Knee orthoses assist patients with impaired gait through the amendment of knee abnormalities, restoration of mobility, alleviation of pain, shielding, and immobilization. The inevitable issues with laborious traditional plaster molding procedures for orthoses can be resolved with 3D printing. However, a number of challenges have limited the adoption of 3D printing, the most significant of which is the proper material selection for orthoses. This is so because the material used to make an orthosis affects its strength, adaptability, longevity, weight, moisture response, etc. This study intends to examine the mechanical, physical, and dimensional characteristics of three-dimensional (3D) printing materials (PLA, ABS, PETG, TPU, and PP). The aim of this investigation is to gain knowledge about these materials’ potential for usage as knee orthosis materials. Tensile testing, Olympus microscope imaging, water absorption studies, and coordinate measuring machine-based dimension analysis are used to characterize the various 3D printing materials. Based on the investigation, PLA outperforms all other materials in terms of yield strength (25.98 MPa), tensile strength (30.89 MPa), and shrinkage (0.46%). PP is the least water absorbent (0.15%) and most flexible (407.99%); however, it is the most difficult to fabricate using 3D printing. When producing knee orthoses with 3D printing, PLA can be used for the orthosis frame and other structural elements, PLA or ABS for moving parts like hinges, PP for padding, and TPU or PP for the straps. This study provides useful information for scientists and medical professionals who are intrigued about various polymer materials for 3D printing and their effective utilization to fabricate knee orthoses.