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Ca(OH)2 action on TNF-alpha and NO release in macrophage culture stimulated by Pseudomonas aeruginosa LPS / Ação do Ca(OH)2 sobre a produção de TNF-alfa e NO de cultura de macrófagos estimulada por LPS de Pseudomonas aeruginosa
Queiroz, Celso Emanuel de Souza; Pappen, Fernanda Geraldes; Lia, Rafael Comelli; Zanin, Fernando Perosa; Leonardo, Renato de Toledo; Carlos, Iracilda Zeponi.
  • Queiroz, Celso Emanuel de Souza; UNESP. Faculty of Dentistry of Araraquara. Graduate Program in Endodontics. Araraquara. BR
  • Pappen, Fernanda Geraldes; UNESP. Faculty of Dentistry of Araraquara. Graduate Program in Endodontics. Araraquara. BR
  • Lia, Rafael Comelli; UNESP. Faculty of Dentistry of Araraquara. Graduate Program in Pathology. Araraquara. BR
  • Zanin, Fernando Perosa; UNESP. Faculty of Dentistry of Araraquara. Graduate Program in Endodontics. Araraquara. BR
  • Leonardo, Renato de Toledo; UNESP. Faculty of Dentistry of Araraquara. Discipline of Endodontics. Araraquara. BR
  • Carlos, Iracilda Zeponi; UNESP. Faculty of Pharmaceutical Sciences. Discipline of Immunology. Araraquara. BR
Rev. odonto ciênc ; 23(3): 225-228, jul.-set. 2008. tab
Article in English | LILACS, BBO | ID: lil-494940
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 Factor-alpha (TNF-alpha) release in mice macrophage culture.

Methods:

For the NO assay, peritoneal exudate cells (PECs) were placed in contact with 25ìg/mL and 50ìg/mL LPS and LPS/Ca(OH)2 suspensions (50ìg/25mg and 25ìg/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 25ìg/mL, and the LPS/Ca(OH)2 suspension was tested in 25ìg/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.
RESUMO

Objetivo:

A ação do hidróxido de cálcio [Ca(OH)2] com o sistema imune e o mecanismo de neutralização das bactérias e seus subprodutos ainda não foi completamente esclarecida. Neste estudo foi avaliada a capacidade do Ca(OH)2 em neutralizar o lipopolissacarídeo (LPS) de Pseudomonas aeruginosa, utilizando-se duas metodologias liberação de Óxido Nítrico (NO) e Fator de Necrose Tumoral Alfa (TNF-alfa) em cultura de macrófagos peritoneais de camundongos.

Metodologia:

No ensaio do NO, as células peritoneais foram expostas a uma solução de LPS (25mg/mL e 50mg/mL); e à suspensão de LPS/Ca(OH)2 em duas concentrações (50mg/ 25mg e 25mg/25mg). Após 8 horas de incubação, foi utilizado reagente de Griess, e a liberação de NO foi quantificada. No ensaio do TNF-alfa, a solução de LPS foi usada na concentração de 25mg/mL e o LPS/Ca(OH)2 a 25mg/25mg. Após 24 horas, as células foram fixadas e coradas com cristal violeta, e os valores de absorbância foram obtidos. Os resultados foram expressos em micromols. Todos os testes foram realizados em triplicata.

Resultados:

A presença de Ca(OH)2 nas duas concentrações avaliadas reduziu significativamente a liberação de NO e TNF-alfa.

Conclusão:

Pode-se concluir que o LPS bacteriano representa um forte estímulo para liberação destas citocinas, mas o hidróxido de cálcio foi capaz de neutralizar este efeito.
Subject(s)

Full text: Available Index: LILACS (Americas) Main subject: Pseudomonas aeruginosa / Calcium Hydroxide / Lipopolysaccharides / Tumor Necrosis Factor-alpha / Macrophages, Peritoneal / Nitric Oxide Type of study: Evaluation studies Limits: Animals Language: English Journal: Rev. odonto ciênc Journal subject: Dentistry Year: 2008 Type: Article Affiliation country: Brazil Institution/Affiliation country: UNESP/BR

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Full text: Available Index: LILACS (Americas) Main subject: Pseudomonas aeruginosa / Calcium Hydroxide / Lipopolysaccharides / Tumor Necrosis Factor-alpha / Macrophages, Peritoneal / Nitric Oxide Type of study: Evaluation studies Limits: Animals Language: English Journal: Rev. odonto ciênc Journal subject: Dentistry Year: 2008 Type: Article Affiliation country: Brazil Institution/Affiliation country: UNESP/BR