Induction of periodontitis and mRNA levels of pro-inflammatory cytokines in gingival tissues. (A) Representative photographs of the maxilla of treated and untreated mice. Scale bar = 1 mm. (B) Schematics of periodontal bone loss measurements. The distances between the cement–enamel junction and the alveolar bone crest (CEJ-ABC) at the distal facial side of the first molar, at the mesial and distal facial sides of the second molar, and at the mesial facial side of the third molar were measured (Mmd2+/+, n = 19; Mmd2V100L/V100L, n = 8; Mmd2A117V/+, n = 8; Mmd2A117V/A117V, n = 8; Mmd2R127P/+, n = 8; Mmd2R127P/R127P, n = 8; Mmd2−/−, n = 20). Measurements of the distances of CEJ-ABC were performed by personnel blinded to sample information. Mmd2+/+ versus Mmd2V100L/V100L; P = 0.9926, Mmd2+/+ versus Mmd2A117V/+; P < 0.0001, Mmd2+/+ versus Mmd2A117V/A117V; P < 0.0001, Mmd2+/+ versus Mmd2R127P/+; P < 0.0001, Mmd2+/+ versus Mmd2R127P/R127P; P < 0.0001, Mmd2+/+ versus Mmd2−/−; P < 0.0001. (C) qPCR analysis of inflammatory genes in the gingival tissues at 24 h after ligation. 18S was used for normalization. (Mmd2+/+, n = 5; Mmd2A117V/+, n = 5; Mmd2A117V/A117V, n = 5; Mmd2R127P/+, n = 5; Mmd2R127P/R127P, n = 5; Mmd2−/−, n = 5). Il6 (Mmd2+/+ versus Mmd2A117V/+; P = 0.0037, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0080, Mmd2+/+ versus Mmd2R127P/+; P = 0.0001, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0080, Mmd2+/+ versus Mmd2−/−; P = 0.0469). Il1b (Mmd2+/+ versus Mmd2A117V/+; P = 0.0142, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0297, Mmd2+/+ versus Mmd2R127P/+; P = 0.0465, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0003, Mmd2+/+ versus Mmd2−/−; P = 0.0370). Ccl3 (Mmd2+/+ versus Mmd2A117V/+; P = 0.0059, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0315, Mmd2+/+ versus Mmd2R127P/+; P = 0.0506, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0003, Mmd2+/+ versus Mmd2−/−; P = 0.0881). (B and C) Data are presented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 with one-way ANOVA with Tukey–Kramer test (B and C) or two-tailed unpaired t test (C, Mmd2+/+ versus Mmd2−/−). ns = not significant. Each dot represents a biologically independent mouse.
Figure 3.

Induction of periodontitis and mRNA levels of pro-inflammatory cytokines in gingival tissues. (A) Representative photographs of the maxilla of treated and untreated mice. Scale bar = 1 mm. (B) Schematics of periodontal bone loss measurements. The distances between the cement–enamel junction and the alveolar bone crest (CEJ-ABC) at the distal facial side of the first molar, at the mesial and distal facial sides of the second molar, and at the mesial facial side of the third molar were measured (Mmd2+/+, n = 19; Mmd2V100L/V100L, n = 8; Mmd2A117V/+, n = 8; Mmd2A117V/A117V, n = 8; Mmd2R127P/+, n = 8; Mmd2R127P/R127P, n = 8; Mmd2−/−, n = 20). Measurements of the distances of CEJ-ABC were performed by personnel blinded to sample information. Mmd2+/+ versus Mmd2V100L/V100L; P = 0.9926, Mmd2+/+ versus Mmd2A117V/+; P < 0.0001, Mmd2+/+ versus Mmd2A117V/A117V; P < 0.0001, Mmd2+/+ versus Mmd2R127P/+; P < 0.0001, Mmd2+/+ versus Mmd2R127P/R127P; P < 0.0001, Mmd2+/+ versus Mmd2−/−; P < 0.0001. (C) qPCR analysis of inflammatory genes in the gingival tissues at 24 h after ligation. 18S was used for normalization. (Mmd2+/+, n = 5; Mmd2A117V/+, n = 5; Mmd2A117V/A117V, n = 5; Mmd2R127P/+, n = 5; Mmd2R127P/R127P, n = 5; Mmd2−/−, n = 5). Il6 (Mmd2+/+ versus Mmd2A117V/+; P = 0.0037, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0080, Mmd2+/+ versus Mmd2R127P/+; P = 0.0001, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0080, Mmd2+/+ versus Mmd2−/−; P = 0.0469). Il1b (Mmd2+/+ versus Mmd2A117V/+; P = 0.0142, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0297, Mmd2+/+ versus Mmd2R127P/+; P = 0.0465, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0003, Mmd2+/+ versus Mmd2−/−; P = 0.0370). Ccl3 (Mmd2+/+ versus Mmd2A117V/+; P = 0.0059, Mmd2+/+ versus Mmd2A117V/A117V; P = 0.0315, Mmd2+/+ versus Mmd2R127P/+; P = 0.0506, Mmd2+/+ versus Mmd2R127P/R127P; P = 0.0003, Mmd2+/+ versus Mmd2−/−; P = 0.0881). (B and C) Data are presented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 with one-way ANOVA with Tukey–Kramer test (B and C) or two-tailed unpaired t test (C, Mmd2+/+ versus Mmd2−/−). ns = not significant. Each dot represents a biologically independent mouse.

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