Real-time quantitative PCR analysis of human gingiva epithelial cells exposed to microbial biofilms

An organotypic gingiva model RHG was constructed by immortalized human gingiva cell lines (keratinocytes and fibroblasts) in vitro. The RHG model was exposed to either control medium or one of the three multi-species oral biofilms (commensal, gingivitis and cariogenic) for 24 hours. We used Qiagen RT² Profiler PCR array to analyse the expression of genes which are related to Toll-like receptor signaling pathway from the unexposed (control) and differently exposed (test) keratinocyte layer of the RHG model. Reference for RHG model and oral biofilms culture: J. K. Buskermolen et al., J Dent Res 97 (2018): 201-208. doi: 10.1177/0022034517729998. qPCR gene expression profiling. For each exposure condition, three (commensal, gingivitis) or two (cariogenic) replicates of RHG samples were used. Equal amount total RNA from each sample was processed to gene expression analysis.