(1) Background: This study aims to compare the effects of 3D-printed splints and conventional manufactured splints on sleep bruxism (SB) EMG activity. (2) Methods: Twenty-six patients (19 M, 7 F, 25.8 ± 2.6 years) were randomly allocated to a study group (3D splints) and a control group (conventional manufactured splints) and followed for a period of three months with night EMG–ECG recordings. Samples of the involved materials were analyzed for nanoindentation. The outcomes of interest considered were the overall SB index, the total amount of surface masseter muscle activity (sMMA), and general and SB-related phasic and tonic contractions. A statistical evaluation was performed with a confidence interval (CI) between 2.5% and 97.5%. (3) Results: Differences between groups with OAs were observed for general tonic contraction (p = 0.0009), while differences between recording times were observed for general phasic contractions (p = 0.002) and general tonic contractions (p = 0.00001). Differences between recording times were observed for the total amount of sMMA (p = 0.01), for general phasic contractions (p = 0.0001), and for general tonic contractions (p = 0.000009) during night recordings without OAs. (4) Conclusions: Three-dimensional splints seem to have a higher impact on SB-related electromyographic activity but not on the overall sleep bruxism index. The more regular surfaces offered by 3D splints could be related to phasic contraction stabilization.