This study is focused on the preparation of doxorubicin-loaded nanopolymersomes (PolyDoxSome) and assessment of the effects of various solvents and process variables on the size and drug loading during preparation of formulation. PolyDoxSome was prepared by nanoprecipitation method using amphiphilic (PEG)₃-PLA copolymer, and the formation of polymersomes was assessed by dynamic light scattering and optical and transmission electron microscopy and evaluated for in vitro release profile and in vitro cytotoxicity. A systematic investigation indicated that solvent composition, order of addition, aqueous phase, copolymer concentration, and external energy input have significant influence on size and dispersity of PolyDoxSome. Under optimized conditions, PolyDoxSome had a size range of 130-180 nm with PDI < 0.2, a zeta potential ∼-8 mV, and a drug loading at ∼11% w/w with an encapsulation efficiency at ∼53% w/w. In vitro release profile of PolyDoxSome at 37 °C demonstrated that doxorubicin release was pH dependent and gave higher release at pH 5.5 in comparison to the release at pH 7.4 (similarity factor, f₂ < 50). PolyDoxSome exhibited enhanced cellular uptake of doxorubicin compared to free doxorubicin solution in MCF-7 cell line and showed a better cytotoxicity of doxorubicin at equivalent dose in nanopolymersomes. In conclusion, size and dispersity were strongly influenced by duration of magnetic stirring and overall composition of organic/aqueous media; however, size and dispersity were retained against different degrees of dilution. PolyDoxSome was able to control the release of doxorubicin in pH dependent manner and effectively deliver the drug in active form to MCF-7 breast cancer cells.
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