|Year : 2019 | Volume
| Issue : 3 | Page : 422-431
In vitro investigation of antimicrobial effects, nanohardness, and cytotoxicity of different glass ionomer restorative materials in dentistry
A Cosgun1, B Bolgul1, N Duran2
1 Department of Paediatric Dentistry, Faculty of Dentistry, University of Hatay Mustafa Kemal, Hatay, Turkey
2 Department of Microbiology, Faculty of Dentistry, University of Hatay Mustafa Kemal, Hatay, Turkey
|Date of Acceptance||11-Dec-2018|
|Date of Web Publication||6-Mar-2019|
Dr. A Cosgun
Address Hatay Mustafa Kemal University, Dental Faculty, Department of Pedodontics, Hatay
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Aims: The aim of this in vitro study was to investigate the antimicrobial effects, nanohardness, and cytotoxicity of different glass-ionomer restorative materials in dentistry. Materials and Methods: The following glass-ionomer restorative materials were used in our study: Argion (VOCO, Germany), Zirconomer (Shofu Inc., Japan), EQUIA Forte (GC, Japan), Fuji II LC capsule (GC, Japan), and Fuji IX GP capsule (GC, Japan). For the cytotoxicity test, a cell culture without release liquids was used as the control group. Microdilution and a disk diffusion test were used to measure the antimicrobial activity. The MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) assay was used to evaluate cytotoxicity. Nanohardness was measured using a nanoindenter device. Results: Our study determined that all restorative materials used in this study inhibited bacterial growth in cultures containing 1 × 102 cfu/ml of the bacterial concentrations (Lactobacillus casei and Streptococcus mutans) and that all restorative materials inhibited fungal growth in the cultures containing <1 × 102 cfu/ml of the fungal strains (Candida albicans). IC50 values obtained for the cytotoxicity test were analyzed using the Chi-square test. After a 72-h incubation period, Zirconomer, EQUIA Forte, Fuji IX GP capsule, and Fuji II LG capsule showed statistically significant lower cell viability values. (P < 0.05). The Kruskal-Wallis analysis was performed on the values obtained from the nanohardness test; the differences between the groups were found to be significant (P < 0.05). Conclusions: All materials tested showed low antimicrobial activity, and the antifungal activity of these materials was found to be lower than their antimicrobial activity. Only Argion showed non-cytotoxic effect. Beginning with the group with the highest nanohardness values and ending with the lowest, the groups are ranked in the following order: Fuji II LC capsule, EQUIA Forte, Argion, Fuji IX GP capsule, and Zirconomer.
Keywords: Antimicrobial activity, cytotoxicity, glass-ionomer cement, nanohardness
|How to cite this article:|
Cosgun A, Bolgul B, Duran N. In vitro investigation of antimicrobial effects, nanohardness, and cytotoxicity of different glass ionomer restorative materials in dentistry. Niger J Clin Pract 2019;22:422-31
|How to cite this URL:|
Cosgun A, Bolgul B, Duran N. In vitro investigation of antimicrobial effects, nanohardness, and cytotoxicity of different glass ionomer restorative materials in dentistry. Niger J Clin Pract [serial online] 2019 [cited 2019 Mar 25];22:422-31. Available from: http://www.njcponline.com/text.asp?2019/22/3/422/253461
| Introduction|| |
Many different kinds of restorative materials are used in pediatric dentistry. One of these materials is glass-ionomer cement (GIC), which is commonly preferred in pediatric dentistry practice.,
Developments in glass-ionomer restorative materials in recent years have enabled physicians to be more conservative to removing caries. Along with the adoption of these conservative approaches, the infected dental tissue repaired with fluoride-releasing restorative material, and thus, it is possible to minimize material loss in the tooth.,, With its fluoride releasing property, glass-ionomer restorative material yields antibacterial effects and supports remineralization. This prevents the formation of secondary caries and reduces microleakage.
Microorganisms play important roles in the onset and development of dental caries. Streptococcus mutans ns) is the primary bacteria responsible for the formation of caries. Numerous studies show that Candida albicans (C. albicans) is a pathogen for dental caries.,, The organic acid and enzymes produced by C. albicans can dissolve the hydroxyapatite of dental hard tissues and degrade the dentinal collagen., Because GICs potentially reduce microleakage by attaching to the tooth structure, inhibit the growth of oral bacteria that result from caries and neutralize the acids produced by these bacteria through ion release, the use of GICs is recommended because of its beneficial antimicrobial effect in cases where protection against caries is necessary.,
Some negative features of earlier GICs restrict their usage in some cases. Some negative features include low wear resistance, short-run time, long hardening time, susceptibility to breakage, structure sensitivity to moisture contamination during hardening, and high levels of microleakage. In recent years, attempts have been made to improve the physical properties of GICs by altering their contents.
Dental hard tissues (such as enamel, dentin, and cementum) are composed of nanoscale structural units, and their mechanical properties, such as hardness and elastic modulus, may vary across cases., Therefore, synthetic biomaterials of a similar nature are required to closely match the properties of natural tissues.
The concentration of ions, such as F −, Al 3+, and Sr 2+, released from glass-ionomer restorative materials determines their toxic potentials. Studies have reported that some glass-ionomer restorative materials cause cytotoxic and genotoxic effects through DNA damage and cell death at high concentrations.,,
| Materials and Methods|| |
This study compared five different glass ionomer restorative materials: Argion (VOCO, Germany), Zirconomer (Shofu Inc., Japan), EQUIA Forte (GC, Japan), Fuji II LC capsule (GC, Japan), and Fuji IX GP capsule (GC, Japan) [Table 1].
We obtained an ethics committee approval (no. 2017/109) from the Hatay Mustafa Kemal University Clinical Research Local Ethics Committee for this study.
Sample preparation methods
All procedures were carried out under aseptic conditions in a laminar airflow chamber (Heal Force, China). Glass coverslips, cement spatulas, and mouth spatulas were packaged separately and sterilized in an autoclave before use. All procedures were performed using sterile gloves.
The manufacturer's recommendations were followed to harden and prepare the samples. [Table 2] shows the application methods of materials and their curing times. An amalgamator device (Zoneray, China) was used to mix materials in capsule form. A light device (Woodpecker, China) was used to light-harden and heat-harden restorative materials.
|Table 2: Forms of application and hardening times of materials in the preparation of samples|
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Preparation of samples for antimicrobial activity and cytotoxicity test
The materials to be evaluated for antimicrobial activity were placed in sterile circular Teflon molds, which were done by using transparent tape on restorative materials and pressing them with glass coverslips to remove excess material and to ensure no air bubbles remained in the sample. The samples of 5 mm diameters and 2 mm depths were prepared and hardened in according to the manufacturer's recommendation [Table 2].
A total of 50 samples, which included 10 samples from each group, were prepared for antimicrobial effectiveness testing, and a total of 60 samples, which included 12 samples from each group, were prepared for the cytotoxicity test. All samples were sterilized through autoclaving at 121°C 1 atmospheric pressure for 15 min.
Preparation of samples for nanohardness and elastic modulus measurement
Five circular grooves were created at equal distances on Bakelite blocks from each other at depths of 2 mm and diameters of 7 mm. Ten Bakelite blocks were used. Fifty grooves to house 10 samples from each group were prepared. The materials used to evaluate nanohardness and elastic modulus were placed in the grooves, and materials were pressed with glass coverslips using transparent tape to remove excess material and to ensure no air bubbles remained in the sample. The samples were prepared and hardened according to the manufacturer's recommendation [Table 2]. The samples placed in light tight containers and kept in distilled water at 37°C for 24 h.
After 24 h of initial hardening, the surfaces of the prepared samples were subjected to aqueous polishing using the FORCIPOL 2V polishing machine (METKON, Turkey) with 600, 1200, and 1500 grit carbide paper. The polished samples were placed in light tight containers and kept in distilled water at 37°C for 24 h before testing was performed.
Nanohardness and elastic modulus were measured using a Hysitron TI 950 TriboIndenter device (Hysitron, USA) with a Berkovich diamond tip. The Bakelite blocks that housed the samples were placed on the device's table. Nanohardness and elastic modulus of the samples were measured by applying 6000 μN of force with the Hysitron TI 950 TriboIndenter device (Hysitron, USA) on five different points of the sample surface to form notches on each prepared sample.
Evaluation of antimicrobial activity
Calculation of minimal inhibitory concentration and minimal bactericidal concentration values
Minimal inhibitory concentration (MIC) values of the prepared samples were calculated using the “broth dilution” method. Bacterial concentrations were prepared in 1 × 106, 1 × 105, 1 × 104, 1 × 103, and 1 × 102 cfu/ml batches with the sample amounts kept constant. They were then exposed to the prepared samples for 72 h. The Lactobacillus casei (L. casei) (ATCC 4646) and S. mutans (NCTC10449) culture strains were reproduced in Mueller-Hinton agar using 24-h incubation, and the C. albicans (ATCC 10231) cultures were reproduced in Sabouraud dextrose agar with 24-h incubation to evaluate MIC and minimal bactericidal concentration (MBC) values. The bacteria (L. casei and S. mutans) and fungus (C. albicans) were measured to 1 × 106 and 1 × 105 cfu/ml, respectively, and distributed into ten sterile tubes in 0.5 ml Mueller-Hinton broth and Sabouraud dextrose broth medium. The bacterial and fungal concentrations were serially diluted to concentrations of 1 × 102 cfu/ml. The tubes were incubated at 37°C for 72 h. The last tube without turbidity was determined to be MIC.
Planting was made on Mueller-Hinton agar for bacterial strains (L. casei and S. mutans) and in Sabouraud dextrose agar for the fungus strain (C. albicans) for the last tube that turbidity was observed in for MBC and for the next two dilutions. The concentration that killed 99.9% of the bacteria was determined to be the MBC. To calculate MBC values, they were evaluated by planting three mediums from each dilution.
Evaluation of cytotoxicity
After 12-, 24-, 48-, and 72-h incubation periods for 12 samples of each group, the groups were each divided into three subgroups that included four samples that would be used to evaluate cytotoxicity using MTT assay.
Cell culture studies
The Vero cells were used to determine the cell cytotoxicity of the previously prepared samples. In all assays, the Roswell Park Memorial Institute (RPMI) 1640 production medium containing 10% fetal bovine serum (FBS), 10 mM 2-[4-(2-hydroxyethyl)-1-piperazine] ethanesulfonic acid (HEPES), 4 mM glutamine, and 100 IU/ml penicillin/streptomycin was used as a cell culture production medium, and the cell cultures were incubated at 37°C in an incubator with 5% CO2 and 95% air. The cells were cultured using inoculation on flat-bottom cell culture plates to reach 1 × 105 cell/ml. Because release liquids were taken at separate times from when the samples with the Vero cells were taken, effects of release liquids on cell viability were evaluated using MTT assay. In addition, the cell culture that did not contain release liquids was used and evaluated as the control group.
Toxicity evaluations of release liquids
The toxic effects of the samples on the cells were evaluated on 12-well plates. In the assays, the samples were evaluated in production media containing 1% FBS at a cell concentration of 1 × 105/ml. Each sample group was removed from the culture vessel surfaces with a solution of versene-trypsin after 24, 48, and 72 h, and the samples were placed in 50 ml centrifuge tubes. Centrifugation was performed at the refrigerated centrifuge of 1500 rpm for 15 min. Cell viabilities and the number of cells collected were determined under a microscope in hemocytometer with 1% trypan blue prepared in 0.9% NaCl.
MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra zolium bromide] assay
The MTT assay detects live cells colorimetrically and quantitatively. In this study, the effects of the release liquids taken at the end of the different sample incubation times on cell proliferation were evaluated using the MTT cell proliferation method. Positive and negative controls were also studied using the MTT assay. The cultures prepared with release liquids were allowed to incubate for one night at 37°C in a 5% CO2 incubator. At the end of incubation, 10 μl MTT was added to each incubator, and the plates were incubated under the same conditions for 4 hours. Absorbance was then measured using a spectrophotometer at 570 nm. Proliferation was conveyed as the ratio of the cells in the wells treated with the synthesized bis-derived compounds in comparison to the cells of the control group. In this spectrophotometric measurement-based method, MTT staining detected the effects of release liquids on cell proliferation. At the end of incubation, cell viability, or the effect of chemical compounds (synthesis) on cells, was determined by a microplate reader using the MTT assay, and IC50 (concentration inhibiting at least 50% of cell proliferation) values were determined. All test procedures and MTT assays were repeated three times.
The data obtained in this study were calculated using the SPSS program (Inc S, SPSS for Windows version 160, Chicago 2007), and P < 0.05 was considered statistically significant.
IC50 values obtained for the cytotoxicity test were analyzed using the Chi-square test.
The statistical normal distribution suitability of nanohardness and elastic modulus values was tested using the Kolmogorov-Smirnov and Shapiro-Wilk methods. Differences between the groups were analyzed using the Kruskal-Wallis analysis method because the obtained values did not show normal distribution. Median, minimum, and maximum values were used as descriptive statistics. The difference between groups was found to be significant (P < 0.05).
A pairwise evaluation (a post hoc multiple comparison test) of the tests was used to determine the origin group from which the difference came from.
| Results|| |
Antimicrobial activity findings of the materials
Although no antimicrobial and no antifungal activity was found in our study at 1 × 106 1 × 105, 1 × 104, and 1 × 103 cfu/ml concentrations for bacterial strains of chemical substances prepared according to McFarland 0.5 against S.mutans, L. casei and at 1 × 105, 1 × 104, 1 × 103, and 1 × 102 cfu/ml concentrations for fungal strains of chemical substances prepared according to McFarland 0.5 against C. albicans, we found that concentrations of prepared chemical substances inhibited bacterial reproduction in the cultures containing 1 × 102 cfu/ml bacterial strains (S. mutans and L. Casei). The fungal concentration at which these chemicals prepared against the C. albicans strain showed activity at <1 × 102 cfu/ml [Table 3].
The results of the disk diffusion tests detected, after 24-h incubation, the absence of inhibition zones at concentrations of 1 × 106 cfu/ml for bacterial strains prepared according to McFarland 0.5 and of 1 × 105 cfu/ml for fungal strains.
Cytotoxicity value findings of the materials
Determination of cell viability using the trypan blue staining method
A solution of 1% trypan blue stain prepared in phosphate buffered saline was used. Following cell incubations, the cells were removed from the culture vessels with 0.25% trypsinization solution, incubated for 15 min at room temperature, stained by adding a 1:1 (v/v) ratio of stain solution, and then examined under a microscope to determine cell viability. We evaluated cell viability using trypan blue, which was performed by means of Thoma lamella.
As shown in [Graphic 1], no significant difference was found in the cell viability of control group cells and the 24-h release water of five different glass-ionomer restorative materials (P > 0.05). Both the cell numbers and MTT assay indicated that the release liquids taken at 24 h were non-toxic.
Similarly, [Graphic 2] shows that the release liquids of five different glass-ionomer restorative materials taken at 48 h did not have statistically significant cell viability compared to the control group cells (P > 0.05).
In [Graphic 3], the effects of the release liquids of five different glass-ionomer restorative materials collected at 72 h on the Vero cell culture are shown. When the cell cultures containing the release liquids taken at the end of 72 h were morphologically evaluated alongside the control group cell lines, no pathological conditions, such as cell rounding, aggregation in cells, enlargement of cell nuclei, or spilling from the cells' surfaces, were found in the cell cultures containing the release liquids of five different restorative materials [Figure 1]. It was determined that the cell numbers in some groups had statistically significant lower results compared to the control group. These differences were found in Zirconomer, EQUIA Forte, Fuji IX GP capsule and Fuji II LC capsule (P < 0.05). Although the release liquids of the glass ionomers taken at 72 h did not produce any pathological differences in cell morphology compared to the control cells that could be detected in an inverted microscope, their viable cell numbers were found to be lower by a statistically significant margin. In other words, the release liquids of these four glass-ionomer restorative materials were determined to possibly lead to a reduction in the number of viable cells associated with the duration.
|Figure 1: Microscopic view of vero cell culture cultivated with restorative materials at the 72 h of incubation|
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The P values of Argion obtained at 72 h were >0.05 when compared to the control group. Argion did not show a toxic effect.
There were no statistically significant differences observed in Zirconomer and EQUIA Forte in terms of their numbers of viable cells (P > 0.05). Similar relationships were found among all the toxic materials (P > 0.05). We can, therefore, conclude that there is no significant difference in terms of cell cytotoxicity among these toxic materials.
Findings on the nanohardness and elastic modulus values of the materials
The median, minimum, and maximum values were determined for the nanohardness and elastic modulus of five different glass-ionomer restorative materials [Table 4]. [Graphic 4] shows the load-displacement curve of five different glass-ionomer restorative materials.
|Table 4: Median, minimum, and maximum values obtained for elastic modulus and nanohardness using the Kruskal-Wallis analysis method|
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According to the test results of elastic modulus, the differences between Fuji IX GP capsule and Argion and between EQUIA Forte and Fuji II LC capsule were determined to be insignificant (P > 0.05), whereas the differences between the other pairwise group matches were found to be significant (P = 0.00).
According to the nanohardness test results, the differences between Fuji IX GP capsule and Argion, between Argion and EQUIA Forte, and between EQUIA Forte and Fuji II LC capsule were determined to be insignificant (P > 0.05), whereas the differences between the other pairwise group matches were found to be significant (P = 0.00).
In comparing all groups, nanohardness and elastic values are ranked from highest to lowest in the following order: Fuji II LC capsule with the highest values, then EQUIA Forte, Argion, Fuji IX GP capsule, and finally, Zircomer with the lowest values. The nanohardness value ranking determined between the groups was found to be statistically significant.
| Discussion|| |
Large-scale studies in modern dentistry have investigated the development of various restorative materials, different methods of treating caries, and early stages of dental restorations.,
In a study carried out by Seppa et al., GICs were reported to have in vitro antibacterial properties. Further, Forss et al. reported that the growth of S. mutans is generally inhibited in vivo by the fluoride released surrounding conventional and silver glass ionomers. The most probable cause of the preventative effect seen in acid production appears to be fluoride production. The fluoride released from the glass ionomers is under the control of pH, and the speed control factors are saliva phosphate and proteins. Shashibhushan et al. reported a positive correlation between the amount of fluoride released and the amount of antibacterial activity. The antibacterial activity of the five different glass-ionomer restorative materials used in our study was found to be low. In protecting against fungal strains, antibacterial activity was found to have a lower value than that of bacterial strain prevention.
Saxsena et al. compared the fluoride release and antimicrobial properties of Zirconomer and Fuji IX. After 48 h, Zircomer showed a statistically significant greater inhibition zone than that of Fuji IX against S. mutans and L.casei, which the authors attributed to the amount of fluoride released. In the same study, neither Zirconomer nor Fuji IX showed any antifungal effects against C. albicans. In our study, Zirconomer and Fuji IX GP capsule showed similar antimicrobial activity against S. mutans and L.casei and demonstrated antifungal activity against C. albicans at lower levels than antimicrobial activity. The differences in these studies' findings may be attributed to the uses of different methodologies and the many other factors influencing the tests.
In studies carried out by Cassanho et al. and Bhavana et al., GICs were reported to have no effects on the growth of C. albicans. Dastjerdie et al. reported that GICs demonstrated a small antifungal effect against C. albicans. All materials used in our study also showed low antifungal activity.
In our study, none of the materials evaluated for cytotoxicity after 24- and 48-h incubation periods showed toxic effects. After 72 h of incubation, Zirconomer, EQUIA Forte, Fuji IX GP capsule, and Fuji II LC capsule showed statistically significant lower cell viability values than the control group. However, the difference between the levels of toxic effects of these toxic groups was not statistically significant (P > 0.05).
No morphological changes, such as cell rounding, granulation, narrowing of cytoplasm, or the dense formation of a nuclear structure, or cytopathological changes were observed in the morphological evaluations of all cultures incubated for 24, 48, and 72 h with the samples of the restorative materials used in our study. Therefore, it can be stated that Zirconomer, EQUIA Forte, Fuji IX GP capsule, and Fuji II LC capsule showed low-toxic effects with statistically significant lower cell viability values after 72 h of incubation than the control group.
Surface hardness is known to negatively correlate with wear on the surface of restorative materials, with lower hardness leading to higher wear. For this reason, we can deduce that Zirconomer, which yielded the lowest hardness value, will wear out more quickly than the other materials we tested. Because of excessive wear, restorations may occasionally need repair when Zirconomer is used. Because of this, it is only recommended for use as a temporary restoration and base material. The surface hardness of Zirconomer, Fuji IX GP EXTRA, and Ketac Molar was evaluated in a study carried out by Asafarial et al., in which Zirconomer showed the lowest hardness value. In a study carried out by Patil et al., the surface hardness of Filtek Z350, Beautiful II, Ketac Molar, Zirconomer, and Compoglass F was evaluated, with Zirconomer again showing the lowest hardness value. These results are all consistent with our own.
Surface hardness tests provide information about the physical properties of restorative materials. Knoop, Vicker, Brinell, and Rockwell are all test methods used to determine the degrees of hardness in different materials. Although these tests perform micro- and macro-hardness measurements, load and displacement procedures have been recorded to yield higher precision in testing nanohardness using very small indentations. Therefore, nanohardness tests tend to be preferred and were used in this study for that reason.
A high elastic modulus is necessary to resist deformation and fracturing in the cusps of teeth,, meaning the elastic modulus is associated with the reliability of restorative dental materials. Particularly, when they are placed in posterior teeth, restorative materials must have sufficient elastic modulus to resist deformation under chewing stresses. The most effective elastic modulus should be similar to the dentine. Therefore, elastic modulus knowledge is important in understanding the clinical behavior of many biomaterials.
The depth sensitive indentation technique, as defined by Oliver-Pharr, has provided a framework for establishing the theoretical relationship between elastic modulus and hardness. In statistical studies carried out, it is stated that the elastic modulus generally increases the function of hardness. The connections established within these statistical studies are also known to vary by the types of materials used. In our study, the elastic modulus and hardness values showed a positive correlation.
Some studies have indicated that the addition of resin to resin-modified GICs does not increase the surface micro-hardness of resin-modified GICs.,, However, in a study carried out by Bala et al., Fuji II LC capsule, which is a resin-modified GIC, showed a higher hardness value than Fuji IX GP capsule, which is a high viscosity GIC. In our study, Fuji II LC capsule, which yielded the highest hardness value in our study, also showed a higher hardness value than Fuji IX GP capsule.
The central light intensity emitted from the light device is greater than the light intensity emitted from the light device surroundings. Therefore, the central part of the resin-containing samples is more polymerized than the surrounding area is. To apply equal light intensity in our study to the entire surface area of the cured material and to provide equal polymerization on the entire surface using the light device with a tip diameter of 8 mm, the samples of 5 mm diameters and 2 mm depths were prepared for antimicrobial activity, and cytotoxicity test and the samples of 7 mm diameters and 2 mm depths were prepared for nanohardness and elastic modulus measurement
| Conclusion|| |
According to the antimicrobial activity test results, all materials used in this study showed low antimicrobial activity. For this reason, we can deduce that these materials may be effective in reducing a small amount of bacteria remaining in the cavity rather than remineralizing the decayed tissue under it. From this, we can conclude that it is necessary to remove the infected decayed tissue from the teeth to restore them with these materials.
Our results showed that Zirconomer, EQUIA Forte, Fuji IX GP capsule, and Fuji II LG capsule exhibited low-cytotoxic effects, but potential cytotoxic effects should still be considered when using these materials.
Fuji II LC capsule and EQUIA Forte can be used in permanent dental restorations with high occlusal loads because these materials exhibited higher hardness values than other materials.
In case of malocclusion and bruxism, using materials with low abrasion resistance are more appropriate than materials with high surface hardness values. Composite resins and GICs can be successfully used instead of amalgams, in deciduous molars showing physiological wear, because amalgams have high-hardness values. For this reason, the use of Argion and Fuji IX GP capsule with low-hardness values is recommended in our study and in the application of class I deciduous molar restorations.
In comparison to other groups, from the data obtained from our study, although Zircomer exhibited the lowest hardness value and the highest toxic value, the difference in the toxic level was not statistically significant. Furthermore, Zirconomer showed antimicrobial effects similar to the other materials. Our data points to a limited area of usage of this material in Pediatric Dentistry.
The present research is in vitro by nature, and therefore, it does not reflect the actual antimicrobial properties in oral cavities. Because of this, further in vivo studies on the aspects we studied are recommended. Further studies should also be conducted to determine the exact mechanism influencing the antimicrobial activity and cytotoxicity of GICs. Moreover, the remineralization potential and fluoride release of GICs should also be evaluated. Long-term clinical trials should be carried out to assess the antibacterial efficacy, cytotoxicity, and longevity of different GICs because practitioners should be equipped with knowledge about different material when selecting which material to use.
Financial support and sponsorship
Mustafa Kemal University Scientific Research Projects Unit project number 17.U.003.
Conflict of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4]