Apoptotic effect of eugenol envolves G2/M phase abrogation accompanied by mitochondrial damage and clastogenic effect on cancer cell in vitro.

BACKGROUND: Eugenol (EUG) is a major phenolic compound present in clove whose anti-cancer properties have been demonstrated previously. These anti-cancer properties may involves the modulation of different mechanisms, including α-estrogen receptor (αER) in luminal breast cancer cells, COX-2 inhibition in melanoma cells or p53 and caspase-3 activation in colon cancer cells. HYPOTHESIS: EUG promotes a burst in ROS production causing cell-cycle perturbations, mitochondria toxicity and clastogenesis triggering apoptosis in melanoma breast- and cervix-cancer cells in vitro. METHODS: Morphological changes were evaluated through the light- and electronic- microscopy. Cell-cycle, ROS, PCNA and Apoptosis was detected by flow cytometry and clastogenicity was evaluated by Comet-assay. RESULTS: The results obtained herein pointed out that EUG promotes, increasing ROS production leading to abrogation of G2/M of phase of cell-cycle, and consecutively, clastogenesis in vitro. In addition, EUG induces Proliferation Cell Nuclear Antigen (PCNA) downregulation and decreasing in mitochondria potential (ΔΨm). Of note, a Bax up-regulation was also observed on cells treated with EUG. All of these findings cooperate in order to induce apoptosis in cancer cells. CONCLUSION: These promising results presented herein shed new light on the mechanisms of action of EUG suggesting a possible applicability of this phenylpropanoid as adjuvant in anti-cancer therapy.

Cytotoxic effects of zoledronic acid on human epithelial cells and gingival fibroblasts.

Bisphosphonate-induced osteonecrosis has been related to the cytotoxicity of these drugs on oral mucosa cells. A previous study showed that 5 µM of zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is the highest concentration of this drug found in the oral cavity of patients under treatment. Therefore, in order to simulate an osteonecrosis clinical condition, the aim of this study was to evaluate the highest concentration of ZA applied on human epithelial cells (HaCaT) and gingival fibroblasts. For this purpose, cells (3 × 10(4) cells/cm2) were seeded in wells for 48 h using complete culture medium (cDMEM). After 48 h incubation, the cDMEM was replaced by fresh serum-free culture medium (DMEM-FBS) in which the cells were maintained for additional 24 h. Then, 5 µM ZA were added to the DMEM-FBS and the cells incubated in contact with the drug for 48 h. After this period, the number of viable cells (trypan blue), cell viability (MTT assay), total protein (TP) production and cell morphology (SEM analysis) were assessed. Data were analyzed statistically by Mann-Whitney, ANOVA and Tukey's test (α=0.05). ZA caused a significant reduction in the number of viable cells and decreased the metabolic activity of both cell lines. However, decrease of TP production occurred only in the epithelial cell cultures. Morphological alterations were observed in both cell types treated with ZA. In conclusion, ZA (5 µM) was cytotoxic to human epithelial cells and gingival fibroblast cultures, which could be associated, clinically, with the development of bisphosphonate-induced osteonecrosis.

Influence of thicknesses of smear layer on the transdentinal cytotoxicity and bond strength of a resin-modified glass-ionomer cement.

This study evaluated the transdentinal cytotoxicity (TC) and the bond strength (BS) of a resin-modified glass-ionomer cement (RMGIC) applied to dentin covered with smear layer (SL) of different thicknesses. Forty dentin discs had thick (TSL) or thin (THSL) smear layer created on their occlusal side. In artificial pulp chambers, MDPC-23 cells were seeded on the pulpal side of the dentin discs and divided into five groups: G1TC: no treatment (control); G2TC: TSL + RMGIC; G3TC: THSL + RMGIC; G4TC: TSL removal + RMGIC; G5TC: THSL removal + RMGIC. After 24 h, cell metabolism and morphology were evaluated by the methyltetrazolium (MTT) assay and by scanning electron microscopy (SEM), respectively. For BS, the following groups were determined: G1BS: TSL removal + RMGIC; G2BS: THSL removal + RMGIC; G3BS: TSL + RMGIC; G4BS: THSL + RMGIC. Shear bond strength was tested to failure in a mechanical testing machine MTS (0.5 mm/min). Statistically significant difference was observed only between the control and experimental groups (Kruskal-Wallis, p<0.05). The metabolic activity of the viable MDPC-23 cells in G2TC, G3TC, G4TC and G5TC decreased by 54.85%, 60.79%, 64.12% and 62.51%, respectively. Mean shear bond strength values for G1BS, G2BS, G3BS and G4BS were 7.5, 7.4, 6.4 and 6.7 MPa, respectively, without significant difference among them (ANOVA, p>0.05). RMGIC presented moderate transdentinal cytotoxic effects. Maintenance or removal of smear layer did not affect the bond strength of RMGIC to dentin substrate.

Correlation between light transmission and permeability of human dentin.

The influence of dentin permeability on transdentinal LED light propagation should be evaluated since this kind of phototherapy may further be clinically used to stimulate the metabolism of pulp cells, improving the healing of damaged pulps. This study evaluated the influence of the dentin permeability on the transdentinal LED light (630 nm) transmission. Forty-five 0.5-mm-thick dentin disks were prepared from the coronal dentin of extracted sound human molars. An initial measurement of transdentinal LED light transmission was carried out by illuminating the discs in the occlusal-to-pulpal direction onto a light power sensor to determine light attenuation. The discs were treated with EDTA for smear layer removal, subjected to analysis of hydraulic conductance, and a new measurement of transdentinal LED light transmission was taken. Spearman's correlation coefficient was used for analysis of data and showed a weak correlation between dentin permeability and light attenuation (coefficient = 0.19). This result indicates that higher or lower dentin permeability does not reflect the transdentinal propagation of LED light. Significantly greater transdentinal propagation of light was observed after treatment of dentin surface with EDTA (Wilcoxon test, p < 0.05). According to the experimental conditions of this in vitro study, it may be concluded that dentin permeability does not interfere in the transdentinal LED light transmission, and that smear layer removal facilitates this propagation.

Influence of the activation mode of a self-etch resin-based luting cement upon the metabolism of odontoblast-like cells.

PURPOSE: To evaluate the cytotoxicity of a self-etch resin-based luting cement, RelyXUnicem (RXU) upon chemical or dual cure and with or without interposition of IPS d.SIGN (IPSD) or IPS Empress II (IPSE) ceramic discs between cement and light source. METHODS: 112 RXU specimens were subjected to different curing conditions and incubated in culture medium (DMEM) to obtain extracts. The following groups were formed: G1: DMEM (control); G2: dual RXU; G3: chemical RXU; G4: dual RXU+IPSD; G5: chemical RXU+IPSD; G6: dual RXU+IPSE; and G7: chemical RXU+IPSE. Cultured odontoblast-like cells were incubated for 24 hours in contact with the extracts. Data from cell metabolism (CM), total protein dosage (TPD) and alkaline phosphatase activity (APA) were obtained and analyzed statistically (alpha = 0.05; Kruskal Wallis and Mann-Whitney tests). Cell morphology was analyzed by SEM. RESULTS: CM and APA were significantly lower in G3 and G7 than in G1 (P<0.05). Significant TPD decrease occurred in G5 and G7 compared to G1 (P<0.05). Only G4 and G6 presented CM changes. RXU caused no cytotoxicity when subjected to dual cure without ceramic interposition. However, mild cytopathic effects were observed after chemical setting without ceramic interposition, and after chemical and dual activation under ceramic discs.