Figure 4
CHD-FA modulates key inflammatory mediators in vitro . An oral epithelial cell line (OKF6) was grown to 90% confluence in 24 well plates for assessing the effect of CHD-FA on the host immune response. Cells were pre-treated with 0.5% (v/v) CHD-FA pH 7.0 for 30 min, washed with PBS and stimulated with the multi-species periodontal biofilm for 4 and 24 h. Untreated controls were also included. Samples were assayed in triplicate and on three separate occasions. RNA was extracted from the 4 h cell lysates, cDNA synthesised and used in the RT2 Profiler to analyse the expression of a panel of pro-inflammatory mediators (A). Duplicate samples from two independent experiments were used in the RT2 Profiler. Data is represented by mean + 95% CI, relative to untreated control. Samples were also assayed for IL-8 gene expression using SYBR® GreenER™ based qPCR (B). Expression of IL-8 is represented relative to housekeeping gene; GAPDH. Retained supernatants were also used to measure IL-8 protein release by ELISA (C). All samples were assayed in triplicate, on three independent occasions. Data represents mean ± SD (*p < 0.05, **p < 0.01).