【摘要】 目的探索真皮脱细胞基质微载体构建可注射软骨的可能性。方法将分离培养得到的软骨细胞与真皮脱细胞基质微载体混合培养24 h后分为两组,分别在三维动态系统(实验组)与静态系统(对照组)下培养。体外培养2周、4周分别进行扫描电镜观察,体外培养4周行活死细胞染色观察。体外培养4周后将软骨细胞微载体复合物植入裸鼠皮下,8周后取材进行大体观察、HE染色和番红固绿染色观察。结果体外培养2周、4周时电镜结果显示,实验组与对照组均发生细胞增殖与细胞外基质的分泌,实验组更明显。体外培养4周,活死细胞染色结果显示,实验组软骨细胞活力较好,与微载体结合更充分,细胞进入微载体内部,而对照组软骨细胞仅黏附于微载体表面。体内培养8周,两组组织块大体观无明显差异;组织学染色显示实验组形成连续软骨细胞陷窝,对照组软骨陷窝呈岛状,实验组形成软骨更成熟。结论真皮脱细胞基质微载体复合软骨细胞可以实现可注射软骨的构建,且动态培养效果优于静态培养。更多还原

【Abstract】 Objective To explore the possibility of construction of injectable cartilage by dermal acellular matrix cytodex.Methods The isolated and cultured chondrocytes were mixed with dermal acellular matrix cytodex and cultured for 24 hours,and then divided into two groups.They were cultured in a three-dimensional dynamic system( experimental group) and a static system(control group),respectively.After cultured in vitro for 2 weeks and 4 weeks,SEM observation was performed respectively.After cultured in vitro for 4 weeks,live-dead cell staining was performed.After 4 weeks of culture in vitro,the chondrocyte cytodex complex was implanted subcutaneously in nude mice.After 8 weeks,samples were collected for general observation,HE staining and Safranin-O staining.Results After cultured in vitro for 2 weeks and 4 weeks,electron microscopy showed that cell proliferation and extracellular matrix secretion occurred both in the experimental group and the control group,especially in the experimental group.After cultured in vitro for 4 weeks,live-dead cell staining showed that the chondrocytes in the experimental group had better vitality and were more fully bound to the cytodex,and the cells entered the cytodex,while the chondrocytes in the control group were bound to the surface of cytodex.After 8 weeks’ culture in vivo,there was no significant difference in the general view of tissue blocks between the two groups.Histological staining showed that continuous chondrocyte lacunae were formed in the experimental group,while the lacunae in the control group was island shaped,indicating cartilage formation in the experimental group was more mature.Conclusion Dermal acellular matrix cytodex composite chondrocytes can be used to construct injectable cartilage in vitro,and the effect of dynamic culture is better than that of static culture.更多还原