Table 1.
The Human Inflammatory Response & Autoimmunity RT² Profiler PCR Array.
This assay profiles 84 key genes involved in autoimmune and inflammatory immune responses. It profiles genes related to inflammatory cytokines and chemokines as well as their receptors and also genes related to the metabolism of cytokines and those involved in cytokine-cytokine receptor interactions.
Table 2.
Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).
MIC and MBC for the ethanolic extract of Malva sylvestris and its chloroform and aqueous fraction against four different periodontopathogens: Aggregatibacter actinomycetemcomitans D7S1, Fusobacterium nucleatum ATCC 25586, Prevotella intermedia ATCC 25611 and Porphyromonas gingivalis ATCC BAA-308. The highest concentration evaluated was 1000 μg/mL and the minus symbol (-) means no inhibitory activity.
Fig 1.
Cytotoxic effect of MSE, CLF and AF on fibroblasts HGF-1 cells. Data are expressed as mean ± SEM using one-way analysis of variance (ANOVA) followed by Dunnet’s multiple comparison tests as compared to non-treated. The level of statistical significance was set at 0.05.
Fig 2.
Cytotoxicity effect in the dual chamber model.
Cytotoxic effects of MSE, chloroform and aqueous fraction on fibroblast HGF-1 and keratinocyte OBA-9 cell lines were found after 24 h hours of invasion by A. actinomycetemcomitans. Data are expressed as mean ± SEM using one-way analysis of variance (ANOVA) followed by Dunnet’s multiple comparison tests as compared to the control group (non-treated). The level of statistical significance was set at 0.05.
Fig 3.
Comparison of colony-forming units.
Comparison of colony-forming units (CFU/mL) among groups treated with MSE, CLF and AF after A. actinomycetemcomitans infection. Data are expressed as mean ± SEM using one-way analysis of variance (ANOVA) followed by Dunnet’s multiple comparison tests as compared to vehicle control. The level of statistical significance was set at 0.05.
Table 3.
The Human Inflammatory Response & Autoimmunity RT² Profiler PCR Array.
Genes in the inflammatory pathway down-regulated by the treatments with MSE, CLF and AF.
Fig 4.
Gene expression analysis of lower chamber co-culture invasion assay.
The expression levels of IL-1alfa, IL-1beta, IL-6, IL-8, IL-19, CD14, PTGS, MMP-1, FOS, BCL2 were evaluated and compared to the control (non-infected cells). Quantification of the relative transcript amounts was performed using qPCR with 50 ng of each cDNA. Data quantification was performed using the 2ΔΔCT method. Statistical analyses included one-way ANOVA followed by Dunnet’s post-hoc tests. A significance level of p < 0.05 (*) indicates differences from the vehicle group.
Fig 5.
Quantification of IL-1alpha, IL-1beta, IL-6, IL-8, IL-10 and GM-CSF in the co-culture supernant after 24h of A. actinomycetemcomitans invasion. Cells were treated with 75μg/mL of MSE and fractions and bacteria inocula were established at 2x106 CFU/mL. Data are expressed as mean±SD, n = 6. Symbols indicate statistical differences (p<0.05, Dunnet’s test). # indicates p<0.05 compared to non-treated group; * indicates p<0.05 compared to vehicle group.