Segmental bone tissue engineering is a highly effective approach for the repair of large bone defects. In this paper, a PCL-based guide film was developed for controlling segmental bone tissue engineering. 3D bioprinting was used to fabricate the PCL/NaCl-based cylindrical films. The effects of the film's thickness and NaCl concentration on the mechanical properties, degradability, swelling behavior, porosity, and cytotoxicity of the samples were investigated. Response surface methodology was employed to study the mechanical behavior using the central composite design (CCD) results showed that increasing the NaCl concentration up to 10% wt. significantly improved the degradability, swelling, and hydrophilicity of the films. It was also indicated that the maximum stiffness of the guide films under vertical loading was almost 5 times more than the maximum stiffness in the horizontal loading direction, but the samples showed greater compressive strength and elongation under horizontal compressive loading. All the evidence indicated that the mechanical properties of the films were more dependent on the film thickness so the thicker films with an 800µm thickness had better mechanical properties in both vertical and horizontal loading. Cytotoxicity assay also approved the non-toxic effect of the PCL films on the MC3T3 osteoblast cell line. Based on the results, the PCL-based films were a suitable candidate to act as a guide for segmental bone tissue engineering.