<p>Femtosecond laser three-dimensional printing has emerged as a transformative multiscale fabrication strategy for organ-on-a-chip systems, enabling hierarchical integration of functional nanomaterials with subcellular precision. This review highlights the synergistic convergence of ultrafast laser processing and nanomaterial engineering, overcoming the limitations of conventional fabrication methods by achieving sub-100-nm resolution via nonlinear optical effects. Key advances include nanoparticle-doped photoresists for mechanically robust, electrically conductive, and optically active microarchitectures, as well as laser-induced self-assembly mechanisms for nanomaterials such as silica nanoparticles, liquid crystals, and carbon nanotubes. The integration of in situ photoreduction, photoexcitation-induced bonding, and programmable optical forces enables precise control over nanomaterial dispersion, interfacial interactions, and multiscale structural integrity. These innovations facilitate the creation of biomimetic vascular networks, three-dimensional tissue scaffolds, and embedded biosensors, while enhancing mechanical strength, stimulus responsiveness, and device functionality. This hybrid fabrication paradigm holds significant potential to advance personalized disease modeling, drug screening, and bioelectronic interfaces via multifunctional organotypic platforms. </p> Graphical Abstract <p></p>