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

  • Celso Emanuel de Souza Queiroz Disciplina de Endodontia - UNESP - Araraquara
  • Fernanda Geraldes Pappen Universidade Estadual Paulista
  • Rafael Comelli Lia Faculdade de Odontologia de Araraquara - UNESP
  • Fernando Perosa Zanin Faculdade de Odontologia de Araraquara - UNESP
  • Renato de Toledo Leonardo Universidade Estadual Paulista
  • Iracilda Zeponi Carlos Faculdade de Ciencias Farmacêuticas - UNESP - Araraquara
Keywords: Peritoneal macrophages, nitric oxide, lipopolysaccharides, tumor necrosis factoralpha

Abstract

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

Author Biographies

Celso Emanuel de Souza Queiroz, Disciplina de Endodontia - UNESP - Araraquara
Doutor em Endodontia - Disciplina de Endodontia - UNESP - Araraquara
Fernanda Geraldes Pappen, Universidade Estadual Paulista
Aluno do curso de Doutorado em Endodontia - UNESP - Araraquara Disciplina de Endodontia Professora de Endodontia da Faculdade de Odontologia UFPel - Pelotas - RS
Rafael Comelli Lia, Faculdade de Odontologia de Araraquara - UNESP
Doutor em Patologia - Faculdade de Odontologia de Araraquara - UNESP
Fernando Perosa Zanin, Faculdade de Odontologia de Araraquara - UNESP
Mestre em Endodontia - Faculdade de Odontologia de Araraquara - UNESP
Renato de Toledo Leonardo, Universidade Estadual Paulista
Prof. da Disciplina de Endodontia - UNESP - Araraquara
Iracilda Zeponi Carlos, Faculdade de Ciencias Farmacêuticas - UNESP - Araraquara
Professora de Imunologia da Faculdade de Ciencias Farmacêuticas - UNESP - Araraquara
Published
2008-07-10
Section
Original Article