Journal: Development (Cambridge, England)

Article Title: Osteogenic capillaries orchestrate growth plate-independent ossification of the malleus

doi: 10.1242/dev.123885

Figure Lengend Snippet: Histological analysis of the mPb. (A) Hematoxylin and eosin (H&E) staining of the mPb. Low and high magnifications are shown. Scale bars: 100 µm in upper panels, 20 µm in lower panels. (B) Immunofluorescence staining of the mPb for osteocalcin (Ocn, green). Sections were counterstained with DAPI (blue). Scale bars: 50 µm. (C) Horizontal section showing H&E staining of the mPb from a 12-week-old female mouse. Scale bar: 100 µm. (D) Phase contrast image corresponding to boxed area in C, with green endothelial cells (arrows) and orange osteocytes (arrowheads).

Article Snippet: Anti-mouse CD31 (PECAM-1) hamster monoclonal antibody (1:500, 2H8, Millipore), anti-mouse Fosl1 rabbit polyclonal antibody (1:50, N-17, Santa Cruz), anti-mouse osteocalcin polyclonal antibody (1:1000, ALX-210-333, Enzo), anti-mouse osteocalcin rat monoclonal antibody (1:200, R21C-01A, Takara), and anti-mouse endomucin rat monoclonal antibody (1:50, V.7C7, Santa Cruz) were used.

Techniques: Staining, Immunofluorescence

Journal: Development (Cambridge, England)

Article Title: Osteogenic capillaries orchestrate growth plate-independent ossification of the malleus

doi: 10.1242/dev.123885

Figure Lengend Snippet: Bone formation around capillaries. (A) Immunofluorescence indicating capillary endothelial cells (CD31-positive, green) and osteoblasts [osteocalcin (Ocn)-positive, red] in the mPb at P21. Scale bar: 50 µm. Right panel: higher magnification of boxed area in left panel. White dots outline malleus. Scale bar: 20 µm. (B) Alizarin Red/calcein double labeling of the mPb. Alizarin Red and calcein were injected at P25 and P28, respectively. Mice were sacrificed at P29. Scale bar: 50 µm. Right panel: higher magnification of boxed area in left panel. Scale bar: 20 µm. (C,D) Calcein was injected intraperitoneally at P21 into female C57BL/6 mice, and animals were analyzed 24 h later. Calcein (green) and endomucin (Emcn) (C, red) or osteocalcin (D, red) labeling in the mPb. White dotted lines in D encircle nuclei of endothelial cells. Scale bars: 20 µm.

Article Snippet: Anti-mouse CD31 (PECAM-1) hamster monoclonal antibody (1:500, 2H8, Millipore), anti-mouse Fosl1 rabbit polyclonal antibody (1:50, N-17, Santa Cruz), anti-mouse osteocalcin polyclonal antibody (1:1000, ALX-210-333, Enzo), anti-mouse osteocalcin rat monoclonal antibody (1:200, R21C-01A, Takara), and anti-mouse endomucin rat monoclonal antibody (1:50, V.7C7, Santa Cruz) were used.

Techniques: Immunofluorescence, Labeling, Injection

Journal: Development (Cambridge, England)

Article Title: Osteogenic capillaries orchestrate growth plate-independent ossification of the malleus

doi: 10.1242/dev.123885

Figure Lengend Snippet: Conditional Fosl1 overexpression antagonizes normal decreases in capillary volume in the mPb. Expression of Fosl1 (red) plus endomucin (Emcn) (A, green) or osteocalcin (Ocn) (B, green) in mPb isolated from 4-week-old male Fosl1 tetON ( Rosa26 -rtTA; TetOP -Fosl1) mice fed food containing dox from P28 to P31. Scale bars: 50 µm in left panel; 20 µm in right panel. White dots outline malleus. (C) Representative μCT images (voxel size, 1.4 µm) of capillaries in the mPb isolated from non-inducible control ( TetOP -Fosl1, top) and Fosl1-inducible Fosl1 tetON ( Rosa26 -rtTA; TetOP -Fosl1, bottom) mice. Mice of both genotypes were fed a dox-containing diet starting from P21 and sacrificed at P56. Scale bars: 100 µm. (D) H&E staining of the mPb in non-inducible control and Fosl1 tetON mice at P56. Capillary area is greater in Fosl1 tetON mice. Arrowheads indicate osteoblasts. Scale bars: 100 µm in left panels; 20 µm in right panels. (E-G) Quantification of capillary volume (E), number (F) and cross-sectional area of capillaries calculated in a plane 150 µm from the distal end (G) of the mPb. Control, n =5; Fosl1 tetON , n =4. ** P <0.01. Data are represented as means±s.e.m.

Article Snippet: Anti-mouse CD31 (PECAM-1) hamster monoclonal antibody (1:500, 2H8, Millipore), anti-mouse Fosl1 rabbit polyclonal antibody (1:50, N-17, Santa Cruz), anti-mouse osteocalcin polyclonal antibody (1:1000, ALX-210-333, Enzo), anti-mouse osteocalcin rat monoclonal antibody (1:200, R21C-01A, Takara), and anti-mouse endomucin rat monoclonal antibody (1:50, V.7C7, Santa Cruz) were used.

Techniques: Over Expression, Expressing, Isolation, Staining

Journal: International Journal of Nanomedicine

Article Title: Nano-Hydroxyapatite Coating Promotes Porous Calcium Phosphate Ceramic-Induced Osteogenesis Via BMP/Smad Signaling Pathway

doi: 10.2147/IJN.S216182

Figure Lengend Snippet: HE histological analysis of in vivo ectopic bone formation ability for BCP ( A ) and nHA-coated BCP ( B ) scaffolds after implanted in back muscles of rabbits for 90 days. Yellow arrows: inflammatory cells; black arrows: bone formation as evidenced by osteocytes settled in bone lacuna. Immunofluorescent staining of osteocalcin (OCN) for BCP ( C ) and nHA-coated BCP ( D ) groups. Green and blue colors represented osteocalcin and DAPI-stained nuclei, respectively.

Article Snippet: The others were subjected to co-staining with DAPI, rabbit-anti-mouse osteocalcin primary antibody, and goat-anti-rabbit fluorescent secondary antibody (Servicebio, China), and visualized under a fluorescence microscope (Leica, Germany) for immunofluorescence analysis.

Techniques: In Vivo, Muscles, Staining