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Background and purpose: Osteosarcoma, as the most common bone cancer has not shown considerable response to current chemotherapy drugs. New nanotechnology-based drug delivery systems seems to be promising in cancer treatment. The aim of this study was to design and synthesize a niosomal nano-carrier for drug delivery of berberine to Saos-2 cell line, as a model of osteosarcoma. Materials and methods: Different niosome formulations composed of cholesterol (25%) and different surfactants (75%) were prepared using thin film method. The resulting niosome with optimal properties was pegylated by adding 5% of DSPE-mPEG2000. Finally, the cytotoxicity of free form of berberine and berberine-containing pegylated niosome in normal fibroblast cell line (HFF) and Saos-2 cell line were evaluated by MTT assay. Results: The niosome made of cholesterol (25%) and Tween 60 (75%) showed the highest drug loading and release rate and was selected as the optimal formulation. Findings showed that the optimized pegylated niosome had high loading efficiency and controlled drug release (79.34% and 66.79%, respectively). Finally, MTT assay showed that the berberine-containing pegylated niosome had more toxicity in Saos-2 cell line than the free form of the drug. Conclusion: The synthesized niosomal nano-carrier had desirable properties such as high loading efficiency and controlled drug release, which can be used as a favorable drug delivery system with low toxicity and high delivery efficiency in order to increase the effect of the drug while reducing its dose. [ABSTRACT FROM AUTHOR]