Aniline blue orange g solution

The maxillae were dissected and fixed with a 4% paraformaldehyde phosphate buffer (Wako) for 3 days. The maxilla blocks were decalcified in Morse's solution at 4 °C for 4 days, followed by dehydration and paraffin embedding. Serial tissue sections with thickness of 8 μm were cut out and stained with haematoxylin-eosin (H&E). The area of newly formed bone and the rate of new cementum length were measured using ImageJ, as described previously [31 (link),32 (link)]. The area of newly formed bone was estimated as the ratio of the area surrounded by the top of the newly formed bone and reference notches on the mesiobuccal and palatal root surfaces. The newly formed cementum was defined as the portion of cementum that was found on the crown side above the notches, and in which Sharpey's fibers were inserted. The rate of new cementum length was defined as the ratio of the new cementum length formed on the root surface facing the defect to the root surface length from notch to notch. Regarding the cell transplant groups, Azan staining was used to confirm the running of Sharpey's fibers. Paraffin-embedded samples were stained with 0.1% Azocarmin G (Wako), Aniline alcohol (Wako), Acetic acid-ethanol (Wako), 5% Phosphotungstic acid (Wako), and Aniline Blue-Orange G Solution (Wako).