Histological analysis
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Histomorphometric analysis was performed at the Hard Tissue Laboratory, Faculty of Medicine, University of Coimbra, Portugal. Bone biopsy samples measuring 6 to 8 mm in length were fixed with 10% formaldehyde (Panreac, Barcelona, Spain) solution buffered at pH 7.4 and stored at 4 °C, until histology analysis of non-decalcified hard tissues with the high-precision Exakt® system (Exakt Technologies, OK, USA). The blocks were sectioned longitudinally in 50–100-μm-thick slices, then stained with toluidine blue and examined with a light microscope (Nikon® Eclipse E600, Tokyo, Japan). The new bone formation, the remaining biomaterial, and the marrow space were quantified in percentages using Bioquant® (Image Analysis Corporation, Nashville, TN).