Objective: To investigate the preclinical feasibility of using bio-airbrush technology for the fully arthroscopic treatment of cartilage defects with a combination of cells and biomaterials. Design: A combination of in vitro, ex vivo, and cadaveric in situ experiments was performed, to evaluate the preclinical feasibility of using bio-airbrush technology for cartilage repair. Cartilage matrix production after airbrushing cells was evaluated. Subsequently, the influence of different spray parameters on successful defect fill was studied. Finally, the surgical feasibility of using bio-airbrush technology was evaluated in situ in cadaveric knees. Results: The most important findings of the presented study are that the matrix producing capacity of the cells was not hampered by airbrushing, air pressure and defect location were important parameters to consider, and using the bio-airbrush was found to be surgically feasible in a CO2-insufflated arthroscopic procedure in cadaveric knees. Conclusions: Bio-airbrush using CO2 arthroscopy would facilitate fully arthroscopic application of (cell laden) two-component in situ gelating biomaterials to cartilage defects, as a treatment for cartilage repair. This would shift the conventional procedures, which are commonly performed using a (mini-)arthrotomy, toward arthroscopy. This study shows that a bio-airbrush can be a clinically feasible instrument for the use in cell transplantations to repair knee cartilage defects. This technology would enable a shift from cartilage repair procedures, which are mostly performed using an arthrotomy, toward a fully arthoscopic approach. In addition, the technology presented could possibly be translated to other fields of research where controlled in situ deposition of cells is required, such as treatment of burn wounds/ulcers/others. Our research has shown that the use of an airbrush is safe, without harming cell viability and performance.