Fibrin gel-based scaffolds have promising potential for microtia reconstruction. Autologous chondrocytes and chondrocyte cell sheets are frequently used seed cell sources for cartilage tissue engineering. However, the aesthetic outcome of chondrocyte-based microtia reconstruction is still not satisfactory. In this study, we aimed to fabricate the chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold for microtia reconstruction. We designed a unique auricular mold that could fabricate a fibrin gel scaffold resembling human auricle anatomy. Primary chondrocytes were harvested from rabbit auricular cartilage, and chondrocyte cell sheets were developed. Chondrocyte-microtissues were prepared from the cell sheets. The mixture of chondrocytes/chondrocyte-microtissues was laden in fibrin gel during the auricular scaffold fabrication. The protrusions and recessed structure in the auricular scaffold surface were still clearly distinguishable. After a one-week in vitro culture, the 3 D structure and auricular anatomy of the scaffold were retained. And followed by eight-week subcutaneous implantation, cartilaginous tissue was regenerated in the artificial auricular structure as indicated by the results of H&E, Toluidine blue, Safranin O, and type II collagen (immunohistochemistry) staining. Protrusions and depressions of the auricular scaffold were slightly deformed, but the overall auricular anatomy was maintained after 8-week in vivo implantation. Extracellular matrix components content were similar in artificial auricular cartilage and rabbit native auricular cartilage. In conclusion, the mixture of chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold showed a promising potential for cartilaginous tissue regeneration, suggesting this as an effective approach for autologous chondrocyte-based microtia reconstruction.

基于纤维蛋白凝胶的支架在小耳畸形重建方面具有广阔的潜力。自体软骨细胞和软骨细胞片层是软骨组织工程中常用的种子细胞来源。然而，基于软骨细胞的小耳畸形重建的美学效果仍然不令人满意。在这项研究中，我们旨在制造用于小耳畸形重建的软骨细胞/软骨细胞-微组织负载纤维蛋白凝胶耳廓支架。我们设计了一种独特的耳廓模具，可以制造类似于人类耳廓解剖结构的纤维蛋白凝胶支架。从兔耳软骨中收获原代软骨细胞，并形成软骨细胞片。从细胞片制备软骨细胞微组织。在耳廓支架制造过程中，软骨细胞/软骨细胞-微组织的混合物充满了纤维蛋白凝胶。耳廓支架表面的突起和凹陷结构仍然清晰可辨。经过一周的体外培养后，支架的 3D 结构和耳廓解剖结构得以保留。然后进行 8 周的皮下植入，如 H&E、甲苯胺蓝、番红 O 和 II 型胶原蛋白（免疫组织化学）染色结果所示，人工耳廓结构中的软骨组织再生。耳廓支架的突起和凹陷有轻微变形，但在体内植入8周后，耳廓的整体解剖结构保持不变。人工耳廓软骨与兔天然耳廓软骨细胞外基质成分含量相似。综上所述，