Two-photon polymerization (2PP) technique is a novel CAD/CAM-based technology allowing the fabrication of any computer-designed 3D structure from a photosensitive polymeric material with a lateral resolution down to 100 nm. The fabrication of highly reproducible scaffold structures for tissue engineering by 2PP is very important for systematic studies of cellular processes and better understanding of in vitro tissue formation. Flexibility of this technology and ability to precisely define 3D construct geometry allow the direct addressing of issues associated with vascularization and patient-specific tissue fabrication. In this chapter, we report on our recent advances in the fabrication of biomedical implants and 3D scaffolds for tissue engineering and regenerative medicine by 2PP technique.