BACKGROUND: The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period. METHODS: Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7. RESULTS: Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs. CONCLUSION The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

At present， there are many difficulties in the diagnosis and treatment of acute vestibular syndrome （AVS）. For example， complex and difficult identification of the cause of disease， uneven diagnosis and treatment levels of clinical doctors， weak humanistic care awareness， lack of communication skills， intrinsic affinity and other reasons， which make it difficult for AVS patients in the process of diagnosis and treatment， and cannot receive timely and effective treatment， resulting in an exacerbation of doctor-patient conflicts. Therefore， it is recommended to explore new paths of AVS diagnosis and treatment work using the humanistic care concept， respect each other between doctors and patients， build a team of medical staff with the value orientation of “humanistic care”， and promote the organic unity of theory and practice of “humanistic care”， with a view to better promoting the implementation of AVS diagnosis and treatment work， helping more patients rebuild confidence， enhancing quality of life， and improving the doctor-patient relationship.