摘要
目的 本研究旨在探索优化脱细胞肝支架的简易脱细胞方案,可为脱细胞肝的下一步应用研究提供良好的生物材料。方法 依次采用0.9%氯化钠注射液500mL冲洗1h、0.5%乙二醇双(2-氨基乙基醚)四乙酸1500mL冲洗3h和1%十二烷基硫酸钠溶液3000mL冲洗大鼠肝脏6h,速度均为120滴/min,用来制备脱细胞肝支架。通过HE染色、扫描电镜和免疫荧光染色、DNA含量检测、X线下显影测试和流动性测试等方式检测脱细胞肝的结构完整性和主要成分。结果 经过10h的灌流,肝脏中的血管清晰可见,HE染色中显示脱细胞肝去除了细胞核,免疫组织化学染色见胶原蛋白I、纤连蛋白和层粘连蛋白表达,扫描电镜显示支架呈疏松多孔的结构,DNA含量显示<50ng/mg,X线测试表明对注射进入的显影剂有良好的显影效果,流动性测试表明脱细胞肝保留了较为完整和通畅的血管系统。结论 本研究所采用的脱细胞方法成功制备了脱细胞肝脏模型,达到了较为理想的脱细胞效果。
Objective To explore and optimize the simple decellularization scheme of decellularized liver scaffolds,which can provide good biomaterials for the next application research of decellularized liver.Methods The rat liver was rinsed with 500ml of 0.9% sodium chloride injection for 1 hour,followed by 1500ml of 0.5% ethylene glycol bis(2-aminoethyl ether) tetraacetic acid for 3 hours,and then with 3000ml of 1% sodium dodecyl sulfate solution for 6 hours,all at a rate of 120 drops per minute,to prepare the acellular liver scaffold.The structural integrity and the preservation of major components within the decellularized liver were assessed through various methodologies including HE staining,scanning electron microscopy,immunofluorescence staining,DNA content analysis,X-ray radiography and fluidity testing.Results After 10 hours of perfusion,the blood vessels in the liver were clearly visible.HE staining showed that the decellularized liver had removed the cell nuclei.Immunohistochemical staining showed the expression of collagen I,fibronectin and laminin.Scanning electron microscopy further confirmed the loose and porous nature of the scaffold.The DNA content was <50 ng/mg.X-ray radiography demonstrated robust imaging development post-injection.Fluidity tests showed that the decellularized liver retained a relatively intact and unobstructed vascular network.Conclusion The decellularization method used in this study successfully prepared a decellularized liver model and achieved a relatively ideal decellularization effect.
作者
陈宁
郭安然
刘安娜
张玲
王彩
邢君
郑敏
李玲
CHEN Ning;GUO An-ran;LI Ling(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning Hubei 437100,China)
出处
《湖北科技学院学报（医学版）》
2024年第4期313-318,共6页
Journal of Hubei University of Science and Technology(Medical Sciences)
基金
湖北省自然科学基金面上项目(2023AFB538,2022CFB755)
国家自然科学基金青年项目(82003308,52303182)
湖北科技学院博士启动基金(BK202118)
湖北科技学院科研创新团队项目(2023T10)。
关键词
脱细胞肝
肝组织工程
脱细胞支架
肝脏
Decellularized liver
Livertissue engineering
Decellularized scaffold
Liver
