Interleukin‐4‐Loaded Gelatin Methacryloyl Hydrogel Promotes Subcutaneous Chondrogenesis of Engineered Auricular Cartilage in a Rabbit Model

ABSTRACT Tissue engineering technology offers a promising solution for ear reconstruction; however, it faces the challenge of foreign body reaction and neocartilage malformation. This study investigates the impact of interleukin‐4 (IL‐4), an anti‐inflammatory factor, on cartilage regeneration of hydrogel encapsulating autologous auricular chondrocytes in a rabbit subcutaneous environment. Initially, we assessed the influence of IL‐4 on chondrocyte proliferation and determined the appropriate concentration using the CCK‐8 test in vitro. Subsequently, we loaded IL‐4 into gelatin methacryloyl (GelMA) hydrogel containing chondrocytes and measured its release profile through ELISA. The constructs were then implanted autologously into rabbits' subcutis, and after 3, 7, 14, and 28 days, cartilage matrix formation was evaluated by histological examinations, and gene expression levels were detected by qRT‐PCR. Results demonstrated that IL‐4 promotes chondrocyte proliferation in vitro, and maximum release from constructs occurred during the first week. In the rabbit subcutaneous implantation model, IL‐4‐loaded constructs (20 ng/mL) maintained a superior chondrocytic phenotype compared to controls with increased expression of anti‐inflammatory factors. These findings highlight IL‐4 as a potential strategy for promoting chondrogenesis in a subcutaneous environment and improving ear reconstruction.