Fig. 3: G3@SeHANs block periodontitis-related TLR9 proinflammatory response in vitro.

a, b Activation of HEK-TLR9 reporter cells by healthy human saliva and periodontitis patient saliva, and activation of HEK-TLR9 reporter cells by healthy human serum and periodontitis patient serum. Data are means ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by assessed by two-tailed Student’s t-test (n = 4 or 5) (a, P = 0.0077; b, P = 0.0002). c, d Activation of HEK-TLR9 reporter cells by periodontitis patient saliva and periodontitis patient serum in the absence or presence of PAMAM-G3 (2 μg/mL) or G3@SeHANs (10 μg/mL) for 24 h. Data are means ± SEM; n = 3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by one-way ANOVA with Tukey’s multiple comparison test (c, P < 0.0001 between Control and Untreated, P = 0.0013 between Untreated and PAMAM-G3, P = 0.0003 between Untreated and G3@SeHANs; d, P < 0.0001 between Control and Untreated, P = 0.0111 between Untreated and PAMAM-G3, P = 0.0333 between Untreated and G3@SeHANs). e Activation of HEK-TLR9 reporter cells by DAMPs from gingival keratinocytes, gingival fibroblasts, mitochondrion, and MAMPs from Porphyromonas gingivalis, or Fusobacterium nucleatum in the absence or presence of PAMAM-G3 (2 μg/mL) or G3@SeHANs (10 μg/mL) for 24 h. Data are means ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by one-way ANOVA with Tukey’s multiple comparison test (n = 4 for mitochondrion DMAPs, n = 3 for the other DAMPs and MAMPs) (Gingival keratinocytes, P < 0.0001 between Control and Untreated, P = 0.0004 between Untreated and PAMAM-G3, P = 0.0003 between Untreated and G3@SeHANs; gingival fibroblasts, P < 0.0001 between Control and Untreated, P = 0.0013 between Untreated and PAMAM-G3, P = 0.0150 between Untreated and G3@SeHANs; mitochondrion, P = 0.0153 between Control and Untreated, P = 0.0031 between Untreated and PAMAM-G3, P = 0.0416 between Untreated and G3@SeHANs; Porphyromonas gingivalis, P < 0.0001 between Control and Untreated, P < 0.0001 between Untreated and PAMAM-G3, P < 0.0001 between Untreated and G3@SeHANs; Fusobacterium nucleatum, P < 0.0001 between Control and Untreated, P = 0.0363 between Untreated and G3@SeHANs). f Activation of HEK-TLR9 reporter cells by cfDNA from periodontitis patient saliva or serum, genomic DNA, mtDNA, and CpG DNA in the absence or presence of PAMAM-G3 (2 μg/mL) or G3@SeHANs (10 μg/mL) for 24 h. Data are means ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by one-way ANOVA with Tukey’s multiple comparison test (n = 4 for mitochondrial DNA, n = 3 for the other DNA) (cfDNA from saliva, P = 0.0158 between Control and Untreated, P = 0.0015 between Untreated and G3@SeHANs; mitochondrial DNA, P = 0.0360 between Control and Untreated, P = 0.0073 between Untreated and PAMAM-G3, P = 0.0123 between Untreated and G3@SeHANs; CpG DNA, P < 0.0001 between Control and Untreated, P < 0.0001 between Untreated and PAMAM-G3, P < 0.0001 between Untreated and G3@SeHANs).