Polylactide (PLA) nanofibers loaded with antioxidants, i.e. vitamin B-6 or pyridoxine (PN) and its analogue pyridoxal (PL), and with hydroxycinnamic acids, i.e. p-coumaric acid (CUM) and caffeic acid (CAF), were prepared by the electrospinning technique. The control consisted of nanofibers loaded with trolox (TRX). Experimentally, these new materials were tested on the inhibition of oxidative DNA damage caused by free radicals initiated by 2,2'-azobis (2-amidinopropane hydrochloride) (AAPH). This damage was assessed in vitro and in vivo by measuring the conversion of supercoiled pUC19 plasmid DNA to open circular and linear forms. It was found that these antioxidants, in solution, could significantly inhibit oxidative DNA damage, and that the antioxidants loaded in the PLA nanofibers maintained in vitro and in vivo their protective role against oxidative DNA damage. Thus, electrospun mats derived from PLA nanofibers loaded with the considered compounds have the capacity to protect DNA against oxidative damage, and appear interesting, for their use in the purification of plasmidic or genomic DNA.