Purpose: To establish a complete digital workflow for the design and manufacture of sports mouthguards and to observe preliminary clinical effects. Materials and Methods: Eighteen healthy participants were included in this study. The self-controlled method was applied, and all participants were provided with two types of mouthguards. Digital mouthguards were designed and milled using CAD/CAM with polyetheretherketone, and conventional mouthguards were fabricated using the vacuum pressure-forming method with ethylene vinyl acetate. The order of wearing was determined using a random number table, and the washout period between was set as 1 month. Degrees of satisfaction in terms of retention, appearance, occlusal comfort, and labial comfort were evaluated. Distribution of occlusal force was tested using the T-scan analysis system. Participants were also asked to choose one mouthguard for future use. Results: The complete workflow of digitally designed and manufactured sports mouthguards was successfully established. No significant difference was noted in retention between the types of mouthguard. The appearance score (P = .025), occlusal comfort score (P = .030), and labial side comfort score (P = .003) of the digital mouthguard group were significantly higher compared to the conventional mouthguard group. T-scan analysis results showed that in centric occlusion, participants exhibited occlusal contact in the second molar alone while wearing conventional mouthguards, while the occlusal force was uniformly distributed with digital mouthguards. Sixteen participants selected the digital mouthguard for future use. Conclusion: Digital design and manufacture of sports mouthguards improved the occlusal design and greatly simplified and optimized the conventional fabrication process.