(a) Schematic illustration of the double emulsion solvent evaporation method used to prepare the Hb-loaded PLGA-NPs (HbNPs). Briefly, the HbNPs were prepared by introducing Hb into the water phase (together with TRE if required) to form a first water-in-oil-emulsion (w1/o). Next, the w1/o emulsion was added to a PVA solution to form a double w1/o/w2 emulsion. Upon removal of the organic solvent, HbNPs were obtained. (b) The effect of several process parameters (i.e., concentrations of protein (Hb), addition of stabilizer (TRE), concentration of polymer (PLGA) and emulsifier (PVA) on the size (i), polydispersity (PDI) (ii), loading capacity (iii), entrapment efficiency (iv) and functionality (v) of the resulting HbNPs was evaluated. Hb: hemoglobin, PLGA: poly(lactide-co-glycolide), PVA: poly(vinyl alcohol), TRE: trehalose.