Danijela Marović

Raman Spectroscopic Assessment of Degree of Conversion of Bulk-Fill Resin Composites - Changes at 24 Hours Post Cure

OBJECTIVE The aim of this study was to determine degree of conversion (DC) of solid and flowable bulk-fill composites immediately and after 24 hours and investigate the variations of DC at surface and depths up to 4 mm. MATERIALS AND METHODS Eight bulk-fill composites (Tetric EvoCeram Bulk Fill [shades IVA and IVB], Quixfil, X-tra fil, Venus Bulk Fill, X-tra Base, SDR, Filtek Bulk Fill) were investigated, and two conventional composites (GrandioSO, X-Flow) were used as controls. The samples (n = 5) were cured for 20 seconds with irradiance of 1090 mW/cm(2). Raman spectroscopic measurements were made immediately after curing on sample surfaces and after 24 hours of dark storage at surface and at incremental depths up to 4 mm. Mean DC values were compared using repeated measures analysis of variance (ANOVA) and t-test for dependent samples. RESULTS Surface DC values immediately after curing ranged from 59.1%-71.8%, while the 24-hour postcure values ranged from 71.3%-86.1%. A significant increase of DC was observed 24 hours post cure for all bulk-fill composites, which amounted from 11.3% to 16.9%. Decrease of DC through depths up to 4 mm varied widely among bulk-fill composites and ranged from 2.9% to 19.7%. CONCLUSIONS All bulk-fill composites presented a considerable 24-hour postcure DC increase and clinically acceptable DC at depths up to 4 mm. Conventional control composites were sufficiently cured only up to 2 mm, despite significant postcure polymerization.

Monomer conversion and shrinkage force kinetics of low-viscosity bulk-fill resin composites

Abstract Objective. To investigate the subsurface degree of conversion (DC) and shrinkage force formation of low-viscosity (flowable) bulk-fill composite materials. Materials and methods. Three flowable bulk-fill resin composites [SureFil SDR flow (SDR; Dentsply DeTrey), Venus Bulk Fill (VB; Heraeus Kulzer) and x-tra base (XB; VOCO)] and one conventional flowable control composite material [EsthetX flow (EX; Dentsply DeTrey)] were tested. The materials were photoactivated for 20 s at an irradiance of 1170 mW/cm2 and the DC (n = 5) was recorded at 0.1-, 1.5- and 4-mm depth using Fourier transform infrared spectroscopy. Shrinkage forces (n = 5) of 1.5-mm-thick specimens were continuously recorded for 15 min using a custom-made stress analyzer. Data were statistically analyzed by ANOVA, Tukey’s HSD and Bonferroni’s post-hoc tests (α = 0.05). Results. SDR generated the significantly lowest shrinkage forces (22.9 ± 1.4 N), but also attained the significantly lowest DC at 1.5-mm depth (67.5 ± 0.8%). The conventional flowable composite EX generated the significantly highest shrinkage forces (40.7 ± 0.7 N) and reached a significantly higher DC (74.4 ± 1.3%) compared to SDR and XB at 1.5-mm depth. The shrinkage force values of VB (29.4 ± 1.1 N) and XB (28.3 ± 0.6 N) were similar (p > 0.05). All materials attained significantly higher DC at 4-mm depth than at the near-surface. Conclusion. The tested low-viscosity bulk-fill materials show lower shrinkage force formation than a conventional flowable resin composite at high levels of degree of conversion up to 4-mm incremental thickness.

Pre-heating of high-viscosity bulk-fill resin composites: Effects on shrinkage force and monomer conversion

OBJECTIVES To investigate the influence of pre-heating of high-viscosity bulk-fill composite materials on their degree of conversion and shrinkage force formation. METHODS Four bulk-fill composite materials (Tetric EvoCeram Bulk Fill-TECBF, x-tra fil-XF, QuixFil-QF, SonicFill-SF) and one conventional nano-hybrid resin composite (Tetric EvoCeram-TEC) were used. The test materials were either kept at room temperature or pre-heated to 68°C by means of a commercial heating device, before being photoactivated with a LED curing unit for 20s at 1170mW/cm(2). Shrinkage forces (n=5) of 1.5-mm-thick specimens were recorded in real-time for 15min inside a temperature-controlled chamber at 25°C (simulating intraoral temperature after rubber dam application) with a custom-made stress analyzer. Degree of conversion (n=5) was determined at the bottom of equally thick (1.5mm) specimens using Fourier transform infrared spectroscopy. Data were analyzed with Students t-test, ANOVA and Tukeys HSD post-hoc test (α=0.05). RESULTS Composite pre-heating significantly increased the degree of conversion of TECBF, but had no effect on monomer conversion of the other materials investigated. For each of the test materials, pre-heated composite generated significantly lower shrinkage forces than room-temperature composite. At both temperature levels, TECBF created the significantly highest shrinkage forces, and QF caused significantly higher shrinkage forces than both XF and TEC. CONCLUSIONS Both the composite material and the pre-cure temperature affect shrinkage force formation. Pre-heating of bulk-fill and conventional restorative composites prior to photoactivation decreases polymerization-induced shrinkage forces without compromising the degree of conversion. CLINICAL SIGNIFICANCE Composite pre-heating significantly reduces shrinkage force formation of high-viscosity bulk-fill and conventional resin composites, while maintaining or increasing the degree of monomer conversion, dependent upon the specific composite material used.

Reinforcement of experimental composite materials based on amorphous calcium phosphate with inert fillers.

OBJECTIVES The aim of this study was to examine the influence of the addition of glass fillers with different sizes and degrees of silanization percentages to remineralizing composite materials based on amorphous calcium phosphate (ACP). METHODS Four different materials were tested in this study. Three ACP based materials: 0-ACP (40 wt% ACP, 60 wt% resin), Ba-ACP (40 wt% ACP, 50 wt% resin, 10 wt% barium-glass) and Sr-ACP (40 wt% ACP, 50 wt% resin, 10 wt% strontium-glass) were compared to the control material, resin modified glass ionomer (Fuji II LC capsule, GC, Japan). The fillers and composites were characterized using scanning electron microscopy. Flexural strength and modulus were determined using a three-point bending test. Calcium and phosphate ion release from ACP based composites was measured using inductively coupled plasma atomic emission spectroscopy. RESULTS The addition of barium-glass fillers (35.4 (29.1-42.1) MPa) (median (25-75%)) had improved the flexural strength in comparison to the 0-ACP (24.8 (20.8-36.9) MPa) and glass ionomer control (33.1 (29.7-36.2) MPa). The admixture of strontium-glass (20.3 (19.5-22.2) MPa) did not have any effect on flexural strength, but significantly improved its flexural modulus (6.4 (4.8-6.9) GPa) in comparison to 0-ACP (3.9 (3.4-4.1) GPa) and Ba-ACP (4.6 (4.2-6.9) GPa). Ion release kinetics was not affected by the addition of inert fillers to the ACP composites. SIGNIFICANCE Incorporation of barium-glass fillers to the composition of ACP composites contributed to the improvement of flexural strength and modulus, with no adverse influence on ion release profiles.

Composite-induced toxicity in human gingival and pulp fibroblast cells

Abstract Objective. The most important requirement for a material to be used in medical applications is its biocompatibility. Dental composite materials come into direct contact with oral tissues, especially gingival and pulpal cells. This study was performed to evaluate possible DNA damage in cells of human origin exposed to dental composites in vitro using a cytogenetic assay. Materials and methods. Two composite resins (Vertise Flow, Kalore) were tested on human gingival and pulp fibroblasts using the acridine orange/ethidium bromide viability staining and alkaline comet assay. Cultures were treated with polymerized composites in two different concentrations (20 mg/ml, 40 mg/ml) for 14 days. Chi-square and Kruskall-Wallis non-parametric test were used for the statistical analysis (p < 0.05). Results. Significant cytotoxicity was observed for 40 mg/ml of Vertise Flow in both cultures, while Kalore (40 mg/ml) showed cytotoxic effect only on human pulp fibroblasts. A significant level of DNA damage was detected for both materials and concentrations, in both cell cultures. Conclusion. If the two cell cultures are compared, the pulp cells were more sensitive to the cyto/genotoxic effects of dental composites. Based on the results, one can conclude that the use of tested materials may cause cellular damage in gingival and pulp fibroblasts in vitro.

Conversion and temperature rise of remineralizing composites reinforced with inert fillers.

OBJECTIVES Remineralizing experimental composites based on amorphous calcium phosphate (ACP) were investigated. The impact of curing time (20 and 40s), curing depth (1, 2, 3 and 4mm) and addition of inert fillers (barium glass and silica) on the conversion and temperature rise during curing were examined. METHODS Five ACP-composites and two control composites were prepared based on the light-curable EBPADMA-TEGDMA-HEMA resin. For temperature measurements, a commercial composite was used as an additional control. Conversion was assessed using FT-Raman spectroscopy by comparing the relative change of the band at 1640 cm(-1) before and after polymerization. The temperature rise during curing was recorded in real-time using a T-type thermocouple. RESULTS At 1mm depth, the ACP-composites attained significantly higher conversion (77.8-87.3%) than the control composites based on the same resin (60.5-66.3%). The addition of inert fillers resulted in approximately 5% lower conversion at clinically relevant depths (up to 2mm) for the curing time of 40s. Conversion decline through depths depended on the added inert fillers. Conversion values higher than 80% of the maximum conversion were observed for all of the ACP-composites at depths up to 3mm, when cured for 40s. Significantly higher total temperature rise for the ACP-composites (11.5-13.1 °C) was measured compared to the control composites (8.6-10.8 °C) and the commercial control (8.7 °C). CONCLUSIONS The admixture of inert fillers represents a promising strategy for further development of ACP-composites, as it reduced the temperature rise while negligibly impairing the conversion. CLINICAL SIGNIFICANCE High conversions of ACP-composites are favorable in terms of mechanical properties and biocompatibility. However, high conversions were accompanied with high temperature rise, which might present a pulpal hazard.

Impedance changes during setting of amorphous calcium phosphate composites.

OBJECTIVES To investigate the electrical properties of experimental light-curable composite materials based on amorphous calcium phosphate (ACP) with the admixture of silanized barium glass and silica fillers. METHODS Short-term setting was investigated by impedance measurements at a frequency of 1kHz, while for the long-term setting the impedance spectra were measured consecutively over a frequency range of 0.05Hz to 1MHz for 24h. The analysis of electrical resistivity changes during curing allowed the extraction of relevant kinetic parameters. The impedance results were correlated to the degree of conversion assessed by Raman spectroscopy, water content determined by gravimetry, light transmittance measured by CCD spectrometer and microstructural features observed by scanning electron microscopy. RESULTS ACP-based composites have shown higher immediate degree of conversion and less post-cure polymerization than the control composites, but lower polymerization rate. The polymerization rate assessed by impedance measurements correlated well with the light transmittance. The differences in the electrical conductivity values observed among the materials were correlated to the amount of water introduced into composites by the ACP filler. High correlation was found between the degree of conversion and electrical resistivity. Equivalent circuit modeling revealed two electrical contributions for the ACP-based composites and a single contribution for the control composites. SIGNIFICANCE The impedance spectroscopy has proven a valuable method for gaining insight into various features of ACP-based composites. Better understanding of the properties of ACP-based composites should further the development of these promising bioactive materials.

Genotoxic biomonitoring of flowable and non-flowable composite resins in peripheral blood leukocytes.

Abstract Objective. Composite restorative materials represent one of the most important groups of materials in contemporary dental practice. However, their incomplete polymerization may lead to monomer-induced genotoxicity. The objective of this study was to evaluate the genotoxicity of three flowable (Filtek Supreme XT Flow, Tetric EvoFlow, Gradia Direct Flo) and three non-flowable dental composite materials (Filtek Z250, Tetric EvoCeram, Gradia Direct Posterior). Materials and methods. Genotoxicity assessment of composite materials was carried out in vitro in human peripheral blood leukocytes using the alkaline single-cell gel electrophoresis technique (comet assay). Prepared materials were eluted in saline solution for 1 h, 1 day and 5 days. Thereafter leukocyte cultures were treated with different concentrations of eluates obtained from each of the tested dental composite materials. Kruskall-Wallis non-parametric test was used for statistical analysis (p < 0.05). Results. The tested materials did not show genotoxic effects after exposure of leucocytes to 1 h eluates. Culture treated with 1 day eluates of all tested materials, only at a highest concentration (10−2), affected the measured cytogenetic parameters. Of all tested materials, only Filtek Z250 and Filtek Supreme XT Flow did not exhibit a genotoxic effect in cultures that were under the influence of 5 day eluates. Conclusion. Tested materials exhibited limited genotoxic activity in peripheral blood leukocytes. Since the effect was observed only in leukocyte cultures treated by 1-day eluates at the highest concentration (10−2) and it decreases in cultures exposed to 5 day eluates, it should not pose a significant risk to the human genome.

Genotoxic potential of dental bulk-fill resin composites

OBJECTIVE To investigate both genotoxicity and hardening of bulk-fill composite materials applied in 4-mm layer thickness and photo-activated for different exposure times. METHODS Three flowable bulk-fill materials and one conventional flowable composite were filled in molds (height: 4mm) and irradiated for 20 or 30s. The top (0mm) and bottom (4mm) specimen surface were mechanically scraped, and eluates (0.01g composite in 1.5ml RPMI 1640 cell culture media) prepared for each material, surface level and irradiation time. Genotoxicity was assessed in human leukocytes using both the alkaline comet assay and cytokinesis-blocked micronucleus assay, and Knoop hardness (KHN) was measured at the top and bottom specimen surface (n=8). RESULTS At both irradiation times, none of the bulk-fill composites significantly affected comet assay parameters used in primary DNA damage assessment or induced significant formation of any of the scored chromatin abnormalities (number of micronuclei, nuclear buds, nucleoplasmic bridges), whether eluates were obtained from the top or bottom surface. Furthermore, no decrease in KHN from the top to the bottom surface of the bulk-fill materials was observed. On the other hand, the conventional composite irradiated for 20s showed at 4-mm depth a significant increase in the percentage of DNA that migrated in the tail and a significant increase in the number of nuclear buds, as well as a significant decrease in KHN relative to the top surface. SIGNIFICANCE Bulk-fill resin composites, in contrast to conventional composite, applied in 4-mm thickness and photo-activated for at least 20s do not induce relevant genotoxic effects or mechanical instability.

Remineralizing amorphous calcium phosphate based composite resins : The influence of inert fillers on monomer conversion, polymerization shrinkage, and microhardness

Aim To determine if the addition of inert fillers to a bioactive dental restorative composite material affects its degree of conversion (DC), polymerization shrinkage (PS), and microhardness (HV). Methods Three amorphous calcium phosphate (ACP)-based composite resins: without added fillers (0-ACP), with 10% of barium-glass fillers (Ba-ACP), and with 10% of silica fillers (Si-ACP), as well as commercial control (Ceram•X, Dentsply DeTrey) were tested in laboratory conditions. The amount of ACP (40%) and the composition of the resin mixture (based on ethoxylated bisphenol A dimethacrylate) was the same for all ACP materials. Fourier transform infrared spectroscopy was used to determine the DC (n = 40), 20 min and 72 h after polymerization. Linear PS and Vickers microhardness (n = 40) were also evaluated. The results were analyzed by paired samples t test, ANOVA, and one-way repeated measures ANOVA with Student-Newman-Keuls or Tukey’s post-hoc test (P = 0.05). Results The addition of barium fillers significantly increased the DC (20 min) (75.84 ± 0.62%) in comparison to 0-ACP (73.92 ± 3.08%), but the addition of silica fillers lowered the DC (71.00 ± 0.57%). Ceram•X had the lowest DC (54.93 ± 1.00%) and linear PS (1.01 ± 0.24%) but the highest HV (20.73 ± 2.09). PS was significantly reduced (P < 0.010) in both Ba-ACP (1.13 ± 0.25%) and Si-ACP (1.17 ± 0.19%) compared to 0-ACP (1.43 ± 0.21%). HV was significantly higher in Si-ACP (12.82 ± 1.30) than in 0-ACP (10.54 ± 0.86) and Ba-ACP (10.75 ± 0.62) (P < 0.010). Conclusion Incorporation of inert fillers to bioactive remineralizing composites enhanced their physical-mechanical performance in laboratory conditions. Both added fillers reduced the PS while maintaining high levels of the DC. Silica fillers additionally moderately improved the HV of ACP composites.