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. Author manuscript; available in PMC: 2020 Jul 5.
Published in final edited form as: Cell Rep. 2020 May 26;31(8):107696. doi: 10.1016/j.celrep.2020.107696

Figure 5. Local Ngf Deletion Impairs Calvarial Bone Defect Repair.

Figure 5.

Ngffl/fl animals were locally injected with Ad-GFP or Ad-Cre virus, followed by creation of a 1.8 mm, circular, full-thickness frontal bone defect.

(A and B) μCT reconstructions of the defect site in a top-down view (above) and coronal cross-sectional images (below) among Ngffl/fl animals locally injected with either (A) Ad-GFP or (B) Ad-Cre. Analysis performed at d28 post-injury. Margins of original defect are indicated by dashed black or red lines. Black scale bar, 500 μm; white scale bar, 200 μm.

(C–G) μCT quantification of bone healing among Ad-GFP- and Ad-Cre-injected mice, including (C) bone volume (BV), (D) bone volume/tissue volume (BV/TV), (E) bone formation area (BFA), (F) bone healing score (score), and (G) defect diameter.

(H and I) H&E stain of coronal cross-section of the healing defect site from (H) Ad-GFP- or (I) Ad-Cre-injected mice. Black arrowheads indicate span of initial defect. Black scale bar, 50 μm.

(J–L) Immunohistochemical staining of TUBB3+ (beta III tubulin) nerve fibers at the defect edge from (J) Ad-GFP- or (K) Ad-Cre-injected mice, appearing green and (L) quantification of TUBB3 immunoreactivity within the calvarial defect. Dashed white lines indicate bone edge.

(M–O) Immunohistochemical staining of CD31+ blood vessels at the defect edge from (M) Ad-GFP- or (N) Ad-Cre-injected mice, appearing red, and (O) quantification of CD31 immunoreactivity within the calvarial defect site. Dashed white lines indicate bone edge.

(P–R) Immunohistochemical staining of OCN+ (osteocalcin) osteoblasts from (P) Ad-GFP- or (Q) Ad-Cre-injected mice, appearing cyan, and (R) quantification of OCN immunoreactivity within the calvarial defect site. Dashed white lines indicate bone edge. White scale bar, 50 μm.

DAPI counterstain appears blue in all images. In graphs, each dot represents a single animal; n = 8 animals per group. †p < 0.05 and ††p < 0.01 in comparison with Ad-GFP control. Data are represented as mean ± SD. See also Figures S3 and S4.