Suppression of TLR2 inhibits T. denticola-stimulated upregulation of MMPs 2, 14, 17 and 28 while exacerbating MMP 11 expression in hPDL cells.
A) RT-qPCR validation of stable gene suppression using shRNA vectors targeted against TLR2 in healthy hPDL cells. Cells transduced with scrambled shRNA vectors were used as a control. Statistical significance was determined using an unpaired t-test. Bars represent ± SEM of mean value (n = 3 clones). *p < .05 versus control. B-F) RT-qPCR for MMP-2, MMP-11, MMP-14, MMP-17 and MMP-28 mRNA expression of scrambled shRNA control and TLR2 shRNA hPDL cells challenged or stimulated with Td-CF522, purified dentilisin or Td-WT at an MOI of 50 and concentration of 1 μg/mL for 2-hours in alpha-MEM media with no supplementation followed by a 22-hour incubation in alpha-MEM media supplemented with 10% FBS, 1% Pen/Strep and 1% Amphotericin B. The expression of each gene was normalized to that of GAPDH. Statistical significance was determined using a Two-Way ANOVA followed by post-hoc Tukey’s multiple comparisons. Bars represent mean ± SEM (n = 3). #p < .05 versus scramble control group. ##p < .01 versus scramble control group. ###p < .001 versus scramble control group. *p < .05 versus TLR2 shRNA equivalent group. ***p < .001 versus TLR2 equivalent shRNA group.