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.