dentine/material interface and also improve their sealing ability through the formation of interfacial layer.

Keywords: bioactivity, Biodentine, eggshell powder, marginal adaptability, mineral trioxide aggregate


How to cite this article:
Beshr KA, Abdelrahim RA. Enhanced bioactivity and interfacial marginal adaptation of mineral trioxide aggregate and Biodentine materials modified by eggshell powder at root dentine. Tanta Dent J 2019;16:136-41

How to cite this URL:
Beshr KA, Abdelrahim RA. Enhanced bioactivity and interfacial marginal adaptation of mineral trioxide aggregate and Biodentine materials modified by eggshell powder at root dentine. Tanta Dent J [serial online] 2019 [cited 2020 Sep 24];16:136-41. Available from: http://www.tmj.eg.net/text.asp?2019/16/3/136/275933


  Introduction Top


Bioactivity is the ability of a material to form apatite carbonate when exposed to body fluids in vivo[1] or simulated body fluid (SBF) in vitro [2]. The ability of the material to mineralize the dental tissue is considered as bioactive material and it is a very important material property that can help in tissue repair and/or regeneration[2],[3]. Ideally, the biomaterial should promote the healing process through the apatite formation at tooth structure/restoration interface[3],[4]. In endodontic, there are many examples of the importance of bioactivity of biomaterials as perforation repair, internal root resorption, vital pulp capping, apexogenesis, and root-end filling[1],[3],[5],[6].

Mineral trioxide aggregates (MTA) cement is a mix of hydrophilic fine powder and distilled water by using 3: 1 powder/liquid ratio [7]. MTA, when mixed with water, will form a hydrogel of both calcium silicate and calcium hydroxide[7],[8]. MTA exhibit initial pH after mixing of 10.2 which increased gradually to reach 12.5 after setting [9]. MTA consider a bioceramic material this can use in human body safely due to its biocompatibility [10]. The bioactivity of MTA cement was proved by the formation of apatite crystal on its surface when exposed to a physiologic solution [11–13].

Biodentine recently used as an alternative to MTA, and, it is also a calcium silicate-based cement which considers the first dentine substitute material because its higher mechanical properties that associated with the higher biological behavior[14],[15]. Biodentine also designed to be the treatment and/or the replacement for dentine in endodontic purpose[15],[16].

Biodentine can stimulate the reparative regeneration of dentine in addition to mineral deposition on dentine surface, that is help in dentine remineralization [17]. Also, Biodentine on setting release calcium hydroxide as MTA[15],[17],[18]. Crystal growth of Biodentine inside the dentinal tubules and/or the possible ion exchanges with dentine tissues can appear at the Biodentine/dentine interface [17–19].

Eggshell powder (ESP) composed mainly from calcium carbonate (98.2%), with other mineral salts as magnesium (0.9%) and phosphate (0.9%); thus, it is considering the main source of calcium [20]. Recently; many clinical studies use it alone or in combination with other materials as tooth re-mineralizing agent or bone repair [21–23].

This study was aimed to assess marginal adaptability, the interfacial layer and the enhanced bioactivity of MTA and/or Biodentine modified with ESP at root dentine/material interface in Hank's Balanced Salt Solution (HBSS) SBF.


  Materials and Methods Top


The materials used in this study included: white ProRoot MTA (Maillefer, Dentsply, Switzerland), Biodentine (Septodont, Paris, France), and ESP.

Eggshell powder preparation

ESP prepared by calcination process [24], to produce pure, pathogenic free, calcium oxide powder [25]. Chicken eggs were cleaned with sterile distilled water, and, then boiled in hot water bath at 100°C for about 10 min to facilitate their internal white membranes removal. The eggshells then, crushed to small particles by using sterile mortar and pestle. The resulting powder then kept in furnace muffle (Thermolyne 47900, Model F4791; Kerper Boulevard, Iowa, USA) at a temperature of 1200°C for about 1 h. The obtained mass from firing crushed again for several times in a sterile mortar and pestle to ensure homogeneous particles size[26] at the National Research Center, Cairo, Egypt.

Specimens preparation

Ten roots were obtained from extracted single-rooted human teeth by removal of the crown at the cemento-enamel junction. Each root was prepared mechanically by using endodontic files to enlarge their canals. Thirteen root dentine segments of about 2 mm thickness were obtained by using low speed, water cooled, diamond saw [3]. The root canal then irrigates by 5% sodium hypochlorite and 17% EDTA to remove the smear layer and dried with paper point [3].

Unmodified MTA and Biodentine materials were mixed according to manufacturer's instructions. Modified MTA and Biodentine groups are prepared by addition ESP to the powder of MTA and Biodentine with proportion of 0.5 and 1% by weight using digital balance (Precisa 205A; Moosmattstrasse, Dietikon, Switzerland), at the Regional Center of Mycology and Biotechnology, Cairo, Egypt, and then mixed according to manufacturer's instructions. The segmented root dentine specimens were randomly categorized into six groups (N = 5) and filled with modified and unmodified materials to simulate the clinical use of materials as root-end filling and/or perforation repair [3].

Specimens grouping

Group I: unmodified MTA.

Group II: MTA modified with 0.5% weight ESP.

Group III: MTA modified with 1% weight ESP.

Group IV: unmodified Biodentine.

Group V: Biodentine modified with 0.5% weight ESP.

Group VI: Biodentine modified with 1% weight ESP.

All specimens were polished with 1000 grit abrasive sand-paper and then immersed in HBSS as a SBF for 30 days. The SBF solution was renewed every 3 days to keep ion concentration at the same level.

Root dentine/material interface examination

At the end of the immersion period, all specimens were examined by Scanning Electron Microscopy (SEM) (JEOL JSM-5500 LV; JEOL Ltd, Japan) by using low vacuum mode after gold coating using SPI-module sputter coater to assess the interfacial layer and marginal adaptability, at the Regional Center of Mycology and Biotechnology, Cairo, Egypt, and Energy Disperse X-ray Spectroscopy (EDX) (Module Oxford 6587 INCA x-sight) attached to SEM at 20KV to measure the Ca/P ratio at the root dentine/material interface, at the Regional Center of Mycology and Biotechnology, Cairo, Egypt.

Statistical analysis

All collected data were analyzed with one-way analysis of variance followed by Tukey's test. Statistical analysis was done with SPSS, version 23, software (IBM Corp., Chicago, Illinois, USA) with the significant level at P value less than or equal to 0.05.


  Results Top


Scanning electron microscopy analysis of dentine/material interface

SEM images of all investigated groups are shown in Fig. 1. MTA group (group III) exhibited the existence of a distinct interfacial layer between the MTA material and root dentine (Fig. 1c) and also showed that the interfacial layer was in intimate contact with root dentine without any gap between the MTA material and dentine. While, in group I and group II, exhibited only separated areas of interfacial layer attached on the dentine surface (Fig. 1a, b).

In Biodentine group VI, the interfacial layer can be clearly seen but with a gap between the Biodentine material and the interface (Fig. 1f). While, in group IV, always showed a gap without the presence of interfacial layer between the Biodentine and dentine surface (Fig. 1d). However, in group V, there were separated areas of interfacial layer attached on the dentine surface (Fig. 1e).

The marginal gap results for MTA and Biodentine showed that there was a statistically significant difference among all investigated groups [Table 1]. For MTA groups, the minimum mean gap at dentine/material interface was for group III (Fig. 1c), followed by group II and group I, respectively. However, for Biodentine groups, the minimum mean gap was at group VI followed by group V and group IV, respectively.
Table 1 Mean±SD of marginal gap values at dentine/material interface among all investigated groups

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Energy disperse x-ray spectroscopy analysis of dentine/material interface

EDXA results of MTA groups showed that the highest Ca/P ratio mean value was in group III, followed by and for group II and group I, respectively. While, EDXA results of Biodentine groups, showed that, the highest Ca/P ratio mean value was for group VI, followed by and for group V and group IV, respectively [Table 2].
Table 2 Mean±SD Ca/P values among all investigated groups

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There was a statistically significant difference among all investigated groups (P ≤ 0.05). Tukey's test showed that there was no statistically significant between MTA groups (group I and group II) and Biodentine group VI and also between group IV and group V.


  Discussion Top


MTA and Biodentine are bioactive materials used in endodontic case needed the formation of hard tissues such as root perforation, apexogenesis, and apexification [27]. Both MTA and Biodentine can induce hard tissue formation via ion release upon undergoing relatively similar hydration process [28]. Calcium hydroxide is the hydration end product of MTA and Biodentine materials[17],[18],[28].

MTA can produce a promising result as root-end filling material due to its good bioactivity [3], marginal adaptability[29],[30], and, ability to stimulate cementogenesis [29]. Regenerative potential of MTA considers the main advantage in addition to its biocompatibility[29],[30].

Generally, the bioactivity of cement-like materials depends mainly on the hydrophilicity of their particles, which enhance their solubility and ion release [5]. The main constituting component of MTA powder is calcium silicate [31], and, can be described as a hydrolytic calcium silicate because its setting depends mainly on hydration[31],[32]. It was found that the increase water/powder ratio of MTA during mixing will lead to an increase in its solubility and calcium ion release[9],[33]. MTA can form and precipitate new hydroxyapatite through releasing calcium ions in high amount upon hydrolysis which could react with extrinsic phosphate ions in body fluid[12],[34],[35]. Hydroxyapatite that formed can improve the chemical bond between dentine and MTA, and also can stimulate remineralization of the dentine [12], in agreement with our results shown in Fig. 1 and [Table 2].

There is a correlation between marginal adaptation and sealing ability at the dentine/material interface of the cement material and this will reflect on their clinical rate of success [36]. MTA also has superior marginal adaptation as root-end filling material when compared with other materials, as Biodentine[12],[19],[37]. That in agreement with our results in Fig. 1 and [Table 1].

Biodentine is also calcium silicate-based cement and it is used as a recent alternative to MTA[14],[15]. Biodentine designed primary as dentine substitute, but it is used also in endodontic for treatment and/or replacement of root dentine [16]. Biodentine remineralization was associated with its alkaline pH, calcium ions release, as well as silicon ions, release through the creation of what called mineral-infiltration zone at dentin/material interface [38].

Although, Biodentine cement has the ability to initiate and propagate the process of mineralization as MTA[37],[39]. Clinical results showed that, the long-term effect of MTA as bioactive material better than Biodentine [40]. That can explain our results as shown in [Figure 1] and [Table 2].
Figure:1 SEM micrographs of material/dentine interface after immersion in SBF for 30 days. (a–c) MTA-dentine; (d–f) Biodentine-dentine. IL, interfacial layer; MTA, mineral trioxide aggregate; SBF, simulating body fluid; SEM, scanning electron microscopy.

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MTA and Biodentine undergo surface dissolution to form an amorphous hydroxy carbonate-apatite in physiologic solution as HBSS [41]. Calcium ion concentration considers a critical factor that can play a significant role in hard tissue regeneration [42]. Hydroxy carbonate-apatite can be formed as a result of CaO-P2O5 crystal formation due to the existence of carbonate (CO32−) and hydroxyl (OH) groups in the physiologic solution [41]. Biodentine is slowly soluble cement due to its hydro-soluble polymer liquid [28]. The low solubility of Biodentine lead to slower calcium ion release rate than MTA[28],[43]. In addition to the faster setting time of Biodentine due to the presence of calcium chloride in its liquid, which could result in a decreased time that available to the interaction between phosphate from HBSS and calcium from Biodentine [44].

The amount of calcium released may be related to the amount of calcium carbonate content [28]. The ESP is a natural calcium carbonate biomaterial which considers an important factor in hard tissue remineralization[28],[45]. So, the addition of ESP can explain the higher Ca/P ratio in modified MTA (group II and group III) and modified Biodentine groups (group V and group VI) in comparison with the unmodified groups (group I and group IV).

Many studies consider that the SEM and EDX are the main devices used to examine the interface at dentine and tricalcium silicate-based materials[12],[13].

HBSS as SBF is the best physiological solution used in bioactivity testing in vitro because it produces a storage medium with calcium and phosphate ions concentrations resample to that of human plasma[2],[46].

Our results show that the Ca/P ratio in both MTA and Biodentine is higher than the stoichiometric Ca/P ratio (1.67) [47]. That indicates the formation of calcium-deficient carbonated apatite at the material/dentine interface[13] [Table 2].

Our results also show that the SEM examination of Biodentine groups revealed frequent microgaps at the interface between dentine and Biodentine cement (Fig. 1d and e), this may be due to the fact that Biodentine is sensitive to moisture exposure during initial setting. Moreover, in this study to simulate the clinical situation all specimens of all groups were soaked immediately in HBSS as SBF, which can hamper the Biodentine setting reaction due to excessive moisture and cause its separation from dentine [3].


  Conclusion Top


Within the limitation of this study, the addition of ESP to tricalcium silicate-based cement as MTA and Biodentine could enhance their bioactivity and improve the marginal adaptation at dentine/material interface.

Recommendation

Other physical and biological properties of the newly formulated MTA and Biodentine cements should be examined in the future studies.

Clinical significance

Biodentine modified with 1% ESP can be used as an alternative to unmodified MTA due to comparable bioactivity which can also create better adaptability through the formation of interfacial layer with root dentine.

Acknowledgements

This work was primarily carried out in Faculty of Dentistry, Endodontic Department, Beni Suef University.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
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