| Summary: | Bacterial nanocellulose (BNC) membranes, with remarkable physical and mechanical properties, emerged as a versatile biopolymeric carrier of bioactive compounds for skin care applications. In this study, BNC membranes were loaded with glycerol (as plasticizer and humectant agent) and different doses (1–3 μg cm<sup>−2</sup>) of an aqueous extract obtained from the hydro-distillation of <i>Eucalyptus globulus</i> Labill. leaves (HDE), for application as sheet facial masks. All membranes are resistant and highly malleable at dry and wet states, with similar or even better mechanical properties than those of a commercial BNC mask. Moreover, the HDE was found to confer a dose-dependent antioxidant activity to pure BNC. Additionally, upon 3 months of storage at 22–25 °C and 52% relative humidity (RH) or at 40 °C and 75% RH, it was confirmed that the antioxidant activity and the macroscopic aspect of the membrane with 2 μg cm<sup>−2</sup> of HDE were maintained. Membranes were also shown to be non-cytotoxic towards HaCaT and NIH/3T3 cells, and the membrane with 2 μg cm<sup>−2</sup> of HDE caused a significant reduction in the senescence-associated β-galactosidase activity in NIH/3T3 cells. These findings suggest the suitability and potential of the obtained membranes as bioactive facial masks for anti-aging applications.
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