Vertical arrays of nanostructures can direct access to cell cytoplasma and probe intracellular molecules. Here, we report an efficient cytosolic delivering platform by simply combining diamond nanoneedle arrays with centrifugation-induced supergravity. We prove that diamond nanoneedle arrays can efficiently deliver drugs and biomaterials into cytosol within several minutes, negotiating the endocytososomal system on cytomembrane. More importantly, we studied the potential influence of the technique on cell metabolism. By detecting the phosphorylated histone variant H2AX (pH2AX) in the nucleus, we prove that the operating process will not lead to DNA double-strand breaks. However, the mechanical disruption can temporarily improve the permeability of the cell membranes. Furthermore, we find that nanoneedle treatment can affect the metabolism in living cells. The treatment can slightly elevate the apoptotic signal in A549 cells. The diamond nanoneedle array treatment can significantly increase the production of reactive oxygen species (ROS) in cells, particularly when working together with anticancer drug molecules. Meanwhile, the activity of cytosolic glucose 6-phosphate dehydrogenase (G6PD) is also raised to counterbalance the elevated ROS content. The result of the marked depolarization of mitochondrial membrane potential suggests that mitochondrial have involved in the intracellular redox reactions and cell apoptosis, which are induced by diamond nanoneedle treatment. Overall, we systematically investigate the potential influence of diamond nanoneedle arrays treatment on the physiology of cells and this study provides a novel understanding on the intracellular delivery mediated by nanoneedles.