This study explores the use of urease-powered nanobots for radionuclide therapy in bladder cancer treatment. Bladder cancer, a significant health concern, is often treated with intravesical drug administration, but this method has limited therapeutic efficacy. To address this, self-propelled nanoparticles or nanobots have been proposed due to their enhanced diffusion and mixing capabilities in urine compared to conventional drugs. The study tested radiolabelled mesoporous silica-based urease-powered nanobots in an orthotopic mouse model of bladder cancer. In vivo and ex vivo results showed enhanced nanobot accumulation at the tumor site, with an eightfold increase in positron emission tomography (PET) signals. Label-free optical contrast confirmed tumor penetration by nanobots using polarization-dependent scattered light-sheet microscopy. Treating tumor-bearing mice with intravesically administered radio-iodinated nanobots resulted in a 90% reduction in tumor size, demonstrating the potential of nanobots as efficient delivery systems for bladder cancer therapy. The study highlights the underexplored potential of nanobots for innovative bladder cancer treatment options.This study explores the use of urease-powered nanobots for radionuclide therapy in bladder cancer treatment. Bladder cancer, a significant health concern, is often treated with intravesical drug administration, but this method has limited therapeutic efficacy. To address this, self-propelled nanoparticles or nanobots have been proposed due to their enhanced diffusion and mixing capabilities in urine compared to conventional drugs. The study tested radiolabelled mesoporous silica-based urease-powered nanobots in an orthotopic mouse model of bladder cancer. In vivo and ex vivo results showed enhanced nanobot accumulation at the tumor site, with an eightfold increase in positron emission tomography (PET) signals. Label-free optical contrast confirmed tumor penetration by nanobots using polarization-dependent scattered light-sheet microscopy. Treating tumor-bearing mice with intravesically administered radio-iodinated nanobots resulted in a 90% reduction in tumor size, demonstrating the potential of nanobots as efficient delivery systems for bladder cancer therapy. The study highlights the underexplored potential of nanobots for innovative bladder cancer treatment options.