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. Author manuscript; available in PMC: 2016 Feb 1.
Published in final edited form as: J Orthop Res. 2014 Oct 28;33(2):174–184. doi: 10.1002/jor.22749

Figure 6.

Figure 6

SDF-1β significantly increases bone area by histomorphometry following BMSC tibial transplantation in irradiation-preconditioned recipients. The standard 2-D bone histomorphometric parameter BV/TV was assessed in the proximal tibiae 4 weeks post-transplantation. (A) Overview of a representative H&E-stained coronal section through the center of the diaphysis (EP: epiphyseal plate, M: marrow, T: trabecular bone, C: cortical bone, 2.5×, bar 500 μm). The 876.9 × 657.1 μm rectangular ROI (20×, bar 20 μm, 1388 × 1040 pixels per image, 576211.68 μm2 or 0.58 mm2 combined total area in 4 identical quadrants) is outlined in dashed lines. (B) One H&E-stained central coronal section 20X ROI quadrant. (C) The same 20X ROI depicting bone tissue in black (Photo-shop wand tool, tolerance: 10). (D) Final black-and-white image mask of the 20X ROI showing bone tissue in black and all other tissue in white. Using ImageJ, the entire image area was calculated as tissue area (T.Ar) and the black-colored area was summarized as bone area (B.Ar; default ImageJ wand tool) to calculate BV/TV. Per definition, BV/TV is numerically identical with the corresponding area/area ratio B.Ar/T.Ar. E) BV/TV in vehicle- and Tet-Off-SDF-1β (±Dox) or Tet-Off-EV BMSC-transplanted tibiae at 1.32 × 107 cells/ml (*p <0.05, ***p <0.0001 BMSC-transplanted vs. vehicle controls; ap <0.05 Tet-Off-SDF-1β (−Dox) vs. Tet-Off-SDF-1β (+Dox) or Tet-Off-EV, n = 3 animals per group).