The present article focuses on the electrical discharge treatment of duplex stainless steel (DSS-2205) to enhance its biological responses and corrosion resistance. Electrical discharge machining (EDM) was executed at different spark energy levels (by varying input factors) and 18 different morphological DSS substrates were prepared according to the L18 experimental array. Further, an electrochemical potentiodynamic test was performed on the unmachined and treated surfaces to investigate the corrosion performance. Moreover, the high corrosion resistance substrates along with untreated specimen were further analyzed for the in-vitro interaction with human red blood cells. The results claimed that the surface treatment by EDM significantly enhances the corrosion resistance (< 90%) as well as the biological responses of DSS alloy. The current of 10A, on-time = 60 µs, off-time = 150 µs, machining with W-Cu electrode in deionized water was witnessed as the best set of parameters in both corrosion and hemocompatibility test. Thus, surface alteration of DSS-2205 by EDM can be believed as a promising technique for improving the corrosion resistance and biological responses that essential for biomedical applications.