The growing need of varieties of products made up of advanced materials, exhibiting better properties, is compelling the material scientists to develop varieties of such materials. Among wide categories, the ones that are able to save human life or make it more comfortable, can be termed as the best materials ever developed, and such materials are known as biomaterials. Although shaping most of these materials as per the needs, puts forth a challenge before the manufacturing engineers, because of their certain mechanical properties. Ti-6Al-4V alloy, an extremely hard material, is also well known for its outstanding mechanical biocompatibility with human body. Such necessities in the midst of difficult circumstances have led to non-traditional machining means, which are known for their highly precise and accurate machining irrespective of the type, hardness and shape to be machined. Electric discharge machining (EDM) is highly capable material-removal means, with some of its more efficient variants such as wire electric discharge machining (WEDM) etc. for more specific purposes. Removing excess material of convoluted and baroque shape, with excellent degree of smoothness and finish, are some of the best features of WEDM. Cryogenic treatment (CT) is another term, known for improving the machining performance in various ways, such as by revamping the cutting tool life and standard and refinement of the mechanically produced surface. In this investigation, the influence of varying pulse on time (T-on), pulse off time (T-off), flushing pressure (FP), wire feed rate (WFR), wire tension (WT), spark gap set voltage (SV) and peak current (PC) of WEDM on the integrity of machined regions (SR) of deep cryogenically treated Ti-6Al-4V alloy has been examined experimentally. The results present a significant fall in roughness of the machined surfaces, which again prove and confirm Ti-6Al-4V alloy as one of the best biocompatible materials for producing biomedical implants, suitable with tissues inside the human body.