Imagescope version 9

Histomorphometric and histological examination was executed by a competent examiner, who was blinded to the experimental settings. Digital images were evaluated with the help of a software program (Imagescope Version 9.1.19.1571; Aperio Technologies, CA, USA). The extraction socket evaluation was done by measuring the total bone height of lingual and buccal calculated from the top of the buccal and lingual alveolar wall to the most apical point of the jaw; buccolingual alveolar bone wall width was calculated at 3 levels A, B, and C perpendicular to the long axis or vertical line (VL) of the extraction socket marked at 1 mm (A), 3 mm (B), and 5 mm (C) infracrestally from HL (highest level) which is the highest point of alveolar bone and perpendicular to VL (Figure 1).

After radiographic micro-CT analysis, each specimen underwent dehydration and embedding. Specimens were fixed in 10% neutral buffered formalin, dehydrated in ethanol, and decalcified using 5% formic acid (Fisher Diagnostics, Fair Lawn, NJ, USA) for 2 weeks. Two fragments were both embedded to show the sagittal section in the paraffin block. Tissue blocks were cut into at 4 μm, and staining was done using hematoxylin and eosin stain. Selective digital images of sections were taken using a digital camera (DP-20; Olympus, Tokyo, Japan; Figure 3). Sections from the center of each block were examined by light microscopy, and images were captured (ImageScope Version 9.1.19.1571; Aperio Technologies, CA, USA). The images were analyzed by SigmaScan Pro (SPSS, Chicago, IL). The amount of new bone as a percentage of the bone defect area was calculated.