The purpose of this study was to assess the technical feasibility of 3-dimensional (3D) super-resolution reconstruction (SRR) of 2D turbo spin echo (TSE) knee magnetic resonance imaging (MRI) and to compare its image quality with conventional 3D TSE sampling perfection with application optimized contrast using different flip angle evolutions (SPACE) MRI. Super-resolution reconstruction 2D TSE MRI and 3D TSE SPACE images were acquired from a phantom and from the knee of 22 subjects (8 healthy volunteers and 14 patients) using a clinical 3-T scanner. For SRR, 7 anisotropic 2D TSE stacks (voxel size, 0.5 × 0.5 × 2.0 mm; scan time per stack, 1 minute 55 seconds; total scan time, 13 minutes 25 seconds) were acquired with the slice stack rotated around the phase-encoding axis. Super-resolution reconstruction was performed at an isotropic high-resolution grid with a voxel size of 0.5 × 0.5 × 0.5 mm. Direct isotropic 3D image acquisition was performed with the conventional SPACE sequence (voxel size, 0.5 × 0.5 × 0.5 mm; scan time, 12 minutes 42 seconds). For quantitative evaluation, perceptual blur metrics and edge response functions were obtained in the phantom image, and signal-to-noise and contrast-to-noise ratios were measured in the images from the healthy volunteers. Images were qualitatively evaluated by 2 independent radiologists in terms of overall image quality, edge blurring, anatomic visibility, and diagnostic confidence to assess normal and abnormal knee structures. Nonparametric statistical analysis was performed, and significance was defined for P values less than 0.05. In the phantom, perceptual blur metrics and edge response functions demonstrated a clear improvement in spatial resolution for SRR compared with conventional 3D SPACE. In healthy subjects, signal-to-noise and contrast-to-noise ratios in clinically relevant structures were not significantly different between SRR and 3D SPACE. Super-resolution reconstruction provided better overall image quality and less edge blurring than conventional 3D SPACE, yet the perceived image contrast was better for 3D SPACE. Super-resolution reconstruction received significantly better visibility scores for the menisci, whereas the visibility of cartilage was significantly higher for 3D SPACE. Ligaments had high visibility on both SRR and 3D SPACE images. The diagnostic confidence for assessing menisci was significantly higher for SRR than for conventional 3D SPACE, whereas there were no significant differences between SRR and 3D SPACE for cartilage and ligaments. The interreader agreement for assessing menisci was substantial with 3D SPACE and almost perfect with SRR, and the agreement for assessing cartilage was almost perfect with 3D SPACE and moderate with SRR. We demonstrate the technical feasibility of SRR for high-resolution isotropic knee MRI. Our SRR results show superior image quality in terms of edge blurring, but lower image contrast and fluid brightness when compared with conventional 3D SPACE acquisitions. Further contrast optimization and shortening of the acquisition time with state-of-the-art acceleration techniques are necessary for future clinical validation of SRR knee MRI.