Selectech hematoxylin and eosin h e staining system

After fixing in 10% phosphate-buffered formalin for 48 hours and μCT imaging, explanted cranial bone samples were submerged in 70% ethanol for long-term storage. Embedding, sectioning, and staining of tissue was performed by the Stephenson Cancer Tissue Pathology Core at the University of Oklahoma Health Sciences Center. Briefly, cranial implants with surrounding host bone were embedded in paraffin and sectioned in the sagittal plane following a standard protocol. Paraffin blocks were cut to a thickness of 4 μm and affixed to glass microscope slides. The SelecTech hematoxylin and eosin (H&E) staining system (Leica Biosystems, Wetzlar, Germany) was performed using a Leica ST5020 multistainer (Leica Biosystems) following the manufacturer’s protocol (n = 5).

Ovine calvarial bone samples for histological analysis were harvested and stored in 10% phosphate buffered formalin for 48 h (n = 5). Embedding, sectioning, and staining were performed using standard methods. Briefly, explanted calvarial bone samples were decalcified using Richard-Allan Scientific^™ Decalcifying Solution (Cat# 8340, ThermoFisher Scientific) at room temperature for 4 days. Decalcified bone was then embedded in paraffin and sectioned in the sagittal plane to a thickness of 4 μm and affixed to glass microscope slides. The SelecTech hematoxylin and eosin (H&E) staining system (Leica Biosystems, Wetzlar, Germany) was performed using a Leica ST5020 multistainer (Leica Biosystems) following the manufacturer’s protocol.