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Objective Silver nanoparticles (AgNPs) are widely used in orthopaedic implants because of their excellent antimicrobial properties. However, the effects of AgNPs on bone cells and osteogenic activity are still poorly understood.Method Here, we investigated the effect of AgNPs on the cell viability, uptake, and osteogenic activity of osteoblast-like cells (MG-63 cells) at low concentrations.Results Our results showed that uptake and retention of AgNPs reduced the cell viability and increased cell membrane penetrability even after termination of exposure of MG-63 cells to AgNPs. In addition, AgNPs induced cell shrinkage, reduced the expressions of ALP, COL-I, OCN and OPG, and enhanced the expressions of Runx2 and RANKL.Conclusion Collectively, our work demonstrated that the cytotoxic effects of low-dose AgNPs on MG-63 cells persisted even after termination of exposure. AgNPs may interfere with bone formation. More attention should be paid to the toxicity of AgNPs during the design of future orthopaedic implants.