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Inducing Temporal and Reversible Autophagy by Nanotopography for Potential Control of Cell Differentiation.

Tuning autophagy has become a new strategy to control cell differentiation in tissue engineering. The nanosized surface is well-known for its ability to interfere with intracellular procedures, while its role in autophagy regulation is unclear. In this study, we found that a nanotube (NT) structure was able to induce enhanced mTOR-independent autophagy in osteoblasts compared to a flat surface. Further analysis revealed that autophagy was temporally promoted by NTs in the initial day contact and it was also reversible by exchanging the substrate nanotopographies. Actin filaments were significantly dispersed and there were numerous filopodia on the leading edge of cells grown on the NT surface. Intracellular Ca2+ was significantly increased on the NT surface. Moreover, the phenomenon was also found on different nanotopographies as well as in different cell lines. These indicated that cell membrane stretching might be the central regulation factor. Finally, we found that the NT surface exhibited enhanced autophagy-dependent osteogenic differentiation efficacy. In addition, the enhancement on NT surface could be remembered. In conclusion, the nanotopographic surface is able to induce temporal, reversible, and memorable autophagy via cell membrane stretching, which may be used as a versatile method to control cell differentiation.