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. Author manuscript; available in PMC: 2015 Oct 1.
Published in final edited form as: J Biomed Mater Res A. 2013 Oct 29;102(10):3429–3438. doi: 10.1002/jbma.35006

FIGURE 2.

FIGURE 2

Determination of optimal timeline for BMP9-induced ectopic bone formation in vivo. (A) Verification of efficient gene transfer mediated by AdBMP9 and AdGFP in Subconfluent C2C12 cells. (B) Demonstration of BMP9-induced ALP activity. Subconfluent C2C12 cells were transduced with AdBMP9 or AdGFP for 4 days. The cells were subjected to histochemical staining of ALP activity. Representative results are shown. (C) Radiographs of ectopic bone formation. BMP9-transduced C2C12 cells with type I collagen carrier were implanted subcutaneously in the flanks of athymic nude mice. The animals were radiographically imaged at 2, 3, and 4 weeks after implantation. Implantation sites are indicated by arrows. Masses formed in the GFP group are circled with dotted lines. Representative results are shown. (D) Histologic evaluation of the retrieved samples from BMP9 or GFP-treated samples. Animals were sacrificed at the endpoint (4wk), masses were retrieved from the implant sites, fixed, decalcified, paraffin-embedded, and subjected to H & E staining. Representative results are shown. ‘b”, osteoid matrix; “c”, injected and undifferentiated cells; magnification, 200x