Nanoengineering offers great opportunities for advanced applications in agricultural and biological fields. Here, we proposed a bio-instructive nanospike patch with the precisely controlled sizes using a sophisticated nanoengineering (e.g., capillary force lithography). To investigate the effect of a developed nanospike, the various types of cells including the mammalian cells and bacterial were cultured. Interestingly, the nanospike can both promote the behaviors including proliferation, differentiation, and cytokine excretion of the mammalian cells and prevent the bacterial biofilm formation by the designed feature sized of the nanospike. More specifically, the mammalian cells with relatively large size were received the proper structural stimulation from the nanopske, while the bacterial membrane were damaged by the structure of the nanospike. Our finding reveal the importance of the nanoscale sizes of the spikes to regulate the behaviors of cells, and the proposed nanospike could be used as new bio-instructive platforms such as food packaging and cell therapy platforms in agricultural and biological fields.