Extracellular matrix (ECM) hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the adequate bioactivity of native matrix. In this study, we developed decellularized cartilage ECM (dcECM) hydrogels from porcine ears innovatively via the main method of enzymatic digestion and verified good biocompatible properties of dcECM hydrogels to deliver chondrocytes and form subcutaneous cartilage in vivo. The scanning electron microscopy and turbidimetric gelation kinetics were used to characterize the material properties and gelation kinetics of the dcECM hydrogels. Then we evaluated the biocompatibility of hydrogels via the culture of chondrocytes in vitro. To further explore the dcECM hydrogels in vivo, grafts made from the mixture of dcECM hydrogels and chondrocytes were injected subcutaneously in nude mice for the gross and histological analysis. The structural and gelation kinetics of the dcECM hydrogels altered according to the variation in the ECM concentrations. The 10 mg/ml dcECM hydrogels could support the adhesion and proliferation of chondrocytes in vitro. In vivo, at 4 weeks after transplantation, cartilage-like tissues were detected in all groups with positive staining of toluidine blue, Safranin O, and collagen II, indicating the good gelation of dcECM hydrogels. While with the increasing concentration, the tissue engineering cartilages formed by 10 mg/ml dcECM hydrogel grafts were superior in weights, volumes, collagen, and glycosaminoglycan (GAG) content compared to the dcECM hydrogels of 1 mg/ml and 5 mg/ml. At 8 weeks after grafting, dcECM hydrogel grafts at 10 mg/ml showed very similar qualities to the control, collagen I grafts. After 12 weeks of in vivo culture, the histological analysis indicated that 10 mg/ml dcECM hydrogel grafts were similar to the normal cartilage from pig ears, which was the source tissue. In conclusion, dcECM hydrogel showed the promising potential as a tissue engineering biomaterial to improve the regeneration and heal injuries of ear cartilage.

细胞外基质 (ECM) 水凝胶具有可注射性、填充不规则形状空间的能力以及天然基质足够的生物活性等优点。在这项研究中，我们通过酶消化的主要方法创新地开发了来自猪耳朵的脱细胞软骨 ECM (dcECM) 水凝胶，并验证了 dcECM 水凝胶具有良好的生物相容性，可输送软骨细胞并形成皮下软骨体内. 扫描电子显微镜和浊度凝胶动力学用于表征 dcECM 水凝胶的材料特性和凝胶动力学。然后我们通过软骨细胞的培养评估了水凝胶的生物相容性体外. 进一步探索 dcECM 水凝胶体内，由 dcECM 水凝胶和软骨细胞的混合物制成的移植物被皮下注射到裸鼠中，用于大体和组织学分析。dcECM 水凝胶的结构和凝胶动力学根据 ECM 浓度的变化而改变。10 mg/ml dcECM 水凝胶可以支持软骨细胞的粘附和增殖体外. 体内, 移植后 4 周，各组均检测到软骨样组织，甲苯胺蓝、番红 O 和 II 型胶原染色呈阳性，表明 dcECM 水凝胶凝胶化良好。虽然随着浓度的增加，与 1 mg/ml 和 5 mg/ml 的 dcECM 水凝胶相比，由 10 mg/ml dcECM 水凝胶移植物形成的组织工程软骨在重量、体积、胶原蛋白和糖胺聚糖 (GAG) 含量方面更优。在移植后 8 周，10 mg/ml 的 dcECM 水凝胶移植物显示出与对照胶原蛋白 I 移植物非常相似的质量。12 周后体内培养后，组织学分析表明，10 mg/ml dcECM 水凝胶移植物与猪耳朵的正常软骨相似，是源组织。总之，dcECM 水凝胶显示出作为组织工程生物材料改善耳软骨再生和愈合损伤的潜力。