Polyurethane (PU) scaffolds were prepared using three-dimensional printing technique. 3D-printed scaffold can be designed with interconnecting pores and uniform porosity for cell ingrowth. Gelatin was used as a chain extender to enhance the mechanical properties as well as promoting attachment, proliferation, and differentiation of chondrocyte to mimic cartilage. The effect of incorporation of gelatin into PU, physicomechanical properties such as surface hardness, tensile strength, and percentage elongation has been investigated. The gelatin chain extension promoted significantly higher initial modulus and yield stress. Results revealed that mechanical properties of PU can be controlled through the structure of gelatin chain extenders and hard segment cross-linking present in the polyurethane scaffold. Further investigations into cell-mediated scaffold will help to identify the suitability of this new biomaterial as scaffolds for soft tissue applications.