软骨细胞三维支架短期固定培养实验研究

目的 :观察兔肋软骨细胞聚羟基乙酸 (polyglycolic acid,PGA)三维支架短期固定培养对软骨细胞利用率、软骨细胞长满支架所需时间及软骨细胞支架复合体厚度等影响。方法 :设 PGA支架固定组和非固定组 ,固定组 :先用鼠尾胶把 PGA支架粘附在盖玻片上 ,再往固定后的 PGA支架上滴加软骨细胞悬液 ,3～ 4天后支架与盖玻片分离。非固定组 :把软骨细胞悬液直接滴加到 PGA支架上进行复合培养。固定组和非固定组软骨细胞 PGA支架复合体培养 2周后分别行同种异体皮下移植 ,3月后取出标本 ,光镜观察软骨形成情况。结果 :固定组中 PGA支架在加培养液过程中 ,不会在培养液内漂动 ,支架内软骨细胞无大量流失现象 ;相反 ,非固定组中 PGA支架在加培养液和移动培养皿过程中 ,会在培养液内飘动或晃动 ,支架内很多软骨细胞重新漂浮至培养液中。固定组 PGA支架内软骨细胞分裂、增殖 1周时 ,软骨细胞已充满 PGA网眼 ;在非固定组 ,2周时 ,软骨细胞才长满 PGA支架内网眼。固定组软骨细胞支架复合体同种异体移植后能形成成片软骨 ;非固定组软骨细胞支架复合体移动后无软骨形成。结论 :软骨细胞 PGA三维支架短期固定培养 ,能有效减少支架内软骨细胞流失 ,缩短软骨细胞长满 PGA支架网眼的时间 ,增加软骨细胞支架复合体厚度 ,有利于组

Experimental study of short-time fixed chondrocytes-PGA constructs on the culture in vitro

Objective:To observe the change of seeding ratio of chondrocytes seeded on the PGA (polyglycolic acid) scaffolds,the time of PGA scaffolds fulled with chondrocytes and thickness of tissue enginering cartilage when chondrocytes PGA constructs was fixed on the coverglass within short time culture in vitro.Methods.PGA scaffold were firstly fixed on the coverglass with rat tail gel in fixation group,then the rib chondrocytes of New Zealand rabbit were seeded on the fixed PGA scaffolds,DMEM medium was added 2h later.The fixed chondrocytes PGA constructs were left coverglass 3 to 4 days later.The chondrocytes were seeded on the PGA scaffolds directly in control group.Chondrocytes PGA constructs in fixed group and control group were implanted allogenically into subcutis of the back of rabbits after 2 weeks culture,the samples were collected 3 months after transplantation and were examined histologically.Results.In the course of adding medium,the chondrocytes PGA constructs fixed on the coverglass did not float and chondrocytes were kept in chondrocytes PGA constructs in fixed group while PGA scaffolds floated in medium and many chondrocytes left into medium from PGA scaffolds in control group.The PGA scaffolds were fulled with chondrocytes after one week culture in fixed group or after two weeks culture in control group.Cartilage was formed by chondrocytes PGA constructs in fixed group while no cartilage were formed in control group.Conclusion:The short time fixed chondrocytes PGA constricts culture in vitro could prompt more chondrocytes seeded on the PGA scaffolds,shorten the time of PGA scaffolds fulled with chondrocytes and make it easy for tissue enginerring cartilage formed by allogenical chondrocytes.