Ca(OH)2 action on TNF-alpha and NO release in macrophage culture stimulated by Pseudomonas aeruginosa LPS

Celso Emanuel de Souza Queiroz, Fernanda Geraldes Pappen, Rafael Comelli Lia, Fernando Perosa Zanin, Renato de Toledo Leonardo, Iracilda Zeponi Carlos


Purpose: Calcium hydroxide [Ca(OH)2] interaction with the immune system to destroy or neutralize bacteria and their by-products is not completely understood. This study evaluated the calcium hydroxide ability to neutralize Pseudomonas aeruginosa lipopolysaccharides (LPS) using two different methods: nitric oxide (NO) stimulation and Tumoral Necrosis Factoralpha (TNF-alpha) release in mice macrophage culture.
Methods: For the NO assay, peritoneal exudate cells (PECs) were placed in contact with 25μg/mL and 50μg/mL LPS and LPS/Ca(OH)2 suspensions (50μg/25mg and 25μg/25mg). After incubation for 8h, Griess reagent was used, and the NO release was quantified. In the TNF-alpha assay the LPS solution was tested in 25μg/mL, and the LPS/Ca(OH)2 suspension was tested in 25μg/25mg concentration. After incubation for 24 hours, cells were fixed and stained with crystal violet. Absorbance values were obtained. Results were expressed in micromols. All tests were performed in triplicate.
Results: The presence of Ca(OH)2 in both tested concentrations significantly reduced NO and TNF-alpha release.
Conclusion: It can be concluded that bacteria LPS is a strong stimulus for NO and TNF-alpha release, but calcium hydroxide can neutralize it.
Key words: Peritoneal macrophages; nitric oxide; lipopolysaccharides; tumor necrosis factor alpha


Peritoneal macrophages; nitric oxide; lipopolysaccharides; tumor necrosis factoralpha

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e-ISSN: 1980-6523

ISSN-L: 0102-9460


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