Human recombinant epidermal growth factor (EGF) promotes cell proliferation as well as skin remodeling and healing. However, it’s unclear if EGF is stable in in vitro cell culture medium treatment conditions. Since EGF’s instability in aqueous solutions has resulted in different kinds of challenges, novel treatment approaches that preserve EGF’s stability and usefulness have been developed. The current study examined the effects of the antioxidants sodium selenite (Se), disodium ethylenediaminetetraacetic acid dihydrate (EDTA), zinc chloride (Zn), and ascorbic acid (AA) on the stability of EGF in cell culture using Dulbecco’s Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute 1640 medium (RPMI), as well as their ability to scavenge free radicals. In vitro , DMEM and RPMI serum free medium with 2.5 µM of EDTA, Se, AA, and Zn antioxidants were used separately at 37˚C for three days to examine the stability of EGF. The stability of EGF was evaluated using reversed phase high-performance liquid chromatography (RP-HPLC). Cells were cultivated for 3 days to further estimate the impact of antioxidants on cell cytotoxicity and proliferation using the MTT assay with NIH-3T3, L929, TF-1, and Sp2/0-Ag14 cell types. The antioxidants EDTA, Se, AA, and Zn at an extracellular concentration of 2.5 µM improved EGF stability in both DMEM and RPMI medium while having no effect on the proliferation of NIH-3T3, L929, TF-1, and SP2/0-Ag14 cells.
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