Fig. 4: Increased host epithelial inflammatory response to prefrailty-associated human biofilms.

After coculture with a normalized amount of biofilm-dispersed bacteria, epithelial cells were collected for RNA extraction. A The heatmap represents scaled relative expression of a targeted list of host response genes. Specifically, these genes are involved in innate inflammatory signaling (IL6, TNF, IL8, IL10, COX2, and BD2), secretion (BD2 and MUC2), and tight junction function (OCLN and CLDN2). The color gradient from blue to red corresponds to the scale of relative mRNA expression. B Scatter plot depicts relative mRNA expression of IL6, TNF, CLDN2 and OCLN for each individual’s and were grouped for each three cohort of individuals. The gender of each donor is indicated using open (male)/filled (female) circles. Dotted line at 1 corresponds to basal mRNA expression in wells without bacteria. Statistical significance was determined by ANOVA followed by Tukey’s for multiple comparisons, where P < 0.05 was considered significant. C Gene expression dataset was ordinated using Partial least squares-discriminant analysis (PLS-DA) with its corresponding loading vectors. Pairwise PERMANOVA revealed statistical differences between the group of 30–70 years and +70 years prefrail (P = 0.007). D The heatmap corresponds to Pearson correlation coefficients between host epithelial gene response and biofilm features. The color gradient from blue to red signifies negative to positive correlations, and * corresponds to Pearson P value < 0.05.