Selenium (Se) an oligoelement for mammals is not considered essential in plant fertilization programs. Its deficiency affects almost one million persons, making it a world health problem and nano-biofortification of cultivation is a strategy to attenuate it. Thus, this study determines the foliar selenium nanoparticles (Se-NPs) application effects on yield, enzymatic and non-enzymatic antioxidants, and their accumulation in lettuce. Therefore, six applications of different Se-NPs concentrations: 0, 1, 5, 10, 15, and 20 mg L-1 were considered. The results showed that the high doses decrease yield, enzymatic and non-enzymatic antioxidants but increase Se concentration in lettuce leaves. The recommended dosage here is of 1 mg L-1 for biomass production and 10 mg L-1 for bioactive compounds and enzymatic activity. The recommended daily consumption of lettuce biofortified with 1 mg of Se-NPs (100 g) provides the recommended daily consumption in adults. Thus, the Se-NPs application is an alternative to increase Se concentration and bioactive compounds in lettuce to influence positively, human nutrition after its consumption.