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. 2020 Aug 6;5(15):e136706. doi: 10.1172/jci.insight.136706

Figure 4. ERM suppresses ligature-induced inflammatory bone loss in a DEL-1–dependent manner.

Figure 4

(A) Experimental design. Periodontal bone loss was induced in WT or Del1–/– mice for 9 days by ligating a maxillary second molar and leaving the contralateral tooth unligated (baseline control). Groups of mice were given ERM (20 mg/kg), JSM (20 mg/kg), PC (20 mg/kg), or ethanol control i.p. every day until the day before sacrifice (day 8). (B) Measurements of bone loss in the indicated groups of LIP-subjected mice (left panel; n = 10 mice/group) and representative images of maxillae from each group (right panel). (C) Bone loss was measured in littermate WT or Del1–/– mice that were subjected to LIP and treated with ERM (20 mg/kg) or ethanol control as shown in panel A (n = 10 mice/group). (D) Numbers of neutrophils in the gingiva of LIP-subjected WT mice treated with ethanol control, ERM (20 mg/kg), JSM (20 mg/kg), or PC (20 mg/kg) as described above (n = 6 mice/group). (E) Relative mRNA expression of the indicated molecules in the gingival tissue from LIP-subjected WT mice treated with ERM, JSM, PC, or ethanol control as above. Data were normalized to Gapdh mRNA and are presented as fold change relative to baseline (unligated control), which was set as 1 (n = 6 mice/group). (F) Numbers of neutrophils in the gingival tissue of LIP-subjected WT or Del1–/– mice treated with ERM (20 mg/kg) or ethanol control (n = 6 mice/group). (G) Determination of the protein and mRNA levels of IL-17, IL-6, and IL-10 in the gingival tissue of LIP-subjected WT or Del1–/– mice, which were treated (or not; ethanol control) with 20 mg/kg ERM as outlined in panel A. Protein concentrations (pg cytokines/mg total protein in tissue lysates are shown) and mRNA expression were determined by ELISA and qPCR, respectively. The mRNA data were normalized to Gapdh mRNA and are presented as fold change relative to vehicle-treated WT mice, which was set as 1 (n = 6 mice/group). (H) Tissue sections from LIP-subjected WT mice were stained for DEL-1, neutrophil elastase and nuclei using DAPI. Scale bars, 100 μm. Data are presented as the mean ± SD. *P < 0.01, ***P < 0.0001 by 1-way ANOVA followed by Tukey’s multiple comparisons test (B–G).