Figure 4
CHX and RSV immunomodulate epithelial cell responses to multi-species biofilm co-culture in vitro. The oral epithelial cell line OKF6-TERT2 was seeded at 1×105 cells/ml in 24 well plates for co-culture with multi-species biofilms. Biofilm were pre treated with 0.2% (v/v) CHX or cells were treated with 0.01% (w/v) RSV for 30 minutes before washing with PBS and then co-cultured with untreated biofilms or cells respectively for 4 and 24 h. Untreated control co-culture were also included. RNA was extracted from the cell lysates at 4 h, cDNA was synthesized and pro-inflammatory gene expression quantified on an RT2 profiler plate (A). Duplicate samples from three independent experiments were used and data are mean ± SD relative to media control. Samples were also assayed for IL-8 relative to housekeeping gene GAPDH at 4 and 24 h using using SYBR® GreenER™ based qPCR (B). Supernatants were also analyzed for exogenous IL-8, measured by ELISA (C). Samples were assayed in triplicate from three independent experiments, data are mean ± SD (**p < 0.01, ***p < 0.001).