Abstract:  Objective · To construct C-shaped cartilage rings by rabbit auricular cartilage-derived chondrocytes combing with both electrospun gelatin/ polycaprolactone(GT/PCL) nanofibrous membranes and 3D printed supporters for repairing tracheal cartilage defects.  Methods · Primary chondrocytes were isolated from rabbit auricular cartilage with methods of trypsin enzyme digestion and collagenase enzyme digestion. After proliferation in vitro, the chondrocytes of passage 2 were harvested for further experiments. Ultrafine composite fibers of GT/PCL were fabricated via electrospinning. The electrospun GT/PCL membranes were tailored into rectangle shape, the length of which is 12 cm and the width is 2.5 cm. Chondrocytes were seeded on membrane at a density of 1×108 cells/mL. Then the membrane were rolled onto a 3D printed supporter of poly(DL-lactide-ε-caprolactone) (PLCL) material to construct a C-shaped cartilage-like complex. After 8 weeks of subcutaneous incubation in vivo, gross inspection and paraffin section staining were applied for evaluation.  Results · After 8 weeks of culture in vivo, mature cartilage-like tissue were formed with open-cylindrical bellow appearance and pecific mechanical property. C-shaped rings arranged at regular intervals on the inner surface of tissue, which were similar to the normal structure of tracheal cartilages. Histological and immunohistological staining showed a large number of typical lacunar structures and extracellular matrix secretions.  Conclusion · It is feasible to construct tissue engineered C-shaped cartilage tissue by combing chondrocytes with GT/PCL membrane and 3D printed PLCL supporter for tracheal cartilage repair.

Key words: electrospun fibrous membrane, 3D printed supporter, tissue engineering, C-shaped cartilage tissue