Potentially toxic elements in municipal sewage sludge can be effectively immobilized during biochar production via pyrolysis. However, the bioavailability of these elements when biochar is applied in soilless cultivation to improve substrate quality has yet to be sufficiently established. In this study, we investigated the chemical speciation and cucumber plant uptake of potentially toxic elements in soilless cultivation when the growth substrate was amended with sewage sludge biochar (0, 5, 10, 15, and 20 wt%). It was found that the addition of 10 wt% biochar was optimal with respect to obtaining a high cucumber biomass and achieving low environmental risk considering the occurrence of hormesis. When the substrate was amended with 10 wt% biochar, cucumber fruit contained lower concentrations of As, Cr, and Zn and smaller bioavailable fractions of As, Cd, Cr, Ni, Cu, and Zn compared with the fruit of control plants, thereby meeting national safety requirements (standard GB 2762–2012, China). Most of the As and Cd taken up by cucumbers accumulated in the leaves and fruit, whereas Cr was found primarily in the roots, and most Ni, Cu, and Zn was detected in the fruit. Importantly, only small proportions of the potentially toxic elements in biochar were taken up by cucumber plants (As: 0.0075%; Cd: 0.038%; Ni: 0.0064%; Cu: 0.0016%; and Zn: 0.0015%). Given that the As, Cd, Ni, and Zn speciation in sewage sludge biochar was effectively immobilized after cultivation, the findings of this study indicate that sewage sludge biochar is a suitable substrate amendment in terms of the risk posed by potentially toxic elements.