Cardiovascular diseases (CVDs) remain one of the most prevalent healthcare problems worldwide, with the ageing of the population causing a significant increase in CVD–related mortalities. Treatment of this group of patients may require a multilateral approach, including pharmaceutical correction, endovascular technologies, and invasive bypass surgeries in patients with severe symptoms. During a bypass surgery, a vascular substitute, or graft, is used to create a new pathway for blood flow, allowing for the restoration of the functional blood supply. Lack of optimal vascular grafts has necessitated evaluating the utility of tissue-engineering methods for the development of new functional surgical conduits, which possess the most physiological properties and exhibit excellent patency. These “ideal” cardiovascular bypass grafts should fit a range of characteristics, including biocompatibility, non-immunogenicity, easy handling, mechanical resistance and compliance, ability to grow, remodel, and self-repair after implantation. Decellularized vascular scaffolds seem to be an attractive therapeutic solution; however, the efficiency of the decellularization (DC) process and the quality of the obtained matrices are dependent on a range of factors. Cytotoxicity of the vascular graft obtained by DC is induced by the presence of DC agents’ residue in the matrix. Identification of the conditions allowing the chemicals’ optimal removal, including the demand to perform repeating washing steps, is mandatory before in vivo evaluation and clinical application.