Day 2 hMSC sheets (n = 3 per group) were fixed in 10% neutral-buffered formalin (NBF) for 24 hours at 4°C before switching to 70% ethanol. One representative femur per group was taken for histology at weeks 4 and 12 after surgery and chosen on the basis of microCT-calculated average bone volumes. Femora were fixed in 10% NBF for 72 hours at 4°C and then transferred to 0.25 M EDTA (pH 7.4) for 14 days at 4°C under mild agitation on a rocker plate, with changes of the decalcification solution every 3 to 4 days. Following paraffin processing, 5-μm mid-sagittal sections were cut using a microtome (Leica Microsystems Inc., Buffalo Grove, IL) and stained with hematoxylin and eosin and Safranin-O/Fast-green. Light microscopy images were captured using an Olympus BX61VS microscope (Olympus, Center Valley, PA) with a Pike F-505 camera (Allied Vision Technologies, Stadtroda, Germany).

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