Mouse bone marrow mesenchymal stem cells (mBMSCs) were used to investigate the osteogenic differentiation of GH-NbBG. Similarly, the mBMSCs were cultured with Control, BG and NbBG extracts. The medium was replaced every 2–3 days. Alkaline phosphatase (ALP) activity was employed to investigate the osteogenic differentiation potential at day 7. The mBMSCs were fixed with 2.5% glutaraldehyde for 15 min and washed with PBS, and incubated in ALP stain working solution (Beyotime, China) for 30 min at room temperature. The reaction was stopped by removing the ALP stain working solution and rinsing with PBS. Stained cells were visualized with light microscope (Observer7, Zeiss, Germany).
The osteogenic differentiation mechanism analysis of GH-NbBG were researched by bioinformatic analysis and RNA sequencing (RNA-seq). In order to verify the correctness of mechanism speculation, the protein expressions of RUNX2 and OCN were analyzed by western bloting and immunofluorescent staining. The detailed experimental procedures were provided in supplementary data.
The osteogenic differentiation mechanism analysis of GH-NbBG were researched by bioinformatic analysis and RNA sequencing (RNA-seq). In order to verify the correctness of mechanism speculation, the protein expressions of RUNX2 and OCN were analyzed by western bloting and immunofluorescent staining. The detailed experimental procedures were provided in supplementary data.