Table 1.
Primers used for qPCR analyses.
Fig 1.
MicroCT parameters illustrate the effects of B2 kinin receptor deletion on alveolar bone (control side) and OTM.
(A) Representative image of the OTM model. (B) Quantification of OTM with corresponding representative images. OTM was determined by calculating the difference in the shortest linear distance between the CEJ of the first and second maxillary molars on the experimental (right) and control (left) sides of the same animal, using a split-mouth design. Yellow dotted squares denote the analyzed area for microarchitectural parameters (control side), and green lines indicate the distance between the first and second molars, highlighting the extent of tooth movement. (C) ABL area (control side). The ABL was determined by calculating the area between the CEJ and the ABC of the first, second, and third molars on the control side. Red arrows indicate the levels of the ABC. (D) Bone Mineral Density (BMD, g/cm−3). (E) Bone Volume/Total Volume (BV/TV, %). (F) Trabecular Thickness (Tb.Th, μm). (G) Trabecular Number (Tb.N, μm−1). (H) Trabecular Separation (Tb.Sp, μm). ** Significant difference from control (p < 0.01). *** Significant difference from control (p < 0.001). Data are expressed as the mean ± SD. Statistical analysis was performed using Student’s t-test (n = 5 animals per group, totaling 10 animals).
Fig 2.
Bone cell counts in the tooth surrounding bone.
(A) Quantification and corresponding representative images of TRAP-positive osteoclasts counted on the maxillary bone in WT and B2R−/− groups on the control side and under OTM. Black arrows indicate osteoclasts. (B) Quantification and corresponding representative images of the bone lining osteoblasts counted on the maxillary bone of WT and B2R−/− mice on the control and OTM side. Black arrows indicate osteoblasts. AB, alveolar bone; R, root; ObN/BPm, osteoblast number/bone perimeter. Scale bar = 100 μm. * Significant difference from control (p < 0.05). *** Significant difference from control (p < 0.001). Data are expressed as the mean ± SD. Statistical analysis was performed using Student’s t-test (n == 5 animals per group, totaling 10 animals).
Fig 3.
Gene expression of biomarkers in alveolar bone, at control and OTM sides.
(A) RANKL. (B) OPG. (C) RANKL/OPG ratio. (D) RANK. (E) BDKRB1. (F) IL-1β. (G) TNF-α. (H) IL-6. * Significant difference from control (p < 0.05). ** Significant difference from control (p < 0.01). Data are expressed as mean ± SD. Statistical analysis was performed using Student’s t-test (n = 5 animals per group, totaling 10 animals).
Fig 4.
Effects of B2R deletion on osteoclast differentiation and activity in vitro.
(A) Representative image of bone marrow cells differentiated into osteoclasts from WT and B2R−/− mice (scale bar = 200 μm). (B) Representative image of resorption pit areas. (C) Count of TRAP-positive osteoclasts from WT and B2R−/− mice. (D) Quantification of resorption areas by osteoclasts derived from WT and B2R−/− mice. (E) MTT assay to assess the viability of cells after seven days of differentiation. * Significant difference from control (p < 0.05). ** Significant difference from control (p < 0.01). Data are expressed as the mean ± SD. Statistical analysis was performed using Student’s t-test (n = 3 animals per group, totaling six animals).
Fig 5.
B2R deletion does not impact the differentiation of osteoblasts in vitro.
(A) Representative image of osteoblast bone marrow cell differentiation of WT and B2R−/− mice. (B) Quantification of areas of mineralization expressed in optical density. Data are expressed as the mean ± SD. Statistical analysis was performed using Student’s t-test (n = 3 animals per group, totaling six animals).