Maxillary Bone Formation Imaging

Following sacrifice, the maxillae were collected, fixed for 48 h in 4% paraformaldehyde and stored in − 80 °C until being dehydrated in 30% sucrose and were embedded in optimal cutting temperature (O.C.T.; Tissue-Tek, Thermo Fisher, Waltham, MA, USA) and cut into serial 8-μm-thick sections. Cryosections were prepared and imaged with a Keyence BZ-X9000 all-in-one Fluorescence Microscope (Itasca, IL, USA) for MSC tracking (n = 4/group). Surface-based bone formation was quantitated using BIOQUANT image analysis software (n = 4/group) (BIOQUANT Image Analysis Corporation, Nashville, TN, USA) [17 (link)].