IADR Abstract Archives
Novel Stannous Toothpaste Detoxifies Inflammatory Markers Induced by Bacterial Endotoxins
Objectives: Determine efficacy of stannous fluoride toothpastes in deactivating NF-κB and in reducing the inflammatory markers activated by the P. gingivitis endotoxin LPS.
Methods:
Supernatant of toothpaste was prepared by mixing 1:4 toothpaste in sterile water and centrifuged for 10 minutes at 4,000g. The supernatant was collected as toothpaste test material and further diluted in the tissue culture media before treating the cells.
HEK-blue cells (Invivogen, hkb-htlr4) were co-incubated with the toothpaste supernatant overnight at 37oC with 5% CO2 in the presence of 1ug/mL ultrapure P. gingivitis LPS (Invivogen, tlrl-ppglps).
NF-κB analysis was performed using HEK-blue detection media per Invivogen’s protocol (Invivogen, hb-det). The quantification of the IL-8 analysis was performed on the cell supernatants using the Enzo IL-8 ELISA kit (Enzo, ADI-900-156). The cell viability analysis was performed using PrestoBlue cell viability reagent per the protocol (Invitrogen, A13262). The concentration of the IL-8 was normalized with the cells treated with the culture medium only. A T test was conducted to calculate the P values with 95% of confidence.
Results: The novel stannous fluoride toothpaste stabilized with nitrate and phosphate is significantly better at reducing ultrapure P. gingivalis LPS-induced NF-κB activation than stannous fluoride toothpastes stabilized with gluconate (p<0.05). Meanwhile the stabilized stannous fluoride toothpaste stabilized with nitrate and phosphate exhibits significantly stronger potency in the reduction of inflammatory markers IL-8 induced by the P. gingivalis LPS compared to two commercial gluconate stabilized stannous fluoride toothpastes (p<0.05).
Conclusions: The novel stannous toothpaste stabilized by nitrate and phosphate exhibit stronger in vitro efficacy in deactivating NF-kB and inhibiting inflammatory markers than two gluconate stabilized stannous fluoride toothpastes.
Methods:
Supernatant of toothpaste was prepared by mixing 1:4 toothpaste in sterile water and centrifuged for 10 minutes at 4,000g. The supernatant was collected as toothpaste test material and further diluted in the tissue culture media before treating the cells.
HEK-blue cells (Invivogen, hkb-htlr4) were co-incubated with the toothpaste supernatant overnight at 37oC with 5% CO2 in the presence of 1ug/mL ultrapure P. gingivitis LPS (Invivogen, tlrl-ppglps).
NF-κB analysis was performed using HEK-blue detection media per Invivogen’s protocol (Invivogen, hb-det). The quantification of the IL-8 analysis was performed on the cell supernatants using the Enzo IL-8 ELISA kit (Enzo, ADI-900-156). The cell viability analysis was performed using PrestoBlue cell viability reagent per the protocol (Invitrogen, A13262). The concentration of the IL-8 was normalized with the cells treated with the culture medium only. A T test was conducted to calculate the P values with 95% of confidence.
Results: The novel stannous fluoride toothpaste stabilized with nitrate and phosphate is significantly better at reducing ultrapure P. gingivalis LPS-induced NF-κB activation than stannous fluoride toothpastes stabilized with gluconate (p<0.05). Meanwhile the stabilized stannous fluoride toothpaste stabilized with nitrate and phosphate exhibits significantly stronger potency in the reduction of inflammatory markers IL-8 induced by the P. gingivalis LPS compared to two commercial gluconate stabilized stannous fluoride toothpastes (p<0.05).
Conclusions: The novel stannous toothpaste stabilized by nitrate and phosphate exhibit stronger in vitro efficacy in deactivating NF-kB and inhibiting inflammatory markers than two gluconate stabilized stannous fluoride toothpastes.