Abstract
Keywords
Introduction
Dental caries is the most prevalent disease in the world, and restorative dental materials are essential during its treatment. 1 The main goal of restorative dental material is to replace the esthetic, biologic, and functional properties of a tooth, principally by being resistant to masticatory forces. For this, it is expected that the material has good physical-mechanical properties, like an adequate modulus of elasticity (ME), flexural strength (FS), diametral tensile strength (DTS), and microhardness.2,3 The most common way to evaluate and compare restorative dental materials is by physical-mechanical testing. 4
Resin composites (RC) are the most popular restorative dental materials, mainly because of the high esthetic demands from patients. 5 RC are formulated with a silane coupling agent that connects inorganic fillers with an organic matrix. Inorganic filler particles are commonly ceramic oxides, quartz, or glass, while the organic matrix can comprise several monomers, such as 2,2-bis[p-(2′-hydroxy-3′-methacryloxypropoxy)phenyl]-propane (BisGMA), trietylenglycoldimethacrylate (TEGDMA), dimethylaminoethylmethacrylate (DMAEMA), and several additives (photoinitiators-camphoroquinone, stabilizers, and inhibitors). 6
It is well known that RC possesses short longevity compared to classical materials like the amalgam; this is principally attributed to polymerization shrinkage and failure in adhesion, but also degradation.7
–10 At first, the wear of RC in the oral environment was attributed entirely to mechanical function.11,12 Subsequently, RC wear was related to chemical degradation, and several investigations have shown the contribution of oral enzymes in their chemical breakdown.8,13
–15 Additionally, it is well known that oral enzyme sources include salivary glands, gingival epithelium, inflammatory responses, and bacteria.14,16
–19 Hundreds of bacterial species (e.g. Streptococcus) are present in the oral cavity; some of them have a crucial role in the development of dental caries and possess a considerable affinity to RC in addition to the ability to produce acids and esterases.20
–22 It has been reported that depending on the strain,
A great variety of RC are available for clinical use. Manufacturers claim improvements every year in handling, color stability, biocompatibility, physical-mechanical properties, and longevity. These improvements are mainly associated with modifications in their composition. The use of different fillers and monomers, their combinations, and proportions would result in distinct features and rates of degradation. However, the degradation impact on the physical-mechanical properties of contemporary RC has not been extensively studied. It is essential to evaluate the possible effect of degradation on its physical-mechanical properties. Then, the objective of this investigation was to measure the ME, FS, DTS, and microhardness of three contemporary RC (Enamel Plus HRi, IPS Empress Direct, and Clearfil AP-X) to establish if degradation by exposure to
Methods
Sample preparation
Three contemporary RC brands with distinct formulations were studied (Table 1). Two stainless-steel molds were fabricated. The first had dimensions of 25 mm in length, 2 mm in width, and 2 mm in height (for ME and FS tests) according to ISO standard 4049/2000. 25 The second had dimensions of 6 mm in diameter and 4 mm in height (for DTS, microhardness tests, and Scanning Electron Microscope (SEM) observation) according to the ANSI/ADA specification No. 27. 26
Characteristics of the resin composites used in this study.
One hundred thirty-eight bar-shaped specimens (46 of each RC brand) and 276 disc-shaped specimens (92 of each RC brand) were manufactured. Each material was inserted and packed inside the molds. Filled molds were then compressed between two glass slides, and finger pressure was applied to extrude the excess and achieve a uniform surface. The material was light-cured, bar-shaped specimens with three consecutive 10 s exposures points by side, producing a partial overlapping. The disc-shaped specimens had one exposure on each side of 1000 mW/cm2 light intensity for 20 s with a Valo cordless LED curing unit (Ultradent Products, South Jordan, Utah, USA). The specimens were extracted from the molds and polished using 1000 and 1200-grit abrasive papers and Sof-Lex discs (3M ESPE, Dental Products, St. Paul, Minneapolis, USA). Specimen dimensions were verified with a digital caliper (Digimatic caliper, Mitutoyo Corp., Tokyo, Japan). All specimens were made in sterile conditions at room temperature (23°C ± 2°C). After each specimen was manufactured, they were immediately settled in sterile distilled water for 1 h, air-dried, and subjected to ultraviolet light for 10 min inside a laminar flow hood before the incubation with or without
The 46 bar-shaped and the 92 disc-shaped specimens of each RC brand were divided into five groups, four of them with 10 specimens and one with six specimens (Figure 1). Groups were named: 24H-DW groups (control groups,

FS (Flexural strength), ME (Modulus of elasticity), DTS (Diametral tensile strength), VHN (Vickers hardness numbers), 24H-DW Group (incubated in distilled water during 24 h), 30D-sBHI (incubated for 30 days in sterile Brain Heart Infusion), 30D-MUT (incubated for 30 days in BHI with
Incubation of the specimens
Each group was incubated in individual glass vials containing 8 mL of distilled water, 8 mL of sterile BHI (Becton, Dickinson Sparks, Maryland, USA), or 8 mL of BHI with 50 µL of an overnight BHI with
Mechanical tests
A computer-controlled universal testing machine (UTM) (CMS Metrology, Model WDW-5Y, Querétaro, Mexico) was used for the mechanical tests. The FS and ME tests were done with the same bar-shaped specimens and tested by the three-point bend test in concordance to ISO 4049:2000. 25 Each specimen was mounted with its edges equidistant from the midline of the UTM. The load was applied at a crosshead speed of 0.5 mm/min until it fractured. Data were collected in Newtons and converted to megapascals (MPa) using the following equation: FS = 3FL/(2BH²), where the maximum load was represented by F, L was the distance between supports (mm), B was the width of the specimen (mm), and H was the height (mm). The ME was determined in gigapascals (GPa) as ME = FL3/4BH3d, where F was the maximum load (N), L was the distance between supports (mm), B was the width of the specimen (mm), H was the height (mm), and d was the deflexion (mm) corresponding to the F load. DTS was determined in the 138 disc-shaped specimens (46 of each RC brand). They were mounted between the compressive plates of the UTM, and a compressive load was applied at a crosshead speed of 1.0 mm/min vertically on the lateral portion of the disc to produce tensile stress perpendicular to the vertical plane. DTS was obtained using DTS = 2P/πDL, where P was the maximum load (N), D was the diameter (mm), and L was the length of the specimen (mm).
Microhardness test
The microhardness test was done with a microhardness tester (CMS Metrology, Model CHV-1, Queretaro, Mexico). One hundred thirty-eight disc-shaped specimens (different from the DTS test) were used, 46 of each brand. A 2.9-N force was applied using a diamond indenter for 15 s. All measures were generated in Vickers hardness number (VHN). The VHN (kgf/mm2) was obtained with the following equation: VHN = 1.854 (Ld2), where L was the applied load (kgf), and d was the mean diagonal length (mm). This was determined from three indentations at different zones on one side of each specimen.
Scanning electron microscopy (SEM)
Three disc-shaped specimens per group chosen at random after performing the microhardness tests were sonicated (Tuttnauar-Ultrasonic cleaner, Tuttnauer, Israel) for 15 min to remove bacteria or bacterial products and were dried at room temperature (20°C). They were mounted on a holder and coated with 4 nm of carbon. The samples were analyzed using a SEM (Hitachi TM1000, Mito City, Japan) operating at 15 kV. SEM images were obtained at least from three different locations and in different magnifications using a backscattering electrons detector.
Statistical analyses
Results were statistically analyzed with two-way ANOVA combined with a post hoc Tukey-Kramer multiple comparisons test using Graph-Pad Instat, version 3.0 (Graphpad Software, San Diego, CA, USA). Statistical significance was set at
Results
Table 2 shows the ME, FS, DTS, and microhardness means and standard deviation of the three tested RC brands after the experimental conditions. None of the RC brands presented changes when comparing the five experimental conditions in the ME, FS, or DTS tests. However, there were some differences in the three mechanical tests (ME, FS, and DTS) when comparing RC brands. In general, Enamel Plus HRi presented the highest FS, DTS, and microhardness means in each tested condition (
Comparison of the three resin composite brands at each of the experimental conditions.
24H-DW: incubated in distilled water during 24 h; 30D-MUT: incubated 30 days in BHI with
Significant difference versus IPS Empress Direct, in the same column.
Significant difference versus Clearfil AP-X in the same column. ANOVA and post hoc Tukey-Kramer multiple comparisons test.
On the other hand, the Enamel Plus HRi and Clearfil AP-X specimens decreased their microhardness at 30 (71.77–64.23 and 55.66–48.26, respectively) and 60 days (70.25–61.67 and 57.41–47.58, respectively) after being exposed to
Figure 2 shows a representative image of each RC brand in each experimental condition. Eroded surfaces were evident in

Scanning electron microscope photographs (8000 X) representatives of the surface of each resin composite brand after each experimental condition. The scale bar represents 10 µm and applies to all figures. Note the erosion on the Enamel Plus HRi and Clearfil AP-X bacteria-incubated specimens mainly due to a decrease in the number of small particles, in some areas a complete lack of them (white arrow) and the presence of the larger ones more easily observed.

Two-and-a-half-dimensional (2.5D) perspectives of the same scanning electron microscope images (8000 X) of the surface of each resin composite brand after each experimental condition shown in Figure 2. The scale bar represents 10 µm and applies to all images. The 30 and 60 day images of the sBHI groups have rotated 180° and were spliced with the MUT groups to appreciate the surfaces better. Note the erosion on the Enamel Plus HRi and Clearfil AP-X bacteria-incubated specimens mainly due to a decrease in the number of small particles, in some areas a complete lack of them (white arrow) and the presence of the larger ones more easily observed. Processed by the 2.5D tool of imaging software Zen 2011, blue edition (Carl Zeiss MicroImaging GmbH, 1997–2011).
Discussion
The quality of a RC is essential for clinical success, and suitable mechanical properties are necessary for supporting occlusal forces during chewing. FS is considered the most representative feature of a RC due to considerable flexural stresses that occur during chewing.27,28 ME describes material rigidity, 29 a low ME could result in higher deformability, which would result in serious failures. 6 While the DTS test could reveal different features for brittle materials that are similar with few or no plastic deformation.30,31
Although mechanical tests are not enough to establish a valid prediction of material performance or long-term success, it is hypothesized that stronger materials better distribute the stress, resist fracture and deformation, have stability, longevity, and a higher probability of success. Then, the evaluation of the physical-mechanical properties of RC are common, but not after simulating a clinical scenario related to degradation. In the present investigation, three contemporary RC were incubated with
Our results showed that the three tested RC brands remained without differences in the ME, FS, and DTS tests regardless of exposure to bacteria; then, the tested hypothesis has to be rejected. This is consistent with the only previous investigation where a mechanical test was done on a laboratory formulated resin incubated with
On the other hand, differences between the RC were present; this could be explained because of their distinct composition.33,34 It has been reported that the filler concentration strongly influences FS and ME. 35 However, their influence is of controversy since some studies have reported that RC with lower filler content (% volume) present low FS, 36 while others report a similar FS in RC with different filler volume. 37 While a correlation between the ME and the percentage of filler by volume 38 or weight 39 has been reported. Also, the content, the filler particles’ size, and shape influence the RC mechanical performance. 39 Regarding the organic matrix, it could also influence mechanical results; it has been reported that BisGMA provides the RC with good mechanical properties. 40 While the presence of TEGDMA in RC formulations has been associated with a significant increase in ME and a decrease in FS. 41 This is consistent with our general results since Clearfil AP-X and IPS Empress Direct showed the lowest FS, both specimens have TEGDMA as a component. At the same time, Clearfil AP-X showed the highest ME.
Most of commercial and model RC contain BisGMA and/or TEGDMA, making them susceptible to degradation, mainly attributed to esterases like CE and PCE.4,17,18 Modify the proportion of monomers, as well as the design of new ones, is the primary strategy of researchers and manufacturers to avoid or at least reduce RC degradation. Attending to this, other monomers are currently used in the composition of contemporary RC, like urethane dimethacrylate (UDMA), 1,4-butandiol-dimethacrylate (1,4-BDMA), or 2,2-bis(4-(2-Methacryl-oxyethoxy)phenyl)propane (BIS-EMA). However, until now, it is unknown which may be the best combination or their exact proportions. Currently, different RC proposals exist. All are manufactured with different monomers and in different proportions; additionally, they have significant differences in their fillers, particle size, and shape that can be confusing factors to establish the best organic matrix option. While this happens, it is necessary to have as much information about the commercial RC that are already being clinically used, allowing the clinician to make the best choice.
Although in this investigation there were no changes in the physical-mechanical tests, SEM images showed that exposure to
Differences in the surface after bacterial exposure may be due to the loss of the organic matrix caused by bacteria-enzyme degradation which causes the remotion of small particles (inorganic and intrinsically stable), and the exposure of the larger ones, which promote surface erosion. 42 This is evident when comparing 24H-DW (with a lot of small particles) versus 60D-MUT (with few small but abundant large particles) SEM images of Enamel Plus HRi and Clearfil AP-X in Figure 2.
Regarding microhardness decrease, it seems to be associated with surface erosion. The RC with evident surface modifications after 30 and 60 days of bacteria exposure (Enamel Plus HRi and Clearfil AP-X) decreased microhardness. On the other hand, the IPS Empress Direct did not show convincing surface modifications, neither showed differences in its microhardness. These differences between RC could be attributed to the fact that IPS Empress Direct was the only one formulated with four different monomers (TEGDMA, BISGMA, UDMA, and BIS-EMA), this combination and their proportion could be blocking the degradation. Until now, there is no information about the possible degradation of UDMA and BIS-EMA by esterases. This association (Surface erosion-Microhardness decrease) could be due to when the organic matrix of the surface is lost, and the small inorganic particles are removed, the large particles remain on the surface, but they are separated by larger areas (which were occupied by the small particles). Therefore, when the indentation is carried out, the tip encounters less resistance on the surface (only that represented by large and separated particles), and a diminished microhardness is registered.
This result does not concur with a previous investigation that reported an increase in surface roughness but no difference in the RC microhardness.
24
Several factors could be involved in this discrepancy, one of the most important is the difference in composition, filler size, and loading of the tested RC since correlations of volume,
43
mass fraction of filler,
44
and hardness have been reported. Another discrepancy could be the bacteria strain chosen in both experiments. While in this investigation, we used
There is an imperious need for more investigation on RC to improve the reduction or blockage of the degradative effect of enzymes. Although this investigation did not simulate the complex mechanisms involved in the complete degradation process that could occur in the mouth, excluding factors like the thermal and other mechanisms of degradation and the chewing process; it is reasonable to state that the degradation observed in two of the RC could be maximized in a clinical situation because once the action of enzymes softens a layer of the RC, the chewing forces will easily expose a new surface layer 45 and the chemical attack can continue. Furthermore, it is well known that surface roughness plays an essential role in biofilm formation46,47 promoting microbial adhesion and plaque retention, 48 allowing more bacteria to attach and colonize the surface, increasing the degradation and promoting more erosion. All these will undoubtedly influence the restoration longevity.
Conclusions
It was demonstrated that
