Bone mineralization was analyzed as previously described (59). L3 to L5 vertebra and femur were collected, fixed in 4% PFA at 4°C for 1 day, and rinsed thoroughly with PBS. The fixed samples were placed in a 50-ml centrifuge tube with styrofoam spacers and loaded into a microCT scanner (vivaCT 80, Scanco Medical, Wayne, PA) and scanned at 55 keV at a pixel resolution of 10.4 μm. The reconstruction of scanned images was performed using microCT Evaluation Program V6.6 (Scanco Medical), followed by generation of radiographs and three-dimensional models using microCT Ray V4.0 (Scanco Medical). Mineral density of the scaffolds was quantified and presented as a percentage of BV/TV based on 100 contiguous slices.

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