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Research Article
- High-plasticity mineral trioxide aggregate and its effects on M1 and M2 macrophage viability and adherence, phagocyte activity, production of reactive oxygen species, and cytokines
- Betânia Canal Vasconcellos, Layara Cristine Tomaz Tavares, Danilo Couto da Silva, Francielen Oliveira Fonseca, Francine Benetti, Antônio Paulino Ribeiro Sobrinho, Warley Luciano Fonseca Tavares
- Restor Dent Endod 2023;48(1):e6. Published online December 29, 2022
- DOI: https://doi.org/10.5395/rde.2023.48.e6
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Abstract
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ePub Objectives This study evaluated the effects of high-plasticity mineral trioxide aggregate (MTA-HP) on the activity of M1 and M2 macrophages, compared to white MTA (Angelus).
Materials and Methods Peritoneal inflammatory M1 (from C57BL/6 mice) and M2 (from BALB/c mice) macrophages were cultured in the presence of the tested materials. Cell viability (MTT and trypan blue assays), adhesion, phagocytosis, reactive oxygen species (ROS) production, and tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β production were evaluated. Parametric analysis of variance and the non-parametric Kruskal-Wallis test were used. Results were considered significant when
p < 0.05.Results The MTT assay revealed a significant decrease in M1 metabolism with MTA-HP at 24 hours, and with MTA and MTA-HP later. The trypan blue assay showed significantly fewer live M1 at 48 hours and live M2 at 48 and 72 hours with MTA-HP, compared to MTA. M1 and M2 adherence and phagocytosis showed no significant differences compared to control for both materials. Zymosan A stimulated ROS production by macrophages. In the absence of interferon-γ, TNF-α production by M1 did not significantly differ between groups. For M2, both materials showed higher TNF-α production in the presence of the stimulus, but without significant between-group differences. Likewise, TGF-β production by M1 and M2 macrophages was not significantly different between the groups.
Conclusions M1 and M2 macrophages presented different viability in response to MTA and MTA-HP at different time points. Introducing a plasticizer into the MTA vehicle did not interfere with the activity of M1 and M2 macrophages.
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Original Article
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IL-1 and TNF-α release in human polymorphonuclear leukocytes after exposure to calcium hydroxide treated
Porphyromonas endodontalis lipopolysaccharide - Chan-Je Park, Dong-Sung Park, Hyeon-Mee Yoo, Tae-Seok Oh, Sung-Sam Lim
- J Korean Acad Conserv Dent 2002;27(5):463-472. Published online September 30, 2002
- DOI: https://doi.org/10.5395/JKACD.2002.27.5.463
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Abstract
PDFPubReaderePub
Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-α from immune cells. Although monocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-α. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)2 may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-α, and it was compared with Escherichia coli LPS.
P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 µg/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)2 at 37℃ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4×106 cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10µg/ml) for 24 hours at 37℃ in 5% CO2 incubator. The supernatants of cells were collected and the levels of IL-1α, IL-1β and TNF-α were measured by enzyme-linked immunosorbent assay.
The results were as follows;
1. The levels of IL-1α, IL-1β, TNF-α from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05).
2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05).
3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05).
4. The levels of all three cytokines released from PMN stimulated with P. endodontalis LPS were significantly lower than those released from PMN stimulated with E. coli LPS (p<0.05).
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