<i>miR-200c</i> delivered using PEI nanoparticles inhibits IL-6, IL-8, and CCL-5 in primary human periodontal ligament fibroblasts.

<p><b>A-C</b>: The transcripts of IL-6 (<b>A</b>), IL-8 (<b>B</b>), and CCL-5 (<b>C</b>) in the cells with <i>miR-200c</i> or empty vector cultured in DMEM supplemented with LPS after 24 hours; <b>D</b> and <b>E</b>: the amounts of IL-6 (<b>D</b>), IL-8 (<b>E</b>), and CCL-5 (<b>F</b>) secreted by the cells with miR-200c or empty vector cultured in DMEM supplemented with LPS after 12 and 32 hrs, respectively. *: p<0.05 vs empty vector with the same amount.</p

<i>miR-200c</i> modulates proinflammatory mediators in human preosteoblasts.

<p><b>A</b> and <b>B:</b> the transcripts of IL-6 <b>(A)</b> and IL-8 (<b>B</b>) in non-treated HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented with LPS at 0, 1, 5 and 10 μg/mL after 24 hours; *:p<0.05 vs non-treated; <b>C:</b> the amounts of IL-8 secreted by HEPM cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented with or without LPS at different time points; *: p<0.05 vs cells with scrambled miRs; <b>D</b> and <b>E:</b> the amounts of IL-6 (<b>D</b>) and CCL-5 (<b>E</b>) secreted by HEPM cells with <i>miR-200c</i> or <i>scrambled miRs</i> cultured in DMEM supplemented with or without LPS after 24 hrs; <b>F:</b> the amounts of OPG secreted by HEPM cells with different <i>miRs</i> cultured in DMEM supplemented with or without LPS after 32 hours. *: p<0.05.</p

<i>miR-200c</i> increases osteogenic biomarkers in human preosteoblasts.

<p><b>A</b> and <b>B</b>: the amounts of the transcript of OCN (<b>A</b>) and calcium content (<b>B</b>) in non-treated HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented <i>β</i>-glycerophosphate and ascorbic acid after 1 and 2 weeks, respectively. *: p<0.05.</p

Enhancement of osteogenic differentiation of human bone marrow MSCs with overexpression of <i>miR-200c</i> using PEI nanoparticles.

<p><b>A</b>: Images of ALP and von-Kossa staining in MSCs overexpressing <i>miR-200c</i>, one and two weeks after treatment with osteogenic medium. <b>B</b> and <b>C</b>: the transcripts of ALP (<b>B</b>) and Runx2 (<b>C</b>) in MSCs overexpressing <i>miR-200c</i>, one week after treatment with osteogenic medium. <b>D</b> and <b>E</b>: Quantitative measurement of ALP levels (<b>D</b>) and calcium content (<b>E</b>) in MSCs overexpressing <i>miR-200c</i>, one and two week after treatment with osteogenic medium. Each measurement was made in triplicate. *: p<0.05.</p

<i>miR-200c</i> overexpression in HEPM cells and the effects on their proliferation.

<p><b>A</b>: Microphotographs of HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> under phase-contrast. Bar = 10μm. <b>B</b>: Fold change of <i>miR-200c</i> expression in non-treated HEPM cells and the cells with <i>miR-200c</i> and scrambled <i>miRs</i>. <b>C</b>: The doubling time of non-treated HEPM cells and the cells in the context of <i>miR</i> infection. **: p<0.01.</p

Intracellular delivery of <i>miR-200c</i> using PEI nanoparticles to human primary periodontal ligament fibroblasts and bone marrow MSCs.

<p><b>A</b>: TEM image of PEI-<i>miR-200c</i> nanoplexes. <b>B</b> and <b>C:</b> Fold change of the transcript of <i>miR-200c</i> in non-treated human periodontal ligament fibroblasts (<b>B</b>) and bone marrow MSCs (<b>C</b>) and the cells transfected with empty vector (EV) (10μg/per well) and <i>miR-200c</i> (1, 5, 10μg/per well).</p

<i>PMIS-200c</i> reduces binding activity of <i>miR-200c</i> to the <i>3’UTR</i> of IL-6, IL-8, and CCL-5 and the function of <i>miR-200c</i>.

<p><b>A-C</b>: Normalized luciferase activities of the <i>3’ UTR</i> IL-6, IL-8, and CCL-5-luciferase reporters and their <i>3’UTR</i>-mutated-luciferase reporters co-treated with <i>miR-200c</i> and <i>PMIS-EV</i> or PMIS-200c at different ratios of concentration. <b>D-F</b>: the transcripts of IL-6 (<b>D</b>), IL-8 (<b>E</b>), and CCL-5 (<b>F</b>) in the cells co-treated with <i>miR-200c</i> and <i>PMIS-EV</i> or <i>PMIS-200c</i> cultured in DMEM supplemented with LPS after 24 hours; *: p<0.05.</p