Evaluation of the effects of three different Mineral Trioxide Aggregate cements on proinflammatory cytokine levels
Project no. 20965 · Status: Kapanmış (Yayın bekleniyor) · 2019
Summary
The aim of this study was to compare the cytotoxicity of ProRoot MTA, MTA Repair HP and Bio MTA+ and to evaluate the effects of these materials on proinflammatory cytokine levels of human monocytes. MTA cements were prepared according to their manufacturers’ instructions, were placed in standard cylinder-shaped moulds and were incubated 24 hours at 370 C. THP-1 (human monocyte cells) cell lines were used for the assays. THP-1 cells were differentiated into macrophages by 24-hour incubation with phorbol 12-myristate 13-acetate (PMA). Then, a group of cells was activated by lipopolysaccharide (LPS). The cytotoxic effects of the samples were evaluated by colorimetric determination of mitochondrial reductase activity in viable cells at 24th, 48th and 72nd hours. In the second phase of the study, supernatants of cells used in cytotoxicity test were utilized to test the proinflammatory potential of the tested materials. IL-1ß, IL-6 and TNF-? levels in supernatants were determined using enzyme linked immunosorbent assay (ELISA) kits. The data were examined with Two way ANOVA, Tukey’s Multiple Comparison Test, Kruskal Wallis test and Dunnett’s Multiple Comparison Test (P=0,05). Macrophage viability values at the end of 72 hours were observed as 95.782 µg/ml**** for ProRoot MTA, 65.274 µg/ml for MTA Repair HP *, 68.012 µg/ml* for Bio MTA+ and 58.767 µg/ml**** for positive control group (P<0,05:*, P<0,01:**, P<0,001:***, P<0,0001:****). In the cytotoxicity analysis involving LPS-treated cells, values observed as 77.101 µg/ml for ProRoot MTA, 69.211 µg/ml for MTA Repair HP, 39.134 µg/ml for Bio MTA+, and 88.438 µg/ml for positive control group. IL-1ß values observed at 72 hours were 9.857 pg/ml in the control group, 30.082 pg/ml* in ProRoot, 22.503 pg/ml** in MTA Repair HP and 58.660 pg/ml* in Bio MTA+ (P <0,05: *, P <0,01: **, P <0,001: ***, P <0,0001: ****). While IL-6 levels were similarly low in all groups, MTA Repair HP statistically significantly suppressed cytokine secretion at 24th hour. The TNF-? values secreted at 72 hours were 6.214 pg/ml for the control group, 23.025 pg/ml** for ProRoot, 7.260 pg/ml for MTA Repair HP and 8.738 pg/ml for Bio MTA+ (P <0,05: *, P <0,01: **, P <0,001: ***, P <0,0001: ****). The presence of LPS increased the amount of the cytokine secretion regardless of cements. At the end of the evaluation period, the survival rates of macrophage cells were sorted in a descending order starting from high to low as ProRoot MTA, Bio MTA+, MTA Repair HP and positive control group, respectively. The survival rates of LPS-treated cells were ranked as positive control group, ProRoot MTA, MTA Repair HP and Bio MTA+. The highest IL-1ß values observed at 72 hour were listed in order as Bio MTA+, ProRoot MTA, MTA Repair HP and the control group, whereas the IL-6 values were ranked from the highest as the control group, ProRoot MTA, MTA Repair HP and Bio MTA+. TNF-? levels were ranged as ProRoot MTA, Bio MTA+, MTA Repair HP, and the control group. The highest IL-1ß values obtained from LPS differentiated cells were aligned as Bio MTA+, MTA Repair HP, ProRoot MTA and the control group. IL-6 values were ranked in a descending order as the control group, ProRoot MTA, MTA Repair HP and Bio MTA+, but TNF-? values were listed in order as Bio MTA+MTA Repair HP, ProRoot MTA and the control group. It is anticipated that MTA Repair HP, which is known to have a very short setting time, will provide an advantage in clinical applications, considering its optimum biocompatibility and anti-inflammatory properties. In order to provide a better understanding of the in vitro results of the present study, further clinical studies are required.