目的探索鱼胶原作为支架材料用于构建组织工程软骨的可行性。方法将鱼胶原经冷冻干燥后制备成多孔海绵状支架材料。取兔耳软骨细胞接种于鱼胶原支架,体外培养2周和4周,验证其细胞相容性和体外促进软骨基质分泌的能力。随后植入裸鼠皮下培养6周,行大体观察、组织学检测、生物力学及生物化学定量检测。结果鱼胶原具有良好的细胞相容性,且在体内、外均能形成软骨特异性细胞外基质。体外培养4周和体内培养6周后,HE染色可见典型的软骨陷窝出现。体内培养6周后,Safranin-0染色显示有蛋白聚糖基质产生。生物力学和生物化学结果表明,新生软骨组织均接近正常软骨组织。结论鱼胶原可作为支架材料用于构建组织工程软骨。

Objective To explore the feasibility of fish collagen in constructing tissue-engineered cartilage. Methods Porous fish collagen was fabricated by lyophilization. Auricular chondrocytes derived from rabbit were seeded into the porous fish collagen. After cultured in vitro for 2 or 4 weeks, the cytocompatibility and the capacity of cartilage-specific extracellular matrix formation based on fish collagen were investigated. Thereafter, the cell-scaffold constructs were implanted into the rude mice subcutaneously. Gross observation, histological observation and quantitative evaluation of neocartilage test were carried out after cultured for 6 weeks. Results Fish collagen possessed satisfactory cytocompatibility, and facilitated cartilage-specific extracellular matrix formation both in vitro and in vivo. Notably, HE staining showed typical mature lacuna after cultured in vitro for 4 weeks or cultured in vivo for 6 weeks. After cultured in vivo for 6 weeks, Safranin-O staining showed abundant proteoglycan, biomechanical and biochemistry analyses indicated that neocartilage was comparable than native cartilage. Conclusion Fish collagen is a promising natural derived material as scaffold in constructing tissue engineering cartilage.