IADR Abstract Archives
Protective Effect of Immunized Mouse Serum on Pyocyanin-induced Macrophage Cell Death
Objectives: Pseudomonas aeruginosa is an important bacterium causing nosocomial infection. It can be found in dental and oral chronic infections. Pyocyanin (PCN) produced by P. aeruginosa plays an important role in bacterial invasion. The macrophage is responsible for defense mechanism toward chronic infection. This study was aimed to investigate the effect of immunized mouse serum to protect macrophage from cell death induced by PCN.
Methods: BALB/c mice were immunized with 50 µg PCN intraperitoneally. The serum was collected from retro-orbital vein one week after the second booster. The intraperitoneal macrophages (1 x 105 cell/well) were cultured for 24 h prior to the stimulation. The cells were pretreated with various concentration of mouse serum (10, 20, 40, 80, 160, 320, 640 times dilution) for 30 min and underwent treated with 50 µg/mL PCN for 24 h at 37oC in the presence of 5% CO2. Percentages of living cells were assessed after MTT assay. The results were analyzed using one-way ANOVA followed by subsequent analysis using LSD Scheffe test at significance level of 95%.
Results: The ANOVA results showed a significant difference between experimental groups. Those results demonstrated that the treatments were affecting cell viability. The results of LSD test indicated significant differences between control and each treatment group, except with the group treated with 640 times dilution serum and 50 µg/mL PCN. Moreover, no significant differences were found between the group treated with serum at 40 times and 80-time dilution, 80 times and 160-time dilution, 160 times and 320-time dilution, 160 times and 640-time dilution, 320 times and 640-time dilution.
Conclusions: It is suggested that immunized mouse serum enhanced macrophage viability under PCN exposure. The protective effect of serum increased as the increasing serum concentration. The 20-time dilution of serum recommended as the effective concentration to inhibit macrophage cell death.
Methods: BALB/c mice were immunized with 50 µg PCN intraperitoneally. The serum was collected from retro-orbital vein one week after the second booster. The intraperitoneal macrophages (1 x 105 cell/well) were cultured for 24 h prior to the stimulation. The cells were pretreated with various concentration of mouse serum (10, 20, 40, 80, 160, 320, 640 times dilution) for 30 min and underwent treated with 50 µg/mL PCN for 24 h at 37oC in the presence of 5% CO2. Percentages of living cells were assessed after MTT assay. The results were analyzed using one-way ANOVA followed by subsequent analysis using LSD Scheffe test at significance level of 95%.
Results: The ANOVA results showed a significant difference between experimental groups. Those results demonstrated that the treatments were affecting cell viability. The results of LSD test indicated significant differences between control and each treatment group, except with the group treated with 640 times dilution serum and 50 µg/mL PCN. Moreover, no significant differences were found between the group treated with serum at 40 times and 80-time dilution, 80 times and 160-time dilution, 160 times and 320-time dilution, 160 times and 640-time dilution, 320 times and 640-time dilution.
Conclusions: It is suggested that immunized mouse serum enhanced macrophage viability under PCN exposure. The protective effect of serum increased as the increasing serum concentration. The 20-time dilution of serum recommended as the effective concentration to inhibit macrophage cell death.