[Neurotrophin-3 enhances the osteogenesis ability of human bone marrow mesenchymal stem cells stimulated by lipopolysaccharide].

Objective To investigate the protective effect of neurotrophin-3 (NT-3) on human bone marrow mesenchymal stem cells (hBMSCs) and its ability to promote the differentiation of hBMSCs into osteoblasts in the inflammatory environment. Methods The cell inflammation model was established by lipopolysaccharide (LPS). The hBMSCs without any stimulation was defined as the inflammatory control group; the hBMSCs stimulated by 100 ng/mL human NT-3 recombinant protein as the NT-3 group; the hBMSCs stimulated by 100 mmol/L pyrvinium pamoate (PP) for 12 hours and then stimulated by 100 ng/mL human NT-3 recombinant protein as the Wnt inhibitor group; the normal cultured hBMSCs as the normal control group. We performed the experiment of osteoblast induction on all groups. CCK-8 assay was used to detect the proliferation of hBMSCs; the fluorescein isothiocyanate labeled annexin V/propidium iodide (Annexin V-FITC/PI) double labeling combined with flow cytometry was used to detect the apoptosis of hBMSCs; ELISA was used to detect the protein levels of runt-related transcription factor 2 (RUNX2) protein and alkaline phosphatase (ALP); and the alizarin red staining experiment was conducted to detect the ability of calcium nodule formation. Results Compared with the inflammatory control group, the proliferative activity of hBMSCs in NT-3 group significantly increased, the apoptosis obviously decreased, and the contents of RUNX2 and ALP, as well as the intensity of alizarin red staining in NT-3 group evidently rose. Compared with the NT-3 group, the proliferative activity of hBMSCs in the Wnt inhibitor group was inhibited, the cell apoptosis was promoted, and the contents of RUNX2 and ALP, as well as the intensity of alizarin red staining in the Wnt inhibitor group were reduced remarkably. Conclusion NT-3 can protect hBMSCs from anti-inflammatory damage and promote the differentiation of hBMSCs into osteoblasts.