|Year : 2020 | Volume
| Issue : 5 | Page : 476-484
Assessment of physical properties of a ZnO/E sealer modified by adding moringa oleifera: An experimental in-vitro study
Engy M Kataia, Maram E Khallaf, Nada Omar, Yousra Aly, Nancy ElShafei
Restorative and Dental Materials Department, National Research Centre, Cairo, Egypt
|Date of Submission||22-Dec-2019|
|Date of Decision||13-Apr-2020|
|Date of Acceptance||14-Apr-2020|
|Date of Web Publication||21-Oct-2020|
Prof. Engy M Kataia
Restorative and Dental Materials Department, National Research Centre, Cairo.
Source of Support: None, Conflict of Interest: None
Aim: In an attempt to move toward exploration of the antibacterial properties of herbal products, this study evaluated the effect of how the addition of Moringa would affect the physical properties of the commercial Endofil sealer. Materials and Methods: An experimental in vitro study was performed using three experimental sealers. There were four groups in this study: Group A (ZnO/E sealer alone), Group B (ZnO/E and Moringa root), Group C (ZnO/E and Moringa leaf), and Group D (ZnO/E and Moringa leaf extract liquid). Measurements were carried out according to International Organization for Standardization (ISO) standardization 6876:2012. Moringa oleifera was added to Endofil’s powder and liquid and the physical properties of the new sealer were tested according to ISO 6876:2012 and compared to those of Endofil alone. One-way analysis of variance (ANOVA) followed by Tukey post hoc test was used to compare between more than two groups in non-related samples. Results: All groups showed varying degrees of shrinkage with Group B having the lowest shrinkage and setting time, whereas Groups A and D experienced the highest shrinkage. All groups experienced solubility; Group B had the highest mass loss and Group D showed the lowest. Group A showed the lowest film thickness. Radiopacity of all groups satisfies the ISO recommendations for root canal sealers. Conclusion: Moringa added to ZnO/E sealer showed acceptable physical properties of the commercial Endofil sealer.
Keywords: Herbal, Moringa Oleifera, Physical Properties, Sealer, Zinc Oxide Eugenol
|How to cite this article:|
Kataia EM, Khallaf ME, Omar N, Aly Y, ElShafei N. Assessment of physical properties of a ZnO/E sealer modified by adding moringa oleifera: An experimental in-vitro study. J Int Oral Health 2020;12:476-84
|How to cite this URL:|
Kataia EM, Khallaf ME, Omar N, Aly Y, ElShafei N. Assessment of physical properties of a ZnO/E sealer modified by adding moringa oleifera: An experimental in-vitro study. J Int Oral Health [serial online] 2020 [cited 2020 Nov 29];12:476-84. Available from: https://www.jioh.org/text.asp?2020/12/5/476/298796
| Introduction|| |
Various techniques have been advocated to fill root canals. The most commonly used is filling with gutta-percha as the core material, combined with root canal sealers or pastes. Because gutta-percha alone is not appropriate for ideal root canal filling due to absence of flow and adhesion to canal walls, using sealers is mandatory to obtain a satisfactory seal., The principle functions of the root canal filling and sealers are sealing off the root canal, entombment of remaining bacteria, and filling irregularities in prepared canals. Root canal sealers must be biocompatible,, antimicrobial, and present satisfactory physiochemical properties,, dimensional stability, slow setting time, and insolubility.
The main causes of failed endodontic treatments are the persistence of intraradicular infections. Even if the eradication of microorganisms from the canal space, or at least reducing their levels to be appropriate to the periapical tissue health are the objectives of endodontic treatment, the presence of bacteria in dentinal tubules after treatment was recorded.
Knowing that complete elimination of microorganisms from the endodontic space is not predictable, sealers’ antimicrobial action may help get rid of residual microorganisms unaffected by the chemo-mechanical preparation. Accordingly, endodontic sealers with good antimicrobial activity help to minimize or stop the growth of microorganisms and aid in the repairing of periapical tissues.
Although during routine endodontic therapy, it is desired that the endodontic sealers remain inside the root canal, sometimes they are accidentally pushed beyond the apical constriction. In fact, they remain in intimate contact with the surrounding soft and hard tissues for an extended period.
Zinc oxide eugenol (ZnO/E) sealers were successfully used in obturation for over 100 years. They possess antimicrobial activity and are the most common among clinicians, especially when used with thermos-plasticized obturation technique. But moderate to severe cytotoxicity and localized inflammation were observed both in soft tissue and in bone if extruded periapically. Lindqvist and Otteskog suggested that cytotoxicity is due to free eugenol liberated from the set material. Moreover, another study showed that ZnO/E sealers also released formaldehyde after setting.
In an attempt to decrease the cytotoxic effect and add to the antimicrobial property of the ZnO/E sealer, the thought of adding herbal antimicrobial was suggested.
Moringa oleifera is an amazing nutritious tree to the extent that it is also called “Tree of Life or Miracle Tree.” It has been widely used for treating bacterial, fungal infections, and inflammation., The reported antibiotic activity of this tree was a subject of serious scientific research.
Moreover, the recently proven antibacterial effect against Enterococcus faecalis without any cytotoxicity, encouraged us to undergo this attempt of adding of Moringa oleifera to Endofil (ZnO/E) sealer, in a trial to decrease its inflammatory and cytotoxic effect and to take the advantage of the antibacterial and anti-inflammatory properties of Moringa. The physical properties of this new sealer were evaluated.
| Materials and Methods|| |
Setting and design
An experimental in vitro study was carried out in the National Research Centre (Dokki, Giza, Egypt) in 2019, for 3 months. Three experimental sealers were prepared by adding M. oleifera to ZnO/E sealer. M. oleifera powder and liquid extracts were obtained from Egyptian Scientific Society of the Moringa Trees, National Research Centre (Dokki, Giza, Egypt). Physical properties (measured according to International Organization for Standardization [ISO] 6876:2012) of the experimental sealers were compared to those of the ZnO/E sealer alone as control.
Grouping of the samples
The study was conducted on the following four groups: Group A: Endofil (Promedica, Neumuenster, Germany) (ZnO/E) sealer alone, Group B: The sealer powder was prepared by adding Moringa leaf powder to ZnO powder in the ratio 2:1 (ZnO and Moringa leaf respectively), Group C: Moringa root powder was added to ZnO powder in the ratio of 2:1, ZnO and Moringa Root, respectively, and Group D: Moringa eugenol liquid was mixed with leaf extract liquid in the ratio 2:1, respectively. For measuring the physical properties of each sealer, the powders and liquids were mixed till required consistency was reached.
Setting time, dimensional stability, solubility, film thickness, and radiopacity were measured according to ISO 6876:2012 as follows,:
Setting time: Test was performed under controlled temperature and humidity: 37°C ± 1°C and 95% ± 5% relative humidity. Molds of dimensions 10 mm diameter × 2 mm thickness were filled with the mixed sealers. For each sealer, three specimens were prepared. After initial cement setting time, a Gilmore needle with an active tip of 1.0 mm diameter and 110g weight was used at 5-min intervals to determine the final setting time. The setting times of a sealer was the time that elapsed from the beginning of mixing to the time when no indentation was detected on specimen surface.
Dimensional stability: Molds made of teflon (dimensions 6 mm diameter × 12 mm height) were used. They were backed up with a glass plate, filled with the mixed sealer, then the other side was also backed with another glass plate. This assembly was then placed in an incubator for three times the setting time. The ends of the molds were ground using 600-grit sandpaper after complete setting. The specimens were then removed from the mold, and their length was measured by a digital caliper (L0). Specimens were stored in distilled water, and the container was stored in incubator for 30 days. At the end of the period, samples were dried using tissue paper and their length was remeasured (L1). This was done three times for each sealer, and the difference in length was recorded as the dimensional change (D) in the following formula: (%) = (L1 − L0)/L0 × 100.
Teflon ring molds with dimensions 20 mm diameter × 1.5 mm height were used for Solubility measurements. Three specimens were prepared for each sealer. Before setting of the sealer, a nylon thread was inserted in it. The specimens were kept in an incubator (at 37°C and 95%) for three times their setting times. After setting, the samples were removed from the molds, and any loose material particles were removed from the surface, using a soft brush. Samples were weighed in an analytical balance with 0.0001g (UMark 210; Bel Engineering, Monza, Italy) precision. The samples were suspended by the nylon thread inside a glassware containing 50 mL of distilled water. Each container was then kept in an incubator for 24h. Samples were then removed and gently washed with distilled water, dried with filter paper, placed in oven for 24h, and then reweighed. This was repeated three times for each sealer. Solubility was determined by calculating the weight loss.
Film thickness: Two glasses of 5-mm thickness were used; 0.5 mL of sealer was placed in the center of one of them and the other was placed centrally over the sealer. After 180 ± 10s post mixing, a load of 150N was applied centrally and vertically on top of the plates. Ten minutes after commencement of mixing, the load was removed, and total thickness of the two plates and the sealer film was measured with a digital caliper. The difference between two measurements showed the film thickness of the materials. This test was repeated three times, and the mean of the three readings was recorded.
Radiopacity: Six specimens were prepared for each material. Specimens with no visible defects were positioned on photo-stimulable phosphor plate (Soredex, Digora, Helsinki, Finland) with the aluminum step wedge of thickness 2–16 mm in 2 mm increments. A dental X-ray system (Progeny, Midmark, Versailles, Ohio), operating at 60kV, 10 mA for exposure time 0.3s with a 30-cm focus-film distance, was used. X-ray images were analyzed using the Digora 1.51 software (Orion Corporation Soredex, Helsinki, Finland). Equal-density areas in the radiographs were identified by the equal-density tool to compare the radiographic density of cements and the radiopacity of the different aluminum step wedge thicknesses. Readings for each sample were recorded and their averages were calculated. Radiopacity values were determined according to the following equation:
where A = radiographic density of the material (RDM) – radiographic density of the aluminum step wedge increments immediately below RDM; B = radiographic density of the aluminum step wedge increments immediately above the RDM – radiographic density of the aluminum step wedge increment immediately below the RDM; 2 = 2 mm increments of the aluminum step wedge.
The data were statistically analyzed with IBM Statistical Package for the Social Sciences (SPSS) Statistics software, version 20.0, for Windows. One-way analysis of variance (ANOVA) followed by Tukey post hoc test was used to compare between more than two groups in non-related samples. The significance level was set at P ≤ 0.05 [Table 1],[Table 2],[Table 3],[Table 4].
|Table 1: The mean, standard deviation (SD) values of physical properties.|
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|Table 2: The descriptive data of analysis of variance test for each method of evaluation|
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|Table 3: The descriptive data of post-hoc test and CI for mean difference between each pair for each method of evaluation.|
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|Table 4: The descriptive data and CI for mean of each group for each method of evaluation.|
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| Results|| |
Setting time results
There was a statistically significant difference between Group A, Group B, Group C, and Group D, where P < 0.001. A statistically significant difference was found between Group A and each of Group B, Group C, and Group D, where P < 0.001, P = 0.022, and P < 0.001, respectively. Also, a statistically significant difference was found between Group B and each of Group C and Group D, where P < 0.001. A statistically significant difference was found between Group C and Group D, where P < 0.001.
The highest mean value was found in Group B, followed by Group C and Group A, whereas the least mean value was found in Group D [Table 1].
Dimensional stability results
There was a statistically significant difference between Group A, Group B, Group C, and Group D, where P < 0.001. A statistically significant difference was found between Group A and each of Group B and Group C, where P < 0.001 and P = 0.003, respectively, whereas no statistically significant difference was found between Group A and Group D, where P = 1. No statistically significant difference was found between Group B and Group C, where P = 0.192, whereas a statistically significant difference was found between Group B and Group D, where P < 0.001. A statistically significant difference was found between Group C and Group D, where P = 0.003.
The highest mean value of shrinkage was found in Group A and Group D, followed by Group C, whereas the least mean value of shrinkage was found in Group B [Table 1].
There was a statistically significant difference between Group A, Group B, Group C, and Group D, where P = 0.002. A statistically significant difference was found between Group A and Group B, where P = 0.014, whereas no statistically significant difference was found between Group A and each of Group C and Group D, where P = 0.070 and P = 0.611, respectively. No statistically significant difference was found between Group B and Group C, where P = 0.649, whereas a statistically significant difference was found between Group B and Group D, where P = 0.003. A statistically significant difference was found between Group C and Group D, where P = 0.012.
The highest mean value was found in Group B, followed by Group C and Group A, whereas the least mean value was found in Group D [Table 1].
Film thickness results
There was a statistically significant difference between Group A, Group B, Group C, and Group D, where P = 0.001. A statistically significant difference was found between Group A and each of Group B and Group C, where P = 0.002, whereas no statistically significant difference was found between Group A and Group D, where P = 0.624. No statistically significant difference was found between Group B and Group C, where P = 0.999, whereas a statistically significant difference was found between Group B and Group D, where P = 0.007. A statistically significant difference was found between Group C and Group D, where P = 0.009.
The highest mean value was found in Group B, followed by Group C and Group D, whereas the least mean value was found in Group A [Table 1].
A statistically significant difference was found between Group A, Group B, Group C, and Group D, where P < 0.001. A statistically significant difference was found between Group C and each of Group A and Group D, where P = 0.001 and P < 0.001, respectively, whereas no statistically significant difference was found between Group C and Group B, where P = 0.173. No statistically significant difference was found between Group B and Group A, where P = 0.066, whereas a statistically significant difference was found between Group B and Group D, where P = 0.007. No statistically significant difference was found between Group A and Group D, where P = 0.606.
The highest mean value was found in Group D, followed by Group A and Group B, whereas the lowest mean value was found in Group C [Table 1].
| Discussion|| |
Proper root canal system disinfection is important for successfulness of endodontic treatment. In spite of the several enhancements attained recently, endodontic chemo-mechanical disinfection techniques used today cannot provide complete canal sterility.
It was sighted that after canal preparation chemo-mechanically, the antimicrobial properties of sealers could control infections and prevent penetration of fluids, which provide nutrition to the remaining microorganisms. Therefore, endodontic sealers are used in root canal therapy to eliminate microorganisms still remaining in canal systems after chemo-mechanical preparation and to prevent their recolonization. Sealers have to be biologically compatible and with dimensional stability, as well as possess a long-lasting antibacterial effect. Sealers containing eugenol are well-known for their antibacterial property, which was higher against E. faecalis than resin and calcium hydroxide–based sealers.
Due to the proven antimicrobial and anti-inflammatory activity of the M. oleifera against oral and root canal bacteria,, and its recently proved antibacterial effect against E. faecalis without any toxicity, this made the authors suggest its safe used as an alternative antimicrobial agent in the root canal therapy; in this study, it was added to ZnO/E sealer in an attempt to dilute its cytotoxic and inflammatory effect on host cells, without weakening its antimicrobial effect on microorganisms.
The importance of using root canal sealers and their essential requirements was reported. Grossman and other authors inspected different properties, including solubility, flow, setting time, and radiopacity of the root canal sealers. That is why in this research, the physical properties (setting time, dimensional stability, solubility, film thickness, and radiopacity) of ZnO/E sealer modified by adding Moringa powder and liquid were evaluated and compared to the ZnO/E sealer alone as a control. The procedures were performed as outlined in the ISO standard 6876:2012.
The amount of Moringa powder or liquid added to the ZnO/E was kept half the amount of the Endofil powder or the liquid, to avoid adversely affecting the physical properties of the original sealer.
According to ISO specifications, sealer’s setting time should differ no more than 10% of what is stated by the manufacturer. Setting time of Endofil was not mentioned by the manufacturer.
Setting time depends on particle size, room temperature, and relative humidity of the sealer. Setting time is clinically important, where it is desirable to have a setting time that is neither too fast nor too slow. The advantages of a slow setting time are that it allows sealer placement in more than one canal as well as the ability to recover gutta-percha from a canal directly after obturation (if correction is necessary). On the contrary, too slow setting time is a disadvantage, because coronal leakage may take place shortly after the completion of root canal treatment. Unset or partially set sealers may allow more rapid diffusion of bacteria or bacterial byproducts, through the obturation.
The results showed that Group B, where the Moringa root powder was added to the powder of ZnO/E took the longest time to set, which was nearly 24h compared to Group A (ZnO/E sealer alone), which was nearly 5h. The shorter setting time of Endofil, was probably because of the absence of calcium hydroxide in its composition, where calcium hydroxide when contacts water delays the setting process. Moringa has vast quantities of calcium, which may be the cause of prolonging the setting time.
Dimensional change determines the percent of shrinkage or expansion of a material after setting. The maximum allowed linear shrinkage is 1%, whereas the maximum allowed expansion is 0.1%. Both shrinkage and expansion are not favorable for a root canal filling material after setting. Shrinkage creates slits and passageways for bacteria and their products, whereas expansion might produce threatening forces, which could cause infractions, leading to dentin fracture. All sealers showed shrinkage more than that considered acceptable by the ISO specification. Group B had the lowest shrinkage of 2.78%, and Groups A and D experienced the highest shrinkage of 9.63%. The contraction presented by the ZnO/E sealer maybe due to the loss of zinc in the immersion solution, which could be related to the continued loss of eugenol from the matrix, these results agreed with the work of Marín-Bauza et al. To the contrary of our results, some literature reported expansion of ZnO/E sealer. From this, we can conclude that the lesser degree of shrinkage occurring with Groups B and C may be due to the lesser percentage of ZnO powder in the mix (where the Moringa powder was added), so decreasing the amount of zinc lost in the immersion solution.
Solubility is an unsatisfactory characteristic for an endodontic sealer because it leads to the release of constituents that may show biological incompatibility and may lead to gap formations that can negatively affect the hermetic seal of the endodontic filling. Solubility and disintegration of endodontic sealers must be as minimal as possible to allow for the creation of a hermetic seal, thus promoting clinical success, because microleakage may occur from the apical third—cervically or vice versa.
ISO 6876:2012 stated that solubility of sealers should not exceed 3% by mass. Solubility results of Groups A and D showed mass loss of 1.45% and 0.65%, respectively, and were within the range approved by the ISO recommendations. On the contrary, Group B showed the highest solubility, which may be due to its delayed setting time, leading to leaching out of its unset components.
On studying the solubilization of different sealers, it was found that the highest solubility was presented by the ZnO/E-based sealer (Endofil) due to the continuously losing eugenol from sealer matrix. It noted that the presence of sodium borate in Endofil aids in increasing its solubility because it is very soluble. The hydrolysis reaction of the hardened zinc eugenolate also contributes to its solubility. Wilson and Batchelor, described the disintegration mechanism of ZnO/E cements as a sequence of continuous loss of eugenol from cement matrix by decomposing the balance between this matrix and eugenol. This may also clarify why group D had the lowest solubility results because the liquid used in mixing was a combination of Moringa extract liquid and eugenol, so the percentage of eugenol in the final mix was less than that in the other three groups. The weight loss of Endofil sealer coincides with the finding of other studies.,
Adequate film thickness is mandatory for a satisfying distribution of the sealer into lateral canals, narrow irregularities, and the apical foramina. According to ISO specification, sealers should have a film thickness not more than 50 μm. Greater film thicknesses are undesirable due to the possibility of interfering with proper gutta-percha cones seating inside root canals during the filling procedures. Thin film thickness sealers are expected to better wet the surface than thick film thickness sealers, providing a better seal. Group A, showed the lowest film thickness of 30 μm, followed by Group D of 50 μm film thickness, with no significant difference between them. Both groups had values that complied with what the ISO specification required. Groups B and C, where the Moringa root and leaf powders were added to the ZnO powder, had the highest film thickness of 150 μm. This could possibly be due to different particle sizes of Moringa powder than those of the ZnO powder.
Radiopacity of endodontic filling materials is important as it makes it possible to distinguish between tooth and surrounding structures. It also aids in the assessment of the root canal filling.
The radiopacity for a root canal sealer should not be less than 3 mmAl. All sealers examined showed radiopacity values above the minimum recommendation of the ISO standards. The high radiopacity of the ZnO/E sealer (Endofil) used in this study is due to its content of barium sulfate, zinc oxide, and bismuth subcarbonate in its powder. That explains the high radiopacity of both Groups A and D in comparison to Groups B and C, which have a significantly lower radiopacity, due to incorporation of Moringa powder to the ZnO/E powder, decreasing the amounts of radiopacifiers in the final mix. This agrees with several studies, stating that the degree of radiopacity of Endofil sealers is affected by the amount of ZnO powder in the final sealer mix.,
In this study, we recommend that the concentration used was 2:1 (ZnO/E to Moringa either powder or liquid, respectively), maybe future studies could investigate the effect of different concentrations of Moringa. The use of Moringa powder with nano-sized particles when combined with a sealer should be considered in future studies. These tests measuring the physical properties of the experimental sealers do not assure that the material is suitable for its purpose, which is why more tests on clinical and biological performances are being carried out now for a full assessment.
ZnO/E sealer modified with moringa showed acceptable physical properties.
Financial support and sponsorship
This work was conducted as a part of a project fundedfrom the National Research Centre, Giza, Egypt.
Conflict of Interest
There are no conflicts of interest.
Data availability Statement
Data can be available on valid request on contacting to corresponding author mail.
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[Table 1], [Table 2], [Table 3], [Table 4]