© 2021 The Textile Institute.Cardiac patch has been proposed as an alternative therapeutic tissue engineering strategy to treat damaged heart for patients who survive the acute myocardial infarction (MI). To that end, studies have focused on designing an ideal cardiac patch assisting myocardium to repair itself. The engineering of three dimensional (3 D) anisotropic knitted scaffolds could be an efficient and effective strategy for cardiac patches. Thus, the polyethylene terephthalate (PET) pile loop knit fabrics were fabricated as 3 D scaffolds, where the values of stiffness (Young’s modulus) of the scaffolds were similar to those reported for the human myocardium. Anisotropic porous knitted scaffolds supported the adhesion and proliferation of murine C2C12 myoblasts cell line in vitro. Scaffolds with a high surface to volume ratio and porosity exhibited superior performance in terms of cell adhesion and proliferation. Furthermore, out of thirteen proinflammatory cytokines, the levels of three cytokines including IL-1α, IL-1β, and IL-11 were observed to be higher in cells seeded on scaffolds coated with fibronectin than those in related control groups. Thus, the proposed knitted structure can potentially be used for tissue engineering and regenerative medicine.