Benchmark platform

The implant-bone samples were fixed in formalin for 2-7 days, decalcified in formic acid for 7 days, and embedded in paraffin. Three-micron sections were cut for immunohistochemistry and stained with CD34, collagen IV, and ETS-related gene (ERG) endothelial markers. ERG was chosen as the preferred antibody because collagen IV also stained the membranes of the fatty cells, making it difficult to interpret. ERG was easier to quantify because it only stains nuclei [32] , compared to CD34 and collagen IV, which stain the membrane and basal lamina [33, 34] .
ERG is a rabbit monoclonal antibody (EPR3864, Ventana Medical Systems) and runs on the Ventana Benchmark platform in a ready-to-use concentration.
NDP.view2 was used to view the scanned slides, and QuPath-0.2.3 was used for quantification of ERG-positive capillaries stained brown by the 3.3 ´-Diaminobenzidine (DAB) reaction.
Positive nuclei and the total number of nuclei were recorded in the 2-mm implant gap area (Figure 4). Unfortunately, the areas containing hydroxyapatite were difficult to section, resulting in lost tissue, which made comparison between groups impossible. Thus, immunohistochemical results were not included in this study.
(FIGURE 4 SHOULD BE PLACED HERE)

IL-13Ra2 expression was estimated immunohistochemically using 4-mm-thick sections taken from TMA slides and an automated Benchmark platform (Ventana Medical Systems, Tucson, AZ, USA) according to the manufacturer's instructions. Briefly, slides were treated with protease for 4 min, and then incubated with anti-IL-13Ra2 mouse monoclonal antibody (1:1000, Abcam, Cambirdge, MA, USA) for 60 min at 37°C. Stainings were visualized using an UltraView universal DAB detection kit (Ventana Medical Systems).
Immunohistochemical expressions of IL-13Ra2 were interpreted semi-quantitatively by two pathologists (Y.K.B. and H.J.K.) under a multiheaded microscope. Immunoreactivity scores (IRSs) were calculated based on staining intensities and extents, as previously described. 17 Staining intensities were assessed as follows: negative (0), weakly positive (1), moderately positive (2), or strongly positive (3), and staining extents were defined as the proportions of positive tumor cells showing cytoplasmic staining and classified as follows: 0% (0), 1%-25% (1), 26%-50% (2), 51%-75% (3), or .75% (4). IRSs were calculated by multiplying intensity and extent scores (minimum score 0 and maximum score 12). For the statistical analysis, a case was classified as positive for IL-13Ra2 expression when its IRS was 1.