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. 2021 Jun 4;10:e62917. doi: 10.7554/eLife.62917

Figure 5. Activation of Hh/Gli1 pathway accelerates mouse meniscus repair.

(A) Schematic representation of the study protocol. WT mice received meniscus injury at 12 weeks of age followed by transplantation of 5000 Gli1+ or Gli1- meniscus cells at the injury site. Knee joints were harvested at 1 and 4 weeks after injury. (B) Representative overview (a–c), safranin O/fast green staining (d–i), and polarizing images (j–l) of mouse knee joints at 4 weeks after injury. Yellow dashed lines outline the overview morphology of injured meniscus. Meniscus is shown attached to tibial plateau. Arrows point to the injury site. Red boxed areas in d-f are shown at high magnification in g-i, respectively. Scale bars, 200 μm. F: femur; T: tibia; MS: meniscus synovial end; ML: meniscus ligamental end. (C) Repair score was evaluated. n = 8 mice/group. (D) Representative confocal images of mouse knee joints at 1 and 4 weeks after injury. Boxed areas in the top panel are shown at a high magnification at the bottom panel. Dashed line outlines meniscus. Scale bars, 200 μm. Blue: DAPI, Red: Td. (E) Schematic representation of the study protocol. Gli1ER/Td mice received Tam injections and meniscus injury at 12 weeks of age (day 0) followed by vehicle and PMA injection. Knee joints were harvested at 1 and 4 weeks after injury. (F) Representative overview (a, b), safranin O/fast green staining (c–f), and polarizing images (g, h) of mouse knee joints at 4 weeks after injury. Yellow dashed lines outline the overview morphology of injured meniscus. Meniscus is shown attached to tibial plateau. Red boxed areas in c and d are shown at high magnification in e and f, respectively. Arrows point to the injury site. Scale bars, 200 μm. (G) Repair score was evaluated. n = 7 mice/group. (H) Representative fluorescence images of vehicle- and PMA-treated mouse meniscus at 1 and 4 weeks after injury. Scale bars, 200 μm. Statistical analysis was performed using one-way ANOVA with Tukey-Kramer post-hoc test for (C) and unpaired two-tailed t-test for (G). Data presented as mean ± s.e.m. ***p<0.001.

Figure 5—source data 1. Raw data for Figure 5C and G.

Figure 5.

Figure 5—figure supplement 1. Hh agonist treatment does not promote joint calcification.

Figure 5—figure supplement 1.

(A) Representative microCT images of WT mouse knee joints at 3 months post surgery as well as vehicle (Veh) or PMA treatment. (B) The volume of calcified meniscus was quantified. n = 5–6 mice/group. (C) Osteophyte volume was quantified. n = 5–6 mice/group. Statistical analysis was performed using unpaired two-tailed t-test. Red arrows indicate the osteophytes. Data presented as mean ± s.e.m.