Effects of nerve growth factor on proliferation, migration, differentiation and apoptosis of mouse jaw bone marrow mesenchymal stem cells

  Background  Nerve growth factor (NGF) is one of the neurotrophic factors, which can promote the growth of central and peripheral neurons and repair the damaged nervous system. It has been proved that NGF can promote the osteogenic differentiation and bone formation of mesoderm-derived long bone marrow mesenchymal stem cells. However, the effect of NGF on the biological characteristics of ectoderm-derived jaw bone marrow mesenchymal stem cells (JBMMSCs) remains unclear.

  Objective  To investigate the effects of NGF on the proliferation, migration, differentiation and apoptosis of JBMMSCs.

  Methods  Mouse JBMMSCs were cultured and identified. The second generation of JBMMSCs were cultured in medium containing 50 ng/mL of NGF for 72 h. CCK-8, qRT-PCR, Western blot and immunofluorescence (IF) assay and other methods were used to detect the cell proliferation and the expression of related markers MCM-2 and Ki67. qRT-PCR and Western blot were used to detect the expression of apoptosis-related markers Bcl-3 and Bax. The migration ability of JBMMSCs was detected by scratch test. Alkaline phosphatase (ALP) staining, qRT-PCR, Western blot, and IF were used to detect the expression of osteogenic related markers.

  Results  CCK-8 results showed that 50 ng/mL of NGF could significantly enhance the OD value of JBMMSCs on day 3 and day 5. Western blot and IF results showed that 50 ng/mL of NGF could significantly enhance the protein levels of Ki67 and MCM-2 in JBMMSCs. The results of scratch test showed that 50 ng/mL NGF could significantly increase the migration rate of JBMMSCs and effectively reduce the scratch area at each time point. And it also could effectively enhance the alkaline phosphatase activity of JBMMSCs and up-regulate the mRNA expression levels of ALP, OCN, RUNX2 and BMP2 and the protein levels of ALP, OCN, BMP2, OPN and RUNX2. In addition, 50 ng/mL of NGF could up-regulate the mRNA and protein expression of Bcl-2 and down-regulate the mRNA and protein expression of Bax.

  Conclusion  It suggests that 50 ng/mL of NGF can effectively promote the proliferation, migration and osteogenic differentiation of JBMMSCs, and inhibit their apoptosis.