TGase-Enhanced Microtissue Assembly in 3D-Printed-Template-Scaffold (3D-MAPS) for Large Tissue Defect Reparation.

Engineering large and functional tissue constructs with complex structures (e.g., external ear or nose) for reparation and reconstruction of tissue defects remains one of the major challenges in regenerative medicine, which demands abundant cell sources, advanced biofabrication schemes, and satisfactory integration with the host for long-term efficacy post implantation. Here the 'Microtissue Assembly in 3D-Printed-template-Scaffold' (3D-MAPS), as a platform technology to rapidly fabricate centimeter-sized functional tissue constructs with complex structures, is developed. 3D-MAPS facilitates bottom-up assembly of large-scale manufactured microtissues within the 3D-printed hollow polymeric templates with pre-defined architectures. The assembly and fusion of 2×106 mesenchymal stem cell-based microtissues within the defined 3D-printed template is further enhanced by addition of a natural protein crosslinker (i.e., transglutaminase (TGase)), and thereby achieves construction of centimeter-sized tissue with high cell viability and mechanical stability in vitro within 30 min. Further in vivo implantation of the 3D-MAPS-fabricated ear-like tissue construct in rabbit models assisted by flap prefabrication technique results in increased structural vascular support and strengthened functional survival. Thus, the TGase-enhanced 3D-MAPS demonstrates its potential and feasibility as a powerful biofabrication platform for tissue engineering application.