RESUMO
Bisphenol A (BPA) may adversely affect human health by inducing oxidative stress and irreversible damage to cells. Bioactive compounds found in some functional foods, individually or in combination, can attenuate the negative effects of BPA exposure; an example is the multi-supplement containing guarana (Gua), selenium (Se), and L-carnitine (LC) -GSC- which has already demonstrated antioxidant, genoprotective, and immunomodulatory activities. This study aimed to determine the effect of GSC and its constituents on oxidative and genotoxic alterations triggered by BPA exposure in the retinal epithelial cell line. The cells exposed to BPA (0.001, 0.01, 0.1, 1, 3, and 10 µM) to determine the lowest concentration required to induce cyto-genotoxicity. ARPE-19 cells were then concomitantly exposed to the selected BPA concentration, GSC, and its components (Gua, 1.07 mg/mL; Se, 0.178 µg/mL; and LC, 1.43 mg/mL). Flow cytometry, biochemical assays, qRT-PCR, genotoxicity, apoptosis, and cellular proliferation. Based on our results, 10 µM of BPA could induce cyto-genotoxic and oxidative alterations. BPA did not alter the Bcl-2/BAX expression ratio but induced Casp3 and Casp8 overexpression, suggesting that apoptosis was induced mainly via the extrinsic pathway. GSC partially reversed the alterations triggered by BPA in ARPE-19 cells. However, Se had unexpected negative effects on ARPE-19 cells. The multi-supplement GSC may attenuate changes in oxidative and genotoxic markers related to exposure of ARPE-19 cells to BPA. our results revealed that the antioxidant, anti-apoptotic, and genoprotective properties of GSC were not universally shared by its individual, once Se did not exhibit any positive impact.
Assuntos
Apoptose , Compostos Benzidrílicos , Carnitina , Estresse Oxidativo , Fenóis , Epitélio Pigmentado da Retina , Selênio , Fenóis/toxicidade , Compostos Benzidrílicos/toxicidade , Humanos , Selênio/farmacologia , Carnitina/farmacologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Linhagem Celular , Paullinia/química , Dano ao DNA/efeitos dos fármacos , Antioxidantes/farmacologia , Células Epiteliais/efeitos dos fármacos , Citometria de Fluxo , Suplementos NutricionaisRESUMO
AIM: To evaluate the in vitro toxicity of irrigating solutions and pharmacological associations used in the pulpectomy of primary teeth. METHODOLOGY: The cell viability (MTT), lipid peroxidation (TBARS), alkaline comet assay and GEMO tests were performed to evaluate the cytotoxicity and genotoxicity of solutions: sodium hypochlorite (1% and 2.5%), 2% chlorhexidine, 6% citric acid and 17% EDTA, which were tested, individually and in association, exposing human peripheral blood mononuclear cells (MTT, TBARS and alkaline comet assay), at 24 and 72 h, and dsDNA (GEMO). After performing the Kolmogorov-Smirnov test, data were analysed by anova followed by Dunnett's post hoc test, and Kruskal-Wallis followed by Dunn post hoc test. A significance level was established at P < 0.05. RESULTS: All irrigating solutions and pharmacological associations reduced cell viability at 24 h (P < 0.05). These reductions were maintained after 72 h, except for EDTA and associations of sodium hypochlorite (1% and 2.5%) with EDTA and of chlorhexidine with EDTA. Lipid peroxidation at 24 h was caused by EDTA and by 2.5% sodium hypochlorite with EDTA; it was also caused at 72 h by sodium hypochlorite (1% and 2.5%) and the three associations with citric acid (P < 0.05). All groups caused DNA damage when assessed by the alkaline comet assay, at 24 h and 72 h (P < 0.05). In the GEMO assay, all groups caused dsDNA damage (P < 0.05), except for chlorhexidine with EDTA. CONCLUSION: All groups showed some level of toxicity. Amongst the main solutions, chlorhexidine presented less cytotoxic potential. EDTA was the least cytotoxic of the auxiliary irrigant solutions, and the association of these two solutions showed the lowest toxicity potential amongst all groups.
Assuntos
DNA/efeitos dos fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Pulpectomia/efeitos adversos , Dano ao DNA , Leucócitos Mononucleares/metabolismo , Irrigantes do Canal Radicular , Dente Decíduo , Testes de ToxicidadeRESUMO
AIM: To evaluate the cytotoxicity, oxidative stress and genotoxicity in vitro of four iodoform pastes and three calcium hydroxide pastes. METHODOLOGY: Peripheral blood mononuclear cells (PBMCs) and pure calf thymus DNA (dsDNA) were exposed to extracts of the pastes. Cytotoxicity was assessed with the MTT assay. Generation of reactive oxygen species (ROS) was evaluated using a DCFH-DA assay, and lipid peroxidation was evaluated using a TBARS assay. Genotoxicity was evaluated using the alkaline comet assay and Genomodifier capacity assay (GEMO). All tests were performed after 24 h and 72 h of cell exposure, except GEMO. After performing the Kolmogorov-Smirnov test, data were analysed by Kruskal-Wallis and Dunn's post-tests, and anova with Dunnett's post-test, with a significance level established at P < 0.05. RESULTS: The MTT assay revealed that chlorhexidine, Maxitrol and neomycin sulphate + bacitracin pastes decreased cell viability after 24 h (P < 0.05). No group was associated with a significant decreased cell viability or lipid peroxidation after 72 h. Calcium hydroxide pastes increased the cell viability levels at both experimental times (P < 0.05). Lipid peroxidation was observed with the exposure of cells to calcium hydroxide pastes after 24 h (P < 0.05). Exposure to chlorhexidine, Guedes-Pinto and calcium hydroxide pastes resulted in a significant increase in ROS after 24 h (P < 0.05), whereas iodoform pastes and Calen thickened with zinc oxide significantly increased the ROS after 72 h (P < 0.05). The comet assay revealed that exposure of the PBMCs to iodoform pastes did not damage DNA at either period of time (P > 0.05). However, chlorhexidine paste caused DNA damage in dsDNA (P < 0.05). Calcium hydroxide pastes caused DNA damage in both tests (P < 0.05). CONCLUSION: The pastes varied in their ability to induce cytotoxicity, genotoxicity and oxidative stress. In general, Guedes-Pinto, Maxitrol and neomycin sulphate + bacitracin pastes exhibited better biocompatibility in vitro.