Steel slag is an industrial solid waste produced during the steelmaking process. To explore the application of steel slag in the agricultural field, the present experiment was carried out to study the effect of substrates with different contents of steel slag on the growth of Hydrangea macrophylla cuttings. The conventional substrate (perlite: vermiculite: peat = 1:1:1) was used as the control (CK), and the treatments were designed as T1 (steel slag: perlite: vermiculite: peat = 1:3:3:3, v/v/v/v), T2 (steel slag: perlite: vermiculite: peat = 1:2:2:2, v/v/v/v), T3 (steel slag: perlite: vermiculite: peat = 1:1:1:1, v/v/v/v), and T4 (steel slag: perlite: vermiculite: peat = 1:0:0:0, v/v/v/v). The results showed that the addition of steel slag significantly increased the substrate’s bulk density, EC, and pH and improved its water retention capacity to a certain extent. There were significant differences among different treatments in morphological indicators, root growth and development, and physiological and biochemical characteristics of cutting seedlings. All traits, including plant height, fresh weight, dry weight, root length, root surface area, root volume, the number of root tips, root activity, and soluble protein content of seedlings grown in T3 were significantly higher than those in other substrates. The results indicated that the appropriate addition of steel slag is helpful to hydrangea cuttings’ growth, and the optimal mixing ratio is steel slag: perlite: vermiculite: peat = 1:1:1:1 (v/v/v/v). This is a significant innovation in applying steel slag in agricultural production.