RÉSUMÉ
OBJECTIVE: To explore the immune cytotoxicity effect and its mechanism of trichloroethylene( TCE) on activated human T cells. METHODS: a) Different concentrations of TCE( 0. 32,0. 63,1. 25,2. 50,5. 00,10. 00 mmol / L)were used to treat activated T cells [activated with cluster of differentiation( CD) 3 and CD28] respectively. Dimethyl sulfoxide( DMSO) was used in the solvent group and the control group used no TCE or DMSO. The survival rate of activated T cells was calculated using CCK-8 assay after being cultured for 24 hours. b) Different concentrations of TCE( 0. 00,2. 50,5. 00 mmol/L) were used to treat activated T cells. The apoptosis of cells was detected using flow cytometry. c) Different concentrations of TCE( 0. 00,0. 32,0. 63,1. 25,2. 50,5. 00 mmol / L) were used to treat activated T cells and the level of cytokines as interleukin( IL)-2 and IL-6 in cell culture supernatant was detected using enzyme linked immunosorbent assay after culturing for 24 hours. d) The control group and TCE treatment group of activated T cells were treated with 0. 00 and 5. 00 mmol / L TCE respectively. Cells were collected after culturing 0,30,60 and 120 minutes. Western Blot was used to detect the protein expression of signal transducers and activators of transcription3( STAT3) and phospho-STAT3( p-STAT3). RESULTS: a) After 24-hour-exposure to TCE,the activated T cell survival rate of 10. 00 mmol / L TCE treatment group were significantly lower than that in the control group and DMSO group( P <0. 05). b) There were no significant differences in cell apoptosis of activated T cells after treatment with 0. 00,2. 50 and5. 00 mmol / L TCE( P > 0. 05). c) In groups treated with different concentrations of TCE( 0. 32,0. 63,1. 25,2. 50,5. 00 mmol / L),the level of IL-2 and IL-6 in the cell culture supernatant of activated T cells were significantly higher than that in the control group( P < 0. 05). With the increasing of TCE exposure doses,the levels of IL-2 and IL-6 significantly increased( P < 0. 01) with dose-effect relationship. Compared with the control group,the levels of IL-17 A,interferongamma and transforming growth factor-beta in cell culture supernatant of activated T cells of the TCE treatment groups were no significant differences( P > 0. 05). d) The expression of p-STAT3 protein was low in the control group at different times. The expression of p-STAT3 protein in TCE treatment group was low at 0 minute,but increased at 30,60,120 minutes. The expression of p-STAT3 protein in TCE treatment group was higher than that in the control group at different time points. The levels of STAT3 total protein in TCE treatment group and the control group were similar at different time points,and were higher than the p-STAT3 proteins. CONCLUSION: TCE at 5. 00 mmol / L had no observed toxic effect on activated T cells. High doses of TCE( ≥10. 00 mmol / L) showed cytotoxic damages to activated T cells,and low doses of TCE( ≤5. 00 mmol / L) could stimulate activated T cells to secrete IL-2 and IL-6. Treatment of TCE at 5. 00 mmol / L on activated T cells could up-regulated the level of p-STAT3.
RÉSUMÉ
O objetivo deste estudo foi avaliar a citotoxicidade de 3 resinas acrílicas (RA) para bases de prótese sendo duas polimerizadas por energia de microondas (Vipi Wave-VW e Nature Cryl-NC) e uma fotoativada (Eclipse-E); e a biocompatibilidade de materiais reembasadores resilientes (MRR), sendo 2 à base de silicone (Ufi Gel P e Sofreliner S), 2 à base de resina acrílica (Durabase Soft e Trusoft) e 2 condicionadores de tecidos (Softone e Coe Comfort) por meio de análises do metabolismo celular, da morfologia celular, do padrão de morte celular, da proliferação celular e da expressão do fator de crescimento TGFß 1, da integrina α5ß1 e de citocinas. A análise das RA foi realizada com células L929; para os MRR, as análises foram realizadas utilizando-se células L929, e/ou HaCaT e/ou RAW 264,7. Os resultados obtidos foram submetidos à análise estatística, realizada ao nível de significância de 5%. Todos os materiais quando testados com eluatos não apresentaram efeitos citotóxicos, e quando testados em contato direto (MTT e azul de alamar) alteraram significativamente a viabilidade celular, tendo sido os menores valores observados para o material NC quando comparado ao E. Para os materiais reembasadores o T, S e C. Maior taxa de necrose e apoptose foram observadas para os eluatos de todos os materiais quando avaliados no período de 24 horas; além disso, foi demonstrado que os materiais avaliados podem desencadear a expressão do TGFß 1, da integrina α5ß1 e de proteínas inflamatórias (fator de necrose tumoral - TNF-α, da citocina IL-1ß e de 5 quimiocinas: CCL2, CCL3, CCL5, CXCL2 e CXCL4). Sendo assim, de acordo com as limitações do presente estudo, pode-se concluir que a resina Nature Cryl resultou em menor viabilidade celular que a resina Eclipse e que os materiais Ufi Gel P e Sofreliner S apresentaram, em geral, melhor bicompatibilidade dentre os materiais reembasadores resilientes avaliados
The aim of this study was to evaluate the cytotoxicity of 3 denture base acrylic resins with different polymerization methods (Eclipse, Vipi Wave and Nature Cryl) and the biocompatibility of soft lining materials, 2 silicone-based (Ufi Gel P and Sofreliner S), 2 acrylic-based (Durabase Soft and Trusoft) and 2 tissue conditioners (Softone and Coe Comfort), by analysing cell metabolism, cell morphology, pattern of cell death, cell proliferation and expression of growth factor TGFß 1, integrin α5ß1 and cytokines. For the acrylic resins, analyses were made with L929 cells, while for the resilient lining materials, analyzes were performed using L929, and/or HaCaT and/or RAW 264.7 cells. Results were statistically analyzed, at a significance level of 5%. The 24 and 48-h eluates from all materials were not cytotoxic to the cells. For the direct contact tests (MTT and Alamar Blue), a significant effect on cell viability was found, with the lowest values observed for materials NC when compared with E and between the reliners Trusoft, Softone and Coe Comfort. For all materials, increased rate of necrosis and apoptosis were observed for the 24 hs eluates. It was also demonstrated that all materials tested can trigger the expression of TGFß 1, α5ß1 integrin and inflammatory proteins (tumor necrosis factor -TNFα, cytokine IL-1ß and 5 chemokines- CCL2, CCL3, CCL5, CXCL2 and CXCL4). Thus, within the limitations of this study, it can be concluded that the denture base acrylic resin Nature Cryl resulted in lower cell viability compared to the Eclipse resin and that the materials Ufi Gel P and Sofreliner S showed better biocompatibility among the soft liner materials evaluated