Figure 1.
Effects of Zoledronate on the course of tibial fracture repair.
(A) Sections through the calluses (green dashed-lines) of PBS- (left) and zoledronate- (right) treated mice were stained with Safranin-O/Fast Green (days 7, 10 and 14 post-fracture) to detect cartilage (arrowsheads) and Trichrome (days 21 and 28 post-fracture) to detect bone (arrows). White arrows indicate the fracture site. Scale bar = 500 µm. (B) Histomorphometric analyses of total callus volume (Top), total cartilage volume (CV, middle) and total bone volume (BV, bottom) on PBS treated and ZA treated-mice at days 7, 10, 14, 21 and 28 post-fracture (n = 6 per group). *p<0.05, **<0.01. Bars represent mean±s.d.
Figure 2.
Mandibular fracture model and effects of zoledronate on the course of mandibular fracture repair.
(A) To create a mandibular fracture, the skin was opened and the mandible was exposed (left). Five holes were drilled (black arrow) and the fracture was created (white arrow) by surgical tweezers. (B) Sections through the PBS- (left) and zoledronate- (right) treated calluses (green dashed-lines) were stained with Safranin-O/Fast Green (day 7 and day 10 post-fracture) to detect cartilage (arrowheads) and Trichrome (day 14, 21, and 28 post-fracture) to detect bone (arrows). White arrows indicate the fracture site. Scale bar = 500 µm. (C) Histomorphometric analyses of total callus volume (TV, Top), total cartilage volume (CV, middle) and total bone volume (BV, bottom) on PBS treated and zoledronate treated-mice at days 7, 10, 14, 21 and 28 post-fracture (n = 6 per group). *p<0.05, **<0.01. Bars represent mean±s.d.
Figure 3.
Effects of Zoledronate on collagen types II and X expression during tibial and mandibular fracture repair.
Immunostaining of collagen type II (A–B, G–H, black arrows) and collagen type X (D–F, J–L, white arrows) near the periosteum within tibial (top) and mandibular (bottom) fracture calluses of PBS (left) and zoledronate (middle and right) treated mice. No staining is observed in negative controls (C and I).
Figure 4.
Effects of zoledronate on osteoclastogenesis during tibial and mandibular fracture repair.
Stereological analyses of TRAP positive cells per area of fracture callus (n = 6 per group) in PBS- and zoledronate-treated mice at day 7 (A) and day 14 (B) post-fracture. **p<0.01. Bars represent mean±s.d.
Figure 5.
Effects of zoledronate on apoptosis during tibial and mandibular fracture repair.
TUNEL and TRAP double staining on ZA-treated tibial (A and B) and mandibular (C–E) fractures at day 5 post-injury. High magnifications (B, D, E) indicate apoptotic cells (black arrows) and TRAP-positive cells (white arrows) within the fracture callus. No TRAP-positive apoptotic cells can be detected at the fracture site. (A, C) Scale bar = 50 µm; (B, D, E) Scale bar = 10 µm.
Figure 6.
Effects of zoledronate on angiogenesis during tibial and mandibular fracture repair.
(A) PECAM immunostaining (black arrows) on PBS- (top) and ZA-treated (bottom) tibial (left) and mandibular (right) fracture calluses. Scale bar = 200 µm. (B) Analysis of the length density of blood vessels within the callus (mm/mm3) and (C) analysis of the surface density of blood vessels within the callus (mm2/mm3) in PBS- and zoledronate-treated mice at day 5 post-fracture (n = 6 per group). *p<0.05. Bars represent mean±s.d.