Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the gene-expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with heat-killed <i>P. gingivalis</i>.

<p>Cells were stimulated with heat-killed <i>P. gingivalis</i> (hk <i>Pg</i>, 10⁸ cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). ^# means significantly different from control group (2^−δδCt = 1). * means significantly different from cells stimulated with heat-killed <i>P. gingivalis</i> only.</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the production of pro-inflammatory mediators by hPdLF in response to stimulation with heat-killed <i>P. gingivalis</i>.

<p>Cells were stimulated with heat-killed <i>P. gingivalis</i> (hk <i>Pg</i>, 10⁸ cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. ^# means significantly different from control group (non-stimulated cells). * means significantly different from group stimulated with heat-killed <i>P.gingivalis</i> only</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the gene-expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with <i>P. gingivalis</i> LPS.

<p>Cells were stimulated with <i>P. gingivalis</i> LPS (<i>Pg</i> LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). ^# means significantly different from control group (2^−▵▵Ct = 1). * means significantly different from cells stimulated with <i>P. gingivalis</i> LPS only.</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the gene-expression levels of pro-inflammatory mediators in hPdLF with silenced VDR in response to stimulation with <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i>.

<p>Gene expression levels of IL-6, IL-8, and MCP-1 were measured using q-PCR in hPdLF after transfection with either VDR siRNA or control siRNA and stimulation with <i>P. gingivalis</i> LPS (A, <i>Pg</i> LPS, 1 µg/ml) or heat-killed <i>P. gingivalis</i> (B, hk <i>Pg</i>, 10⁸ cells/ml) in the presence or in the absence of 25(OH)D₃ or 1,25(OH)₂D₃. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells. ^# means significantly different from control group (2^−▵▵Ct = 1). * means significantly different from cells stimulated with heat-killed <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i> only.</p

Gene expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with <i>E. coli</i> LPS, <i>P. gingivalis</i> LPS, and heat-killed <i>P. gingivalis</i>.

<p>Cells were stimulated with <i>E. coli</i> LPS (1 µg/ml), <i>P. gingivalis</i> LPS (0.1–1 µg/ml), or heat-killed <i>P. gingivalis</i> (10⁷–10⁸ cells/ml) for 24 h. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control).</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the production of pro-inflammatory mediators by hPdLF in response to stimulation with <i>P. gingivalis</i> LPS.

<p>Cells were stimulated with <i>P. gingivalis</i> LPS (<i>Pg</i> LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. ^# means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with <i>P.gingivalis</i> LPS only</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the production of pro-inflammatory mediators by hPdLF with silenced VDR.

<p>Cells were transfected with either VDR siRNA or control siRNA and stimulated with <i>P. gingivalis</i> LPS (<i>Pg</i> LPS, 1 µg/ml) or heat-killed <i>P. gingivalis</i> (hk <i>Pg</i>, 10⁸ cells/ml) for 24 h in the presence or in the absence of 25(OH)D₃ or 1,25(OH)₂D₃. ^# means significantly different from control group (2^−▵▵Ct = 1). * means significantly different from cells stimulated with either <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i> only</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the production of pro-inflammatory mediators by primary hPdLC in response to stimulation with <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i>.

<p>Cells were stimulated with <i>P. gingivalis</i> LPS (<i>Pg</i> LPS, 1 µg/ml) or heat-killed <i>P. gingivalis</i> (hk <i>Pg</i>, 10⁸ cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. ^# means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with <i>P.gingivalis</i> LPS or heat-killed <i>P. gingivalis</i> only.</p

Effect of 25(OH)D₃ and 1,25(OH)₂D₃ on the gene-expression levels of pro-inflammatory mediators in primary hPdLC in response to stimulation with <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i>.

<p>Cells were stimulated with <i>P. gingivalis</i> LPS (<i>Pg</i> LPS, 1 µg/ml) or heat-killed <i>P. gingivalis</i> (hk <i>Pg</i>, 10⁸ cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D₃ or 1,25(OH)₂D₃. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). Data are presented as mean±SEM of six different donors. ^# means significantly different from control group (2^−▵▵Ct = 1). * means significantly different from cells stimulated with <i>P. gingivalis</i> LPS or heat-killed <i>P. gingivalis</i> only.</p

Effect of nicotine and <i>P. gingivalis</i> LPS on the proportion of viable HUVECs.

<p>HUVECs were stimulated with nicotine (10 µM-10 mM) and/or <i>P. gingivalis</i> LPS, and the proportion of viable cells was measured using a flow cytometry apoptosis assay after 4 (A), 24 (B), and 72 (C) h. Viable cells were those negative for annexin V and propidium iodide. Data are presented as mean ±SD of three independent experiments. * – significantly lower compared to control group, p<0.05.</p