NLRP12 negatively modulates inducible nitric oxide synthase (iNOS) expression and tumor necrosis factor-α production in Porphyromonas gingivalis LPS-treated mouse macrophage cell line (RAW264.7).

OBJECTIVE: The aim of the present study is to investigate the participation of NLRP12 in Porphyromonas gingivalis LPS-activated mouse macrophages. METHODS: NLRP12-depleted mouse macrophages were stimulated with P. gingivalis LPS (1 µg/ml.). At indicated time points, the treated cells were lysed and the supernatant from treated cells was collected. Gene and protein expression of NLRP12 and iNOS were determined by RT-PCR and immunoblotting, respectively. The level of TNF-α production in the supernatant of the activated cells was determined by ELISA. RESULTS AND CONCLUSION: NLRP12 was upregulated in response to stimulation with P. gingivalis LPS. In addition, when NLRP12 was depleted in P. gingivalis LPS-treated macrophages, an increase in TNF-α production and iNOS expression were observed when compared to those of the control cells, indicating that NLRP12 downregulates the inflammatory cytokine and antimicrobial molecule production in the macrophages.

Induction of inducible nitric oxide synthase (iNOS) in Porphyromonas gingivalis LPS-treated mouse macrophage cell line (RAW264.7) requires Toll-like receptor 9.

OBJECTIVE: The aim of this study is to investigate the involvement of TLR9 in the regulation of iNOS expression and nitric oxide (NO) production in Porphyromonas gingivalis LPS-treated mouse macrophages. METHODS: Mouse macrophage cell line (RAW264.7) was transfected with siRNAs against TLR9 and then stimulated with P. gingivalis LPS. At indicated time points, the activated cells were lysed. Gene and protein expression of iNOS were determined by RT-PCR and immunoblotting, respectively. The level of nitric oxide (NO) production in the supernatant of the activated cells was determined by Griess reaction assay. RESULTS AND CONCLUSION: Depletion of TLR9 in mouse macrophages demonstrated the markedly decreased iNOS gene and protein expression by P. gingivalis LPS compared to those of the wild-type or control siRNA transfected cells. In consistent with these results, the level of NO secretion was also significantly diminished in TLR9-depleted cells after challenged with P. gingivalis LPS. These results indicate that TLR9 involves in the regulation of the iNOS expression and the NO secretion in P. gingivalis LPS-treated macrophages.

Efficacy of antiseptics for rubber dam sterilization prior to endodontic treatment.

PURPOSE: This study investigated the efficacy of various antiseptics for disinfection of rubber dams used during endodontic treatment, the duration of disinfection effectiveness, and the disinfection protocol employed by dental schools in Thailand. METHODS: The efficacy of 10% povidone-iodine, 1.5% tincture iodine and 70% ethyl alcohol in eliminating Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) on the rubber dam was investigated. Time duration of disinfection was evaluated at 0, 30, 60, and 120 min. The two-step disinfection method adopted at Thai dental school was examined. Independent t-test or Kruskal-Wallis followed by a Dunnett's test was used for statistical analysis. RESULTS: Among the three antiseptics, 10% povidone and 1.5% tincture iodine eradicated the microorganisms completely, whereas 70% ethyl alcohol did not achieve a statistically significant decrease. The duration of sterilization effectiveness was 120 min for 10% povidone-iodine, but bacteria were eliminated only at 0 min by 1.5% tincture iodine. The results also indicated that the two-step protocol scarcely eliminated the microorganisms. CONCLUSION: The best antiseptic for rubber dam disinfection is 10% povidone-iodine, which remains effective for 120 min. The two-step protocol typically practiced in Thai dental schools needs to be updated. The use of 10% povidone-iodine alone is adequate for complete eradication of E. faecalis and C. albicans.

Effects of Litsea cubeba Essential Oil Incorporated into Denture Soft Lining Materials.

The antimicrobial properties, cell cytotoxicity and surface hardness of soft lining materials (GC soft liner, Viscogel and Coe comfort) incorporated with various concentrations of Litsea cubeba essential oil (LCEO) were evaluated. The minimum inhibitory concentrations of LCEO against Candida albicans and Streptococcus mutans were 1.25% v/v and 10% v/v, respectively. However, when LCEO was incorporated into the three soft lining materials (GC soft liner, Viscogel and Coe comfort), 10% v/v and 30% v/v of LCEO could inhibit the growth of C. albicans and S. mutans, respectively. The extracts of soft lining materials with 10% and 30% v/v LCEO, 2% chlorhexidine, 30% v/v nystatin and no additive were used for cytotoxicity tests on a human gingival fibroblast cell line. There was no significant difference in cell viability in all groups with additives compared to the no additive group (p > 0.05). Surface hardness increased significantly between 2 h and 7 day incubation times in all groups, including the controls (p < 0.05). A higher LCEO concentration had a dose-dependent effect on the surface hardness of all soft lining materials (p < 0.05). However, the surface hardness of materials with additive remained in accordance with ISO 10139-1. LCEO could be used as a natural product against oral pathogens, without having a negative impact on soft lining materials.