Journal: Bone
Article Title: A Xenograft Model to Evaluate the Bone Forming Effects of Sclerostin Antibody in Human Bone Derived from Pediatric Osteogenesis Imperfecta Patients
doi: 10.1016/j.bone.2019.115118
Figure Lengend Snippet: Immunohistochemistry with fluorescence (IHC-F) following removal from the host at 2 and 4 weeks treated (SclAb) and untreated. Human osteogenesis imperfecta (OI) implants were dual IHC-F stained to probe for the presence of Osterix (Osx; green) and human mitochondria (hMito; red) (A-D) and on serial sections, Sclerostin (yellow) and hMito (red) (E-H). In all cases, hMito was used to indicate donor derived cells and Osx or sclerostin primary antibody (validated sensitivity to both mouse and human antigens) were used to probe all instances of expression (both host and donor). Zoomed insets (1) depict lining cells expressing Osx (A-D) and osteocytes expressing sclerostin (E-H). Representative Hematoxylin and Eosin stained bone acquired at baseline (I). Images were acquired at 40x (50 μm scale bar). DAPI= nuclear stain (blue). Panel represents data from one OI patient (OI 6, Type III/IV OI) who yielded cortical-derived bone samples (I), hematoxylin and eosin (H&E) stained implant acquired at 20x with a 250 μm scale bar.
Article Snippet: Sections were incubated with the primary anti-hMito antibody (MAB1273, EMD Millipore) at a 1:200 dilution and either a primary polyclonal rabbit anti-Osx antibody (ab22552, Abcam; 1:400) or primary polyclonal rabbit anti-sclerostin antibody (bs-10200r, Bioss; 1:200) overnight at 4°C.
Techniques: Immunohistochemistry, Fluorescence, Staining, Derivative Assay, Expressing
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