Influence of novel nanoscaffolds on biological behaviors of neural stem cells and the related gene expression

Abstract

BACKGROUND: The scaffold made by nanotechnology may have a similar surface structure to basement membrane of natural organism, which can effectively regulate the behavior of seed cells. OBJECTIVE: To observe the effects of parallel and staggered nanofiber membranes on the proliferation and differentiation of neural stem cells. METHODS: The parallel and staggered nanofiber membranes were prepared using electrostatic spinning technology with type I collagen as raw material. Neural stem cells of newborn rats were seeded on the surface of two kinds of nanofiber membranes. Cell culture alone was used as control. Cell proliferation was detected by MTT assay. Cell proliferation cycle was detected by flow cytometry. Cell differentiation rate was detected by immunohistochemistry. The gene expression of Bcyclin D1 aned CDK2 was detected by real-time quantitative polymerase chain reaction. RESULTS AND CONCLUSION: (1) The absorbance values of cell proliferation of parallel arrangement group and staggered arrangement group at 1, 3, 5, 7 and 9 days were higher than that of the control group (P < 0.05). The absorbance value of cell proliferation of parallel arrangement group was higher than that of staggered arrangement group at 3, 5, 7 and 9 days (P < 0.05). (2) Cell differentiation rate in the parallel arrangement group and the staggered arrangement group was higher than that in the control group (P < 0.05), and that in the parallel arrangement group was higher than that in the staggered arrangement group (P < 0.05). (3) Immunohistochemistry showed that there was no significant difference in neuronal differentiation rate among the three groups (P > 0.05). (4) Real-time quantitative polymerase chain reaction demonstrated that the expression of Bcyclin D1 and CDK2 was higher in the parallel arrangement group and staggered arrangement group was higher than in the control group (P < 0.05); and the expression was higher in the parallel arrangement group than in the staggered arrangement group (P < 0.05). (5) The results confirmed that parallel and staggered nano tissue engineering can promote the proliferation of neural stem cells, but has no significant effect on cell differentiation, and can regulate the biological behavior of neural stem cells from the level of gene expression.